App LC Ms Pesticides Applicationperkin

ANALYTICAL
SOLUTIONS FOR
PESTICIDES
LC/MS Pesticides Application Notes Compendium

Introduction
The increasing demand for organic products, evolving global
regulations, and improving accessibility of agricultural hubs to global
markets results in pressure on food safety precautions. Translating,
in part, to increased pesticide residue analysis demand. Improving
throughput to meet demand can be accomplished with both
increases in number of laboratories offering testing, as well as with
migrating existing methods and technologies to novel alternatives to
leverage their efficiency gains. More targets per analysis, with fewer
instruments, with less strain on laboratory operation.
Leading this transition for decades now has been mass
spectrometry, largely due to the analytical power it provides
scientists. Further, the coupling of mass spectrometry to
chromatographic techniques has vastly increased the number and
confidence of pesticide analysis workflows today. Commonplace
are procedures covering hundreds of target analytes with results
available in tens of minutes all with the shared vision of improving
global food safety.
Collected within this compendium are applications developed on
PerkinElmer’s QSight LC/MS/MS with the goal of assisting you
achieve an efficient, green, and easy-to-operate pesticide analysis
laboratory or procedure. A wide breadth of matrices are covered,
including: cannabis, fruits and vegetables, tea, grains, and more.
Additionally, QSight’s Dual Source Technology allows for seamless
incorporation of APCI into analysis methods, resulting in reduced
dependence on gas chromatographic approaches and potentially
allowing for the consolidation of an analysis to only one instrument.
For all searching to streamline their pesticide workflows or to those
rearing to get started exploring the world of pesticide analysis, we
believe this is a must read.
Table of Contents
Multi-residue Analytical Method for the Confirmation................ 3 LC/MS/MS Analytical Method for Pesticide Residue................ 46
and Quantification of 500+ Pesticides in Fruit and Vegetables in Cannabis Flower as Defined by Florida Testing Requirements
Analysis of Multi-Residue Pesticides in Rice by LC/MS/MS..... 21 “No Dilute” Just Shoot: Robustness of a QSight....................... 55
LC-ESI-MS/MS for Low Level Pesticide Residue Analysis in Wine
Analysis of Target Pesticide Residues in Berries........................ 8
with LC/MS/MS Coupled with a QuEChERS Sample Preparation Analysis of near 400 Pesticides in Tea Via LC/MS/MS:........... 59
Simple Sample Preparation and APCI to Improve
Direct Analysis of Glyphosate and Similar Polar....................... 32
Analyte Coverage
Pesticides in Oatmeal by UHPLC-MS/MS
Quantitation of Pesticide and Mycotoxin Residues in............... 76
Analysis of 213 Pesticide Residues in Grapes by..................... 36
Cannabis as Definedby AZ, MI, OK, CO, OR, NV and WA State
LC/MS/MS with Time-Managed MRM
Recreational Cannabis Regulations
Detection of Fipronil and its Metabolites in Eggs..................... 42
by LC/MS/MS
2
APPLICATION NOTE
Liquid Chromatography /
Mass Spectrometry
Authors:
Maria Laura Pati, Nicola Barbieri,
Alfredo Fantastico, Piero Pontrelli
S.A.Mer. Servizio Analisi Chimico-Merceologiche
Bari, Italy
Aristide Ganci
Roberto Troiano
PerkinElmer, Inc.
Milan, Italy
Ignazio Garaguso
PerkinElmer, Inc.
Rodgau, Germany
Multi-residue Analytical
Method for the Confirmation Introduction
Pesticides are a group of compounds
and Quantification of 500+ containing hundreds of listed listed
substances, most of which are
Pesticides in Fruit and Vegetables regulated by governmental agencies.
Their function is to prevent, destroy,
or control harmful organisms or diseases, as well as protect plants or plant products during
production, storage and transport. Pesticides are primarily utilized in the agricultural sector, and
contain one or more active substances. From the point of application, pesticides can be transported
through various media, and ultimately be deposited on plants and animals humans consume. While
some of these compounds have not been found to be harmful, others may have toxic properties to
humans and animals, as well as pose a danger to our environment and ecosystems.
The European Commission (EC) has set maximum residue limits (MRLs) for pesticide residues in or
on food and feed of plant and animal origin, as detailed in legislative framework Regulation (EC)
396/2005.1 MRLs vary for given pesticides and food products, but generally, the MRLs are set at
0.01 mg/kg for many fruits and vegetables. For certain pesticides and matrices, different legally
permitted concentrations have been set, mostly ranging from 0.001 – 100 mg/kg.2 For pesticides
not listed in the regulation, a default MRL of 0.01 mg/kg applies.1
3
Rapid multi-residue analytical methods are needed to confirm Table 1. LX50 UHPLC Parameters.
and quantitate the wide range of regulated pesticides, which are Column C18, 100x4.6 mm, 2.7 µm
often present at very low levels in sample matrices with different
chemistries. Based on regulatory requirements in recent years, Solvent A: 9 mM ammonium formate in water-
Mobile acetonitrile (90:10, v/v) + 0.1 % formic acid
liquid chromatography coupled to tandem mass spectrometry
Phase Solvent B: 1 mM ammonium formate in acetonitrile-
(LC/MS/MS) has become the technique of choice to perform water (90:10, v/v) + 0.1 % formic acid
confirmatory analysis on these compounds. Owing to its ability
Step Time [min] Flow Rate [ml/min] %A %B
to detect hundreds of compounds in a single analysis, LC/MS/MS
1 0 0.4 100 0
offers unmatched sensitivity and selectivity based on large 2 10 0.4 0 100
numbers of multiple reaction monitoring (MRM) transitions. Gradient
3 15 0.4 0 100
4 16 0.4 100 0
For pesticide residue analysis in food, the European Commission’s
5 20 0.4 100 0
General-Directorate for Health and Food Safety (DG SANTE) has
published a guidance document on analytical quality control, Injection
10 µL
which details the performance requirements for the validation of Volume
analytical methods which should be followed by laboratories. The Column oven 40 °C
Temperature
most recent guidance was published under SANTE/12682/2019,3 Autosampler 10 °C
and includes procedures on acceptable retention time deviation,
Table 2. QSight ESI Source Parameters.
ion ratio (quantifier/qualifier ratio) range, and selectivity criteria to
ensure reliable quantification results. Ionization Mode ESI with Polarity Switching
Drying Gas 120
For sample preparation in the SANTE/12682/2019 method,
QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) is the HSID Temperature 200 °C
most commonly followed procedure and is based on extraction Nebulizer Gas 350
with acetonitrile, followed by a dispersive solid phase clean-up Spray Voltage 5000V / -4800V
step.4 As acetonitrile is compatible with LC/MS/MS, this approach
Source Temperature 340 °C
has emerged as the method of choice for pesticide analysis in
Detection Mode Time-managed MRM™
fruits and vegetables.5–7
In this application note, a fast, sensitive, and selective multi- Standard Preparation
residue method, utilizing a QSight® 220 triple quadrupole mass Pesticide standards were obtained from Lab Instruments (Bari, Italy).
detector coupled to the LX50 ultra-high performance liquid Repeatability and recovery were evaluated by fortifications of
chromatography (UHPLC) system, is described for the analysis pesticides prior to sample extraction at 10 μg/kg, 50 μg/kg and
of 503 pesticides in fruit and vegetable matrices. 100 μg/kg, measured in replicates (n=10). Calibration curves were
built by five levels of standards prepared in a neat solution
Experimental
(acetonitrile). Limit of detection (LOD) and limit of quantification
Hardware/Software (LOQ) values were determined based on signal-to-noise (S/N) values
Chromatographic separation of pesticides was conducted on a of 1:3 and 1:10, respectively. Two MRM transitions were monitored
PerkinElmer UHPLC system, with subsequent analyte determination for each pesticide. A procedural internal standard (IS), triphenyl
achieved utilizing a PerkinElmer QSight 220 triple quadrupole mass phosphate (TPP), was used for quantification of the obtained
detector with a dual ionization source. All instrument control, data results, accounting for various sources of errors throughout all
acquisition and data processing were performed using stages in the method, as well as possible matrix effects.
Simplicity 3Q™ software.
Sample Preparation
Instrumental Parameters Samples were obtained from local farms, and prepared according to
The UHPLC separation method and MS/MS source parameters are an established procedure (method EN 15662) using a QuEChERS
shown in Table 1 and Table 2, respectively. A full list of the multiple based extraction technique (PerkinElmer part number: N9306901),
reaction monitoring (MRM) mode transitions of the studied followed by a clean-up step (PerkinElmer part number: N9306920).
pesticides are shown in Table 3. The table includes both positive The latter, depending on the vegetable matrix to be analyzed, can
(460) and negative (43) analytes measured in ESI mode using fast include the use of 45 mg of porous graphitic carbon (PGC) for highly
polarity switching times to avoid losing data points. To facilitate pigmented fruit and vegetables. In short, 10 g of sample was
method development, the MS analysis parameters are homogenized and placed in a 50 ml tube. 100 µL of IS are added to
automatically generated by selecting the pesticides of interest from the tube to arrive at a final concentration of 50 μg/kg, accounting
the built-in compound library in the time-managed-MRM module for both extraction recoveries, as well as matrix effects in the mass
of the Simplicity 3Q software. Depending on expected peak width, spectrometry analysis. 10 mL of acetonitrile was added, and the
the cycle time of the instrument is set, and dwell times are solution was shaken and centrifuged. An aliquot of the supernatant
optimized accordingly by the built-in algorithm. was transferred to the clean-up tube, shaken and centrifuged. The
supernatant was transferred to a vial and 10 μL of extract was injected
directly onto the QSight 220 LC/MS/MS system for quantification.
24
Results
A total of 503 pesticides were investigated and analyzed in this influence the quality and precision of the measured data. In this
study (Table 3) and measured in three matrices - apple, orange and work, automated time managed MRM experiments were used, and
lettuce - to cover a range of both fruit and vegetable based samples an optimal dwell time calculation was automatically performed using
of varying composition and acidity. Orange was chosen as an acidic a software integrated algorithm to obtain 9-12 data points across
matrix, while apple as a neutral one for the group of fruits, and the peak for reliable and accurate quantification. Data points
lettuce as a vegetable representative. Figure 1 shows an example of selected amongst early, mid and late eluting peaks are shown in
overlayed MRM transitions for selected pesticides and their Figure 2 as representative examples for all measured analytes.
chromatographic separation. Limit of Detection
High resolution peak acquisition requires enough sampling points LOD and LOQ values were calculated by considering the signal of
across the peaks of interest in order to obtain a precise and quantifier ions of matrix-matched standards with signal-to-noise
reproducible result. MRM utilizes the retention time of measured ratio (S/N) > 3 and S/N > 10, respectively. The LOD values for 97%
compounds, and overlapping windows ensure that only the of the 503 measured pesticides are equal or below 1 ng/ml, as
compounds eluting at that time are measured. Dwell times of each detailed in Figure 3. The LOQ values for 90% of the 503 compounds
transition and the associated acquisition speed influence the data are equal or below 2 ng/ml, and thus easily meet the required MRL
points across the peak. Indeed, both factors are relevant and values as set by the EC 396/2005, as shown in Figure 4.
Figure 1. Example of overlayed peaks showing separation of described pesticides (Quantifier MRM) at 10 ng/ml in neat solution.
Figure 2. Representative examples for data points selected amongst peaks during early, mid and late eluting compounds in the chromatographic run.
Figure 3. Distribution of LOD for tested pesticides. 96 % of all 503 pesticides tested had Figure 4. Distribution of LOQ for tested pesticides. 80 % of all 503 pesticides tested had a
a LOD of equal or below 1 ppb with a S/N > 3. Only 1% of all pesticides had an LOD of LOQ of equal or below 1 ppb with a S/N > 10. Only 4% of all pesticides had an LOQ of
above 5 ppb. above 5 ppb.
35
Linearity QuEChERS-method” in foods of plant origin. An overlay of validated
The calibration curves for all compounds were created from their pesticides at 10 µg/kg in lettuce matrix is shown in Figure 6.
LOQ up to 100 ng/ml in neat solution with good linear correlation
SANTE/12682/2019 Method Performance
of R2>0.99 and 1/x weighing factor. TPP was used as the IS for all
For pesticide analysis in the EU, identification criteria in
pesticides. Selected examples are shown in Figure 5, which are
SANTE/12682/2019 requires the retention time and ion ratio from at
representative of results achieved for all the measured analytes.
least two MRM transitions per compound to be within acceptable
Method Validation tolerance limits. Two MRM transitions were monitored for each
Initially, a set of 144 pesticides were selected for method validation pesticide - one for the quantifier ion and one for the qualifier ion. The
utilizing the same matrices described in this work (apple, orange product ion ratios were monitored to ensure that they were within a
and lettuce) as for the total analysis of pesticides. This validation is 30% tolerance windows of the expected, when comparing standards
planned to be extended soon on a more extensive set of pesticides. in neat solution to pesticides in the studied matrix. Similarly, for
The method described here follows the UNI EN 15662:20188 norm, retention times, the difference between compounds in a neat solution
which entails a “multimethod for the determination of pesticide was compared to pesticides in the studied matrix, all of which were
residues using GC- and LC-based analysis following acetonitrile within the limit of 0.1 minutes for all pesticides monitored.
extraction/partitioning and clean-up by dispersive SPE - Modular
Figure 5. Representative examples for calibration curves (neat solution with IS TPP) for Alachlor, Dinoterb, Phosmet and Zoxamide, x-axis shows concentration in ng/ml, y-axis
shows peak area ratio of respective analyte and IS.
Figure 6. Method validation - overlay of 144 pesticides (Quantifier MRM) at 10 μg/kg in lettuce matrix.
46
Method Recovery and Reproducibility Intra-day reproducibility was calculated on n=10 replicate injections of
Method recovery in the three selected matrices at all spiking levels the three spiking levels and sample matrixes. A graph summarizing
(10, 50 and 100 μg/kg) for most validated pesticides fell within the the results is shown in Figure 8. At the 10 μg/kg spiking level in the
range of 70-120%, which is accepted by SANTE guidelines for lettuce, apple and orange samples, RSD values below 10% were
method validation. Detailed recovery values are illustrated in Figure 7. achieved for 77%, 71% and 67% of all validated pesticides,
In the apple matrix, the 10 μg/kg calculated values for selected respectively. At the 50 and 100 μg/kg spiking levels, values were
pesticides are both over- and under-estimated, possibly due to signal 98% and 99% in the lettuce sample, 82% and 98% in the apple
enhancement and suppression effects. To account for these and 95% and 98% in the orange sample. All of the validated
instances, a matrix-matched calibration could be done to further pesticides meet the SANTE guidelines, where reproducibility values
improve these results, as well as the use of an internal standard, as are defined to be ≤20%.
routinely employed in this method.
Figure 7. Distribution of pesticide recoveries (average of n=10 measurements) for percentage of total pesticides at three different spiking levels (10, 50 and 100 µg/kg) in three different
matrices (lettuce, apple and orange). Most of the compounds have recoveries in the required range of 70-120%, especially at 50 and 100 µg/kg.
Figure 8. Distribution of RSD (n=10) value ranges (0-10%, 10-20% and >20%) for percentage of total pesticides at three different spiking levels (10, 50 and 100 µg/kg) in three different
matrices (lettuce, apple and orange).
5 7
Figure 9. Positively identified and quantified pesticides in various fruit and vegetable samples at varying concentration, *values obtained through extrapolation. Number identify
the relative food matrix: 2794= lettuce; 2795=endive; 2760=black chickpeas; 2803=olives.
Tests on Other Food Matrices

As part of routine farming processes, different food matrices, such The targeted pesticides were determined with typical LOQs at or
as grapes, bananas, oranges, peaches, and apricots are typically below 1 ppb, which is well below the MRL set by the EC 396/2005.
sampled and analyzed for pesticides. In order to verify reliability of All pesticide calibration linearities are better than 0.99 in the
the developed method, several of these matrices were also testing working range of LOQ-100 ng/ml. SANTE guidelines were met for
utilizing the method described herein. Samples (including lettuce, the 144 validated pesticides in terms of recovery in the required
endive, black chickpeas and olives) were analyzed using the range of 70-120%, as well as RSD values below 20%, ion ratios
described method, and selected pesticides were positively identified within 30% tolerance windows, and retention time deviation
and quantified. Figure 9 displays chromatograms of common between neat and matrix matched solution less than 0.1 min.
pesticides found when routinely analyzing these types of samples. The QSight 220 LC/MS/MS system employed in this work provides a
robust and reliable platform for reproducible results in compliance
Conclusion with regulatory limits. Utilizing the PerkinElmer system, confirmation
In this application note, we present a single sample preparation and quantification of hundreds of pesticide analytes in fruit and
and development of a multi-residue pesticide analysis using vegetable matrixes was measured in positive and negative polarity
our QSight 220 triple quadrupole mass detector for rapid, switching mode, allowing for greater efficiency and higher
reliable results. This method is suitable for laboratories wanting to laboratory throughput. Further, the automated time-managed MRM
comply with the most stringent of pesticide regulations. The algorithms optimize dwell times for fast acquisition rates without
method described allows for the identification and quantification compromising data quality.
of 500+ pesticides in various food and vegetable matrices.
6
8
Table 3. MRM transition, collision energy (CC) and LOD/LOQ data for pesticides measured in this work (validated pesticides are marked in bold).
LOD LOQ
ESI Compound Parent Quantifier CE EV CCL2 Qualifier CE EV CCL2
(ng/ml) (ng/ml)
- 2,4,5-T 253 194.5 14 -10 40 158.5 42 -10 96 ≤0.5 ≤0.5
- 2,4,5-TP 267/269 195 23 -15 40 197 23 -16 48 ≤0.5 ≤2.0
- 2,4-D 219 160.8 15 -10 40 125.1 37 -10 72 ≤0.5 ≤0.5
- 2,4-DB 247/249 127 12 -39 36 129 12 -39 36 ≤0.5 ≤2.0
- 2,4-Dimethylaniline 122 107 -26 29 -28 77 -48 7 -28 ≤1.0 ≤5.0
+ 2,4-Dimethylphenylformamide 150 107 -28 28 -32 106 -43 25 -40 ≤0.5 ≤0.5
+ 3-Hydroxycarbofuran 238 163 -23 28 -48 181 -16 24 -36 ≤0.5 ≤0.5
+ 3-Phenoxybenzoic acid 215 171 -22 10 -40 153 -16 13 -44 ≤5 >5.0
+ 4,4-Dichlorobenzophenone 250.9 139 -31 21 -48 111 -59 7 -80 ≤5 >5.0
+ Acephate 184.1 143 -21 8 -40 125 -35 8 -62 ≤1.0 ≤5.0
+ Acetamiprid 223.2 126.1 -25 10 -52 73 -87 10 -100 ≤0.5 ≤0.5
+ Acetochlor 270 148 -31 31 -48 133.1 -54 29 -72 ≤0.5 ≤1.0
+ Acibenzolar-s-Methyl 211 136 -42 20 -41 140 -36 11 -35 ≤0.5 ≤2.0
- Acifluorfen 359.9 316 15 -30 52 286 23 -29 64 ≤1.0 ≤5.0
+ Aclonifen 265.1 248.1 -25 20 -68 182.1 -39 20 -56 ≤0.5 ≤0.5
+ Acrinathrin 559.3 181.2 -63 20 -88 208.2 -23 20 -80 ≤0.5 ≤0.5
+ Alachlor 270 162 -29 25 -56 238 -16 25 -44 ≤0.5 ≤0.5
+ Aldicarb 208.2 116.1 -12 10 -32 89.1 -28 10 -36 ≤0.5 ≤1.0
+ Aldicarb-Sulfone 223.1 148 -15 25 -78 86 -15 26 -37 ≤0.5 ≤2.0
+ Aldicarb-Sulfoxide 207.1 132.1 -13 25 -32 89.1 -19 25 -38 ≤0.5 ≤0.5
+ Allethrin 303.1 135 -26 10 -44 91 -51 18 -52 ≤1.0 ≤5.0
+ Allidochlor 174 98 -18 24 -32 41 -51 24 -36 ≤0.5 ≤1.0
+ Ametoctradin 276 149 -50 4 -80 176 -50 2 -76 ≤0.5 ≤0.5
+ Ametryn 228 186 -25 30 -50 68 -64 31 -48 ≤0.5 ≤0.5
+ Aminocarb 209 137 -24 10 -44 152 -10 10 -44 ≤0.5 ≤0.5
+ Amisulbrom 466/468 227 -40 15 -76 229 -32 4 -84 ≤0.5 ≤0.5
+ Amitraz 294 122 -45 12 -60 163 -26 33 -48 ≤0.5 ≤2.0
+ Amitrole 85 58 -29 27 -24 43 -34 36 -20 ≤0.5 ≤1.0
+ Ancymidol 257 81 -42 29 -48 135 -37 26 -56 ≤0.5 ≤0.5
- Anilazine 273/275 35 120 -25 92 35 125 -29 96 ≤0.5 ≤2.0
+ Atraton 212 170 -25 34 -36 100 -39 32 -52 ≤0.5 ≤0.5
+ Atrazine 216 174 -23 36 -40 176 -24 35 -36 ≤0.5 ≤0.5
+ Avermectin 890.8 145.1 -60 25 -128 305.3 -40 25 -128 ≤5.0 >5.0
+ Azadiractin 743.5 725.5 -35 52 -140 665.5 -48 39 -156 ≤1.0 ≤5.0
+ Azamethiphos 325 139 -28 16 -52 183 -14 26 -56 ≤0.5 ≤0.5
+ Azinphos-Methyl 318.1 124.9 -27 10 -26 260.8 -12 10 -11 ≤0.5 ≤2.0
+ Azoxystrobin 404.1 344.1 -35 25 -71 372.1 -19 25 -57 ≤0.5 ≤0.5
+ Benalaxyl 326.2 91.1 -15 25 -46 294 -10 25 -42 ≤0.5 ≤0.5
+ Bendiocarb 224 167 -30 25 -65 109 -35 25 -72 ≤0.5 ≤1.0
+ Benodanil 324 203 -35 45 -76 76 -107 38 -152 ≤0.5 ≤0.5
+ Benoxacor 260/262 149 -28 30 -52 149 -25 31 -48 ≤0.5 ≤1.0
+ Bensulfuron-Methyl 411 119 -51 1 -68 149 -50 3 -92 ≤0.5 ≤0.5
+ Bensulide 398.1 158 -30 16 -64 77 -72 15 -76 ≤0.5 ≤0.5
+ Bensultap 432 150 -44 32 -76 290 -24 32 -72 ≤0.5 ≤0.5
- Bentazone 239.1 174.8 24 -22 64 132.1 35 -30 68 ≤0.5 ≤0.5
79
Table 3. continued...
LOD LOQ
(ng/ml) (ng/ml)
+ Bentiavalicarb Isopropyl 382.1 196.8 -16 10 -68 179.9 -30 10 -68 ≤0.5 ≤0.5
+ Benzoximate 364.1 105 -53 1 -64 198.9 -19 7 -52 ≤0.5 ≤0.5
- Bialaphos 322 94 41 -38 60 134.2 41 -11 64 ≤1.0 ≤5.0
+ Bifenazate 301.2 170.1 -27 8 -52 198.1 -14 6 -44 ≤0.5 ≤0.5
+ Bifenthrin 440.4 165.2 -116 20 -128 181.2 -38 20 -64 ≤0.5 ≤2.0
+ Bispyribac Sodium 431 413 -40 7 -65 275 -30 19 -60 ≤0.5 ≤0.5
+ Bitertanol 338 269 -13 25 -45 70 -46 25 -75 ≤0.5 ≤1.0
+ Bixafen 414 394 -10 32 -12 266 -29 14 -72 ≤0.5 ≤0.5
+ Boscalid 343.1 140 -29 25 -28 307 -26 25 -25 ≤0.5 ≤0.5
+ Bromacil 261 188 -40 25 -62 205 -20 25 -44 ≤0.5 ≤1.0
- Bromadiolone 525/527 250 46 -60 116 250 47 -60 120 ≤0.5 ≤0.5
- Bromoxynil 276 81 78 -25 36 79 88 -25 45 ≤0.5 ≤0.5
+ Bromuconazole 376 70 -29 25 -28 159 -6 25 -5 ≤0.5 ≤2.0
+ Bupirimate 317 108 -38 3 -60 166 -32 5 -60 ≤0.5 ≤0.5
+ Buprofezin 306.1 145 -24 10 -52 201 -18 10 -52 ≤0.5 ≤0.5
+ Butachlor 312.1 238.1 -18 7 -44 57.1 -55 26 -60 ≤0.5 ≤0.5
+ Butocarboxim 208 166 -21 31 -36 72 -34 32 -40 ≤0.5 ≤0.5
+ Butoxycarboxim 240 166 -17 8 -36 106 -18 5 -44 ≤0.5 ≤0.5
+ Butralin 296 222 -29 20 -64 240 -20 10 -48 ≤0.5 ≤0.5
+ Butylate 218 156 -25 25 -45 57 -60 16 -68 ≤1.0 ≤5.0
+ Cadusafos 271.1 131.1 -30 25 -54 158.9 -18 25 -43 ≤0.5 ≤0.5
+ Cafenstrole 351 100 -34 15 -52 72 -55 14 -60 ≤0.5 ≤0.5
+ Carbaryl 202 145 -26 8 -32 127 -41 6 -40 ≤0.5 ≤0.5
+ Carbendazim 192.2 132.1 -51 25 -86 160 -56 18 -40 ≤0.5 ≤0.5
+ Carbofuran 222.2 123 -46 35 -90 165 -33 30 -65 ≤0.5 ≤0.5
+ Carbophenothion 343 199 -7 21 -6 157 -14 25 -6 ≤0.5 ≤0.5
+ Carboxin 236 93 -77 26 -116 87 -67 16 -36 ≤0.5 ≤2.0
+ Carfentrazone-Ethyl 412 346 -28 25 -66 366 -22 25 -61 ≤0.5 ≤0.5
+ Chlorantraniliprole 482/484 450.8 -28 24 -76 452.8 -26 15 -88 ≤0.5 ≤0.5
+ Chlorbromuron 292.8 204 -26 31 -56 182 -26 30 -56 ≤0.5 ≤0.5
- Chlordecone 506.7 35 135 -50 96 426.6 24 -50 108 ≤0.5 ≤1.0
+ Chlorflurazuron 539.8 383 -29 43 -100 158 -31 40 -96 ≤0.5 ≤0.5
+ Chloridazon 222.1 104 -51 10 -300 77.1 -110 42 -76 ≤0.5 ≤0.5
+ Chlormequat 122 63 -29 28 -24 58.1 -42 31 -28 ≤0.5 ≤0.5
+ Chlorotoluron 213/215 142 -31 29 -48 72 -43 31 -48 ≤0.5 ≤0.5
+ Chloroxuron 291 218 -18 25 -45 72 -31 18 -84 ≤0.5 ≤0.5
+ Chlorpyrifos-Ethyl 352 97 -102 29 -252 200 -74 16 -164 ≤0.5 ≤1.0
+ Chlorpyrifos-Oxon 335.9 308 -16 27 -52 280 -24 29 -64 ≤0.5 ≤0.5
+ Chlorsulfuron 358 167 -28 9 -56 141 -31 11 -56 ≤0.5 ≤0.5
+ Chlorthiamid 206/208 188.9 -18 3 -40 191 -24 10 -44 >5.0 >5.0
+ Chlorthiophos 361 333 -17 20 -56 305 -22 18 -64 ≤0.5 ≤1.0
+ Chlothianidin 250 132 -70 7 -70 169.1 -54 7 -60 >5.0 >5.0
+ Cinidon-Ethyl 394 348 -35 31 -100 107 -41 31 -60 ≤0.5 ≤2.0
+ Cinosulfuron 414 215 -23 20 -64 183 -26 20 -64 ≤0.5 ≤0.5
- Clethodim 358 238 39 -23 64 266 25 -23 60 ≤0.5 ≤0.5
108
LOD LOQ
(ng/ml) (ng/ml)
+ Clodinafop-Propargyl 349.9 91 -39 18 -64 266 -23 19 -56 ≤0.5 ≤0.5
+ Clofentezine 303.1 102 -62 5 -80 137.9 -24 10 -48 ≤0.5 ≤0.5
+ Clomazone 242.1 127 -48 16 -60 125 -45 10 -48 ≤0.5 ≤0.5
+ Clopyralid 192 174.1 -15 10 -36 146.1 -28 10 -60 ≤5 >5.0
+ Cloquintocet-mexyl 336 192 -44 20 -72 238 -25 20 -60 ≤0.5 ≤0.5
+ Coumaphos 363 307 -32 26 -188 227 -50 8 -64 ≤0.5 ≤2.0
+ Crimidine 172 107 -37 21 -44 136 -28 36 -36 ≤0.5 ≤2.0
+ Crotoxyphos 332 127 -54 15 -68 105 -95 5 -128 ≤0.5 ≤0.5
+ Crufomate 292 108 -35 40 -60 236 -23 40 -60 ≤0.5 ≤0.5
+ Cyanazine 241 96 -22 2 -68 214 -13 6 -48 ≤0.5 ≤0.5
+ Cyanophos 244 125 -31 20 -44 212 -22 20 -48 ≤1.0 ≤5.0
+ Cyantraniliprole 475 444 -36 25 -76 286 -33 25 -88 ≤0.5 ≤1.0
+ Cyazofamid 325/327 108 -24 17 -48 108 -27 5 -40 ≤0.5 ≤0.5
+ Cycloate 216 154 -16 10 -44 83 -23 9 -40 ≤0.5 ≤0.5
+ Cycloheximide 282.2 265.2 -18 36 -44 247.2 -20 31 -44 ≤3 >5.0
+ Cycloxydim 326 180 -48 34 -85 280 -20 16 -92 ≤0.5 ≤0.5
+ Cycluron 199.3 89.1 -45 3 -60 72 -57 7 -24 ≤0.5 ≤0.5
+ Cyflufenamide 413 241 -25 13 -72 295 -8 28 -76 ≤0.5 ≤2.0
+ Cymiazole 219 144 -42 37 -64 171 -36 5 -48 ≤0.5 ≤0.5
+ Cymoxanil 199 111 -29 6 -40 128 -18 8 -32 ≤0.5 ≤1.0
+ Cyproconazole 292 70 -73 6 -68 125 -60 6 -44 ≤0.5 ≤0.5
+ Cyprodinil 226 77 -67 45 -48 93 -45 36 -60 ≤0.5 ≤0.5
+ Cyprosulfamide 375 254 -20 36 -56 135 -51 38 -72 ≤0.5 ≤0.5
+ Cyromazine 167 85 -31 25 -44 68 -31 25 -44 ≤0.5 ≤0.5
+ Daimuron 269.2 119 -33 24 -44 151 -20 28 -40 ≤0.5 ≤0.5
+ Daminozide 161 44 -65 14 -52 143 -15 5 -32 ≤0.5 ≤0.5
+ Dazomet 163 90 -14 6 -32 120 -16 17 -28 ≤0.5 ≤0.5
+ Deet 192.1 119.1 -25 31 -36 91.1 -41 33 -44 ≤0.5 ≤0.5
+ Demeteon-S-methylsulfoxide 247 125 -45 25 -80 169 -41 25 -80 ≤0.5 ≤2
+ Demeton-S 259 89 -40 6 -52 61 -96 11 -124 ≤0.5 ≤0.5
+ Demeton-S-methyl 231 61 -43 4 -36 89 -20 7 -32 ≤0.5 ≤2
+ Demeton-S-Methylsulfone 263 120.8 -22 20 -44 168.9 -22 19 -48 ≤0.5 ≤0.5
+ Desmedipham 301.2 182.1 -37 22 -88 136.1 -46 27 -90 ≤0.5 ≤0.5
+ Dialifos 394 187 -15 6 -60 208 -38 5 -60 ≤0.5 ≤0.5
+ Di-Allate 269.9 109 -48 8 -72 86 -21 6 -44 ≤0.5 ≤0.5
+ Diazinon 305 97 -130 22 -136 169 -59 22 -136 ≤0.5 ≤2.0
+ Dichlofenthion 315 259 -19 74 -62 287 -10 24 -54 ≤0.5 ≤0.5
+ Dichlormid 208 98 -23 20 -36 140 -16 27 -36 ≤1.0 ≤5.0
+ Dichlorophenylmethylurea 221 164 -22 25 -45 162 -22 25 -45 ≤0.5 ≤0.5
+ Dichlorophenylurea 205/207 162 -20 23 -44 163.9 -22 25 -45 ≤0.5 ≤2.0
- Dichlorprop 233 125 43 -10 36 161 25 -10 40 ≤0.5 ≤1.0
+ Dichlorvos 221 79 -38 25 -56 109.1 -22 25 -42 ≤0.5 ≤2.0
+ Dicrotophos 238 127 -54 16 -76 112 -48 25 -80 ≤0.5 ≤0.5
+ Diethofencarb 268 180 -25 25 -48 226 -16 25 -40 ≤0.5 ≤0.5
+ Difenoconazole 406.2 337 -23 19 -22 251.1 -37 25 -36 ≤0.5 ≤0.5
911
LOD LOQ
(ng/ml) (ng/ml)
+ Difenoxuron 287.2 72 -51 2 -60 123.2 -41 3 -56 ≤0.5 ≤0.5
+ Difenzoquat 249 130 -55 23 -60 193 -37 29 -56 ≤0.5 ≤0.5
+ Diflubenzuron 311 141.2 -67 7 -108 158.2 -19 10 -44 ≤0.5 ≤0.5
+ Dimefuron 339 256 -21 28 -56 167 -29 24 -56 ≤0.5 ≤0.5
+ Dimepiperate 264 119 -32 25 -55 146 -26 2 -132 ≤0.5 ≤0.5
+ Dimethachlor 256 148 -35 7 -60 224 -21 23 -44 ≤0.5 ≤0.5
+ Dimethenamid 276 168 -31 12 -64 244 -17 5 -48 ≤0.5 ≤0.5
+ Dimethoate 230.1 125 -33 25 -52 199.1 -13 25 -34 ≤0.5 ≤0.5
+ Dimethomorph 388.2 165.1 -41 25 -40 301.1 -27 25 -26 ≤0.5 ≤0.5
+ Dimetilan 241.1 196 -12 22 -36 72 -58 5 -64 ≤0.5 ≤0.5
+ Dimoxystrobin 327.1 205 -18 22 -56 116 -53 24 -68 ≤0.5 ≤0.5
+ Diniconazole 326 159 -41 5 -84 70 -29 34 -56 ≤0.5 ≤0.5
+ Dinitramine 323.1 194.9 -53 15 -100 289.3 -25 9 -64 ≤1.0 ≤5.0
+ Dinocap 382.1 116 -30 6 -80 180 -54 10 -96 ≤0.5 ≤0.5
- Dinoseb 239.1 192.5 31 -20 84 134.1 62 -20 96 ≤0.5 ≤0.5
- Dinoterb 239.1 206.5 33 -38 84 175.7 49 -37 108 ≤0.5 ≤0.5
+ Dioxacarb 224 123 -23 11 -36 167 -13 8 -32 ≤0.5 ≤0.5
+ Diphenamid 240.1 91 -59 25 -52 134 -26 29 -48 ≤0.5 ≤0.5
+ Disulfoton 275 121 -39 10 -56 233 -18 8 -44 ≤0.5 ≤2.0
+ Disulfoton-Sulfone 307 171 -50 20 -100 153 -60 22 -108 ≤0.5 ≤1.0
+ Disulfoton-Sulfoxide 291.1 212.9 -6 10 -48 185 -9 9 -4 ≤0.5 ≤0.5
+ Ditalimfos 300.1 130.1 -66 12 -40 148.1 -33 10 -28 ≤0.5 ≤0.5
- Dithianon 295.8 220 35 -20 92 238 25 -6 64 ≤0.5 ≤2.0
+ Dithiopyr 402 272 -41 20 -100 354 -25 36 -64 ≤0.5 ≤0.5
+ Diuron 233 160 -37 1 -68 72 -48 2 -56 ≤0.5 ≤0.5
+ DMST 215 79.1 -55 25 -71 106 -20 25 -40 ≤0.5 ≤1.0
- DNOC 197 122.1 39 -35 64 137 24 -35 36 ≤0.5 ≤0.5
+ Dodemorph 282.8 98 -39 32 -48 116 -27 39 -48 ≤0.5 ≤0.5
+ Dodine 228.3 60 -31 46 -44 57 -33 46 -40 ≤0.5 ≤0.5
+ Doramectin 916.4 593.3 -20 20 -20 331 -45 20 -136 ≤1.0 ≤5.0
+ Emamectin-Benzoate bA 886.5 126.3 -82 47 -172 158.6 -46 22 -232 ≤1.0 ≤5.0
+ Emamectin-Benzoate bB 873/874 158 -59 27 -148 158 -67 21 -164 ≤1.0 ≤5.0
+ Epoxiconazole 330 101 -124 14 -100 121 -64 24 -84 ≤0.5 ≤0.5
+ Eprinomectin Ba 936 490.2 -71 87 -240 352.2 -77 92 -252 ≤1.0 ≤5.0
+ EPTC 190 86 -19 17 -32 128 -15 20 -32 ≤5 >5.0
+ Esprocarb 266 71 -21 5 -40 91 -52 22 -48 ≤0.5 ≤0.5
+ Etaconazole 328.2 123 -69 26 -52 159.1 -34 26 -52 ≤0.5 ≤0.5
+ Ethidimuron 265.1 113.9 -46 25 -80 208 -31 25 -50 ≤0.5 ≤0.5
+ Ethiofencarb 226.2 164 -13 25 -33 107 -31 25 -50 ≤0.5 ≤0.5
+ Ethion 385 171 -26 9 -52 199 -15 7 -56 ≤0.5 ≤0.5
+ Ethiprole 397.1 255.5 -48 2 -84 351 -22 1 -68 ≤0.5 ≤0.5
+ Ethirimol 210 98 -62 40 -32 140 -43 29 -92 ≤0.5 ≤0.5
+ Ethofumesate 287.1 259 -12 30 -44 121 -19 32 -44 ≤0.5 ≤1.0
+ Ethoprofos 243 97 -44 12 -100 131 -68 17 -88 ≤0.5 ≤0.5
+ Ethoxyquin 218.1 160.1 -50 42 -56 174.1 -37 40 -60 ≤0.5 ≤2.0
1210
LOD LOQ
(ng/ml) (ng/ml)
+ Etoxazole 360.1 304 -24 5 -60 141 -44 5 -68 ≤0.5 ≤0.5
+ Famoxadone 392 238 -24 14 -56 331 -15 10 -60 ≤0.5 ≤1
+ Famphur 325.9 93 -46 35 -56 217 -29 32 -60 ≤0.5 ≤0.5
+ Fenamidone 312 65 -18 25 -47 92 -40 25 -62 ≤5.0 >5.0
+ Fenamiphos 304 202 -85 10 -72 217 -52 14 -40 ≤0.5 ≤0.5
+ Fenamiphos-Sulfone 336 188 -34 24 -64 266 -28 28 -60 ≤0.5 ≤0.5
+ Fenamiphos-Sulfoxide 320 171 -41 26 -84 108 -65 28 -100 ≤0.5 ≤0.5
+ Fenamiphos-Sulphone 336 188 -36 22 -72 266 -27 20 -60 ≤0.5 ≤0.5
+ Fenazaquin 307 147 -28 10 -27 161 -23 10 -22 ≤0.5 ≤0.5
+ Fenbuconazole 337 70 -108 8 -92 125 -119 8 -52 ≤0.5 ≤1.0
+ Fenbutatinoxid 517/519 461 -34 66 -100 463 -33 60 -88 ≤0.5 ≤0.5
+ Fenfuram 202 120 -22 30 -36 109 -34 31 -44 ≤0.5 ≤0.5
+ Fenhexamid 302 97 -32 26 -56 55 -70 28 -68 ≤0.5 ≤0.5
+ Fenobucarb 208 152 -12 25 -32 95 -19 25 -38 ≤0.5 ≤0.5
+ Fenothiocarb 254 107 -35 25 -57 160 -14 25 -38 ≤0.5 ≤0.5
+ Fenoxanil 329/331 302 -18 23 -52 304 -19 21 -56 ≤0.5 ≤0.5
+ Fenoxaprop 334.1 65.1 -66 29 -104 288 -21 35 -64 ≤0.5 ≤2.0
+ Fenoxaprop-P-Ethyl 362 121 -41 25 -72 288 -23 25 -56 ≤0.5 ≤0.5
+ Fenoxycarb 302 116 -16 25 -44 88 -35 25 -61 ≤0.5 ≤0.5
+ Fenpiclonil 236.9 140 -53 28 -72 202 -29 29 -60 ≤0.5 ≤1.0
+ Fenpropathrin 350.1 97 -24 3 -4 125 -11 8 -52 ≤1.0 ≤5.0
+ Fenpropidin 274 117 -61 5 -60 147 -30 4 -80 ≤0.5 ≤0.5
+ Fenpropimorph 304 117 -79 45 -78 147 -38 43 -37 ≤0.5 ≤0.5
+ Fenpyroximate 422.2 135.1 -47 26 -76 366.1 -25 15 -68 ≤0.5 ≤0.5
+ Fensulfothion 309 173 -25 5 -60 157 -25 3 -64 ≤0.5 ≤0.5
+ Fensulfothion-oxon 293 265 -14 27 -84 237 -24 32 -40 ≤0.5 ≤0.5
+ Fensulfothion-Sulfone 325 269 -16 25 -58 191 -26 25 -62 ≤0.5 ≤1.0
+ Fenthion 279 169 -22 25 -48 247 -15 25 -41 ≤0.5 ≤0.5
+ Fenthion-Sulfoxide 295.1 279.7 -41 25 -84 108.9 -38 25 -67 ≤0.5 ≤0.5
+ Fenuron 165.1 46.1 -19 2 -24 72 -29 3 -28 ≤0.5 ≤0.5
- Fipronil-Sulfone 450.8 414.9 23 -33 76 282 37 -34 88 ≤0.5 ≤0.5
+ Flamprop-isopropyl 364 105 -22 12 -56 77 -63 6 -56 ≤0.5 ≤0.5
+ Flamprop-M-Isopropyl 364 77 -113 8 -128 105 -44 3 -56 ≤0.5 ≤0.5
+ Flonicamid 230.1 174 -20 25 -41 203.1 -20 25 -41 ≤1.0 ≤5.0
+ Florasulam 360 192 -24 20 -56 129 -66 19 -80 ≤0.5 ≤0.5
- Fluazifop 326 225.3 36 -11 104 253.2 17 -11 60 ≤5.0 >5.0
+ Fluazifop-Butyl 384 328.1 -23 5 -60 282 -28 6 -72 ≤0.5 ≤0.5
+ Fluazifop-P-Butyl 384 328 -23 38 -64 282 -27 38 -64 ≤0.5 ≤0.5
- Fluazinam 463 398 25 -20 92 416 25 -20 104 ≤0.5 ≤0.5
- Flubendiamide 680.8 274 23 -46 104 254 37 -48 100 ≤0.5 ≤0.5
+ Flubenzimine 417 77 -82 9 -78 397 -17 9 -66 ≤0.5 ≤0.5
+ Flucarbazone Sodium 419 137 -44 53 -56 152 -23 50 -60 ≤1.0 ≤5.0
- Fludioxonil 247.1 126.1 42 -32 60 180 41 -31 56 ≤0.5 ≤0.5
+ Flufenacet 364.1 152.2 -19 9 -52 194.1 -9 2 -60 ≤0.5 ≤0.5
+ Flufenoxuron 489.1 141.2 -95 26 -128 158.2 -31 13 -80 ≤0.5 ≤0.5
1113
LOD LOQ
(ng/ml) (ng/ml)
+ Flumetralin 422 107 -120 12 -136 143 -66 16 -72 ≤0.5 ≤0.5
+ Flumetsulam 326 109 -80 34 -88 129 -35 32 -60 ≤0.5 ≤0.5
+ Fluopicolide 385 147 -51 25 -84 175 -31 25 -66 ≤0.5 ≤0.5
+ Fluopyram 397 173 -72 19 -68 208 -89 15 -116 ≤0.5 ≤2.0
+ Fluorochloidone 313 269 -47 45 -96 270 -31 45 -60 ≤0.5 ≤0.5
+ Fluorodifen 329 99 -76 3 -88 125 -49 19 -56 ≤0.5 ≤0.5
+ Fluoxastrobin 459.1 188.1 -37 25 -79 427.1 -18 25 -62 ≤0.5 ≤0.5
+ Fluquinconazole 376 307 -34 25 -68 349 -26 25 -61 ≤0.5 ≤0.5
+ Fluridone 330.1 259 -56 19 -172 310 -31 15 -124 ≤0.5 ≤1.0
+ Flurochloridone 312 145 -65 32 -72 292 -30 22 -56 ≤0.5 ≤1.0
+ Fluroxypyr 255 179 -42 11 -92 181 -31 13 -80 ≤0.5 ≤0.5
+ Fluroxypyr Mepthyl 367 209 -18 5 -8 255 -6 8 -4 ≤0.5 ≤2.0
+ Flurtamone 334 178 -65 34 -64 247 -37 24 -128 ≤0.5 ≤0.5
+ Flusilazole 316 165 -33 25 -60 247 -25 25 -53 ≤0.5 ≤0.5
+ Flutolanil 324 242 -28 8 -64 262 -17 9 -56 ≤0.5 ≤0.5
+ Flutriafol 302 95 -73 10 -72 123 -42 25 -41 ≤0.5 ≤1.0
+ Fluxapyroxad 382 362 -20 22 -80 342 -31 27 -80 ≤0.5 ≤0.5
- Fomesafen 436.9 286 31 -42 76 195 63 -46 112 ≤0.5 ≤0.5
+ Fonofos 247 137 -51 1 -56 109 -30 5 -36 ≤0.5 ≤1.0
- Forchlorfenuron 246/248 127 21 -24 36 129 26 -24 44 ≤0.5 ≤0.5
+ Formentanate 222.2 120 -41 25 -58 165 -26 25 -45 ≤1.0 ≤5.0
+ Fosthiazate 284.1 104 -74 5 -40 227.9 -37 5 -88 ≤0.5 ≤2.0
+ Fuberidazole 185.1 156 -38 3 -64 65 -51 1 -36 ≤0.5 ≤0.5
+ Furathiocarb 383 252 -18 24 -17 195 -26 10 -25 ≤0.5 ≤0.5
+ Griseofulvin 353 215 -27 37 -56 165 -36 34 -68 ≤0.5 ≤0.5
+ Halofenozide 331.1 275 -19 25 -49 104.9 -26 25 -56 ≤0.5 ≤1.0
+ Halosulfuron-Methyl 435 100 -85 17 -116 83 -98 9 -116 ≤0.5 ≤1.0
+ Haloxyfop 362.1 91 -39 31 -68 315.9 -22 33 -56 ≤0.5 ≤1.0
+ Haloxyfop-Ethotyl 434 288 -74 10 -236 316 -20 10 -40 ≤0.5 ≤0.5
+ Haloxyfop-Methyl 376.1 90.9 -65 9 -56 316 -36 5 -60 ≤0.5 ≤1.0
+ Heptenophos 251 109 -70 14 -132 127 -56 14 -132 ≤0.5 ≤2.0
+ Hexaconazole 314 159 -90 9 -280 70 -94 36 -128 ≤0.5 ≤0.5
- Hexaflumuron 459/461 174.7 56 -15 108 176.7 56 -18 120 ≤0.5 ≤0.5
+ Hexaninone 253 171 -13 1 -52 71 -35 16 -48 ≤0.5 ≤0.5
+ Hexythyazox 353.1 168 -38 9 -68 227.7 -20 9 -52 ≤0.5 ≤0.5
+ Imazalil 297 201 -24 4 -52 159 -30 24 -56 ≤0.5 ≤0.5
+ Imazamethabenz-Methyl 289.2 89 -114 30 -108 144 -79 29 -48 ≤0.5 ≤0.5
- Imazamox 304 260 18 -10 48 217 31 -10 48 ≤0.5 ≤0.5
+ Imazapyr 262.2 202.2 -34 2 -64 217.2 -26 2 -64 ≤0.5 ≤0.5
+ Imazaquin 312 224 -28 32 -168 267 -20 12 -68 ≤0.5 ≤1.0
+ Imazethapyr 290.2 86.1 -51 15 -32 177 -36 4 -60 ≤0.5 ≤0.5
+ Imazosulfuron 413 156 -31 20 -64 153 -17 10 -60 ≤0.5 ≤2.0
+ Imibenconazole 411 171 -27 39 -64 125 -55 39 -68 ≤0.5 ≤0.5
+ Imidacloprid 256.1 175 -29 13 -52 209 -21 13 -44 ≤0.5 ≤1.0
+ Imidacloprid Impurity 381 300 -32 15 -60 126 -57 5 -72 ≤0.5 ≤0.5
1412
LOD LOQ
(ng/ml) (ng/ml)
+ Indanofan 341 187 -20 20 -48 175 -33 20 -52 ≤0.5 ≤0.5
+ Indoxacarb 528 56 -68 22 -67 203 -55 24 -54 ≤0.5 ≤1.0
+ Iodosulfuron Methyl Sodium 507.9 141 -55 20 -80 167 -35 20 -80 ≤0.5 ≤2.0
- Ioxynil 369.8 215 41 -31 80 127 43 -25 84 ≤0.5 ≤0.5
- Ioxynil Methyl 385 179 30 -50 76 155 39 -41 72 ≤0.5 ≤1.0
+ Ipconazole 334 125 -45 42 -60 70 -24 7 -56 ≤0.5 ≤0.5
+ Iprobenfos 306 145 -69 55 -112 91 -60 28 -52 ≤0.5 ≤2.0
+ Iprovalicarb 321.3 203.1 -13 10 -12 119 -45 10 -44 ≤0.5 ≤0.5
+ Isazofos 314 162 -21 58 -54 120 -31 38 -54 ≤0.5 ≤0.5
+ Isocarbamid 186 130 -16 19 -36 87.1 -23 22 -36 ≤0.5 ≤0.5
+ Isocarbophos 291 213 -13 15 -44 185 -19 15 -48 ≤0.5 ≤0.5
+ Isofenphos-Methyl 332 273 -10 5 -48 231 -25 5 -52 ≤0.5 ≤0.5
+ Isoprocarb 194 95 -14 19 -36 137 -6 9 -36 ≤0.5 ≤1.0
+ Isoprothiolane 291 189 -34 9 -64 231 -16 9 -48 ≤0.5 ≤0.5
+ Isoproturon 207.2 165.2 -36 3 -56 72 -61 4 -76 ≤0.5 ≤0.5
+ Isopyrazam 360 244 -34 40 -64 340 -23 38 -60 ≤0.5 ≤0.5
+ Isoxaben 333.3 107.1 -120 26 -88 165.1 -79 25 -116 ≤0.5 ≤0.5
+ Isoxaflutole 360.1 220.1 -51 25 -64 250.9 -28 25 -56 ≤0.5 ≤1.0
+ Isoxathion 314 170 -20 18 -52 105 -25 5 -48 ≤0.5 ≤0.5
+ Kresoxim-Methyl 314.1 222 -8 4 -52 235 -12 12 -40 ≤0.5 ≤2.0
+ Lenacil 235 136 -49 25 -88 153 -37 25 -70 ≤0.5 ≤0.5
+ Leptophos 412.8 77 -74 10 -72 171 -46 5 -68 ≤0.5 ≤2.0
+ Linuron 249 182 -21 12 -52 159.9 -28 12 -40 ≤0.5 ≤0.5
- Lufenuron 509/511 325.9 28 -27 80 327.9 31 -20 84 ≤0.5 ≤0.5
+ Malaoxon 315.1 99.1 -47 10 -46 127.1 -18 10 -17 ≤0.5 ≤0.5
+ Malathion 331.2 99 -24 25 -55 127 -22 25 -53 ≤0.5 ≤0.5
- Maleic Hydrazide 111 82 24 -20 24 42 69 -27 28 ≤1.0 ≤5.0
+ Mandipropamid 412.2 125 -60 11 -80 328.1 -19 11 -64 ≤0.5 ≤0.5
- MCPA- 199/201 141 18 -11 36 143 80 -11 36 ≤0.5 ≤0.5
- MCPB 227/229 141 30 -10 40 143 29 -10 52 ≤0.5 ≤2.0
- MCPP- 213.1 64 74 -10 36 141 16 -10 40 ≤0.5 ≤0.5
+ Mecarbam 330 97 -45 25 -78 227 -30 25 -70 ≤0.5 ≤0.5
+ Mefenacet 299 120 -31 2 -50 148 -15 2 -46 ≤0.5 ≤0.5
+ Mefluidide 311 121 -52 30 -80 135 -36 25 -60 ≤0.5 ≤0.5
+ Mepanipyrim 224 77 -107 14 -84 106 -57 13 -44 ≤0.5 ≤0.5
+ Mepiquat 114.2 58.1 -37 6 -28 98.1 -38 20 -28 ≤0.5 ≤0.5
+ Mepronil 270 91 -84 20 -44 119 -68 10 -44 ≤0.5 ≤0.5
- Meptyldinocap 295.1 192.6 39 -38 112 134.1 73 -41 128 ≤0.5 ≤0.5
+ Merphos 299 153 -22 11 -52 243 -19 22 -48 ≤0.5 ≤0.5
+ Mesosulfuron-Methyl 504 139 -85 30 -108 182 -38 21 -80 ≤0.5 ≤0.5
+ Mesotrione 340.3 104.1 -42 31 -56 227.8 -28 31 -68 ≤0.5 ≤2.0
+ Metaflumizone 507 287 -42 17 -344 178 -59 18 -104 ≤0.5 ≤2.0
+ Metalaxyl 280.2 192.3 -25 5 -44 220.2 -18 5 -48 ≤0.5 ≤0.5
+ Metalaxyl-M 280 160 -39 22 -64 220 -19 22 -52 ≤0.5 ≤0.5
+ Metamitron 203.1 104.2 -50 26 -32 175 -30 24 -48 ≤0.5 ≤0.5
1315
LOD LOQ
(ng/ml) (ng/ml)
+ Metazechlor 278 210 -41 10 -76 134 -88 10 -112 ≤1.0 ≤5.0
+ Metconazole 320 125 -107 18 -76 70 -104 28 -104 ≤0.5 ≤1.0
+ Methabenzthiazuron 222 150 -84 9 -40 165 -53 9 -68 ≤0.5 ≤2.0
+ Methidathion 303.1 85.1 -34 25 -61 145 -16 25 -45 ≤0.5 ≤2.0
+ Methiocarb 226.2 121.1 -28 2 -44 169.1 -16 8 -36 ≤0.5 ≤0.5
+ Methiocarb-Sulfone 258.1 200.9 -32 25 -88 122 -45 25 -67 ≤0.5 ≤1.0
+ Methiocarb-Sulfoxide 242.1 122.1 -39 5 -52 185 -16 2 -36 ≤0.5 ≤0.5
+ Methomyl 163.1 87.9 -9 8 -28 105.9 -12 5 -28 ≤0.5 ≤0.5
+ Methoprene 311 237 -15 7 -44 81 -55 7 -80 >5.0 >5.0
+ Methoprotryne 272 240 -19 32 -52 198 -23 24 -68 ≤0.5 ≤0.5
+ Methoxyfenozide 369 133 -41 25 -73 149 -27 25 -60 ≤0.5 ≤0.5
+ Metobromuron 259 148 -21 2 -44 170 -29 4 -52 ≤0.5 ≤0.5
+ Metolachlor 284.1 176.1 -36 18 -60 252.1 -20 19 -52 ≤0.5 ≤0.5
+ Metominostrobin 285.1 238.1 -13 30 -44 195.9 -20 30 -45 ≤0.5 ≤0.5
+ Metosulam 418 140 -83 38 -120 175 -31 36 -64 ≤0.5 ≤0.5
+ Metoxuron 229 156 -34 32 -52 72 -40 31 -44 ≤0.5 ≤0.5
+ Metrafenone 409 227 -35 16 -76 209 -23 16 -60 ≤0.5 ≤0.5
+ Metribuzin 215 89 -24 25 -43 131 -28 25 -47 ≤0.5 ≤1.0
+ Metsulfuron-Methyl 382 199 -33 16 -68 167 -30 16 -60 ≤0.5 ≤0.5
+ Mevinphos 225 193 -11 5 -36 127 -30 2 -44 ≤0.5 ≤0.5
+ Mexacarbate 223.2 151 -40 18 -60 166.2 -19 19 -36 ≤0.5 ≤0.5
+ Milbemycin A3 511.3 153 -40 24 -84 493.3 -15 18 -84 ≤1.0 ≤5.0
+ Milbemycin A4 525.3 161.1 -50 22 -92 507.4 -18 10 -80 ≤1.0 ≤5.0
+ Molinate 188 126 -17 3 -40 83 -26 19 -36 ≤1.0 ≤5.0
+ Monolinuron 215.1 99 -45 25 -78 126.1 -32 25 -54 ≤0.5 ≤0.5
+ Monuron 199 126 -36 21 -56 72 -38 21 -40 ≤0.5 ≤0.5
+ Morpholine 88 44 -25 25 -20 70 -19 23 -20 ≤1.0 ≤5.0
+ Myclobutanyl 289 125 -44 19 -60 70 -23 20 -44 ≤0.5 ≤0.5
+ Napropamide 272.2 171.1 -29 25 -52 129.1 -23 15 -40 ≤5.0 >5
+ Naptalam 292 149 -47 9 -212 144 -46 0 -76 ≤0.5 ≤2.0
+ Neburon 275 114 -17 20 -48 88 -22 24 -48 ≤0.5 ≤0.5
+ Nereistoxin 150 61 -38 11 -40 105 -22 24 -32 ≤0.5 ≤0.5
+ Nicosulfuron 411 106 -60 12 -90 182 -35 16 -72 ≤0.5 ≤0.5
+ Nicotine 163 84 -24 1 -30 132 -18 2 -38 ≤0.5 ≤1.0
+ Nitenpyram 271.2 237.2 -28 6 -48 126.1 -44 3 -64 ≤0.5 ≤0.5
+ Nitralin 346 262 -18 25 -45 304 -20 20 -55 ≤1.0 ≤5.0
+ Norflurazon 304 160 -50 28 -102 284 -40 25 -80 ≤0.5 ≤0.5
- Novaluron 491 155.9 24 -19 76 471.1 17 -15 80 ≤0.5 ≤1.0
+ Nuarimol 315 81 -99 11 -100 252 -45 11 -216 ≤5.0 >5
+ Octhilinone 214 84 -62 28 -52 102 -21 29 -36 ≤0.5 ≤0.5
+ Ofurace 282 160 -31 24 -52 254 -15 24 -48 ≤0.5 ≤0.5
+ Omethoate 214.1 124.9 -31 10 -30 182.9 -17 11 -16 ≤0.5 ≤0.5
+ Orbencarb 258 99.8 -41 25 -80 125 -46 15 -100 ≤0.5 ≤0.5
+ Oryzalyn 347 305 -13 19 -56 288 -11 19 -56 ≤1.0 ≤5.0
+ Oxadiargyl 341 258 -17 33 -56 223 -23 18 -60 ≤5.0 >5.0
1614
LOD LOQ
(ng/ml) (ng/ml)
+ Oxadiazon 362 220 -33 12 -68 177 -44 12 -72 ≤0.5 ≤0.5
+ Oxadixyl 279 133 -22 1 -40 219 -9 7 -52 ≤0.5 ≤1.0
+ Oxamyl 237 220 -7 11 -36 72 -50 11 -56 ≤0.5 ≤0.5
+ Oxycarboxin 268.1 146.9 -43 2 -92 174.9 -25 3 -48 ≤0.5 ≤0.5
+ Paraoxon-Ethyl 276 94 -38 25 -58 220 -34 25 -60 ≤0.5 ≤0.5
+ Pebulate 204 57 -25 27 -32 128 -16 28 -36 ≤0.5 ≤2.0
+ Penconazole 284 159 -89 27 -328 70 -73 19 -92 ≤0.5 ≤0.5
+ Pencycuron 329.3 99 -133 28 -196 125.1 -113 15 -104 ≤0.5 ≤2.0
+ Pendimethalin 282 194 -26 6 -48 212 -7 2 -44 ≤0.5 ≤1.0
+ Penoxsulam 484 195.1 -39 53 -88 164.2 -49 40 -100 ≤0.5 ≤0.5
+ Penthiopyrad 360 256 -28 20 -60 276 -19 20 -60 ≤0.5 ≤0.5
+ Permethrin-cis 408 153 -66 10 -72 183 -51 4 -72 ≤1.0 ≤5.0
+ Permethrin-trans 408 153 -50 17 -8 183 -50 7 -72 ≤1.0 ≤5.0
+ Pethoxamid 296 250 -17 12 -44 131 -32 7 -48 ≤0.5 ≤0.5
+ Phenmedipham 301.2 168 -15 31 -44 136 -34 31 -44 ≤0.5 ≤0.5
+ Phenthoate 321 79 -65 25 -92 163 -16 25 -47 ≤0.5 ≤0.5
+ Phorate-Sulfone 293 115 -28 25 -55 96.9 -33 25 -59 ≤0.5 ≤2.0
+ Phosalone 368 111 -58 25 -57 182 -26 13 -25 ≤0.5 ≤0.5
+ Phosmet 318 76.9 -71 25 -95 160 -21 25 -50 ≤0.5 ≤0.5
+ Phoxim 299 129 -22 17 -44 77 -66 17 -72 ≤0.5 ≤0.5
- Picloram 239 123 33 -9 40 195 15 -10 36 >5.0 >5.0
+ Picolinafen 377 145 -76 20 -96 238 -39 20 -68 ≤0.5 ≤0.5
+ Piperonyl-Butoxide 356.2 119 -67 12 -96 177.2 -25 1 -52 ≤0.5 ≤0.5
+ Piperophos 354 142.9 -47 17 -60 170.9 -32 14 -60 ≤0.5 ≤0.5
+ Pirimicarb 239 182 -43 27 -76 72 -66 14 -88 ≤0.5 ≤0.5
+ Pirimicarb-Desmethyl 225 168 -22 29 -36 72 -39 31 -40 ≤0.5 ≤0.5
+ Pirimiphos-Ethyl 334 198 -46 20 -56 182 -59 22 -160 ≤0.5 ≤0.5
+ Pirimisulfuron-Methyl 469 199 -34 22 -80 254 -28 18 -76 ≤0.5 ≤0.5
+ Probenazole 224 196 -21 24 -48 41 -58 19 -60 ≤1.0 ≤5.0
+ Prochloraz 376/378 308 -19 3 -60 310 -19 3 -64 ≤0.5 ≤0.5
+ Procymidone 284.3 67 -50 37 -48 256.1 -23 28 -48 ≤2 >5
+ Prodiamine 351 267 -25 5 -96 250 -38 4 -64 ≤0.5 ≤2.0
+ Promecarb 208.1 151.1 -13 40 -28 109.1 -25 40 -36 ≤0.5 ≤2.0
+ Prometon 226.4 142 -32 41 -48 184 -25 27 -48 ≤0.5 ≤0.5
+ Prometryn 242 200 -18 3 -60 158 -23 4 -64 ≤0.5 ≤0.5
+ Propachlor 212 94 -60 25 -88 170 -38 25 -60 ≤0.5 ≤1.0
+ Propamocarb 189.2 74 -35 25 -50 102.2 -23 25 -39 ≤0.5 ≤0.5
+ Propaphos 305 203 -34 40 -52 221 -24 42 -52 ≤0.5 ≤0.5
+ Propaquizafop 444 371 -22 27 -68 100 -25 30 -68 ≤0.5 ≤0.5
+ Propargite 368.2 175 -22 1 -52 231.1 -13 7 -52 ≤0.5 ≤0.5
+ Propazine 230 188 -25 19 -52 146 -32 32 -56 ≤0.5 ≤0.5
+ Propham 180 120 -24 5 -40 138 -9 5 -32 ≤5.0 >5.0
+ Propiconazole 342 69 -26 25 -58 159.1 -42 25 -72 ≤0.5 ≤0.5
+ Propisochlor 284 212 -24 27 -48 224 -15 10 -44 ≤0.5 ≤1.0
+ Propoxur 210 168 -11 25 -30 111 -21 25 -39 ≤0.5 ≤1.0
1517
LOD LOQ
(ng/ml) (ng/ml)
+ Propyzamide 256 173 -34 18 -33 190 -20 12 -19 ≤0.5 ≤0.5
+ Proquinazid 373 289 -31 3 -80 331 -20 1 -60 ≤0.5 ≤0.5
+ Prosulfocarb 252.2 128.1 -26 25 -50 91 -31 25 -67 ≤0.5 ≤1.0
- Prothioconazole 342 125 38 -38 68 100 34 -38 64 ≤1.0 ≤5.0
+ Prothoate 286 143 -39 1 -56 227 -15 3 -48 ≤0.5 ≤0.5
+ Pymetrozine 218 105 -37 34 -56 79 -62 39 -68 ≤0.5 ≤0.5
+ Pyracarbolid 218 97 -70 14 -84 125 -62 8 -48 ≤0.5 ≤0.5
+ Pyraclofos 361 138 -57 40 -76 111 -98 39 -120 ≤0.5 ≤0.5
+ Pyraclostrobin 388.1 163 -37 25 -71 194 -17 25 -53 ≤0.5 ≤0.5
+ Pyrasulfotole 363 220 -61 28 -100 251 -34 29 -76 ≤0.5 ≤0.5
+ Pyrethrin I 329.2 143 -29 16 -56 161.1 -16 16 -48 ≤0.5 ≤0.5
+ Pyrethrin II 373.2 133.1 -32 11 -56 161.1 -17 13 -56 ≤0.5 ≤0.5
+ Pyributicarb 331.1 190 -25 3 -56 181 -24 3 -56 ≤0.5 ≤0.5
+ Pyriftalid 319 179 -43 5 -56 139 -39 41 -84 ≤0.5 ≤0.5
+ Pyriproxyfen 322 185 -33 9 -64 96 -20 11 -48 ≤0.5 ≤0.5
+ Pyrmethanil 200.1 183.1 -33 13 -40 107.1 -35 18 -48 ≤0.5 ≤0.5
+ Pyroquilon 174.1 117 -43 34 -56 132 -32 39 -44 ≤0.5 ≤0.5
+ Pyroxsulam 435 258 -32 51 -72 195 -38 32 -68 ≤0.5 ≤0.5
+ Quinalphos 299 96.9 -113 3 -52 162.9 -48 11 -44 ≤0.5 ≤0.5
+ Quinclorac 244 226 -26 11 -64 224 -23 23 -40 ≤0.5 ≤1.0
+ Quinmerac 222 204 -25 14 -52 141 -50 5 -76 ≤0.5 ≤0.5
+ Quinoxyfen 308 162 -64 8 -76 197 -44 5 -72 ≤0.5 ≤0.5
+ Quizalofop Ethyl 375 301 -16 32 -68 299 -15 30 -76 ≤0.5 ≤0.5
+ Quizalofop-P 345 163 -50 2 -84 299 -24 20 -56 ≤0.5 ≤2.0
+ Quizalofop-P-Ethyl 375 301 -26 29 -64 299 -26 37 -72 ≤0.5 ≤0.5
+ Rimsulfuron 432 325 -20 25 -72 139 -66 25 -76 ≤1.0 ≤5.0
+ Saflufenacil 501 349 -38 29 -96 198 -67 31 -112 ≤0.5 ≤0.5
+ Secbumeton 226 100.2 -65 30 -36 170 -41 19 -32 ≤0.5 ≤0.5
+ Sethoxydim 328.3 178 -26 25 -55 282 -16 25 -46 ≤0.5 ≤0.5
+ Siduron 233 94 -51 30 -36 137 -35 27 -68 ≤0.5 ≤0.5
+ Silthiofam 268 139 -26 34 -48 252 -18 4 -48 ≤0.5 ≤0.5
+ Simazine 202 132 -22 25 -50 124 -22 4 -42 ≤0.5 ≤0.5
+ Simeconazole 294 135 -29 32 -52 70 -51 32 -64 ≤0.5 ≤0.5
+ Simetryn 214 96 -24 3 -72 124 -19 5 -48 ≤0.5 ≤0.5
+ S-Metolachlor 284.1 176.2 -35 7 -48 252.2 -21 15 -48 ≤0.5 ≤0.5
+ Spinosyn A 732.5 142.1 -53 38 -112 98 -108 44 -176 ≤0.5 ≤0.5
+ Spinosyn D 746.5 142.1 -54 36 -128 98 -105 47 -216 ≤0.5 ≤0.5
+ Spirodiclofen 411 229 -28 8 -72 209 -25 19 -64 ≤0.5 ≤0.5
+ Spiromesifen 371 255 -36 6 -68 273 -25 6 -56 ≤0.5 ≤1.0
+ Spirotetramat 374 330 -21 48 -56 216 -45 48 -68 ≤0.5 ≤0.5
+ Spiroxamine 298.5 100 -45 27 -48 144 -28 26 -48 ≤0.5 ≤0.5
+ Sulcotrione 329 69 -76 39 -80 139 -26 38 -56 ≤0.5 ≤1.0
+ Sulfallate 224 88 -32 15 -36 116 -17 15 -36 ≤0.5 ≤2.0
- Sulfentrazone 385 198.6 45 -15 132 306.3 31 -20 112 ≤0.5 ≤0.5
- Sulfluramid 525.9 219 32 -44 116 169 35 -44 96 ≤0.5 ≤0.5
1816
LOD LOQ
(ng/ml) (ng/ml)
+ Sulfosulfuron 471 261 -27 18 -76 211 -31 4 -76 ≤0.5 ≤0.5
+ Sulfotep 323 171 -19 0 -284 97 -90 11 -176 ≤5.0 >5.0
+ Sulprofos 323 247 -16 23 -48 219 -23 25 -52 ≤0.5 ≤0.5
+ Tebuconazole 308 125 -65 19 -132 70 -92 19 -104 ≤0.5 ≤0.5
+ Tebufenozide 353.1 133.1 -21 25 -58 297.1 -13 25 -47 ≤0.5 ≤0.5
+ Tebupirimfos 319 153 -41 27 -68 277 -21 29 -48 ≤0.5 ≤0.5
+ Tebutam 234 192 -21 32 -44 91 -47 21 -48 ≤0.5 ≤0.5
+ Tebuthiuron 229.1 116 -36 29 -56 172.1 -21 29 -40 ≤0.5 ≤0.5
- Teflubenzuron 379/381 195.5 36 -13 68 197.5 28 -10 64 ≤0.5 ≤0.5
- Tembotrione 439 226 50 -20 100 403 25 -22 76 ≤0.5 ≤0.5
+ Temephos 466.9 405 -21 38 -68 419 -25 40 -76 ≤0.5 ≤0.5
+ Tepraloxydim 342.2 250.2 -20 21 -56 166.2 -34 25 -60 ≤0.5 ≤0.5
+ Terbufos-Sulfone 289 57 -40 15 -80 103 -28 14 -56 ≤0.5 ≤0.5
+ Terbufos-Sulfoxide 305 187 -46 14 -96 131 -45 25 -80 ≤0.5 ≤0.5
+ Terbumeton 226.1 114.1 -23 4 -68 170.1 -16 3 -52 ≤0.5 ≤0.5
+ Terbuthylazine 230.1 174.1 -24 25 -48 96 -38 27 -56 ≤0.5 ≤0.5
+ Terbuthylazine Desethyl 202 104 -33 15 -40 146 -15 24 -36 ≤0.5 ≤1.0
+ Tetraconazole 372 70 -99 36 -140 159 -135 29 -200 ≤0.5 ≤2.0
+ Tetrametrhin 332 135 -73 6 -64 164 -37 1 -64 ≤0.5 ≤0.5
+ Thiabendazole 202.1 131 -32 25 -49 174.9 -42 25 -58 ≤0.5 ≤0.5
+ Thiachloprid 253.1 99.1 -69 15 -88 126.1 -33 26 -52 ≤0.5 ≤0.5
+ Thiadiazuron 221 94 -18 26 -40 102 -23 29 -36 ≤0.5 ≤0.5
+ Thiamethoxam 292 181 -33 12 -52 211 -18 12 -44 ≤0.5 ≤0.5
+ Thiazopyr 397 335 -38 49 -104 377 -30 48 -84 ≤0.5 ≤0.5
+ Thifensulfuron-methyl 388 205 -32 25 -54 167 -23 24 -56 ≤0.5 ≤0.5
+ Thiobencarb 258/260 125 -44 5 -52 127 -38 10 -52 ≤0.5 ≤0.5
+ Thiodicarb 355 107.9 -90 13 -144 87.9 -85 12 -112 ≤0.5 ≤0.5
+ Thionazin 249 193 -21 22 -40 97 -38 9 -48 ≤0.5 ≤1.0
+ Thiophanate-Methyl 343.1 93.1 -78 25 -84 151 -31 12 -56 ≤0.5 ≤0.5
+ Tiocarbazil 280.2 99.9 -18 22 -48 91 -52 5 -52 ≤0.5 ≤0.5
+ Tralkoxydim 330 138 -30 22 -60 284.1 -18 17 -52 ≤0.5 ≤0.5
+ Triadimefon 294.1 225.1 -18 9 -48 197.1 -21 9 -48 ≤0.5 ≤0.5
+ Triadimenol 296/298 70 -49 11 -44 70 -49 11 -44 ≤0.5 ≤0.5
+ Tri-Allate 304 86 -24 18 -56 143 -34 22 -60 ≤0.5 ≤2.0
+ Trichlorfon 257 220.8 -13 24 -48 108.9 -22 24 -48 ≤0.5 ≤2.0
- Triclopyr 254/256 196 21 -37 48 198 20 -35 36 ≤0.5 ≤0.5
- Triclosan 289 35 62 -6 40 35 56 -6 48 ≤0.5 ≤0.5
+ Tricyclazole 190 136 -40 44 -64 163 -27 42 -52 ≤0.5 ≤0.5
+ Trietazine 230 132 -30 25 -60 99 -35 13 -56 ≤0.5 ≤0.5
+ Trifloxystrobin 409 206 -20 25 -59 186 -26 25 -64 ≤0.5 ≤0.5
+ Triflumizole 346 73 -17 15 -60 278 -5 15 -8 ≤1.0 ≤5.0
+ Triflumuron 359.1 139 -57 25 -76 156.2 -23 9 -56 ≤0.5 ≤0.5
+ Triforine 389.8 98 -35 37 -64 215 -30 36 -68 ≤0.5 ≤2.0
+ Trimethylsulfonium 77 47 -36 18 -24 62 -20 19 -16 ≤0.5 ≤0.5
+ Trinexapac-Ethyl 253 207 -18 27 -40 69 -39 27 -48 ≤0.5 ≤1.0
1719
LOD LOQ
(ng/ml) (ng/ml)
+ Triticonazole 318 125 -53 41 -72 70 -62 38 -56 ≤0.5 ≤0.5
+ Tritosulfuron 446 221 -28 33 -72 195 -30 34 -76 ≤0.5 ≤2.0
+ Valifenalate 399 116.1 -38 23 -56 155.1 -47 25 -64 ≤0.5 ≤0.5
+ Vamidothion 288 118 -80 10 -100 146 -60 15 -88 ≤0.5 ≤2.0
+ Vernolate 204.1 86 -20 24 -32 128 -16 25 -36 ≤0.5 ≤0.5
+ Warfarin 309 251 -28 29 -64 163 -19 29 -44 ≤0.5 ≤0.5
+ Zoxamide 336/338 186.9 -31 18 -30 188.7 -33 18 -32 ≤0.5 ≤0.5
References
1. European Commission (EU). Regulation EC No 396/2005 on 6. Mol, H. G. J.; Zomer, P.; García López, M.; Fussell, R. J.; Scholten,
Maximum Residue Levels of Pesticides in or on Food and Feed of J.; de Kok, A.; Wolheim, A.; Anastassiades, M.; Lozano, A.;
Plant and Animal Origin. 2006, 1881 (February 1998), 1 – 5. Fernandez Alba, A. Identification in Residue Analysis Based on
Liquid Chromatography with Tandem Mass Spectrometry:
2. EU Pesticides database - European Commission https://ec.europa.
Experimental Evidence to Update Performance Criteria. Analytica
eu/food/plant/pesticides/eu-pesticides-database/public/?event=ho
Chimica Acta 2015, 873, 1–13.
mepage&language=EN (accessed Aug 12, 2020).
https://doi.org/10.1016/j.aca.2015.03.007.
3. European Commission (EU). Method Validation Procedures
7. QuEChERS https://www.quechers.com/index.
for Pesticide Residues Analysis in Food and Feed;
php?nav1o=3&nav2o=0&nav3o=0 (accessed Aug 12, 2020).
SANTE/12682/2019.
8. UNI EN 15662:2018 http://store.uni.com/catalogo/uni-
4. Anastassiades, M.; Lehotay, S. J.; Štajnbaher, D.; Schenck,
en-15662-2018?___store=en&josso_back_
F. J. Fast and Easy Multiresidue Method Employing Acetonitrile
to=http%3A%2F%2Fstore.uni.com%2Fjosso-security-check.
Extraction/Partitioning and “Dispersive Solid-Phase Extraction”
php&josso_cmd=login_optional&josso_partnerapp_host=store.
for the Determination of Pesticide Residues in Produce.
uni.com&___from_store=it (accessed Aug 12, 2020).
Journal of AOAC INTERNATIONAL 2003, 86 (2), 412–431.
https://doi.org/10.1093/jaoac/86.2.412.
5. Stachniuk, A. LC-MS/MS Determination of Pesticide Residues in
Fruits and Vegetables. In Bioactive Molecules in Food; Mérillon,
J.-M., Ramawat, K. G., Eds.; Reference Series in Phytochemistry;
Springer International Publishing: Cham, 2019; pp 2137–2161.
https://doi.org/10.1007/978-3-319-78030-6_82.
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122220 PKI
20
A P P L I C AT I O N N O T E
Liquid Chromatography/
Mass Spectrometry
Authors:
Jingcun Wu
Josh Ye
Erasmus Cudjoe
Feng Qin
Shixin Sun
PerkinElmer, Inc.
Woodbridge, Ontario, Canada
Analysis of Multi-Residue
Pesticides in Rice by LC/MS/MS Introduction
Rice is one of the most commonly
consumed foods in the world. A
variety of pesticides have been used in rice production to control pests, weeds and diseases to
increase crop yield. Pesticides applied in rice crops are often country/region specific due to the
differences in legislation, weather and production system. Pesticide residue in rice not only affects
the quality of the rice, but also threatens the health of general consumers. To prevent health
risks, it is important to monitor the presence of pesticides and regulate their levels in rice. Several
countries including the United States, China, Brazil, India, Japan and European Union (EU) have
established maximum residue levels (MRLs) of pesticides for food and feed including rice.1-3 The
EU MRLs for pesticide residues in rice mostly range from 10 µg/kg to 8000 µg/kg depending on
the pesticide.1 To determine low levels of pesticides in rice, highly sensitive, selective and accurate
analytical methods are needed. Due to the large number of pesticides potentially used in rice
production, the use of multi-residue methods capable of determining many pesticides in one single
run is the most efficient approach. Traditionally, pesticide residues were analyzed mainly by gas
chromatography/mass spectrometry (GC/MS) methods,4, 5 but GC is not a suitable technique for
ionic and polar compounds, especially for compounds that are thermally labile in the GC injection
port. Liquid chromatography tandem mass spectrometry (LC/MS/MS) has become the method of
choice for pesticide analysis due to its high selectivity and sensitivity as well as its suitability for a
wide range of compounds in various sample matrices.6-10
21
QuEChERS extraction method has been widely applied for Table 1. LC Method and MS Source Conditions.
analysis of multi-residue analytes in food samples including LC Conditions
rice.4,8,9,10 In this study, a fast, sensitive and selective multi- Brownlee, SPP Phenyl-Hexyl,
LC Column
residue method has been developed for analysis of over 200 100 x 2.1 mm, 2.7 μm
pesticides in rice samples by coupling a modified QuEChERS Mobile Phase A 5 mM ammonium formate in water
extraction method with LC/MS/MS. Using time-managed- Mobile Phase B 5 mM ammonium formate in methanol
MRM™ in the QSight® triple quadrupole mass spectrometer, Start at 10% mobile phase B and hold it for
the optimum dwell time of multiple MRM transitions can be Mobile 1 min., then increase B to 95% in 15 min. and
Phase Gradient keep at 95% B for 2 min. Finally equilibrate the
generated automatically for the targeted analytes. This not only
column at initial condition for 3 min.
saves time in method development but also improves data quality
Column Oven Temperature 40 °C
and analytical performance, as demonstrated in this study by the
Auto Sampler Temperature 15 °C
results of multi- residue pesticide analysis in rice samples.
Injection Volume 1.0 µL
Experimental MS Source Conditions
ESI Voltage (Positive) 5000 V
Hardware/Software
Chromatographic separation of pesticides was conducted by ESI Voltage (Negative) -4000V
a PerkinElmer UHPLC System and analyte determination was Drying Gas 140
achieved using a PerkinElmer QSight 220 triple quadrupole Nebulizer Gas 350
mass detector with a dual ionization source. All instrument Source Temperature 325 °C
control, data acquisition and data processing was performed HSID Temperature 200 °C
using Simplicity 3Q™ software. Detection mode Time-managed MRM™
Method
Sample Preparation Results and Discussion
Pesticide standards were obtained from ULTRA® Scientific (North Analytical Challenges for Multi-residue Pesticides
Kingstown, RI). Rice samples were purchased from local grocery Analysis in Food Samples
stores in Ontario, Canada. Different rice samples such as brown Since the pesticides tested in this study contain both polar
rice, black rice and white rice (including Jasmine, Basmati and and non-polar compounds, to extract all the analytes from
Calrose) as well as two brands of organic rice samples were sample matrices, acetonitrile, an organic solvent, was
tested. These rice samples were originally produced in Thailand, used. However, the reverse phase LC method used aqueous
Vietnam, India, Italy and the U.S. Rice samples were prepared mobile phase at the beginning of the LC run to retain the
according to a published procedure with minor modifications polar compounds on the column. Injecting a larger volume of
using QuEChERS kits (AOAC 2007.01 method) without dispersive organic solvent such as an acetonitrile sample extract on the LC
SPE clean-up.10 One (1) µL of extract was injected directly onto the would lead to poor chromatographic peaks for early eluting
QSight LC/MS/MS system for quantification. polar compounds. To overcome this problem, small sample
volume was injected in this study.
An organic brown rice sample was used as a controlled blank
matrix. Recoveries from the rice sample matrix were evaluated by Traditional MRM method development is not suitable for analysis
fortifications of pesticides at concentrations of 10 and 100 µg/kg. of a large number of analytes such as hundreds of pesticide
Calibration curves were built by eight levels of standards prepared residues in a single run. It is both time-consuming and labor
in a neat solution (acetonitrile) and in the rice sample matrix intensive to input all the mass transitions to a method manually.
(matrix-matched calibration). Matrix effects were evaluated by In addition, the dwell time for each transition cannot be optimized
comparing the slopes of calibration curves obtained from the easily by traditional method. Therefore, a time-managed-MRM
neat solution and rice sample matrix. To reduce false positives was applied for method development in this study to improve
and negatives, at least two MRM transitions were monitored for efficiency, data quality and method performance.
each pesticide. LOQs (limits of quantification) were calculated Sample matrix effect is the main concern for LC/MS/MS method
based on a minimum S/N of 10 for both transitions.12 development, especially for food analysis due to the diversity
LC Method and MS Source Conditions and complexity of food sample matrices. To overcome sample
The LC method and MS source parameters are shown in Table 1. matrix effects, several approaches have been used, such as
A partial list of the multiple reaction monitoring mode (MRM) sample dilution, use of stable isotope internal standards, matrix-
transitions of the studied pesticides are shown in Table 2. The matched calibration, standard addition, sample clean-up, use
acquisition MS method is generated automatically by selecting of high efficiency columns for improved separation, and the use
the pesticides of interest from the built-in compound library in of alternative ionization sources.11 In this study, sample matrix
the time-managed-MRM module of the Simplicity software, effects were evaluated by comparing the slopes (X) of calibration
including both positive and negative analytes. curves obtained from standards prepared in solvent (neat
2
22
solution) with slopes (Y) obtained from standards prepared Table 2. MRM Transitions (partial list of the 213 pesticides studied).
in the rice sample matrix. Sample matrix effect (%) can Compound Name Polarity Q1 Mass Q2 Mass CE EV CCL2
be calculated by the percentage difference between the Acephate Positive 184.1 143.1 -12 25 -29
slopes, i.e. (Y-X) × 100/X. When the percentage of the Acephate-2 Positive 184.1 125.1 -25 25 -41
difference between the slopes of the two curves is Acetamiprid Positive 223.2 126.1 -30 25 -49
positive, there is a signal enhancement effect, whereas Acetamiprid-2 Positive 223.2 99.1 -56 25 -73
a negative value indicates signal suppression effect. As Azoxystrobin Positive 404.1 372.1 -18 25 -57
shown in Table 3 and Figures 1 and 2, sample matrix Azoxystrobin-2 Positive 404.1 344.1 -34 25 -71
effects are compound dependent. For example, some Buprofezin Positive 306.2 201.1 -18 25 -47
pesticides, such as acephate and propiconazole, showed Buprofezin-2 Positive 306.2 116.2 -24 25 -52
signal enhancement (positive values), while others, such Chlorantranilprole Positive 484 452.8 -20 25 -66
as chlorpyriphos and tricyclazole, showed ion suppression Chlorantranilprole-2 Positive 484 285.8 -18 25 -65
(negative values). As shown in Table 3, sample matrix Chlorpyriphos Positive 350 198 -20 25 -53
effects for most of the pesticides studied are less than Chlorpyriphos-2 Positive 350 97 -32 25 -64
20% and thus, calibration curves built from neat solutions Clothianidin Positive 250.1 169.1 -16 25 -39
could be used for their quantification without significant Clothianidin -2 Positive 250.1 132.2 -26 25 -48
error according to EU regulation.12 However, significant Cumyluron Positive 303.1 185 -20 25 -48
ion suppression effects were observed for chlorpyriphos Cumyluron-2 Positive 303.1 125 -43 25 -69
(-55%) and tebuconazole (-18%). Therefore, to overcome Fenbutatin-oxide Positive 519.3 197 -67 25 -112
matrix effects and reduce variations in analytical results, Fenbutatin-oxide-2 Positive 519.3 350.9 -50 25 -97
matrix-matched calibrations were used in this study for Fenobucarb Positive 208 152 -12 25 -32
quantification of all analytes. Fenobucarb-2 Positive 208 95 -19 25 -38
Method Performance Fluopyram Positive 397 173 -35 25 -71
All calibration curves built from both the neat solution Fluopyram-2 Positive 397 145 -70 25 -103
and rice sample matrix (matrix-matched calibration) Halofenozide Positive 331.1 275 -18 25 -49
showed good linearity (0.1 to 200 ng/mL) with correlation Halofenozide-2 Positive 331.1 104.9 -25 25 -56
coefficient (R²) larger than 0.99 (see Figures 1 and 2 for Imazalil Positive 297.1 201 -25 25 -52
typical examples of calibration curves). Imazalil-2 Positive 297.1 159.2 -31 25 -58
Imidachloprid Positive 256.2 175.2 -26 25 -49
The recoveries of pesticides were evaluated by spiking
Imidachloprid-2 Positive 256.2 209 -18 25 -42
the analytes to the samples at two concentration levels
Isoprothiolane Positive 291.1 231 -16 25 -44
of 10 and 100 μg/kg, respectively. As shown in Table 3,
Isoprothiolane-2 Positive 291.1 189 -28 25 -54
the recoveries of analytes ranged from 70% to 120%
Malathion Positive 331.1 127.1 -22 25 -53
with RSD < 20% for most of the pesticides studied.
Malathion-2 Positive 331.1 99.1 -24 25 -55
The limits of quantification (LOQs) were determined by Methamidophos Positive 142 124.9 -20 25 -32
taking into account the signals of both quantifier and Methamidophos-2 Positive 142 94.1 -20 25 -32
qualifier ions (S/N > 10 for both) and ensuring that the Piperonyl butoxide Positive 356.2 177 -13 25 -47
product ion ratios were within 20% tolerance windows Piperonyl butoxide-2 Positive 356.2 119 -37 25 -69
of the expected.12 Most of the tested pesticides have Pirimiphos-methyl Positive 306.1 164.1 -28 25 -56
LOQs ranging from 0.5 to 20 µg/kg, which are well Pirimiphos-methyl-2 Positive 306.1 108.1 -40 25 -67
below the EU MRLs. Profenophos Positive 375 304.8 -50 25 -75
Profenophos-2 Positive 375 346.8 -42 25 -113
Propiconazole Positive 342.1 159.1 -42 25 -72
Propiconazole-2 Positive 342.1 69.1 -26 25 -58
Tebuconazole Positive 308 70 -30 25 -58
Tebuconazole-2 Positive 308 125 -50 25 -76
Thiamethoxam Positive 292 181 -28 25 -54
Thiamethoxam-2 Positive 292 211 -18 25 -45
Triazophos Positive 314.1 161.9 -22 25 -51
Triazophos-2 Positive 314.1 118.9 -50 25 -76
Tricyclazole Positive 190 163 -28 25 -44
Tricyclazole-2 Positive 190 136 -36 25 -51
Trifloxystrobin Positive 409 186 -26 25 -64
Trifloxystrobin-2 Positive 409 206 -20 25 -59
Fludioxonil Negative 246.6 125.9 40 -25 60
Fludioxonil-2 Negative 246.6 179.9 39 -25 60
3
23
A B
C D
Figure 1. Calibration curves for acephate (A), chlorpyriphos (B), propiconazole (C) and tricyclazole (D) obtained from standards prepared in neat solutions
(analyte concentrations range from 0.1 to 200 ng/mL).
A B
C D
Figure 2 . Calibration curves for acephate (A), chlorpyriphos (B), propiconazole (C) and tricyclazole (D) obtained from standards prepared in rice sample matrix
(analyte concentrations range from 0.1 to 200 ng/mL).
4
24
Table 3. Results of retention time, recovery, reproducibility (%RSD), matrix effect and linearity for the most commonly detected pesticides in rice samples.
Retention Time % Recovery(%RSD) % Recovery (%RSD) Matrix Effect Correlation Coefficient
Pesticide
(min) at 10 µg/kg at 100µg/kg (%) (R2)
Acephate 1.88 101.1 (11.8) 81.9 (4.3) 14.0 0.9997
Acetamiprid 8.15 106.5 (2.6) 98.7 (2.3) 2.7 0.9996
Buprofezin 15.05 103.3 (2.9) 98.8 (3.5) -3.1 0.9996
Chlorpyriphos 15.54 109.6 (10.4) 98.7 (5.0) -55.0 0.9991
Clothianidin 6.70 105.7 (5.9) 111.2 (8.6) 17.0 0.9995
Cumyluron 12.74 98.9 (7.2) 96.1 (2.5) -2.6 0.9984
Fenbutatin-oxide 16.90 69.5 (18.6) 78.8 (12.7) 13.1 0.9997
Fenobucarb 11.20 101.6 (2.9) 94.8 (1.9) 2.6 0.9976
Fluopyram 13.00 104.8 (3.6) 101.1 (3.1) -2.7 0.9991
Halofenozide 12.26 89.4 (15.2) 88.3 (11.4) -4.4 0.9980
Imazalil 14.33 89.6 (13.6) 95.3 (4.1) -6.1 0.9996
Imidacloprid 7.57 77.5 (10.8) 112.2 (7.9) -5.7 0.9991
Isoprothiolane 13.01 111.5 (2.7) 101.1 (2.3) -0.4 0.9983
Malathion 13.25 92.0 (12.0) 86.0 (4.3) -9.9 0.9995
Methamidophos 1.41 82.8 (10.1) 76.4 (14.3) 13.3 0.9978
Piperonyl Butoxide 15.26 106.0 (5.0) 105.2 (3.4) -6.3 0.9977
Pirimiphos-methyl 14.71 107.5 (3.7) 98.8 (5.3) -0.1 0.9997
Profenophos 14.82 110.7 (6.9) 103.0 (6.5) -2.5 0.9988
Propiconazole 14.32 106.6 (7.1) 98.3 (2.8) 1.5 0.9994
Tebuconazole 13.72 102.2 (6.9) 104.2 (5.5) -18.9 0.9993
Thiamethoxam 6.43 116.4 (10.0) 114.0 (14.9) 1.9 0.9991
Triazophos 13.46 117.8 (5.7) 99.5(3.0) 2.7 0.9979
Tricyclazole 9.27 84.2 (5.8) 80.7 (7.8) -7.5 0.9998
Trifloxystrobin 14.91 106.7 (2.4) 106(4) -5.8 0.9991
Sample Analysis
The developed method was applied for the analysis
of pesticide residues in different food samples,
including eleven rice samples; one wheat sample
and one veggie straw sample. Figure 3 showed
the overlapped MRM chromatograms of pesticides
identified and quantified from a brown rice sample.
Table 4 lists the pesticide residues determined in the
eleven rice samples and the EU MRLs in
µg/kg (NA*; some pesticides that are not included
in the EU MRLs list were also determined by this
method). As shown in Table 4, many of the pesticides
identified from sample 4 (S4) and sample 10 (S10)
are quite similar because these two rice samples
were produced from the same region, which indicates
that pesticides applied to rice crops during production Figure 3. Pesticides determined from a brown rice sample (S10): thiamethoxam (1), clothiani-
din (2), imidacloprid (3), acetamiprid (4), tricyclazole (5), isoprothiolane (6), triazophos (7),
are country or region specific due to the regulation tebuconazole (8), imazalil (9), propiconazole (10), profenophos (11), trifloxystrobin (12),
and weather conditions in that region. buprofezin (13), and chlorpyriphos (14).
Conclusion sample extraction utilized in this study demonstrated good recovery

A LC/MS/MS method for multi- residue pesticides (70-120%) and reproducibility (RSD <20%) for most pesticides. The
analysis in rice was developed by coupling a UHPLC developed method showed excellent linearity with R2 > 0.99 for all the
system to a QSight 220 triple-quad mass spectrometer. studied pesticides in rice matrix. A number of pesticide residues were
The method can be applied for the analysis of over identified and quantified from eleven rice samples with concentrations at
200 pesticides in rice with LOQs well below the limits or below the EU MRLs. This LC/MS/MS method has also been applied for
set by regulatory agencies. The time-managed-MRM other food analyses such as wheat and veggie strews samples with good
module has simplified the creation of MS method performance. The method presented here can be easily adapted for multi-
with optimum dwell time for monitoring a large analyte screening and quantification, providing a single method for more
number of analytes in food samples. The QuEChERS cost-effective analysis of pesticides in rice and other food samples.
5
25
Table 4. Pesticide residues determined from eleven rice samples (S1 to S11), in μg/kg. References
Pesticide S1 S2 S3 S4 S5 S6 S7 S8 S9 S10 S11 MRL 1. Commission Regulation (EC) 396/2005 on
Acephate 2.0 10 maximum residue levels of pesticides in or on food
Acetamiprid 0.3 0.8 10 and feed of plant and animal origin, J. Eur. Union.
Buprofezin 9.1 46.5 500 L70/1 (2005).
Chlorpyriphos 0.5 0.3 1.4 8.7 50
2. U.S. Environmental Protection Agency, Electronic
Clothianidin 7.0 3.0 500
code of federal regulation: Title 40: part
Fenobucarb 4.1 NA*
180-tolerance and exemptions for pesticide
Fluopyram 0.5 10
chemical residues in Food. http://www.ecfr.gov/cgi-
Halofenozide 5.0 NA*
bin/text-idx?c=ecfr&tpl=/ecfrbrowse/
Imazalil 1.4 2.5 4.6 2.6 1.6 50
Title40/40cfr180_main_02.tpl
Imidacloprid 2.8 1.1 9.2 1500
3. China National Standard GB 28260-2011. 2011.
Isoprothiolane 4.4 9.3 2.9 14.7 5000
Maximum residue limits for 85 pesticides in food,
Malathion 1.8 2.2 8000
Ministry of Health of the People’s Republic of China.
Methamidophos 0.5 10
Piperonyl Butoxide 0.6 1.3 0.8 NA* 4. X. Hou, M. Han, X. Dai, X-F. Yang and S. Yi, A
Pirimiphos-methyl 1.4 500 multi- residue method for the determination of
Profenophos 5.2 10 124 pesticides in rice by modified QuEChERS
Propiconazole 8.3 8.4 6.7 4.1 18.1 1500 extraction and GC-MS/MS. Food Chemistry, 2013,
Tebuconazole 5.9 5.2 0.9 12.0 1000 138, 1198-1205.
Thiamethoxam 10.6 11.0 10 5. M. Kirchner, E. Matisova, S. Hrouzkova, and J. D.
Triazophos 0.6 0.5 17.6 20 Zeeuw, Possibilities and limitations of quadrupole
Tricyclazole 16.4 5.8 7.6 20.6 0.6 40.2 1000 mass spectrometric detector in fast gas
Trifloxystrobin 1.6 5000 chromatography. J. Chromatogr. A, 2005, 1090
NA*: pesticides not listed in the EU MRLs database, but can be determined by this method. (1-2), 126–132.
6. J. Wu, Quantitative Method for the Analysis of Tobacco-Specific Nitrosamines in Cigarette Tobacco and Mainstream Cigarette Smoke
by Use of Isotope Dilution Liquid Chromatography Tandem Mass Spectrometry, Anal. Chem., 2008, 80 (4), 1341–1345.
7. K. Zhang, M.R. Schaab, G. Southwood, E.R. Tor, L.S. Aston, W. Song, B. Eitzer, S. Majumdar, T. Lapainus, H. Mai, K. Tran, A.
El-Demerdash, V. Vega, Yanxuan Cai, J.W. Wong, A.J. Krynitsky, and T.H. Begley, A Collaborative Study: Determination of Mycotoxins
in Corn, Peanut Butter, and Wheat Flour Using Stable Isotope Dilution Assay (SIDA) and Liquid Chromatography–Tandem Mass
Spectrometry (LC-MS/MS), J. Agric. Food Chem. 2017, 65 (33), 7138-7152.
8. A. Wilkowska and M. Biziuk, Determination of pesticide residues in food matrices using the QuEChERS methodology, Food
Chemistry, 2011, 125, 803-812.
9. L. Pareja, A.R. Fernandez-Alba, V. Cesio and H. Heinzen, Analytical methods for pesticide residues in rice, Trends in Anal. Chem.
2011, 30 (2), 270-291.
10. L. Pareja, V. Cesio, H. Heinzen and A.R. Fernandez-Alba, Evaluation of various QuEChERS based methods for the analysis of
herbicides and other commonly used pesticides in polished rice by LC-MS/MS, Talanta, 2011, 83, 1613-1622.
11. A. J. Krynitsky, J. W. Wong, K. Zhang and H. Safarpour, Focus on Food Analysis: Important considerations regarding matrix effects
when developing reliable analytical residue methods using mass spectrometry, LCGC North America, 2017,Vol. 35, No. 7, 444-451.
12. European Commission, SANCO. 2015. Guidance document on analytical quality control and method validation procedures for
pesticides residues analysis in food and feed, SANTE/11945/2015 https://ec.europa.eu/food/sites/food/files/plant/docs/pesticides_mrl_
guidelines_wrkdoc_11945.pdf.
PerkinElmer, Inc.
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Mass Spectrometry
Authors:
Josh Ye, PhD
Feng Qin, PhD
Sharanya Reddy, PhD
Frank Kero, PhD
PerkinElmer, Inc.
Waltham, MA
Analysis of Target Pesticide Residues in Introduction

Berries with LC/MS/MS Coupled with Pesticides are widely used in
agriculture to protect plants
a QuEChERS Sample Preparation from a variety of pests and to
increase productivity. However,
the extensive use of pesticides can pose a health risk to humans and this has led to
worldwide stringent regulations, for maximum allowable limits for these residues in
foods. Among the routinely used testing methods, LC/MS/MS has become the method
of choice, due to its high sensitivity, reliability and accuracy.
In the present study, a unique laminar flow UPLC-ESI-MS/MS triple quad mass
spectrometer was used to identify and quantitate 40 pesticides in four brands of non-
organic berries. The QuEChERs extraction method proved both rapid and reliable for
extracting pesticide residues in the heavily pigmented berry samples.
27
Experimental Solvents, Standards and Sample Preparation
Hardware/Software Berry samples were obtained from a local grocery store in
Ontario, Canada. Pesticide standards were obtained from ULTRA
Chromatographic separation was conducted by a PerkinElmer
Scientific® (North Kingstown, RI). All solvents, reagents and
Altus® A-30 UPLC® System and detection was achieved using a
diluents used were HPLC grade.
PerkinElmer QSight™ 220 MS/MS detector with dual ionization
source. All instrument control, data acquisition and data processing Samples were prepared using Supra-d™ QuEChERS kits (AOAC
was performed using the Simplicity 3Q™ software platform. 2007.01 method). Briefly, samples were homogenized using a
blender at high speed, and 10 g of homogenized samples were
Method parameters
weighed and transferred to a 50 mL extraction tube containing
The LC method and MS source parameters are shown in Table 1. 6 g of MgSO4 and 1.5 g of sodium acetate. 10 mL of cold
Table 1. LC Method and General MS Conditions. acetonitrile was then added and vortexed until the salt was
completely mixed. The solution was then centrifuged at 3500 rpm
LC Method for 5 minutes. 4 mL of supernatant was then transferred to a
15 mL clean-up tube (AOAC 2007.01 Clean-up Kits), vortexed
Column: PerkinElmer Brownlee Phenyl-Hexyl column, 2.7 µm, 2.1 x 100 mm for 3 minutes and centrifuged for 5 minutes.
Mobile Phase: Solvent A: 5 mM ammonium formate in water

0.1 mL of supernatant was transferred to a 1.5-mL centrifuge
tube, diluted 10-fold with mobile phase A and then centrifuged
Solvent B: 5 mM ammonium formate in methanol at 4000 rpm for 5 minutes. The resulting supernatant was
transferred to a 1.5 mL LC vial for direct LC/MS/MS analysis.
Flow rate
Time (min) %A %B
(ml/min)
Optimizing MS/MS Parameters
1 Initial 90 10 0.3
Source parameters, including gas flows, source temperature and
2 1 90 10 0.3 position settings, were optimized to achieve the best sensitivity.
3 15 5 95 0.3 The Q1 and Q2 quadrupole peak widths were set at 0.7 amu.
4 17 5 95 0.3 Multiple reaction monitoring mode (MRM) transitions are listed
5 17.1 90 10 0.3
in Table 2.
6 20 90 10 0.3 Table 2. Optimized compound-dependent MS parameters for tested pesticides

(partial list).
Precursor
Oven Temp.: 40 ºC Compound Product 1 CE1 Product 2 CE2
Ion
Atrazine 216.1 174.1 15 132.0 20

Injection Volume: 20 µL
Azoxystrobin 404.1 372.1 18 344.1 34

General MS Conditions
Bifenazate 301.1 198.0 16 170.0 20
ESI voltage: 5000 V
Boscalid 343.0 307.0 25 140.0 28
Drying gas: 120
Cyprodinil 226.0 93.0 48 108.0 34
HSID Temp: 200 °C
Flonicamid 230.1 203.1 20 174.0 20
Entrance voltage: 30 V
Hexythiazox 353.0 228.0 20 168.0 34
Source Temp: 325 °C

Pyraclostrobin 388.0 194.0 16 163.0 36
Nebulizer gas: 350 Pyrimethanil 200.0 107.0 33 82.0 32
Detection Mode: MRM Mode Thiamethoxam 292.0 211.0 18 181.1 28
2
28
Results and Discussion Recoveries of the pesticides were determined by spiking 10 µg/kg of
Figure 1 shows example chromatograms of the pesticides pesticides in three different berry samples (raspberry, blackberry
analyzed in MRM mode at 1 ng/mL. All of the tested pesticides and blueberry) in triplicates. Both the mean recovery and
were detected with good signal to noise even at concentrations reproducibility (RSD) were determined for each berry/analyte
well below the regulatory limits. combination. The recoveries were between 70 and 115% with a
RSD of <20% for the berry/analyte combinations.
The method showed excellent linearity (R2 ≥ 0.99) over three
orders of concentration (0.1-100 ng/mL for most analytes). Some Berry samples bought in local grocery stores were tested for
of the calibration curves are shown in Figure 2. pesticide residues using the developed method. Figures 3-5 show
the chromatograms of berry samples with positive hits for the
The limit of quantification (LOQ) was 0.1 ng/mL for most of the
target pesticides. The calculated concentrations of those pesticides
analytes, which is well below the regulatory limits of 10 ng/mL.
are listed in Table 3, which ranged from 4.5 to 447.3 μg/kg.
Figure 1. MRM chromatogram of pesticides at 10 ng/mL in neat solution.
Figure 2. Calibration curves for hexythiazox (a), pyrochlostrobin (b), and thiamethoxam (c) with 6 injections at each concentration level (ng/mL).
Figure 3. Pesticides identified and quantified from brand A blueberry. The pesticides are flonicamide (a), thiamethoxam (b), pyrimethanil (c), and bifenazate (d). 3
29
Figure 4. Pesticides identified and quantified from brand B blueberry. The pesticides are boscalide (a), cyprodinil (b), pyrochlostrobin (c), and hexythiazox (d).
Figure 5. Pesticides identified and quantified from brand D blackberry. Only hexythiazox was detected.
Table 3. Summary of Pesticides found in berry samples.
Pesticide Concentration (µg/kg)

Pyraclostrobin 11.8
Flonicamid 9.7
Blueberry (Brand A) Bifenazate 12.6
Thiamethoxam 9.5
Pyrimethanil 11.8
Cyprodinil 16.8
Pyraclostrobin 23.2
Blueberry (Brand B)
Boscalid 51.1
Hexythiazox 4.5
Blueberry (Brand C) N/A --
Blackberry (Brand D) Hexythiazox 447.3
4
30
Conclusions These results demonstrated this methods applicability and
A LC/MS/MS method for multi-pesticide residue analysis in effectiveness in detecting and quantitating pesticides lower than
berries was developed using a PerkinElmer Altus UPLC® system 10 parts per billion (ppb), per regulatory limits set by the EU
coupled to a QSight 220 triple-quad mass spectrometer. directive 91/414/EEC.
The simple/routine sample preparation approach used in this

work provides the following advantages: 1. dilution of the
QuEChERS extract with water makes the sample extract more
compatible with typical reversed phase separation, leading
to reduced solvent effects; 2. dilution also helps to reduce
any potential matrix effects, leading to more accurate and
reproducible results.
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31
Mass Spectrometry
Authors:
Li-Zhong Yang, Zhuo Man, Xiangdong Zhou
PerkinElmer, Inc.
Shanghai, China
Feng Qin
PerkinElmer, Inc.
Bolton, Canada
Direct Analysis of Glyphosate

and Similar Polar Pesticides in Introduction
Glyphosate (N-(phosphonomethyl)
Oatmeal by UHPLC-MS/MS glycine), an organophosphorus
compound, is used to kill weeds
(e.g. annual broadleaf weeds and
grasses) that compete with crops. Since its introduction to market approximately
40 years ago, glyphosate has become one of the world’s most widely used herbicides
due to its relatively low toxicity in comparison with other herbicides towards mammals.
The adoption of glyphosate by farmers intensified after the introduction of genetically
engineered “glyphosate tolerant” crops, such as corn and soybeans, that can withstand
glyphosate treatment unlike the weeds the herbicide is meant to destroy. Like other
pesticides, glyphosate is directly administered to food products and can come in contact
with both food workers and the environment, resulting in the bio burden of exposure
in uncontrolled regional populations. As a registered herbicide product under a number
of regulatory organizations, glyphosate has been considered nontoxic with minimal
risk to human health with persistent exposure at trace levels. However, recent
toxicity evaluations by different organizations have put glyphosate at the center of a
dispute. The World Health Organization's (WHO) International Agency for Research
on Cancer classified it as “probably carcinogenic to humans” in March of 20151.
However, in November of 2015, the European Food Safety Authority (EFSA) published
a report claiming that there was no scientific evidence linking glyphosate to cancer2.
32
Independent of the dispute in the scientific community, federal The mass spectrometer was equipped with an electrospray
regulations have been established by food authorities in several ionization source operating in negative ion mode. The mass
countries. The typical maximum residual level for glyphosate is spectrometer source conditions are shown in Table 2:
between 0.05 to 500 mg/kg, but may vary depending on the
Table 2. Mass spectrometer source conditions.
food commodity.
Parameter Setting
Glyphosate is a very polar compound with high solubility in water
and low solubility in most organic solvents. These properties mean Dry Gas 150
that these compounds do not retain well on conventional C18 LC Nebulizer Gas 220
columns and non-polar GC columns. Therefore, the derivatization Heating Gas Temp 500 °C
with fluorenylmethyloxycarbonyl chloride (FMOC-Cl) is a common
Electrospray Voltage -4500 V
procedure to improve extraction and separation of glyphosate
and other related compounds with LC and GC based methods.
MRM settings for each analyte were optimized by infusing neat
These methods based on derivatization are labor-intensive, time-
standard solutions. The parameters for each analyte’s MRM
consuming and less reproducible.
transition are listed in Table 3. The dwell time for each MRM
There is a growing need to develop a method for analysis was set at 30 ms.
of glyphosate and other related polar compounds without
derivatization. Recently, the EU Reference Laboratories (EURL) Table 3. Optimized MRM settings.
published two methods that can directly analyze glyphosate Compound Transitions m/z EV /V CE /eV
(GLY), its metabolite, aminomethylphosphonic acid (AMPA),
167.6/62.9* 35
and glufosinate (GLU) without derivatization. One method used (GLY) -19
167.6/149.6 14
an ion exchange column with a long run time (23 min), while the
second method utilized a Hypercarb column, which requires a 109.7/63.0* -30
(AMPA) -27
special priming/reconditioning procedure and showed significant 109.7/78.9 37
chromatographic peak tailing3. Our study reports a 12 minute 179.6/63.0* 53
LC/MS/MS method with an amino-based column to analyze (GLU) -20
179.6/84.9 27
glyphosate and other related polar compounds in underivatized * Quantifier ion
states, with exceptional selectivity and sensitivity.
Sample Preparation
Experimental
1.0 g of oatmeal sample was weighed into a centrifuge tube,
A PerkinElmer Altus® A-30 UPLC® system was used with a 10 mL of water/ acetonitrile (V/V, 2/1) was added to the tube
PerkinElmer QSight™ 210 triple quadrupole mass spectrometer. and the mixture was then shaken/vortexed for one minute,
Instrument control, data acquisition and processing was ultra-sonicated for 15 minutes and centrifuged for five minutes
performed using the PerkinElmer Simplicity 3Q™ software. at 6000 rpm. The recovered supernatant was filtered through
The LC method conditions are provided in Table 1. a 0.22 μm nylon membrane filter for LC/MS/MS analysis. To
avoid possible interaction between analytes and glass surfaces,
Table 1. LC method.
plastic sample vials were used during the analysis and samples
were analyzed immediately after preparation.
Column Shodex NH2P-50 2D column, 2.0 x 150 mm, 5 μm
A: 5 mM ammonium acetate (pH11.0) in water; Standards Calibration Solutions
Mobile Phase
B: acetonitrile
Matrix matched calibration standards were prepared by
Flow Rate 0.25 mL/min. adding different levels of analytes (5.0, 10.0, 100.0, 200.0
Oven Temp. 35 ºC and 500.0 ng/mL, respectively) in oatmeal matrix extract.
Injection Volume 10 µL
Mobile Phase
Time(min)
A (%) B (%)
0.00 20 80
Gradient Conditions 2.00 20 80

2.01 80 20
8.00 80 20
8.01 20 80
12.00 20 80
2
33
Results and Discussion
Figure 1 shows typical MRM chromatograms for the three analytes spiked to 10 ng/mL (0.1 mg/kg) in oatmeal extract. All three analytes
were well retained on the column and showed good peak shape and signal to noise. GLY and AMPA were eluted at very similar
retention times due to their similar chemical structure. GLU was baseline separated from the other analytes. No matrix interferences,
which can affect peak integration, were observed.
A B C
Figure 1. MRM Chromatograms of GLY (A), AMPA (B), and GLU (C) spiked at 10 ng/ml in oatmeal extract.
It is commonly known that LC/MS/MS, especially when working in Figure 2 shows the calibration curves for GLY, AMPA and GLU. Good
ESI mode, is susceptible to matrix effects, affecting quantitational linear correlation coefficients (R2≥0.997) were obtained between
accuracy. In this study, signal intensities of standards in neat solution concentrations of 5 to 500 ng/mL (0.05-5 mg/kg in real sample).
were compared with those of standards in matrix-matched solution For the 5 ng/mL calibrant, the signal-to-noise ratios (S/N) for GLY,
at different concentration levels to calculate matrix effects (ME). An AMPA and GLU were 432, 165, and 325, respectively. From these
ME value of less than 100% indicates matrix suppression, whereas values, the limits of quantitation (LOQs; S/N ≥ 10) were calculated
an ME value larger than 100% indicates matrix enhancement. As to be 0.12, 0.30 and 0.15 ng/mL. As the EU has set the maximum
seen in Table 4, both GLY and AMPA show matrix suppression, residue limit (MRL) for glyphosate in oatmeal at 20 mg/kg, the
while GLU shows matrix enhancement. Using matrix-matched method developed in this study easily meets this requirement.
standards, one can often compensate for matrix effects, which
may allow for good quantitational accuracy without the use of Table 4. Matrix effect result in oatmeal matrix.
internal standards. Therefore, calibration curves were generated by Compound GLY AMPA GLU
running matrix-matched calibration standards as described in the
Matrix effect (%) 67.3 87.3 107.6
experimental section.
A B C
Figure 2. Calibration curves for GLY (A), AMPA (B) and GLU (C) in oatmeal extract, respectively
3
34
Table 5. Linear dynamic range, regression coefficients, LOQ and S/N at LOQ level for analytes. Conclusion
Range S/N at LOQ in matrix In this study, we reported a rapid, sensitive and reliable
Compound R2
(ng/mL) 5 ng/mL (S/N ≥ 10) Ng/mL 12 min LC/MS/MS method that allowed direct analysis of
GLY 5-500 0.999 432 0.12 GLY, AMPA, and GLU in oatmeal without derivatization.
AMPA 5-500 0.997 165 0.30 The sample preparation method was a simple water/
acetonitrile extraction, which showed good recoveries
GLU 5-500 0.999 325 0.15
and minimal matrix effects for all three compounds. The
calibration curves for three analytes exhibited good linearity
Recovery of the analytes was evaluated at concentrations of 0.05 and over three orders of magnitude with calibration fit of R2
1 mg/kg. All recoveries were satisfactory, with mean values ranging greater than 0.997. The LOQs for glyphosate and other
from 85% to 130%, and relative standard deviations less than 13% for related polar compounds were much lower than the EU’s
all three analytes (Table 6). MRL of 20 mg/kg in oatmeal.
Table 6. Recovery of the analytes from oatmeal sample at different concentration levels.
Spiked Level (50 μg/kg) Spiked Level (1 mg/kg)

References
Compound
Recovery /% Recovery /% Recovery /% RSD/% 1. http://monographs.iarc.fr/ENG/Monographs/vol112/
mono112-09.pdf. Accessed on Aug 2nd, 2016
GLY 118 8.74 85 6.91
2. Conclusion on the peer review of the pesticide risk
AMPA 130 11.8 94 8.72
assessment of the active substance glyphosate. EFSA
GLU 122 12.9 96 1.93
Journal 2015; 13(11):4302.
3. Quick Method for the Analysis of numerous Highly
Polar Pesticides in Foods of Plant Origin via LC-MS/MS
involving Simultaneous Extraction with Methanol
(QuPPe-Method). http://www.crl-pesticides.eu/userfiles/
file/EurlSRM/meth_QuPPe-PO_EurlSRM.pdf. Accessed
on Aug 2nd, 2016
PerkinElmer, Inc.
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35
Mass Spectrometry
Authors:
Josh Ye, Jingcun Wu, Feng Qin, Shixin Sun,
Avinash Dalmia, Wilhad Reuter, Sergey Rakov,
Jamie Foss and Frank Kero
PerkinElmer, Inc.
Waltham, MA
Analysis of 213 Pesticide

Residues in Grapes by LC-MS/ Introduction
The Grape crop is one
MS with Time-Managed MRM of the most important
fruit crops consumed
in the world. Grapes are consumed both as fresh and as processed
products, such as wine, jam, juice, jelly, grape seed extract, raisins,
vinegar and grape seed oil. A large variety of pesticides are used in
grape production throughout its growing season to control pests and
diseases in vineyards and to increase crop yield. Pesticide residue is a
major concern for the stakeholders of the grape industry, due to more
and more stringent regulations and safety standards in most countries. It
is also a concern for the general consumers, due to increased demand
for safer products. Therefore, to prevent health risks, it is important to
monitor the presence of pesticides and regulate their levels in grapes.
36
Table 3. Optimized MRMs and compound-dependent parameters for selected pesticides.
Compound Name Polarity Q1 Mass Q2 Mass CE EV CCL2
Acetamiprid Positive 223.2 126.1 -30 25 -49
Acetamiprid-2 Positive 223.2 99.1 -56 25 -73
Azoxystrobin Positive 404.1 372.1 -18 25 -57
Azoxystrobin-2 Positive 404.1 344.1 -34 25 -71
Boscalid Positive 343.0 307.0 -25 25 -57
Boscalid-2 Positive 343.0 140.0 -28 25 -60
Chlorantranilprole Positive 484.0 452.8 -20 25 -66
Chlorantranilprole-2 Positive 484.0 285.8 -18 25 -65
Chlorpyriphos Positive 350.0 97.0 -32 25 -64
Chlorpyriphos-2 Positive 350.0 198.0 -20 25 -53
Clofentezine Positive 303.0 138.0 -28 25 -56
Clofentezine-2 Positive 303.0 102.0 -50 25 -75
Cyprodinil Positive 226.0 93.0 -48 25 -66
Cyprodinil-2 Positive 226.0 77.0 -34 25 -53
Diafenthiuron Positive 385.2 329.1 -26 25 -62
Diafenthiuron-2 Positive 385.2 278.1 -44 25 -78
Difenoconazole Positive 406.2 251.1 -32 25 -69
Difenoconazole-2 Positive 406.2 111 -76 25 -109
Difenoconazole-3 Positive 406.2 272.1 -22 25 -60
Dimethomorph Positive 388.2 301.1 -26 25 -62
Dimethomorph-2 Positive 388.2 165.1 -40 25 -75
Fenhexamid Positive 302.0 97.0 -32 25 -59
Fenhexamid-2 Positive 302.0 55.0 -60 25 -84
Fludioxonil Negative 246.6 125.9 40 -25 60
Fludioxonil-2 Negative 246.6 179.9 39 -25 60
Fluopyram Positive 397.0 173.0 -35 25 -71
Fluopyram-2 Positive 397.0 145.0 -70 25 -103
Imidachloprid Positive 256.2 209.0 -18 25 -42
Imidachloprid-2 Positive 256.2 175.2 -26 25 -49
Pyrimethanil Positive 200.0 107.0 -33 25 -50
Pyrimethanil-2 Positive 200.0 82.0 -32 25 -49
Pyraclostrobin Positive 388.0 194.0 -16 25 -53
Pyraclostrobin-2 Positive 388.0 163.0 -36 25 -71
Spinosad -1 Positive 732.6 142.0 -42 25 -111
Spinosad -2 Positive 732.6 98.1 -100 25 -163
Spirotetramat Positive 374.2 330.1 -21 25 -56
Spirotetramat-2 Positive 374.2 216.1 -45 25 -78
Spirotetramat-3 Positive 374.2 302.1 -23 25 -58
Spiroxamine Positive 298.3 144.2 -30 25 -57
Spiroxamine-2 Positive 298.3 100.2 -50 25 -75
Spinetoram Positive 748.4 142.1 -42 25 -113
Spinetoram-2 Positive 748.4 98.1 -100 25 -165
Trifloxystrobin Positive 409.0 186.0 -26 25 -64
Trifloxystrobin-2 Positive 409.0 206.0 -20 25 -59
Tebuconazole Positive 308.0 70.0 -30 25 -58
Tebuconazole-2 Positive 308.0 125.0 -50 25 -76
3
37
Analytical Challenges for Testing Multi-residues of
Pesticides from Food Samples
Traditional MRM method development is not suitable for
analysis of a large number of analytes such as pesticide residues
in a single run. This is not only because it is time-consuming
and labor intensive to manually entering all the mass transitions
into a method, but also because the dwell time for each
transition cannot be optimized easily. Therefore, the time-
managed-MRM feature in Simplicity software is especially
helpful in this regard, as this approach results in better data
quality by generating an optimum dwell time for each MRM. Figure 1. Example of MS method for 500 MRMs for 213 analytes generated by the
Figure 1 shows an example of a method generated using time- time-managed MRM module of the Simplicity Software.
managed-MRM in this study.
Linearity and Sample Matrix Effect
Sample matrix effects are still the main concern for LC-MS/MS, Calibrations were performed by preparing and running seven
especially for food analysis due to the diversity and complexity concentration levels of analytes standards in both neat solution
of food sample matrices. To overcome sample matrix effects, (pure solvent) and grape sample matrix (matrix-matched calibration).
numerous tools have been widely applied to LC-MS/MS method Example calibration curves for some of the most frequently found
development, such as sample dilution, use of stable isotope pesticides in grapes in this study are shown in Figure 2. Overall, for
internal standards, sample matrix-matched standard calibration, all analytes, calibration results showed good linearity over three orders
standard addition method, sample clean-up, use of high of magnitude (0.1 − 200 μg/L), with regression coefficient (R2 ≥ 0.98)
efficiency UHPLC column for better separation, and the use for most of the analytes in both neat solution and grape matrix.
of alternative ionization sources.13 The most common sample Quantitative precision (%RSD) for all analytes at 10 and 100 μg/L (not
matrix effect is discussed in details in the following section. shown), were all found to be between 1.2 to 15.1% (average of five
replicate injections).
Figure 2. Example calibration curves for some of the most frequently found pesticides in grapes: boscalid, cyprodinil, fenhexamid and pyrimethanil.
4
38
Possible sample matrix effects were evaluated by comparison Analyte Recovery, Limit of Quantification and Sample Results
of the responses (peak area in this case) of analytes obtained The percent recoveries of pesticides were evaluated at a
from neat solution and those obtained from grape sample concentration level of 100 μg/kg in two different samples: brand
matrix (spiked at the same concentration, 100 ppb in this case). A non-organic and brand G organic grapes. The recoveries of
When the percentage ratio value is greater than 100%, there analytes were between 75% to 114%, with an RSD < 10%
is a signal-enhancement, whereas a value of less than 100% for most analytes in the studied matrices. An overlay of the total
indicates signal-suppression. As illustrated in Table 3, no significant ion chromatograms (TIC) for an organic grape sample fortified
ion suppression effect was found for the studied compounds, at 100 μg/kg before and after the QuEChERS sample preparation
except for diafenthiuron, which showed some ion suppression. is shown in Figure 3.
The limits of quantification (LOQs) were determined based on
the signal to noise ratio of ≥10 for the quantifier transitions of
Table 3. Example matrix effects for the pesticides identified from the grape samples
in this work. all analytes. The identity of each pesticide residue is confirmed
Peak Area Peak Area by ensuring that the product ion ratios (qualifier vs. quantifier)
Matrix
Pesticides (In Neat (In Matrix were within 30% tolerance windows of the expected ratio.14
Effect
Solution) Spiked Solution)
The majority of the tested pesticides have a LOQ of ≤ than 1 μg/L
Acetamiprid 1024279 1068162 104.3 in grape matrix with a 1 µL direct injection.
Boscalid 696490 734080 105.4
The developed method was applied for the analysis of pesticide
Chlorantraniliprole 197766 190834 96.5 residues in a few brands of grapes. Example chromatograms for
Cyprodinil 453221 464004 102.4 the positively identified pesticides in sample brand B and F are
Diafenthiuron 957108 692523 72.4 shown in Figure 4 and 5, respectively. For brand F, it should be
Difenoconazole 1260903 1344640 106.6
noted that both green and red grape samples were analyzed.
The determined pesticide concentrations from these samples are
Dimethomorph 435381 366000 84.1
summarized in Table 4, along with the corresponding LOQ and
Fenhexamid 373104 411145 110.2 EU maximum residue limit (MRL) values for these pesticides.
Fludioxonil 293046 322095 109.9
Fluopyram 2281108 2379392 104.3
Imidacloprid 305674 304391 99.6
Pyrimethanil 366881 381018 103.9
Pyraclostrobin 1682524 1764409 104.9
Spinosad 666859 698195 104.7
Spirotetramat 517179 485990 94.0
Spiroxamine 1273651 1354261 106.3
Spinetoram 641735 665530 103.7
Trifloxystrobin 2132708 2219020 104.0
Tebuconazole 653128 620261 95.0 Figure 3. An overlay of the total ion chromatograms (TIC) for an organic grape
sample fortified at 100 μg/kg before (red) and after (green) the sample preparation.
# Pesticide Residues RT (min)

1 Acetamiprid 8.03
2 Boscalid 12.76
3 Chlorantraniliprole 12.36
4 Cyprodinil 13.48
5 Fenhexamid 12.63
6 Fludioxonil 12.16
7 Pyrimethanil 11.82
8 Spinetoram 17.40
9 Spirotetramat 13.43
10 Trifloxystrobin 14.85
11 Tebuconazole 13.59
Figure 4. Chromatogram and list of pesticides positively identified in brand B grape sample.
5
39
# Pesticide Residues RT (min)
1 Boscalid 12.76
2 Cyprodinil 13.48
3 Fenhexamid 12.63
4 Fludioxonil 12.16
5 Pyrimethanil 11.82
6 Spirotetramat 13.43
7 Spinosad 17.01
8 Spinetoram 17.40
8
Figure 5. Chromatogram and list of pesticides positively identified in brand F grape samples (green for green grape and red for red grape).
Table 4. Summary results for the positively identified pesticide residues in grapes in μg/kg.
Pesticide Brand A Brand B Brand C Brand D Brand E Brand F* Brand G MRL LOQ
Acetamiprid 17 31 500 0.1
Boscalid 1244 888 2294 1139 1438 (800) 1 5000 0.2
Chlorantraniliprole 29 20 0.2
Cyprodinil 93 51 3 169 (435) 3000 0.2
Diafenthiuron 18 3000 0.2
Difenoconazole 29 27 3000 0.1
Dimethomorph 14 58 3000 0.2
Fenhexamid 319 17 777 1018 340 (553) 15000 1
Fludioxonil 213 66 11 195 (336) 5000 0.2
Fluopyram 1 1500 0.1
Imidacloprid 2 1000 0.2
Pyrimethanil 5 66 271 966 (451) 5000 1
Pyraclostrobin 180 1000 0.2
Spinosad 27 57 46 500 0.2
Spirotetramat 7 7 3 87 (61) 2000 1
Spiroxamine 12 11 600 0.2
Spinetoram 47 6 29 47 (54) 500 0.1
Trifloxystrobin 69 14 4 3000 0.2
Tebuconazole 204 500 0.2
* Initial value is for green grape sample and the value in parenthesis is for red grape sample.
6
40
Conclusions
A LC-MS/MS method for multi-residue pesticides analysis in 6. M. Kirchner, E. Matisova, S. Hrouzkova, and J. D. Zeeuw,
grapes was developed by coupling a UHPLC system to a QSight Possibilities and limitations of quadrupole mass spectrometric
220 triple quadrupole mass spectrometer. This method can be detector in fast gas chromatography. J. Chromatogr. A,
applied for the determination of pesticide residues in grapes, 2005, 1090(1-2), 126–132.
with LOQs well below the limits set by regulatory boards. 7. J.Wu, Quantitative Method for the Analysis of Tobacco-
The time-managed-MRM module in the Simplicity software was Specific Nitrosamines in Cigarette Tobacco and Mainstream
effectively used in this study for monitoring 213 pesticide residues Cigarette Smoke by Use of Isotope Dilution Liquid
in grapes using the QSight™ LC-MS/MS system. This feature has Chromatography Tandem Mass Spectrometry, Anal. Chem.,
simplified the creation and optimization of MS methods for 2008, 80 (4), 1341–1345.
monitoring a large number of analytes in food samples. 8. K. Zhang, M.R. Schaab, G. Southwood, E.R. Tor, L.S. Aston,
The QuEChERS sample preparation method utilized in this W. Song, B. Eitzer, S. Majumdar, T. Lapainus, H. Mai, K. Tran,
study demonstrated good recovery (75−114%) and excellent A. El-Demerdash, V. Vega, Yanxuan Cai, J.W. Wong, A.J.
quantitative reproducibility (RSD<10%) for most pesticides. Krynitsky, and T.H. Begley, A Collaborative Study:
Determination of Mycotoxins in Corn, Peanut Butter, and
The developed LC-MS/MS method showed good linearity, with Wheat Flour Using Stable Isotope Dilution Assay (SIDA) and
LOQ ≤ 1 µg/kg for most of the 213 pesticides in grape matrix. Liquid Chromatography–Tandem Mass Spectrometry (LC-MS/
A number of pesticide residues were identified and quantified MS), J. Agric. Food Chem. 2017, 65 (33), 7138-7152.
with concentrations greater than 1 µg/kg in all the non-organic
9. K. Zhang, J.W. Wong, P. Yang, K. Tech, A.L. DiBenedetto, N.
brands of grapes. However, for the organic grapes tested, only
S. Lee, D.G. Hayward, C.M. Makovi, A.J. Krynitsky, K.
one pesticide (boscalid) was detected at a concentration level
Banerjee, L. Jao, S. Dasgupta, M.S. Smoker, R. Simonds, and
of 1 µg/kg.
A. Schreiber, Multi residue Pesticide Analysis of Agricultural
The same LC-MS/MS method has also been applied successfully Commodities Using Acetonitrile Salt-Out Extraction,
to other fruit analyses, such as berries, orange and grapefruit, Dispersive Solid-Phase Sample Clean-Up, and High-
all with good performance. These results demonstrated the Performance Liquid Chromatography–Tandem Mass
method’s applicability and effectiveness in detecting and Spectrometry J. Agric. Food Chem. 59, 2011, 7936–7946.
quantifying pesticide residues in fruit samples.
10. K. Banerjee, D.P. Oulkar, S. Dasgupta, S. B. Patil, S. H. Patil,
References R. Savant and P. G. Adsule, Validation and uncertainty
analysis of a multi-residue method for pesticides in grapes
1. Commission Regulation (EC) 396/2005 on maximum residue
using ethyl acetate extraction and liquid chromatography–
levels of pesticides in or on food and feed of plant and
tandem mass spectrometry, J. Chromatogr. A, 2007, 1173,
animal origin, J. Eur. Union.L70/1 (2005).
98–109.
2. US Environmental Protection Agency, Electronic code of
11. S. Grimalt and P. Dehouck, Review of analytical methods for
federal regulation: Title 40: part 180-tolerance and
the determination of pesticide residues in grapes, J.
exemptions for pesticide chemical residues in Food. http://
Chromatogr. A, 2016, 1433,1–23.
www.ecfr.gov/cgi-bin/text-idx?c=ecfr&tpl=/ecfrbrowse/
Title40/40cfr180_main_02.tpl. 12. P. Cabras and A. Angioni, Pesticide Residues in Grapes,
Wine, and Their Processing Products, J. Agric. Food Chem.,
3. China National Standard GB 28260-2011. 2011. Maximum
2000, 48 (4),967–973.
residue limits for 85 pesticides in food, Ministry of Health of
the People’s Republic of China. 13. A. J. Krynitsky, J. W. Wong, K. Zhang and H. Safarpour,
Focus on Food Analysis: Important considerations regarding
4. APEDA (2006), Regulation of export of fresh grapes from India
matrix effects when developing reliable analytical residue
through monitoring of pesticide residues, Amendments in
methods using mass spectrometry, LCGC North America,
grape RMP –2007, Amendment-5 (Revised Annexure – 7&11).
2017,Vol. 35, No. 7, 444-451.
5. European Committee for Standardization, Foods of plant
14. Document SANTE/11813/2017 on Analytical Quality Control
origin-Multi-residue methods for the gas chromatographic
and Method Validation Procedures for Pesticide Residues
determination of pesticide residues, EN 12393-1 (2008).
Analysis in Food and Feed.
PerkinElmer, Inc.
940 Winter Street
P: (800) 762-4000 or
(+1) 203-925-4602
www.perkinelmer.com
013885_01 PKI
41
APPLICATION NOTE
Mass Spectrometry
Authors:
Yufeng Gao
Zhenpeng Zhen
GuangZhou Sugarcane Industry Research Institute

Guangzhou, China
Lizhong Yang
Chengyuan Cai
PerkinElmer, Inc.
Shanghai, China
Joshua Ye
PerkinElmer, Inc.
Toronto, Canada
Detection of Fipronil
and its Metabolites in Introduction
A recent recall of contaminated eggs in
Eggs by LC/MS/MS Europe has generated a serious health
concern for fipronil poisoning1, 2. Fipronil is
a highly toxic insecticide and it is used to
protect crops. The U.S. EPA has classified fipronil as a group C carcinogen3. Therefore, it is
not allowed anywhere near animals in the food production chain, specifically around
chickens. Traditionally, fipronil is measured by GC/MS or HPLC, however, its metabolites are
not included. According to (EU) No.1127/2014, the maximum residue allowance level
of fipronil (including its metabolites) in egg is 5 μg/L4. In this study, we describe a fast and
robust analytical method for identifying fipronil and its metabolites with the PerkinElmer
QSight® 200 series LC/MS/MS triple quadrupole instrument. The results demonstrate
excellent recovery, linearity and reproducibility and superior sensitivity for this method. The
three-step sample preparation procedure is quick and easy, and the LC/MS/MS
analysis time can be carried out within six minutes with great separation for all analytes.
42
Experimental Conditions
Sample Preparation LC Method Parameters
5.00 g of homogenized egg was weighed and transferred to a The starting conditions for the gradient flow were (60/40, A/B)
50 mL tube. Then the sample was extracted with 25 mL of with a 0.4 mL/min flow rate. Detailed LC conditions and time
acetonitrile. For better extraction, the samples were first vortexed program are shown in the Table 1.
for one minute, then sonicated for five minutes. Subsequently, 2 g
MS Method Parameters
of NaCl was added to the sample tube, votexed for one minute
MRMs for fipronil and its metabolites are optimized for the QSight
and then centrifuged at 6000 rpm for three minutes. The prepared
220 LC/MS/MS, CE values and source parameters are listed in
sample was finally filtered through a 0.22µm filter then injected
Table 2 and 3, respectively.
onto a PerkinElmer Brownlee® SPP C18 column and analyzed by a
QSight 220 LC/MS/MS system.
Table 1. LC parameters.
Mobile Phase Solvent A: Water

Solvent B: Acetonitrile
Flow Rate
Time (min) %A %B
(mL/min)
1 0 0.4 60 40
2 2.8 0.4 5 95
3 4.0 0.4 5 95
4 4.1 0.4 60 40
5 6.0 0.4 60 40
Column Brownlee SPP C18, 2.1x100 mm, 2.7μm
Oven 35 ºC
Table 2. MS/MS parameters. Results

Precursor Fragment CE Figure 1 shows example of chromatography for each analyte
Compound
(m/z) (m/z) (eV)
at 0.05 μg/L level. Excellent symmetric peak and good
386.8 281.8 44 separation were observed.
Fipronil Desulfinyl
386.8 350.8 20
This LC/MS/MS method showed excellent linear range over three
418.8 261.8 39 orders of magnitude (0.05 – 10 μg/L) with good regression
Fipronil Sulfide
418.8 382.8 21 coefficient (R2 ≥ 0.998) for all analytes. Typical matrix-matched
434.8 249.8 36 calibration curves for all four analytes are shown in Figure 2.
Fipronil
434.8 329.8 24 Recovery was also studied by matrix-spiked experiment at
450.8 243.8 66 concentration level of 0.5 and 5.0 µg/L. The recovery obtained was
Fipronil Sulphone between 92.7% – 114.5% with RSD < 5% for fipronil and its
450.8 281.8 37
metabolites as shown in Table 4.
Table 3. MS source parameters.

Ionization Source ESI negative mode
Source Voltage – 4500 V
Dyring Gas Setting 100
Nebulizer Gas Setting 180
Source Temperature 450 °C
2
43
Fipronil Desulfinyl Fipronil Sulfide
Fipronill Fipronil Sulphone
Figure 1. Matrix spiked chromatograms at 0.05 μg/L level for fipronil desulfinyl, fipronil sulfide, fipronil, and fipronil sulphone.
Figure 2. Selected matrix-matched calibration curves from 0.05 to 10 μg/L for fipronil desulfinyl, fipronil sulfide, fipronil, and fipronil sulphone.
44 3
Table 4. Recovery and RSD at spiked concentration level of 0.5 and 5.0 µg/L.
Spiked Level 0.5 μg/L Spiked Level 5.0 μg/L
Compound
Recovery (%) RSD (%) Recovery (%) RSD (%)
Fipronil Desulfinyl 102.5 3.7 98.6 1.5
Fipronil Sulfide 106.8 4.3 103.9 3.0
Fipronil 107.5 3.4 113.8 2.6
Fipronil Sulphone 114.5 3.2 92.7 2.4
Conclusion References
The LC/MS/MS methodology of detecting fipronil and its 1. http://www.bbc.com/news/world-europe-40824819
metabolites in eggs, namely, fipronil sulphone, fipronil sulfide, 2. http://news.sina.com.cn/o/2017-08-04/doc-ifyiswpt5354785.shtml
and fipronil desulfinyl was developed using a PerkinElmer QSight
220 LC/MS/MS triple quadrupole instrument. This methodology 3. "Fipronil Technical Fact Sheet, National Pesticide Information
showed excellent linearity from 0.05 to 10 μg/L with good Center"; http://npic.orst.edu/factsheets/archive/fiptech.html
regression coefficient (R2 ≥ 0.998) for all analytes. The simple 4. http://eur-lex.europa.eu/legal-content/EN/TXT/?qid=150218046899
three-step sample preparation was demonstrated by excellent 3&uri=CELEX:32014R1127
recovery of between 93 – 115% and RSD of less than 5% at both
0.5 and 5.0 μg/L spiked concentration levels. The method presented
here provides excellent resolution and sensitivity (at least 100 times
lower than that of EU MRL) for the quantification of fipronil and its
metabolites in eggs and meets the analytical needs for food
safety laboratories.
PerkinElmer, Inc.
940 Winter Street
P: (800) 762-4000 or
(+1) 203-925-4602
www.perkinelmer.com
013713_01 PKI
45
APPLICATION NOTE
Technology Line 1
Technology
Mass Spectrometry
Line 2
AUTHORS
AUTHORS
Avinash Dalmia1
John Smith Cai2
Sheng-Suan
Research Scientist
Saba Hariri3
PerkinElmer,
** conclusionInc.,
orShelton, CT text
other key USA **
Erasmus Cudjoe2
John
Jacob Smith
Approx.Jalali
2202Characters
Research Scientist
Toby Astill3
PerkinElmer,
Feng Qin3 Inc., Shelton, CT USA
John Smith
1
PerkinElmer,
Research Inc., Shelton, CT USA
Scientist
2
PerkinElmer,Inc.,
PerkinElmer, Inc.,Shelton,
Waltham,CTMA
USAUSA
3
PerkinElmer, Inc., Woodbridge, ON Canada
LC/MS/MS Analytical
Method for Pesticide
Residue in Cannabis Introduction
Flower as Defined by Since there are no federal regulations for
Florida Testing pesticide analysis in cannabis, individual

states define and regulate the use of
Requirements pesticides for the production of cannabis
intended for both medicinal and/or
recreational purposes. Currently each state has developed its own regulations
outlining the list of pesticides to monitor and the maximum residue limits or
acceptable limits for each pesticide. Among these states, Oregon, California and
Arizona have set acceptable levels for 59, 66 and 56 pesticides, respectively, in
cannabis (1-3). Recently, Florida imposed a requirement to test for 67 pesticide
residues in cannabis products. Florida’s list expanded on the California list of
66 by adding one more pesticide, and lowered acceptable limits for a few of the
pesticides relative to California’s limits. In addition to pesticide residues, Florida
also requires cannabis products to be tested for 5 mycotoxins (4 aflatoxins and
ochratoxin A) (4). Mycotoxin contamination can occur during the cultivation or
storage of cannabis. Like pesticides, these mycotoxins are toxic and pose a
serious health risk to consumers. As a result, testing for the levels of pesticide and
mycotoxins in cannabis is important to ensure health of consumers and quality
control in the State of Florida.
46
LC/MS/MS Analytical Method for Pesticide Residue in Cannabis Flower as Defined by Florida Testing Requirements…
A number of methods described in literature for pesticide • Place the tube on multi-tube vortex mixer and allow it to vortex
analysis in cannabis use time-consuming and costly sample for 10 minutes.
preparation methods that include multiple sample clean up
• Centrifuge extract in tube for 10 minutes at 3000 rpm.
steps using QuEChERS with dSPE or SPE. This approach lends
itself to poor sample preparation recoveries for some of the • Filter the solvent into a 5 mL glass amber vial using 0.22
pesticides due to their affinity to the stationary phase, and also micron nylon syringe-filter and cap it.
requires the lab to use of both LC/MS/MS and GC/MS/MS
• Label the bottle with the sample ID.
instruments to complete the analysis of all of the pesticides
(5-9). This approach is problematic for today’s busy compliance • Transfer 0.5 mL of extracted sample into a 2 mL HPLC vial
cannabis labs due to increased consumable costs, complexity, and dilute it with 0.49 mL of LC/MS grade acetonitrile and mix
and slower turnaround time of analysis resulting from the multi- it. Spike 10 µL of internal standard solution from a PerkinElmer
step clean up, preparation and associated maintenance on two One Pesticide420™ ISO17034 CRM Reagent Kit.
instruments. This application note outlines a new workflow with
LC Method and MS Source Conditions
one instrument platform (LC/MS/MS) method that is ideal for
simultaneous analysis of pesticide and mycotoxins residue in The LC method and MS source parameters are shown in Table 1.
cannabis. This application note demonstrates the performance
of a LC/MS/MS ESI and APCI method with a single solvent Table 1: LC & MS Method Conditions.
extraction step to meet Florida regulations for 67 pesticides and LC Conditions
5 mycotoxins in complex cannabis flower matrix in place as of
LC Column PerkinElmer Quasar SPP Pesticides
the date of this publication.
(4.6 × 100 mm, 2.7 µm)
Mobile Phase Gradient An 18 min. (this time includes both
Experimental analysis time and column equilibration
Materials and Reagents time) LC/MS/MS method with optimized
gradient using an ESI source with polarity
The PerkinElmer One Pesticide 420™ ISO17034 CRM Reagent Kit switching was used for separation and
was used for preparation of pesticide and mycotoxins standards analysis of 61 out of 67 pesticides and
at different concentrations for collecting validation data with a 5 mycotoxins residues at low levels in a
cannabis matrix. A fast 6 min. LC/MS/
LC/MS/MS method. To compensate for matrix effects observed
MS method with short gradient, optimum
in complex cannabis samples, 30 internal standards, all of which
mobile phase composition and an APCI
are included in the PerkinElmer kit, were used to improve method source in negative ion mode was used for
recovery and accuracy of quantitation. measurement of 3 out of 67 pesticides. A
fast 10 min. LC/MS/MS method with an
Hardware/Software ESI source in negative ion mode was used
for analysis of the remaining 3 pesticides.
Chromatographic separation was conducted on a PerkinElmer
QSight® LC/MS/MS LX50 UHPLC system, while detection was Column Oven Temperature 30 ºC
achieved using a PerkinElmer QSight 420 MS/MS detector with a
dual ionization ESI and APCI source, which operate independently Auto sampler Temperature 20 ºC
with two separate inlets. All instrument control, data acquisition Injection Volume 3.0 µL for the LC/MS/MS method using an
and data processing were performed using the Simplicity 3Q™ ESI source with polarity switching. 10 µL
software platform. for the LC/MS/MS method with an APCI
source. 6.0 µL for the LC/MS/MS method
Sample Preparation Method using an ESI source in negative ion mode.
Below is the step by step sample preparation procedure with

MS Source Conditions for ESI Source & APCI Source
10-fold dilution:
ESI Voltage (Positive) +5500 V
• Take approximately 5 grams of cannabis flower as a ESI Voltage (Negative) -4200 V
representative of each sample batch and grind it finely using a APCI Corona Discharge -5 µA
grinder (e.g., OMNI Bead Ruptor 96).
Drying Gas 150 arbitrary units
• Measure 1 gram of sample and place it into 50 mL Nebulizer Gas 350 arbitrary units
centrifuge tube.
Source Temperature (ESI 290 ºC
• Add 5 mL of LC/MS grade acetonitrile to the tube and cap it. 18 min. method)
www.perkinelmer.com 2 47
Table 1: LC & MS Method Conditions. Continued ... 67 pesticides and 5 mycotoxins. Three of the pesticides (captan,
MS Source Conditions for ESI Source & APCI Source methyl parathion and pentachloronitrobenzene (PCNB) were
measured with a 6 minute APCI source method. The remaining
Source Temperature 290 ºC
(ESI 18 min. method) three pesticides (chlorfenapyr, chlordane and cyfluthrin) were
measured using a 10 minute ESI source method in negative
Source Temperature 350 ºC
ion mode. Figure 1 shows MRM chromatograms with excellent
(APCI 6 min. method)
signal to noise ratio for a representative set of challenging
Source Temperature 150 ºC pesticides (MGK-264 and abamectin) and mycotoxins
(ESI 10 min. method)
(Aflatoxin-B1 and Ochratoxin-A) spiked at low levels in a range of
HSID Temperature 200 ºC 5-25ppb in the cannabis flower, respectively, using the 18 minute
(ESI 18 min. method)
LC/MS/MS method with ESI source and polarity switching. For
HSID Temperature 200 ºC determining the limits of quantitation, cannabis extracts were
(APCI 6 min. method)
fortified with different low levels of pesticides and mycotoxins.
HSID Temperature 150 ºC Florida’s acceptable limits, method limits of detection (LODs),
(ESI 18 min. method) method limits of quantification (LOQs), ratio of acceptable limit
Detection mode Time-managed MRM™ to method LOQ, and ratio of acceptable limit to method LOD for
each of the pesticides and mycotoxins in cannabis flower are
Results and Discussion summarized in Tables 2 and 3. The LOQs were determined by
measuring the signal to noise ratio for cannabis flower matrix
Detectability and Reproducibility spiked standards. The LOQ is the lowest concentration for spiked
The LC/MS/MS method with dual ESI and APCI sources cannabis matrix at which S/N = 10 or higher is measured for
analyzes the 67 pesticides and 5 mycotoxins in cannabis flower quantifier ion. The LODs were estimated by dividing the LOQ by
as required by Florida regulations with three separate injections a factor of 3.3, since LOD and LOQ are estimated based on S/N
using the same instrument platform with a total run time of 34 of 3 and 10, respectively. Florida regulations require chemical
minutes. A diverter valve enables quick, automated switching of methods to achieve LOQ and LOD for target analytes of at least
mobile phase eluent to the APCI source from the ESI source and one-half and one-tenth of the acceptable limit, respectively. As
vice versa to facilitate the fast analysis using this method without demonstrated in Tables 2 and 3, the ratio of acceptable limits
requiring the time consuming step of switching APCI and ESI ion to LOQ and LOD for each analyte was higher than 2 and 10,
sources in LC/MS/MS systems with a single source. A 18 minute respectively, meeting Florida regulations for all of 67 pesticides
LC/MS/MS method with polarity switching analyzed 61 out of and 5 mycotoxins.
Figure 1: MRM chromatograms of a representative set of pesticides and mycotoxins: (a) MGK-264, (b) abamectin, (c) ochratoxin-A, and (d) aflatoxin-B1, spiked at level of 5, 25, 5
and 5 ppb, respectively, in a cannabis flower matrix using the 18 minute LC/MS/MS method with an ESI source.
48
www.perkinelmer.com 3
Table 2: This table lists the pesticides and their acceptable limits for cannabis flower currently required by the Florida state regulations. It also shows method LOQ, ratio of
acceptable limit to LOQ, method LOD and ratio of acceptable limit to LOD for each pesticide in cannabis flower using an LC/MS/MS method with ESI and APCI sources. Note:
The pesticides analyzed using the 10 minute LC/MS/MS method with an ESI source are indicated using an asterisk (*). The pesticides analyzed using an LC/MS/MS with an
APCI source are indicated by a dagger (†). The rest of compounds that are not marked with either an asterisk or a dagger were determined using the 18 minute LC/MS/MS
method with an ESI source.
Acceptable Limit/ Acceptable Limit/ Acceptable Limit/

Pesticide LOQ/ppb LOD/ppb
ppb LOQ LOD
Abamectin 100 25 4 7.5 13.3
Acephate 100 10 10 3.0 33.3
Acequinocyl 100 5 20 1.5 66.6
Acetamiprid 100 5 20 1.5 66.6
Aldicarb 100 5 20 1.5 66.6
Azoxystrobin 100 5 20 1.5 66.6
Bifenazate 100 5 20 1.5 66.6
Bifenthrin 100 5 20 1.5 66.6
Boscalid 100 5 20 1.5 66.6
Captan† 700 50 14 15.0 46.6
Carbaryl 500 5 100 1.5 333.0
Carbofuran 100 10 10 3.0 33.3
Chlorantraniliprole 1000 5 200 1.5 666.0
Chlordane* 100 25 4 7.5 13.3
Chlorfenpyr* 100 5 20 1.5 66.6
Chlormequat chloride 1000 10 100 3 333.3
Chlorpyrifos 100 10 10 3.0 33.3
Clofentezine 200 5 40 1.5 133.2
Coumaphos 100 5 20 1.5 66.6
Cyfluthrin* 500 100 5 30.0 16.7
Cypermethrin 500 50 10 15.0 33.3
Daminozide 100 10 10 3.0 33.3
Dimethomorph 200 5 40 1.5 133.2
DDVP (Dichlorvos) 100 10 10 3.0 33.3
Diazinon 100 10 10 3.0 33.3
Dimethoate 100 10 10 3.0 33.3
Ethoprophos 100 10 10 3.0 33.3
Etofenprox 100 5 20 1.5 66.6
Etoxazole 100 5 20 1.5 66.6
Fenoxycarb 100 5 20 1.5 66.6
Fenpyroximate 100 5 20 1.5 66.6
Fenhexamid 100 5 20 1.5 66.6
Fipronil 100 10 10 3.0 33.3
Flonicamid 100 5 20 1.5 66.6
Fludioxonil 100 10 10 3.0 33.3
Hexythiazox 100 5 20 1.5 66.6
Imazalil 100 10 10 3.0 33.3
Imidacloprid 400 5 80 1.5 266.4
Kresoxim‐methyl 100 10 10 3.0 33.3
49
Table 2: Continued ...

Pesticide LOQ/ppb LOD/ppb
ppb LOQ LOD
Malathion 200 5 40 1.5 133.2
Metalaxyl 100 5 20 1.5 66.6
Methiocarb 100 5 20 1.5 66.6
Methomyl 100 10 10 3.0 33.3
Methyl parathion† 100 10 10 3.0 33.3
Mevinphos 100 5 20 1.5 66.6
Myclobutanil 100 5 20 1.5 66.6
Naled 250 5 50 1.5 166.5
Oxamyl 100 5 20 1.5 66.6
Paclobutrazol 100 5 20 1.5 66.6
PCNB† 150 10 15 3.0 50
Permethrins 100 25 4 7.5 13.3
Phosmet 100 5 20 1.5 66.6
Piperonyl Butoxide 3000 5 600 1.5 2000
Prallethrin 100 10 10 3.0 33.3
Propiconazole 100 10 10 3.0 33.3
Propoxur 100 5 20 1.5 66.6
Pyrethrins 500 10 50 3.0 166.5
Pyridaben 200 5 40 1.5 133.2
Spinetoram 200 10 20 3.0 66.6
Spinosad 100 10 10 3.0 33.3
Spiromesifen 100 10 10 3.0 33.3
Spirotetramat 100 5 20 1.5 66.6
Spiroxamine 100 5 20 1.5 66.6
Tebuconazole 100 5 20 1.5 66.6
Thiacloprid 100 5 20 1.5 66.6
Thiamethoxam 500 5 100 1.5 333.0
Trifloxystrobin 100 5 20 1.5 66.6
Table 3: This table lists the mycotoxins and their acceptable limits for cannabis flower as currently required by Florida state regulations. It also shows method LOQ, ratio of
acceptable limit to LOQ, method LOD and ratio of acceptable limit to LOD for each mycotoxin in cannabis flower using the 18 minute LC/MS/MS method with an ESI source.

Mycotoxin LOQ/ppb LOD/ppb
ppb LOQ LOD
Ochratoxin A 20 5 4 1.5 13.3
Aflatoxin B1 20 1 20 0.3 66.6
Aflatoxin B2 20 2.5 8 0.75 26.6
Aflatoxin G1 20 1 20 0.3 66.6
Aflatoxin G2 20 2.5 8 0.75 26.6
50 www.perkinelmer.com 5
Analysis of Non-Polar Pesticides Using an LC/MS/MS with an Evaluation of a Novel LC/MS/MS Method with the ESI Source
APCI Source in Negative Ion Mode for Non-Polar Pesticides
Some of the pesticides (methyl parathion, captan and PCNB) in In the past, we utilized a LC/MS/MS method with either an APCI
cannabis regulated by Florida are traditionally analyzed using source in negative ion mode or an ESI source in positive ion
a GC/MS/MS with an electron impact ionization source, since mode for analysis of non-polar compounds such as chlordane,
these pesticides have low proton affinity, which results in either chlorfenapyr, cyfluthrin and others (10-11). In order to meet
low or no signal with the ESI source in positive ion mode. To Florida’s stringent requirements of LOD and LOQ of one-tenth
achieve the required sensitivity for these three pesticides, the and one-half of the acceptable limit in cannabis for these
selected MRMs were optimized with an APCI source. Figure pesticides, we developed a novel LC/MS/MS method with an
2 shows MRM chromatograms for three pesticides (methyl ESI source in negative ion mode and optimum mobile phase
parathion, captan and PCNB) analyzed in a range of 10-50 ppb composition. Figure 3 shows low detection limits for some of
in the cannabis flower using a LC/MS/MS method with an APCI the pesticides (chlorfenapyr, cyfluthrin and chlordane) analyzed
source. LOQ for these analytes in cannabis flower was in the in low range of 5-100 ppb in the cannabis flower using this novel
range of 10-50 ppb, consistent with Florida regulations. LC/MS/MS method.
(a) (a)
S/N = 27 S/N = 20
(b) (b)
S/N = 18 S/N = 20
(c) (c)
S/N = 20 S/N = 25
Figure 2: MRM chromatograms of non-polar pesticides: (a) methyl parathion, (b) Figure 3: MRM chromatograms of non-polar pesticides: (a) chlorfenapyr, (b) chlordane
PCNB and (c) captan, spiked at a level of 10, 10 and 50 ppb, respectively, in a and (c) cyfluthrin, spiked at level of 10, 25 and 250 ppb, respectively, in a cannabis flower
cannabis flower matrix using the LC/MS/MS method with an APCI source. matrix using the 10 minute LC/MS/MS method with an ESI source in negative ion mode.
Analysis of Pesticides in a Cannabis Matrix Using Two Sample Preparation Extraction Efficiency
Different Modes of Ionization
Different groups have developed complex sample preparation
Apart from monitoring two transitions for unambiguous methods using either solvent extraction or QuEChERS extraction
identification of all of the pesticides in a cannabis matrix, we followed by SPE (solid phase extraction) or d-SPE with different
were able to measure some of the more challenging pesticides in sorbents (5-8). A number of analytes such as daminozide,
a cannabis matrix with two different LC/MS/MS methods using imazalil, spirotetramat, pyridaben and few others showed poor
different forms of ionization with either ESI or APCI sources recoveries with such intricate, expensive and time consuming
to increase our confidence in measuring these pesticides in sample preparation methods. Recently, another group developed
a challenging cannabis matrix with another complementary a solvent extraction method followed by an evaporation step
method. For example, a number of pesticides, such as chlordane, to carry out solvent exchange with initial LC mobile phase
chlorfenapyr, fipronil, fludoxinil, acequinocyl, captan, methyl for extraction of pesticides from a hemp matrix. The extra
parathion and abamectin, were measured with both ESI and evaporation step in this sample preparation method resulted
APCI sources. Figure 4 shows the response for acequinocyl in low recoveries for some of pesticides such as abamectin,
spiked at a level of 10 ppb in a cannabis matrix using both ESI fenpyroximate and others due to their precipitation (9). Due
and APCI ion sources. In this case, the APCI source method to low recoveries of some of pesticides with time consuming,
could qualify as an orthogonal method for confirmation of expensive, and difficult sample preparation methods such as
acequinocyl in cannabis samples. Although the ESI source is QuEChERS, solvent extraction with evaporation, d-SPE and SPE
slightly more sensitive for analysis of acequinocyl in some of to extract pesticides and mycotoxins from cannabis matrix,
strains of cannabis flower matrix, we observed that the APCI we used a simple acetonitrile based solvent extraction method
source is a better option for analysis of this pesticide in some other for extraction to get good recovery with high throughput. The
strains of cannabis flower matrix and cannabis concentrates fortified cannabis flower samples were used to determine
which show higher matrix interference and ion suppression for pesticides and mycotoxin extraction efficiency. The cannabis
this compound with an ESI source in comparison to an APCI flower samples were tested to confirm the absence of pesticides
source. Two of the pesticides, cyfluthrin and cypermethrin, were before they were spiked. Five cannabis flower samples were
measured with two different ESI source methods by ionizing spiked at 2 levels (low and high) of the 67 pesticides (100
them differently in positive and negative ion mode. and 1000 ppb) and 5 mycotoxins (10 and 100 ppb) standard.
The extraction efficiency of all of the 67 pesticides and five
mycotoxins at two different levels were within an acceptable
(a) range of 80-120% with RSD less than 20% for five cannabis
S/N = 20 flower samples. The sample extraction efficiency was calculated
by taking the ratio of signal of analyte in pre-spiked to post-
spiked extract and multiplying this number by 100.
Signal in prespiked Extract * 100

Sample Preparation Extraction Efficiency (%) =
Signal in postspiked Extract
Internal Standards to Compensate for Sample Matrix Effects

and Improve Overall Recovery or Accuracy
(b)
A cannabis flower matrix can cause severe matrix effects such
S/N = 14
as ion suppression or enhancement for a number of pesticides
and mycotoxins. The ion suppression/enhancement effects
were determined by checking signal for spiked pesticides and
mycotoxins at a fixed concentration in a cannabis flower matrix
and solvent standard. The calculation was performed by taking
the difference between the signal of the analyte in the post
spiked cannabis flower extract and clean solvent, and dividing
it by the signal in clean solvent. Among the 66 analytes (61
pesticides and 5 mycotoxins) measured with the ESI source,
most analytes showed signal suppression effects. Since a
Figure 4: MRM chromatograms of acequinocyl spiked at a level of 10 ppb in a
cannabis flower matrix using the LC/MS/MS method with an ESI source (a) and an cannabis flower matrix is very hydrophobic, significant matrix ion
APCI source (b). suppression effects of < -20% were observed mainly at higher
elution time for analytes. For those analytes measured with the others can get co-extracted with trace levels of pesticides and
APCI source, ion suppression effects greater than 20 % were can cause matrix interference if they have the same retention
only observed for 1 out of 3 analytes. For the three analytes time and MRM transition as pesticide. To improve the selectivity
measured with new method using an ESI source in negative of pesticides analysis in cannabis flower, it is necessary to have
ion mode, ion suppression effects greater than 20 % were multiple transitions for certain compounds in order to find a
observed for all three analytes. A solution containing 30 internal transition that does not have matrix interference. For example,
standards from a PerkinElmer One Pesticide420TM ISO17034 Figure 6a shows an overlay for a spiromesifen response in a
CRM Reagent kit was spiked in both solvent standards used for blank cannabis matrix and a cannabis matrix spiked with 10
generation of calibration curves for quantitation and cannabis ppb of spiromesifen using MRM transition displaying matrix
samples to improve the accuracy of quantitative analysis interference from a compound isobaric to this pesticide in
by compensating for the matrix ion suppression effects and cannabis. Therefore, as shown in Figure 6b, another MRM
correcting for any inaccuracies in sample injection in LC. The transition was determined to reduce matrix interference.
overall recovery (accuracy) of the LC/MS/MS method takes into
account both the recovery from sample preparation as well as
ion suppression effects and is given by following equation.spiked
extract and multiplying this number by 100. (a)
Spiromefisen
Measured Concentration
Overall Recovery (%) = * 100
Spiked Concentration
Figure 5 shows a comparison of the overall recoveries of 61

pesticides and 5 mycotoxins analyzed using an ESI source with
and without an internal standard. The data illustrates that the
addition of internal standards resulted in overall recovery in
range of 80-120% for 61 pesticides and 5 mycotoxins with an ESI
source. Finally, the overall recoveries in a range of 80-120% with
RSD less than 20% were achieved for all 67 pesticides (at a level (b)
of 1000 ppb) and all 5 mycotoxins (at a level of 100 ppb) with the Spiromefisen
addition of 30 internal standards to a cannabis flower matrix for
the LC/MS/MS methods with ESI and APCI sources
Figure 6: (a) Overlay of the response of a blank cannabis matrix (Red) and
spiromesifen (Green) spiked at a level of 10 ppb in cannabis flower matrix with
MRM transition showing matrix interference, and (b) overlay of the response of a
blank cannabis matrix (Red) and spiromesifen (Green) spiked at level of 10 ppb in
a cannabis flower matrix with MRM transition showing minimized matrix interference.
Conclusions
Figure 5: The overall recovery (accuracy) % for analysis of 66 analytes (61 pesticides This application note describes an LC/MS/MS methodology
and 5 mycotoxins) calculated with and without using an internal standard for the 18 with dual ESI and APCI sources for analysis of the pesticides
minute LC/MS/MS method with an ESI source.
and mycotoxins residues regulated by Florida in cannabis
samples. The method LOD and LOQ for each analyte meets
LC/MS/MS Method with Optimum MRM Transitions to
Florida’s stringent requirements in place as of the date of this
Minimize Matrix Interference for Challenging Analytes in
publication, of at least one-tenth and one-half of the acceptable
Cannabis Matrices
limit, respectively, in a cannabis flower matrix. The sample
Cannabis is a complex plant and is comprised of more than 500 preparation method obtained good extraction recovery for all
different compounds, including phytocannabinoids and terpenes of pesticides and mycotoxins. A number of internal standards
(12). Some of its constituents like cannabinoids, terpenes and were added to the cannabis matrix to get overall recovery in
LC/MS/MS Analytical Method for Pesticide Residue in Cannabis Flower as Defined by Florida Testing Requirements
the range of 80-120%, improving the accuracy of quantitation 4. Florida state regulations for pesticide and mycotoxin analysis
by compensating for ion suppression effects. The LC/MS/ in cannabis flower, accessed at https://www.flrules.org/
MS method was optimized to reduce matrix interference for a gateway/ruleNo.asp?id=64ER20-9.
number of pesticides from a cannabis flower matrix.
5. K. K. Stenerson and G. Oden, Cann. Sci. & Tech., 1(1),
Highlights of this Application 48-53 (2018).
• Analysis of all of pesticides and mycotoxins currently regulated 6. J. Kowlaski, J. H. Dahl, A. Rigdon, J. Cochran, D. Laine and G.
by Florida in cannabis using an LC/MS/MS with ESI and APCI Fagras, LCGC, 35(5) 8-22 (2017).
sources. 7. X. Wang, D. Mackowsky, J. Searfoss and M. Telepchak, LCGC,
• A method that fulfills Florida’s current strict requirement that 34(10), 20-27 (2016).
LOD and LOQ for each analyte should be at least one-tenth 8. J. R. Moulins, M. Blais, K. Montsion, J. Tully, W. Mohan, M.
and one-half of the acceptable limit, respectively, in a cannabis Gagnon, T. McRitchie, K. Kwong, N. Snider and D. R. Blais, J.
matrix. AOAC Int.,101(6),1948-1960 (2018).
• Reduces time for procurement and preparation of pesticides, 9. N. Michlig, S. J. Lehotay, A. R. Lightfield , H. Beldomenico , M.
mycotoxins and internal standards using a PerkinElmer One R. Repetti , Journal of Chromatography A (2021), doi: https://
Pesticide420TM ISO17034 CRM Reagent kit. doi.org/10.1016/j.chroma.2021.462097.
• The internal standards (available from a PerkinElmer One 10. A. Dalmia, E. Cudjoe, T. Astill, J. Jalali, J.P. Weisenseel, F.
Pesticide420TM ISO17034 CRM Reagent kit) are added to Qin, M. Murphy, and T. Ruthenberg, Cannabis Science and
compensate for ion suppression effects to improve overall Technology 1(3), 38-50 (2018).
recovery and accuracy of quantitation.
11. A. Dalmia, C. Johnson, S. Hariri, J. Jalali, E. Cudjoe, J.
• Optimized MS and LC method to minimize matrix interference Kingstad and F. Qin, Current Trends in Mass. Spec., 18(3),
from a complex cannabis flower matrix. 22-29 (2020).
• Dual source, ESI/APCI, removes the need for two instrument 12. S. N. Atapattu and K. R. D. Johnson, J. Chromatogr. A, 1612
platforms in a lab. (2020) 460656.
References
1. Exhibit A, Table 3. Pesticide analytes and their action levels.
Oregon Administrative Rules 333-007-0400; Oregon/gov/oha,
effective 5/31/2017.
2. Arizona state regulations for pesticide and mycotoxin

analysis in cannabis accessed from https://www.azdhs.
gov/documents/licensing/medical-marijuana/az-medical-
marijuana-rules.pdf.
3. Chapter 5. Testing Laboratories Section 5313 Residual

Pesticides, Bureau of Marijuana Control Proposed Text of
Regulations, CA Code of Regulations, Title 16, 42, pp 23-26.
PerkinElmer, Inc.
940 Winter Street
P: (800) 762-4000 or
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401812 PKI
54
Mass Spectrometry
Authors:
Josh Ye, Feng Qin, Frank Kero, Craig Young,
Jason Weisenseel, Jamie Foss
PerkinElmer, Inc.
Downers Grove, IL
“No Dilute” Just Shoot:

Robustness of a QSight Introduction
Traditional analysis by chromatography and mass
LC-ESI-MS/MS for Low spectrometry often requires sample cleanup to minimize
matrix effects and to avoid contamination of the ion
Level Pesticide Residue source in the mass spectrometer. However, sample
Analysis in Wine preparation is usually labor intensive and requires trained

analysts with specialized skills. Strategies to redesign the
front end of mass spectrometers to minimize source
contamination and thereby avoid the need for extensive sample cleanup, led to the invention of
a hot surface induced desolvation (HSID™) interface1. The PerkinElmer QSight™ LC/MS/MS mass
spectrometer contains the HSID interface coupled to a Laminar flow ion guide™, both of which
prevent accumulation of contamination along the ion path making it a very sensitive and
maintenance free instrument.
In this study, we used the QSight LC/MS/MS system to evaluate the potential of eliminating
sample preparation for trace level pesticide analysis in a complex sample such as wine2.
We injected undiluted red and white wine samples into the mass spectrometer and studied
reproducibility in analysis of the spiked pesticides over 200 injections. The instrument
showed excellent reproducibility with minimal signal drift during the duration of the study
(over a week), confirming the robustness of the QSight mass spectrometer.
55
What is the HSID Interface and How Does it Work? Experimental
The HSID apparatus is a multiorthogonal channel interface directly Hardware/Software
heated up to 300 °C that is present immediately after the Chromatographic separation was conducted by a PerkinElmer
sampling orifice in the source and connects the orifice to the Altus® A-30 UPLC® System and detection was achieved using a
Laminar flow ion guide of the QSight mass spectrometer. PerkinElmer QSight 220 MS/MS detector with a dual ionization
Unlike traditionally designed interfaces, the HSID with its source. All instrument control, data acquisition and data
multi-channels orthogonal to each other (Fig. 1) produces processing were performed using the Simplicity 3Q™ software
turbulent and Laminar flow and disrupts the free jet expansion of platform. The mobile phase flow rate was at 0.5 mL/min.
the sample ions. The orthogonal channels prevent neutrals from Method Parameters
entering the mass spectrometer reducing chemical noise, and MS settings are shown in Table 1 and Table 2, respectively.
any solvated charged clusters entering the HSID are entrained Source parameters including gas flows, source temperature and
and desolvated in the hot flow of gas, further contributing to position settings were optimized for maximum sensitivity. The
reduction in chemical noise. quadrupole peak widths (Q1 and Q2) were set at 0.7 amu.
The ions from the HSID interface are gently transferred by gas Example compound-dependent parameters for the partial list
flow to the Laminar flow ion guide™, which is not subject to the of MRM transitions are listed in Table 3.
traditional axial fields, but is at zero potential. The ion guide has
multiple pumping stages to generate several pressure regions Table 1. MS source settings.
from the sample interface to the mass analyzer. In these regions, ESI voltage 5000 V
pressure gradually drops, creating a well-defined flow pattern Drying gas 120
along the ion path enabling ions to be gently extracted into the
HSID Temp 200 °C
analyzer. Both the HSID and laminar flow ion guide prevent
accumulation of contamination along the ion path making the Entrance voltage 30 V
QSight maintenance free. Among many benefits of the HSID Source Temp 325 °C
interface include high sensitivity due to an inherent reduction
Nebulizer gas 350
in chemical background (i.e. S/N, reduced N) and the ability
to perform analysis at high LC flow rate (3 mL/min) without Detection Mode MRM Mode
reduction in signal.
Table 2. Optimized compound-dependent parameters for selected pesticides.
Collision
Name Precursor Fragment Type
Energy
Dimethenamid 276.1 244.0 Quantifier 18
Dimethenamid 276.1 168.0 Qualifier 30
Benthiavalicarb-
382.1 180.0 Quantifier 38
isopropyl
Benthiavalicarb-
382.1 197.0 Qualifier 24
isopropyl
Pyriproxyfen 322.0 96.0 Quantifier 22
Pyriproxyfen 322.0 185.0 Qualifier 30
Standards and Samples

Argentina wine samples, including a bottle of Cabernet
Sauvignon (Cabernet) and a bottle of Pinot Grigio (Pinot), were
purchased from a local grocery store. The samples were fortified
to 10 ng/mL of pesticide mixed standards obtained from ULTRA
Scientific® (North Kingstown, RI). Then, 10 µL of spiked samples
(no further sample preparation and no dilution) were injected for
robustness and reproducibility analysis. Linearity and LOQs of the
Figure 1. Schematic of an HSID interface for laminar flow tandem mass spectrometry. analytes were also evaluated. Matrix-matched standards were
prepared at 0.1, 1, 10, and 100 ng/mL levels by diluting the
pesticide mix standards stock solution with blank Cabernet and
Pinto wine samples.
2
56
Robustness Study
Commercial wine samples (Pinot and Cabernet) were fortified (a) Pinot
to 10 ng/mL with a pesticides mixed standard. These fortified
samples were then tested with 200 repeat injections over the
course of 7 days. Three pesticides (Dimethenamid, Benthiavalicarb-
isopropyl and Pyriproxyfen) were selected to demonstrate the
stability of the system. Summarized plot of peak area versus
injection number for these analytes in spiked pinot and cabernet
are presented in Figure 2. As observed, the trace remains flat
across the 200 injections, and peak area reproducibility (CV,
calculated as relative standard deviation) was ~5%, indicating
excellent performance stability during analysis.
(b) Cabernet
Comparison of Calibration Curves Before and
After 200 Injection Study
Matrix-matched representative calibration curves for quantitative
and qualitative ions for Benthiavalicarb-isopropyl, Dimethenamid,
and Pyriproxyfen after the wine injection study was performed
are shown in Figure 3 (Pinto) and 4 (Cabernet). The selected
pesticides have a linear dynamic range of 0.5 to 100 ng/mL and
are identical to the calibration curves generated prior to wine
injections. Linear regression coefficients were obtained for all
the pesticides with R2 >0.992. There is no maximum residue limit
(MRL) of pesticides set for wine yet, according to the EU regulation. Figure 2. Peak area of three pesticides vs injection number for spiked (a) Pinot Grigio and
However, the limit of quantitation (LOQ) at level of low part per (b) Cabernet Sauvignon samples.
billion or less is considered sufficient.
Figure 3. Pinot matrix-matched calibration curves for Benthiavalicarb-isopropyl, Dimethenamid, and Pyriproxyfen.
3
57
Figure 4. Carbernet matrix-matched calibration curves for Benthiavalicarb-isopropyl, Dimethenamid, and Pyriproxyfen.
Conclusion References
A “no-dilute-just-shoot” approach was presented in this study 1. Flow Characteristics of a Laminar Flow Interface for
to demonstrate the advantages of an HSID interface on a LC-MS/MS. PerkinElmer Tech Note 0129893.
PerkinElmer QSight 220 LC/MS/MS system. The orthogonal 2. Meglioli M, Kero F, Ye J, Young C, Reddy S “No dilute”
design of the interface, and the turbulent and laminar flows just shoot LC-ESI-MS/MS : feasibility and robustness of
used for ion transportation provide maximum protection for a maintenance-free source for applications in low-level
the MS instrument from being contaminated. Over the two pesticide residue analysis "The Proceedings of the 64th
matrix variables x200 continuous injections of wine samples ASMS Conference on Mass Spectrometry and Allied
without any cleanup steps, the instrument performance Topics, San Antonio, TX, June 2016.
remained consistent. Peak area CVs are ~5%, and peak shape
and height are nearly the same. Linearity and LOQs of the
calibration curves for all analytes using matrix-matched standards
was found to be maintained after >200 injections. This study
suggests that it is possible to increase the lab productivity and
reduce cost on QSight LC/MS/MS system by injecting samples that
are prepared with minimal cleanup steps.
PerkinElmer, Inc.
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013032_01 PKI
58
APPLICATION NOTE
Liquid Chromatography/Mass
Spectrometry
AUTHORS
Alexander Kasperkiewicz
Feng Qin
Feng Qin
PerkinElmer, Inc.
Bolton, ON, Canada
Avinash Dalmia
Thomas
ThomasDillion
Dillion
PerkinElmer, Inc.
Shelton, CT, USA
Analysis of near 400

Pesticides in Tea Via
LC/MS/MS: Simple Introduction
Sample Preparation As the most consumed beverage globally,
and APCI to Improve the global tea crop has immense cultural and
economic importance.(1) As with any crop,
Analyte Coverage growers use pesticides to ensure high harvest
yields and quality resulting in consumer
exposure via the brewing process (most notably for polar analytes such as
neonicotinoids and organophosphates).(2–4) Pesticides are frequently found in
teas, with 41% of samples reported with at least one residue detected and 10%
of samples with a residue above the maximum regulatory limit according to the
2019 European Union report on pesticide residues in food. (5) In some regions a
positivity rate of over 6% with a mean concentration of 138 ng/g can be found for
chlorpyrifos, which was recently banned in the EU due to increased neurological
damage risk in children.(6,7) Regulatory and testing laboratories face a challenge
as consumer preference in organic products increase, pesticide screening lists
broaden, and global tea trade and consumption grow. These laboratories rely on
mass spectrometry techniques such as GC/MS/MS and LC/MS/MS to quantify
pesticide targets, often requiring both instrument platforms to fully cover their
target lists.(8) As demonstrated in past publications, the atmospheric pressure
chemical ionization (APCI) source in QSight’s Dual Source technology can
substitute for GC/MS/MS for the analysis of some non‑polar compounds which
can reduce cost and complexity for routine analysis laboratories.(9–11)
59
Analysis of near 400 Pesticides in Tea Via LC/MS/MS: Simple Sample Preparation and APCI to Improve Analyte Coverage
Along the same efficiency vein, sample preparation techniques (New Haven, CT, USA). Internal standards were sourced from
for tea analysis are often based around the Quick, Easy, the ONE Pesticide420 Reagent Kit (PerkinElmer, Inc., Shelton,
Cheap, Effective, Robust, Safe (QuEChERS) extraction CT, USA), containing 29 deuterated pesticide analogues. For
techniques, solid‑phase extraction, or both.(12) These sample sample extraction, 15 mL centrifuge tubes, a high‑capacity
preparation approaches add complexity, cost, and time for vortex mixer (DVX‑2500, VWR International LLC, Mississauga,
minimal to no improvement in extraction efficiency and matrix ON, CA), and a refrigerated centrifuge (5702 R, Eppendorf
effects as compound panels are scaled up to hundreds or Canada Ltd, Mississauga, ON, CA) were used. Black tea,
thousands of targets.(13,14) Leveraging QSight’s StayClean chamomile tea, herbal blend tea, and catnip matrix were
hardware design, which allows for the transfer of ions from purchased from a local organic specialty store in Toronto,
the source to analyzer region via laminar flow with no ion Ontario, Canada. The tea was ground prior to extraction using
guides or funnels, simpler solvent extraction approaches a consumer‑grade personal blender and sieved to a particle
can be applied with constant instrument performance and size of < 1 mm. All solvents and mobile phase additives used
minimal downtime. were LC‑MS grade.
In the following work, a simple solvent extraction approach Analytical Protocols

was coupled with a LC/MS/MS instrument with dual ESI and
APCI source for the analysis of approximately 400 pesticides A solvent extraction sample preparation procedure (illustrated
extracted from black tea (345 compounds analyzed via ESI in Figure I) with internal standard dilution was employed:
and 50 compounds determined via APCI). Excellent method 1 ± 0.01 g of the ground and sieved tea sample was weighed
performance (defined as accuracy of 70 – 120 % and precision into a 15 mL centrifuge tube. For pre‑spiked recovery
of < 20%) was achieved for over 93 % of analytes, including experiments, pesticide standards were spiked into the solid
compounds normally analyzed by GC/MS/MS such as fluralins, matrix with the samples placed on a high‑capacity vortex
chlormephos, chlorfenson, thiometon and others.(8) shaker pulsing at 1500 rpm for 1 hour. Following agitation,
10 µL of the internal standard mix was added along with
5 mL of acetonitrile + 0.1% formic acid. After extraction
solvent addition, samples were agitated again for 30 minutes
at 1500 rpm. Following extraction, the samples were
centrifuged for 5 minutes at > 3 × 103 RCF, with 1 mL of the
extract filtered through a 0.2 µm filter into a 2 mL amber vial
for LC/MS/MS analysis.
Solvent calibrants with stable isotope labelled internal standards

were used for calibration, with 9 calibration standards prepared
in neat solvent spanning the range 0.1 – 250 ng/mL. The
accuracy and precision of the method were determined at the
10 ng/g and 100 ng/g level (spiked in tea samples) across 4
sample replicates. Limits of quantification (LOQs) were reported
as the lowest concentration level which met accuracy qualifiers
within 70 ‑ 120%, RSD < 20%, and signal to noise ratio (S/N) of
greater than 10 (for the quantifier transition) requirements. For
Figure 1: Analytical procedure employed for the analysis of pesticides from tea: analytes with accuracy values outside of the range of 70 ‑ 120%
1) 1 g of tea sample measured into a 15 mL centrifuge tube along with internal
standards, 2) 5 mL of acetonitrile with 0.1% formic acid added, 3) sample agitated LOQs were determined as the lowest point in the calibration
for 30 minutes during extraction, 4) sample centrifuged before 5) filtration and
analysis via LC/MS/MS with ESI and APCI methods.
curve with a S/N of greater than 10.
Matrix effects were determined by comparing slopes between

Experimental a calibration curve made in neat solvent and tea extract (with
Materials stock standards spiked into solvent and extract)—reported as
% signal suppression/enhancement (% SSE). Absolute matrix
All experiments were carried out using a QSight LX50 UHPLC
effects determined in the additional tea matrices of chamomile,
coupled to a QSight 420 triple quadrupole mass spectrometer
herbal blend, and catnip were determined by comparing peak
(PerkinElmer, Inc., Shelton, CT, USA) via a divert valve. All
areas between a post‑extract spike and solvent‑only standards
instrument control, method development, and data processing
at 10 ng/mL and 100 ng/mL, expressed as %SSE.(15)
were completed using the Simplicity 3Q software. All pesticide
standards were sourced in custom mixes from AccuStandard
Instrumental Conditions the ESI method, and 100 MRM transitions were monitored
for 50 compounds in the APCI method along with internal
Instrumental parameters for both APCI and ESI methods
standards (shown in Figure II). For both LC methods, a
are shown in Table I. A Time Managed MRM (TM‑MRM)
100 mmX4.6 mm I.D., 2.7 µm Quasar SPP C18 column
method was built for both APCI and ESI compounds. In total,
(PerkinElmer, Inc., Shelton) was used.
690 MRM transitions were monitored for 345 compounds in
Table 1. LC/MS/MS Instrumental Conditions for ESI and APCI Methods
LC Conditions
Mobile Phase A
ESI LC/MS grade Water + 0.1% formic acid + 2 mM ammonium formate
APCI LC/MS grade Water
Mobile Phase B
ESI LC/MS grade Methanol + 0.1% formic acid + 2 mM ammonium formate
APCI LC/MS grade Methanol
For the ESI method, the 19 minute run had initial conditions of 5% B at 0.8 mL/min for a 0.5 minute hold, with a
ramp to 50% B by 4 minutes, followed by a ramp to 100% B by 17.5 minutes, with a 1.5 minute re-equilibration
period at initial conditions.
Gradients used
For the APCI method, the 12 minute run had initial conditions of 30% B at 0.8 mL/min for a 0.5 minute hold,
followed by a ramp to 95% B by 8 minutes, with a hold for 2 minutes before a 2 minute re-equilibration at initial
conditions.
Column Oven Temperature 40 °C
Sample Tray Temperature 5 °C
Injection Volume
ESI 3 µL
APCI 10 µL
MS Conditions
Positive ESI
Voltage + 5100 V
Negative ESI
Voltage -4500 V
Negative APCI -3 µA
Current
Drying Gas 150 arbitrary units
Nebulizer Gas 350 arbitrary units
Source Temperature
ESI 315 °C
APCI 250 °C
HSID Temperature
ESI 200 °C
APCI 180 °C
Detection Mode MRM
Results and Discussion chamomile, 95 % in the herbal blend, and 85 % in catnip.(15) The
APCI method performed similarly with 100 % of compounds in
LC/MS/MS Method Performance black tea displaying negligible matrix effects, 94% in chamomile
Due to the high sample processing load of routine testing and the herbal blend, and 89% in catnip. The low matrix effects
facilities, several method development decisions were made to across multiple herbal tea products suggests the method
meet their high sample throughput requirements. The pursuit developed can be adapted to analyze several types of tea with
of a protocol based around solvent extraction, a commercially a trivial revalidation.
available internal standard mixture, and internal standard
calibration resulted in an easy‑to‑execute analysis procedure.
Negligible matrix effects (80 < SSE < 120%) were observed
across 94% (371 compounds) of the compound panel.
The remaining 6% of compounds were either suspected to be
degrading in the extract (thiocyclam and diafenthiuron) (16,17) or
displayed signal suppression or enhancement greater than 20%
(such as dodemorph and chlormephos, respectively). These
compounds with matrix effects greater than 20% demonstrated
the need to incorporate internal standards into the calibration/
sample preparation procedure to improve method accuracy as
well as to provide flexibility to expand the protocol to additional
matrices and compounds in the future. Internal standards
were added before extraction to correct for both losses in
the sample preparation procedure along with matrix effects
during the analysis step. Excellent analytical figures of merit
were observed for most of the analytes under study (displayed
for all compounds of interest in Table II) with %SSE in black
tea ranging from 161 to 69%. Black tea has been referenced Figure 2: TIC of 345 and 50 pesticides analyzed using ESI and APCI methods,
respectively.
as a particularly difficult matrix in terms of suppression by
co‑extraction of matrix‑sourced interferences via QuEChERS
and other dilute‑and‑shoot methods, however extensive
suppression was not observed in our implementation.(18–20)
Further Matrix Effects Investigation
Following the promising method validation results in black tea,

an extension of the matrix effects investigation was pursued
to investigate the suitability of this approach in other herbal
teas. An absolute matrix effects study was designed to explore
method performance between black tea, chamomile, an
herbal blend (containing chamomile, spearmint, lemongrass,
blackberry leaves, orange blossoms, hawthorn, and rosebuds),
and catnip.(15) Samples were extracted following the procedure
in Figure I and the extracts were spiked at 10 and 100 ng/mL
with the pesticides under study. Peak areas were compared
between the post‑extract spikes and solvent‑only samples,
expressed as % SSE. Summarized results can be found in
a scatter plot in Figure III for all compounds and matrices.
The robustness of the separation method to matrix effects
continued in the additional tea matrices tested. For the ESI
method, 97% of analytes were found to have negligible matrix Figure 3: Summary of select method validation data, with limits of quantification
(LOQs) under 5 ng/g for approximately 80% of compounds under study (A) along
effects (defined as 85 % < % SSE < 115 %) in black tea, 93% in with individual compound data points and corresponding box and whisker plots
for matrix effects (%SSE), accuracy, and precision at the 10 ng/g validation point
(for the compounds of which LOQ < 10 ng/g) (B). Absolute matrix effects are
summarized for APCI at 100 ng/mL for 50 compounds in (C) and for ESI at 10 ng/g
for 342 compounds in (D).
LOQs Compared with Pesticide Maximum Regulatory

Limits (MRLS) 5 ng/g tea sample
Blank tea sample

For a method developed to be fit‑for‑purpose in the regulatory
environment it needs to meet regulatory performance levels.
A complete comparison of all LOQs determined to MRLs was
not possible as not all compounds are regulated for use on Chlormephos
tea consistently between locals. The most comprehensive

and strict regulatory list of pesticides in tea was found via
the European Commission. Of the 395 compounds validated,
227 compounds have been assigned an EU‑harmonized MRL 10 ng/g tea sample
and are authorized for use on tea products. Upon comparison 20 ng/g tea sample
of method LOQs, it was found that 172 of the 227 compounds 50 ng/g tea sample
had MRLs at least 10‑fold greater than method LOQs (over 80% Benfluralin
of analytes). The LOQs of the remaining 55 compounds were

found to be less or equal to the MRL for all but azadirachtin
and cyhalothrin.
Use of APCI to Reduce ESI Workload and Figure 4: Comparisons of APCI-LC/MS/MS chromatograms for chlormephos
(top) and benfluralin (bottom). Extracts from tea samples compared with differing
Expand Method Coverage concentration levels and blank matrix injections.
APCI has been shown to be less susceptible to matrix effects Conclusion

than ESI for a variety of bioanalytical and agrochemical
analysis.(22) Thus, the addition of APCI to the protocol With routine labs tasked with the analysis of larger and larger
provides two primary benefits: the ability to monitor previously target lists, and with greater numbers of samples requiring
GC/MS/MS‑only compounds (such as pentachloronitrobenzene, analysis, we deployed simple sample preparation protocols
chlordane, fenitrothion, chlorfenapyr, anthraquinone) as well as such as solvent extraction to meet the need of high sample
the flexibility to monitor compounds via APCI due to improved throughput with lower cost. In this work, we demonstrate
sensitivity compared to ESI (such as phthalimide‑class how a practical simple solvent extraction of black tea can
compounds, parathion, fluralin‑class compounds, and be employed to achieve the analysis of a large multi‑residue
fenchlorphos) due to a different ionization mechanism or panel of pesticides, with low limits of detection (with 319 of
a reduction in matrix effects. In this work, samples were 395 analytes displaying LOQs of 5 ng/g or below), minimal
analyzed for the bulk of the analyte target list during the ESI matrix effects (94% of compounds with negligible %SSE
method followed by a repeated sequence/injection for APCI in black tea), and excellent method performance (93% of
analysis of the 50 analytes which were not suitable/optimal compounds with accuracies within 70 ‑ 120% and RSD
for ESI. Low LOQs of 5 ng/g were achieved using APCI < 20%). Prior successes in cannabis matrices along with
for compounds traditionally analysed via GC/MS such as work presented herein have suggested simple, acetonitrile
fluralin‑class compounds (benfluralin, profluralin), fenitrothion, extraction conditions and high‑flow rate LC parameters allow
chlorfenson, fenchlorphos oxon, dicloran, bromophos ethyl, for practical workflows for pesticide analysis. Additionally,
and dichlofenthion. It is worth mentioning that the dual source APCI analysis allowed for traditionally GC‑only compounds
setting for QSight LC/MS/MS allows automatic LC method and (such as pentachloronitrobenzene, chlormephos, and others)
MS probe switching without any manual intervention, which to be included on this LC pesticide panel, reducing support
greatly reduces operation time and cost. Leveraging APCI has instrument workload, or potentially eliminating the need for
vastly simplified pesticide analysis in certain applications and complementary GC‑MS protocol for multi‑residue pesticide
the work completed aims to broaden prior success.(9–11) quantification. The work was made possible using hardware
features unique to QSight, namely Dual Source technology
for easy APCI method integration, rapid detector polarity
switching, and the StayClean source allowing for simple sample
preparation with minimal instrument downtime. We hope to
extend these concepts to a plethora of food matrices of varying
difficulties and compositions.
Table 2. Analytical figures of merit for all analytes of interest extracted from black tea along with ionization method used.
10 ng/g (n = 4)* 100 ng/g (n = 4)

Compound Method LOQ [MRL]± (ng/g) RT (min) Accuracy (%) Precision (%) Accuracy (%) Precision (%)
THPI APCI 10 2.57 89 2.6 91 1.8
Phthalimide APCI 20 3.22 ND ND 91 3.5
Chlormephos APCI 5 5.74 121 3.3 121 1.8
Dicloran APCI 5 [50] 6.03 100 5.8 100 2.6
Captan APCI 20 [100] 6.25 ND ND 62 12.6
o-Phenylphenol APCI 20 6.28 ND ND 105 5.7
Methyl parathion APCI 5 [50] 6.44 103 6.8 103 3.2
Folpet APCI 50 [100] 6.85 ND ND 99 4.8
2, 4, 6-Trichloropenol APCI 5 6.87 98 21.7 100 1.4
Fenitrothion APCI 5 [50] 6.90 103 7.1 105 2.7
Etridiazole APCI 5 [50] 6.93 110 6.0 89 5.8
Fenson APCI 10 7.07 100 9.2 103 14.7
Thiometon APCI 5 7.10 73 2.1 65 1.0
Anthraquinone APCI 10 [20] 7.28 107 19.2 97 6.0
Fenchlorphos oxon APCI 5 [100] 7.29 105 10.1 106 4.6
Chlorothalonil APCI 50 [50] 7.33 ND ND 111 14.8
Chlozolinate APCI 10 [50] 7.35 120 14.1 139 6.2
Vinclozin APCI 10 [50] 7.39 109 20.2 103 4.9
Parathion APCI 5 7.46 103 12.4 106 4.4
Endosulfan APCI 10 7.65 98 21.2 100 4.4
Chlorfenson APCI 5 [100] 7.70 106 4.4 99 3.0
Prothiophos APCI 5 7.89 98 6.1 104 3.3
Chlorobenzilate APCI 20 [100] 8.01 ND ND 100 9.9
Tetradifon APCI 5 [50] 8.01 98 14.4 102 4.4
Bromocyclen APCI 20 8.03 ND ND 101 7.3
Dinobuton APCI 5 8.12 106 8.1 99 9.3
Chlorfenapyr APCI 10 8.16 104 10.7 102 6.7
Oxyfluorfen APCI 5 [50] 8.29 104 16.4 105 7.0
Chloropropylate APCI 50 8.30 ND ND 107 20.4
Oxychlordane APCI 50 8.31 ND ND 107 2.7
Nitrothal-isopropyl APCI 5 8.32 108 3.1 104 6.6
Tetrachloronitrobenzene APCI 5 8.32 100 11.8 97 7.5
Fenchlorphos APCI 50 8.51 ND ND 104 29.2
Bromopropylate APCI 20 [50] 8.52 ND ND 103 11.3
Pentachloroaniline APCI 10 8.53 95 12.7 103 5.2
Ethalfuralin APCI 5 [10] 8.53 102 14.3 100 6.8
Pentachlorocyanobenzene APCI 5 8.57 122 14.7 99 2.3
Heptachlorepoxide APCI 100 8.58 ND ND 92 20.2
10 ng/g (n = 4)* 100 ng/g (n = 4)

Bromophos APCI 5 8.59 94 21.3 106 4.9
Iodofenphos APCI 5 8.61 113 8.4 91 7.1
Benfluralin/Trifuralin APCI 5 [100] 8.73 109 17.4 107 6.0
Flumetralin APCI 20 [50] 8.73 ND ND 102 3.7
Profluralin APCI 5 8.78 107 7.9 104 2.1
Chlordane APCI 50 8.93 ND ND 102 18.2
Pentachloronitrobenzene APCI 5 9.01 117 10.2 102 2.9
Bromophos ethyl APCI 5 [50] 9.14 94 4.4 100 5.3
Dichlofenthion APCI 5 9.19 91 6.1 99 3.2
Pentachloroanisole APCI 100 9.43 ND ND 92 16.9
Pentachlorobenzene APCI 100 9.43 ND ND 98 8.0
Hexachlorobenzene APCI 20 10.21 ND ND 92 4.6
Cyromazine ESI 100 [100] 1.71 ND ND 5 11.6
Desphenyl chloridazon ESI 10 2.10 73 6.9 62 5.5
Methamidphos ESI 5 [50] 2.54 99 1.6 95 0.5
Thiocyclam ESI 10 2.61 25 7.0 22 2.8
Formetanate ESI 1 [50] 2.96 31 7.9 32 3.5
Acephate ESI 1 [50] 3.09 106 2.1 102 1.3
Propamocarb ESI 5 [50] 3.20 33 4.3 32 2.3
Pymetrozine ESI 5 [100] 3.27 36 6.9 35 4.7
Omethoate ESI 5 [50] 3.47 114 0.7 103 1.9
Dinotefuran ESI 5 3.63 137 2.0 110 1.7
Aldicarb sulfoxide ESI 5 [50] 3.71 104 10.1 101 2.4
Aldicarb sulfone ESI 5 [50] 3.77 117 6.0 110 3.1
Nitenpyram ESI 10 3.80 74 4.4 69 1.2
Oxamyl ESI 5 [50] 3.98 110 1.5 105 2.3
Benomyl/Carbendazim ESI 1 [100] 4.01 95 3.3 90 1.2
Flonicamid ESI 1 [100] 4.14 93 4.0 87 1.2
Methomyl ESI 5 [50] 4.23 115 2.7 112 4.6
Demeton-S-methyl-sulfone ESI 1 4.24 124 1.4 119 1.7
Oxydemeton-methyl ESI 1 [50] 4.29 114 3.1 108 2.4
Thiamethoxam ESI 5 4.32 111 1.1 106 1.8
Monocrotophos ESI 1 [50] 4.47 120 2.1 111 2.3
Thiabendazole ESI 5 [50] 4.51 71 2.1 69 3.0
Fuberidazole ESI 5 [50] 4.65 98 5.1 90 1.2
Dicrotophos ESI 1 4.81 96 2.1 93 2.0
Clothianidin ESI 5 4.85 78 5.3 73 1.3
Imidacloprid ESI 5 4.85 105 4.5 98 3.2
10 ng/g (n = 4)* 100 ng/g (n = 4)

Ethiofencarb sulfoxide ESI 1 4.86 110 4.1 109 1.6
Trichlorfon ESI 5 [50] 5.01 101 5.9 99 1.9
3-hydroxycarbofuran ESI 1 [50] 5.06 110 6.9 104 2.8
Dimethoate ESI 5 [50] 5.08 111 2.3 109 1.7
Methiocarb sulfoxide ESI 1 [100] 5.11 113 2.8 111 2.2
Metamitron ESI 5 5.16 102 4.4 95 2.5
Sulfoxaflor ESI 5 [20] 5.23 110 5.1 104 4.5
Methiocarb sulfone ESI 5 [100] 5.26 113 2.1 109 2.8
Acetamiprid ESI 1 [50] 5.26 113 2.3 110 2.2
Oxycarboxin ESI 1 [50] 5.34 111 5.4 108 3.0
Cymoxanil ESI 1 [100] 5.36 111 6.7 106 4.2
Ethirimol ESI 1 [50] 5.49 41 5.2 40 3.5
Thiacloprid ESI 5 5.60 113 2.8 108 1.9
Mevinphos ESI 1 [20] 5.68 111 2.9 104 4.7
Metoxuron ESI 1 5.85 113 3.0 110 1.7
Butocarboxim ESI 50 5.88 ND ND 120 18.6
Aldicarb ESI 1 [50] 5.94 111 5.3 108 2.8
Formothion ESI 5 [50] 5.95 114 11.0 106 2.7
Pirimicarb ESI 1 6.01 114 2.8 111 1.9
Methyl paraoxon ESI 1 [50] 6.08 109 3.7 107 3.0
Terbufos oxon sulfone ESI 5 6.20 115 3.4 111 3.3
Metolcarb ESI 1 6.22 112 5.9 110 1.7
Oxadixyl ESI 5 [20] 6.22 93 4.3 90 3.5
Tricyclazole ESI 5 [50] 6.28 88 3.6 84 2.3
Phosphamidon ESI 5 [20] 6.30 109 2.8 103 2.3
Thiophanate-methyl ESI 5 [100] 6.36 93 4.4 91 4.5
Triasulfuron ESI 5 [50] 6.40 115 3.9 109 3.6
Bentazon ESI 1 6.46 110 4.9 107 1.7
Azamethiphos ESI 1 6.49 114 4.5 112 1.9
Bendiocarb ESI 5 6.61 111 2.3 106 3.7
Metribuzin ESI 5 [100] 6.64 115 5.5 110 2.5
Dichlorvos ESI 5 [20] 6.64 102 3.9 102 1.0
Bromacil ESI 5 6.64 111 2.8 106 2.0
Baygon ESI 5 [100] 6.65 105 2.3 104 3.3
Spirotetramat-mono-OH ESI 5 [50] 6.65 111 5.7 103 2.2
Carbofuran ESI 1 [50] 6.76 126 2.0 120 2.3
Simazine ESI 5 6.78 109 3.3 102 0.8
Ofurace ESI 5 6.80 110 2.4 105 1.7
10 ng/g (n = 4)* 100 ng/g (n = 4)

Terbacil ESI 5 6.88 114 7.2 108 3.8
Dichlormid ESI 5 6.90 114 6.5 107 2.1
Carboxin ESI 1 [100] 7.03 94 5.5 86 3.0
Imazalil ESI 5 [50] 7.04 57 3.8 52 3.5
Carbaryl ESI 5 [50] 7.05 105 11.5 100 2.0
Cyantraniliprole ESI 5 7.12 113 7.2 103 3.1
Terbumeton ESI 5 7.16 103 2.0 97 2.3
Monolinuron ESI 5 [50] 7.19 113 3.9 104 2.6
Fenthion sulfone ESI 5 [50] 7.19 103 2.5 102 3.1
Fenthion sulfoxide ESI 5 [50] 7.24 114 3.0 109 2.5
Fluometuron ESI 1 [50] 7.27 114 2.8 107 2.4
Ethiofencarb ESI 1 7.31 94 6.1 88 4.3
Brominal ESI 5 [50] 7.36 106 5.0 102 3.5
Azadirachtin ESI 50 [10] 7.49 ND ND 93 14.9
Ethoxyquin ESI 5 [100] 7.51 86 5.9 92 1.3
Chlorotoluron ESI 5 [50] 7.54 110 2.4 103 2.2
Fosthiazate ESI 5 7.55 104 3.4 98 2.7
Propham ESI 10 [50] 7.59 111 9.6 103 4.0
Spirotetramat-enol ESI 5 [100] 7.62 95 2.1 89 3.0
Disulfoton sulfone ESI 5 [50] 7.63 112 1.9 107 3.8
Phorate sulfone ESI 5 7.66 104 6.4 94 1.6
Phorate sulfoxide ESI 5 7.71 109 3.9 102 2.8
Disulfoton sulfoxide ESI 5 7.72 113 3.8 106 1.2
Thiodicarb ESI 5 [50] 7.75 106 3.9 97 1.7
Ametryn ESI 1 7.75 118 2.9 111 3.2
Flutriafol ESI 5 7.76 104 1.5 97 5.2
Paraoxon ESI 5 7.78 117 1.5 110 2.6
Dodemorph ESI 10 [10] 7.86 47 20.5 52 7.2
Forchlorfenuron ESI 1 [50] 7.91 107 1.5 102 2.0
Atrazine ESI 1 [100] 7.93 111 2.5 106 2.5
Diuron ESI 5 [50] 7.96 107 4.3 101 1.7
Isocarbophos ESI 1 8.02 113 1.6 103 3.3
Fenpropidin ESI 5 [50] 8.04 71 6.2 69 2.9
Metazachlor ESI 5 [100] 8.05 110 1.7 103 2.2
Isoproturon ESI 1 [50] 8.09 114 1.6 108 3.0
Lenacil ESI 1 [100] 8.12 106 1.9 101 2.2
Desmedipham ESI 5 [50] 8.14 100 5.8 94 2.9
Norflurazon ESI 1 8.20 116 2.8 110 2.3
10 ng/g (n = 4)* 100 ng/g (n = 4)

Propachlor ESI 5 8.20 110 6.1 105 1.8
Bentazon methyl ESI 10 8.21 104 8.6 101 2.5
Spirotetramat-keto-hydroxy ESI 5 [100] 8.24 104 7.5 93 5.6
Fenpropimorph ESI 5 [50] 8.32 80 5.2 75 1.8
Triforine ESI 5 [50] 8.32 101 10.1 97 2.7
Naled ESI 5 8.34 109 3.6 104 2.1
Methidathion ESI 5 [100] 8.35 96 3.4 91 5.4
Fensulfothion ESI 5 8.37 113 4.1 106 1.6
Metalaxyl ESI 5 8.38 117 3.8 106 3.9
DEET ESI 20 8.40 ND ND 112 2.3
Chlorantraniliprole ESI 5 [20] 8.44 106 3.4 96 0.9
Pyrimethanil ESI 1 8.45 109 3.6 100 1.9
Methacrifos ESI 10 [50] 8.50 105 7.8 99 3.6
Azinphos methyl ESI 20 [50] 8.54 119 13.5 93 6.2
Phosmet ESI 5 [100] 8.58 119 7.3 115 3.9
Spiroxamine ESI 5 [50] 8.64 76 2.4 73 2.1
Flumioxazin ESI 5 [100] 8.64 108 20.5 99 3.7
Tridemorph ESI 50 [50] 8.64 ND ND 66 12.5
Saflufenacil ESI 5 [30] 8.72 113 7.8 103 4.2
Propanil ESI 5 [50] 8.77 108 4.9 107 3.1
Linuron ESI 5 [50] 8.77 104 0.9 103 2.0
Clomazon ESI 5 8.83 116 1.9 112 2.6
Terbufos sulfone ESI 5 8.87 100 3.7 94 3.8
Baycarb ESI 1 8.90 116 1.0 106 2.4
Dichlorprop ESI 100 [100] 8.93 ND ND 95 12.1
Demeton-S ESI 1 8.94 101 6.1 94 4.8
Diethofencarb ESI 1 [50] 8.94 110 5.4 105 1.7
Prometryne ESI 5 8.97 116 1.2 111 1.6
Ethiprole ESI 5 9.00 112 2.4 104 3.7
Ethofumesate ESI 5 9.01 108 4.5 98 3.6
Terbufos sulfoxide ESI 5 9.09 113 7.5 102 2.0
Fenamidone ESI 5 [50] 9.11 117 1.8 109 3.4
Methiocarb ESI 1 [100] 9.13 101 3.7 94 2.4
Terbutryn ESI 5 9.14 109 3.3 100 0.9
Fludioxonil ESI 5 9.14 99 5.2 102 4.9
Azoxystrobin ESI 1 9.19 114 3.6 108 2.7
Fluazifop-P ESI 5 9.27 108 11.2 103 8.4
Furalaxyl ESI 1 9.30 122 3.1 114 2.9
10 ng/g (n = 4)* 100 ng/g (n = 4)

Boscalid ESI 5 9.34 106 5.6 100 4.2
Trimidal ESI 5 9.37 96 3.0 95 3.5
Chlorpropham ESI 10 [50] 9.39 108 6.9 100 3.8
Flutolanil ESI 1 [50] 9.39 118 1.2 112 3.5
Terbuthylazine ESI 5 [50] 9.41 118 2.1 110 1.4
Mandipropamid ESI 5 9.42 112 4.9 103 4.1
Pronamide ESI 5 9.45 109 3.7 102 2.4
Paclobutrazole ESI 5 9.48 105 5.0 95 5.7
Promecarb ESI 5 9.49 117 4.9 103 2.6
Malathion ESI 5 9.56 105 2.5 102 4.4
Isoxaben ESI 1 [20] 9.56 116 1.8 113 3.5
Mepronil ESI 5 [50] 9.60 111 2.9 109 2.2
Fluopicolide ESI 5 9.62 110 2.7 107 2.5
Benthiavalicarb-isopropyl ESI 1 [50] 9.70 107 1.7 105 3.7
Isoprothiolane ESI 5 [10] 9.73 109 3.5 106 1.2
Captafol ESI 100 [100] 9.77 ND ND 114 5.6
Myclobutanil ESI 5 [50] 9.82 98 4.6 96 2.9
Dimethomorph ESI 5 [50] 9.86 103 2.7 101 3.6
Methoxyfenozid ESI 5 [50] 9.87 104 7.0 102 6.1
Triadimenol ESI 5 [50] 9.95 98 11.7 96 4.7
Triadimefon ESI 5 [50] 9.95 103 3.7 104 2.4
Bifenazate ESI 1 [100] 9.95 116 3.1 110 4.0
Diphenylamine ESI 10 [50] 9.96 117 6.7 113 4.2
Triazophos ESI 5 [20] 9.97 109 2.3 105 1.6
Cyproconazole ESI 5 9.99 97 3.7 95 1.7
Fluopyram ESI 1 10.01 114 4.9 110 3.1
Isazophos ESI 1 10.01 110 5.2 109 3.1
Procymidone ESI 5 [50] 10.02 116 10.6 99 6.2
Fenpyrazamine ESI 5 10.06 110 6.1 103 2.2
Bupirimate ESI 1 [50] 10.06 108 6.2 103 2.5
Pyridaphenthion ESI 1 10.08 120 1.5 115 2.4
Fenhexamid ESI 5 [50] 10.10 102 8.2 103 2.8
Molinate ESI 5 [50] 10.11 111 1.6 108 3.5
Ditalimfos ESI 5 [10] 10.12 116 3.4 120 3.6
Fluquinconazole ESI 5 [50] 10.12 117 11.1 108 2.6
Mepanipyrim ESI 1 [50] 10.13 118 1.3 115 2.9
Pyrifenox ESI 5 10.16 106 2.4 105 4.0
Oryzalin ESI 20 [50] 10.18 ND ND 105 5.4
10 ng/g (n = 4)* 100 ng/g (n = 4)

Flufenacet ESI 5 [50] 10.19 107 4.6 111 3.0
Iprovalicarb ESI 5 [50] 10.19 107 10.1 107 4.5
Tetraconazole ESI 5 [20] 10.20 116 9.2 115 3.3
Azinphos ethyl ESI 10 [50] 10.22 119 4.5 107 7.4
Mecarbam ESI 5 [50] 10.26 109 3.9 109 3.8
Indaziflam ESI 1 10.26 115 5.3 115 3.7
Cyazofamid ESI 5 [50] 10.32 109 9.2 109 4.3
Cyprodinil ESI 5 10.34 102 5.0 103 3.3
Diflubenzuron ESI 5 [50] 10.36 106 7.4 109 2.3
Fipronil ESI 1 [5] 10.47 111 3.9 115 4.8
Spirotetramat ESI 5 [100] 10.53 109 3.3 106 3.7
Fenarimol ESI 5 [50] 10.56 107 11.0 106 2.5
Iprodione ESI 10 [50] 10.56 108 10.7 106 3.8
Fenbuconazole ESI 5 [50] 10.60 108 9.8 109 5.0
Acetochlor ESI 5 [50] 10.60 105 9.0 108 4.0
Napropamide ESI 5 [50] 10.61 111 4.3 113 4.5
Epoxiconazole ESI 5 [50] 10.62 114 5.7 113 4.6
Uniconazole ESI 5 10.63 108 6.8 102 4.3
Alachlor ESI 5 [50] 10.66 104 7.2 105 4.3
Ethoprophos ESI 5 [20] 10.67 106 3.4 109 3.5
Metolachlor ESI 10 [50] 10.81 121 5.4 116 2.5
Fenoxycarb ESI 5 [50] 10.82 114 4.4 106 3.1
Fenamiphos ESI 5 [50] 10.82 115 5.8 112 3.0
Quizalofop ESI 5 10.82 102 4.1 92 6.1
Picoxystrobin ESI 1 [50] 10.84 107 3.9 103 5.5
Flusilazole ESI 1 [50] 10.85 111 6.3 109 2.3
Phenthoate ESI 5 10.91 118 11.8 105 2.6
Flubendiamide ESI 10 [20] 10.93 113 6.8 105 3.2
Tebufenozide ESI 5 [50] 10.95 106 9.2 101 0.6
Rotenone ESI 5 [20] 10.97 111 6.1 107 6.0
Clozolinate ESI 10 [50] 11.00 111 15.8 97 8.8
Carfentrazone-ethyl ESI 10 [100] 11.00 106 9.4 93 5.2
Dichlobutrazol ESI 5 11.00 113 10.1 107 4.9
Tetrachlorvinphos Z ESI 10 11.01 112 10.9 105 1.9
Bromophos-methyl ESI 50 11.01 ND ND 103 3.8
Haloxyfop ESI 10 11.02 117 7.6 104 3.3
Sulfotep ESI 5 [50] 11.06 108 9.0 107 3.2
Kresoxim-methyl ESI 5 [50] 11.08 113 17.6 104 3.6
10 ng/g (n = 4)* 100 ng/g (n = 4)

Isofenphos-methyl ESI 10 11.08 115 9.7 106 4.5
Quinalphos ESI 5 [50] 11.08 122 5.1 114 3.0
Penthiopyrad ESI 1 11.08 120 7.7 116 3.0
EPTC ESI 5 [50] 11.10 108 8.7 105 1.7
Tolylfluanid ESI 5 [100] 11.11 107 7.6 103 3.6
Bromuconazole ESI 5 [50] 11.13 111 5.9 104 4.4
Fenthion ESI 5 [50] 11.13 116 4.5 110 2.1
Dodine ESI 10 [50] 11.15 20 16.9 20 9.1
Tebuconazole ESI 5 11.18 108 6.5 106 3.0
Pyraflufen-ethyl ESI 5 [50] 11.21 116 8.8 111 4.5
Prothioconazole ESI 10 [50] 11.23 81 17.3 67 8.8
Etrimfos ESI 5 11.24 107 11.6 103 3.9
Zoxamide ESI 5 [50] 11.28 109 5.4 106 2.2
Endosulfan sulfate ESI 5 11.29 112 3.9 110 4.0
Fonofos ESI 5 11.33 112 4.4 105 1.4
Penconazole ESI 5 [50] 11.34 102 10.3 98 2.5
Famoxadon ESI 20 11.37 119 12.4 98 7.0
Triflumuron ESI 5 [50] 11.46 108 10.7 104 2.3
Diazinon ESI 5 [50] 11.53 115 6.7 111 3.3
Chlorfenvinphos ESI 5 [50] 11.55 108 10.5 106 2.5
Hexaconazole ESI 5 [50] 11.56 106 7.2 103 1.9
Oxadiargyl ESI 5 [50] 11.59 113 4.0 101 3.2
Pirimiphos-methyl ESI 1 [50] 11.59 123 6.3 117 3.2
Metaconazole ESI 5 [50] 11.60 101 6.4 99 3.3
Spinosad A ESI 5 [100] 11.65 88 12.6 79 2.7
Phorate ESI 5 11.66 106 5.4 100 3.3
Benalaxyl ESI 1 [50] 11.66 118 6.2 112 2.3
Phosalone ESI 5 [50] 11.66 113 8.0 106 2.7
Cyflufenamide ESI 5 [50] 11.69 120 8.2 111 4.1
Clofentezine ESI 5 [50] 11.70 107 7.0 107 2.2
Pyraclostrobin ESI 1 11.72 113 5.5 108 3.4
Tolclofos-methyl ESI 5 [50] 11.72 108 4.9 105 1.7
Propiconazole ESI 5 [50] 11.73 108 6.8 102 3.7
Diclofop ESI 5 11.78 109 11.9 101 2.9
Prochloraz ESI 5 [150] 11.81 111 8.7 104 3.0
Bitertanol ESI 5 [50] 11.82 110 10.5 104 3.0
Isofenphos ESI 5 11.84 110 8.3 108 4.1
Metrafenone ESI 5 [50] 11.88 113 10.0 107 2.9
10 ng/g (n = 4)* 100 ng/g (n = 4)

Disulfoton ESI 5 [50] 11.90 98 14.7 102 7.3
Thiobencarb ESI 5 [50] 11.92 111 6.1 110 2.8
Pyrazophos ESI 1 [50] 11.94 115 8.4 109 3.4
Pencycuron ESI 1 [100] 11.94 107 6.0 103 2.7
Dialifos ESI 5 11.95 113 10.6 105 3.5
Diniconazole ESI 5 [50] 11.95 102 5.1 100 2.0
Chlorpyrifos-methyl ESI 5 [10] 11.99 107 5.6 103 2.2
EPN ESI 5 12.17 107 3.1 103 2.5
Haloxyfop-methyl ESI 5 [50] 12.18 115 3.9 112 3.0
Hexaflumuron ESI 10 12.19 106 5.2 101 1.4
Indoxacarb ESI 10 12.22 108 6.8 101 4.5
Spinosad D ESI 5 [100] 12.26 81 3.3 77 8.1
Diflufenican ESI 5 [50] 12.27 112 2.2 106 2.5
Trifloxystrobin ESI 5 [50] 12.34 115 0.9 109 2.7
Novaluron ESI 10 [10] 12.36 104 2.2 100 2.3
Spintoram J ESI 5 [100] 12.39 81 2.8 75 1.9
Difenoconazole ESI 5 12.40 104 2.9 99 4.8
Cadusafos ESI 1 [10] 12.42 110 0.9 108 2.8
Cycloate ESI 5 12.44 103 2.5 103 2.2
Fluotrimazole ESI 1 12.46 113 2.8 110 2.8
Triflumizole ESI 5 [100] 12.46 105 3.2 103 2.9
Chinomethionate ESI 10 12.66 92 4.9 93 2.5
Clethodim ESI 5 12.66 91 2.5 88 3.8
Cycloxydime ESI 10 12.71 73 2.7 75 4.9
Prosulfocarb ESI 5 12.71 111 2.7 104 2.8
Cyflumetofen ESI 5 12.73 98 5.5 99 2.7
Teflubenzuron ESI 5 12.81 106 7.6 99 2.7
Haloxyfop-ethoxyethyl ESI 5 [50] 12.83 110 2.6 106 1.7
Profenofos ESI 5 12.90 107 5.5 105 2.8
Fluazinam ESI 5 12.91 109 3.0 109 2.9
Spintoram L ESI 5 [100] 12.91 73 12.1 77 1.5
Fluazifop-p-butyl ESI 1 12.95 118 2.7 113 2.6
Diclofop methyl ESI 5 12.96 104 2.5 108 4.3
Metaflumizone ESI 20 [100] 12.99 104 15.5 92 3.3
Terbufos ESI 5 [10] 12.99 109 9.7 112 5.1
Benfuracarb ESI 1 [50] 13.02 113 3.4 110 2.6
Buprofezin ESI 1 [50] 13.02 112 2.6 110 1.4
Pirimiphos-ethyl ESI 1 13.07 112 4.6 113 1.8
10 ng/g (n = 4)* 100 ng/g (n = 4)

Furathiocarb ESI 5 [50] 13.12 113 1.9 110 1.2
Tetramethrin ESI 10 13.12 109 4.4 105 2.2
Lufenuron ESI 10 13.15 102 8.4 105 1.6
Sethoxydim ESI 5 13.17 98 3.2 99 0.8
Propaquinzafop ESI 5 13.21 110 6.3 107 1.0
Oxadiazon ESI 5 [50] 13.21 108 7.0 106 2.9
Dichlofenthion ESI 5 13.24 108 8.2 108 1.6
Tebufenpyrad ESI 5 [50] 13.24 113 1.4 114 1.2
Emamectin-benzoate ESI 10 [20] 13.25 84 8.9 76 3.8
Ethion ESI 5 13.27 107 4.8 109 2.4
Tolfenpyrad ESI 5 13.33 85 9.7 85 3.3
Pyriproxyfen ESI 1 13.37 108 5.9 106 1.4
Piperonyl butoxide ESI 1 13.41 108 4.9 107 2.2
Butachlor ESI 5 13.43 107 8.2 105 1.7
Bioallethrin ESI 10 13.47 113 6.9 104 2.4
Chlorpyrifos ESI 5 [10] 13.47 99 7.0 94 9.7
Hexythiazox ESI 5 13.53 104 3.0 104 1.5
Pendimethalin ESI 5 13.57 101 1.3 103 2.9
Flufenoxuron ESI 5 13.66 107 3.2 100 2.7
Quinoxyfen ESI 5 [50] 13.75 105 2.6 103 1.3
Flucythrinate ESI 20 [50] 13.76 ND ND 98 11.6
Carbophenothion ESI 5 13.77 107 5.3 107 1.5
Propargite ESI 5 13.96 113 5.3 111 3.4
Butralin ESI 5 [50] 14.01 110 6.6 110 1.1
Etoxazole ESI 5 14.05 138 1.7 138 2.3
Amitraz ESI 5 [100] 14.07 105 8.7 80 8.0
Chlorfluazuron ESI 10 14.11 105 16.1 104 4.6
Spiromesifen ESI 5 14.12 117 9.0 108 3.8
Fenpropathrin ESI 5 14.14 112 7.9 108 4.2
L-Cyhalothrin ESI 20 [10] 14.17 135 10.1 114 4.3
Cyfluthrin ESI 100 [100] 14.22 ND ND 109 10.0
Diafenthiuron ESI 5 14.24 19 10.3 10 27.2
Spirodiclofen ESI 5 [50] 14.48 109 3.9 107 2.7
Proquinazid ESI 1 [50] 14.49 121 0.8 123 3.1
Deltamethrin ESI 10 14.50 105 14.2 111 2.7
Cypermethrin ESI 20 14.51 ND ND 114 7.4
10 ng/g (n = 4)* 100 ng/g (n = 4)

Esfenvalerate ESI 20 [100] 14.61 ND ND 106 1.9
Acrinathrin ESI 20 [50] 14.61 ND ND 109 6.1
Fenpyroximate ESI 1 14.62 112 2.4 112 2.9
Meptyldinocap ESI 5 [100] 14.66 121 0.9 120 3.0
tau-Fluvalinate ESI 5 [50] 14.82 102 6.4 100 4.1
Pyridaben ESI 1 [50] 14.95 126 2.7 116 3.0
Permethrin ESI 5 15.00 108 4.1 100 3.9
Tralomethrin ESI 100 15.10 ND ND 96 16.4
Abamectin ESI 20 [50] 15.22 ND ND 108 7.2
Pyridate ESI 1 15.37 100 3.1 96 1.3
Carbosulfan ESI 5 [50] 15.37 131 1.7 117 2.8
Fenazaquin ESI 1 15.43 114 3.4 111 1.2
Etofenprox ESI 5 [50] 15.51 108 3.9 102 3.3
Bifenthrin ESI 5 15.58 111 3.2 104 1.8
Pyridalyl ESI 1 [50] 15.99 92 1.4 93 1.5
Fenbutatin oxide ESI 5 [50] 16.50 92 9.4 83 6.7
Acequinocyl ESI 5 [20] 16.88 83 6.9 87 1.7
*
Compounds with LOQs above 10 ng/g were expressed as Not Detected (ND) for the 10 ng/g validation level.
±
Maximum regulatory limit (MRL) taken from EU Pesticides Database (v2.2) under Teas (Product Code: 0610000), February 2022
References 12. G. Chen, P. Cao, and R. Liu, Food Chem. 125(4),

1406–1411 (2011).
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resources/docs/APP_Canadian‑Cannabis‑Pesticide.pdf>
PerkinElmer, Inc.
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75
APPLICATION NOTE
QSight 420 LC/MS/MS
AUTHOR
Avinash Dalmia
PerkinElmer, Inc.
Shelton, CT USA
Kevin Smith or other key text **
** conclusion
Napro Research
Approx. 220 Characters
Sacramento, CA USA
Toby Astill
PerkinElmer, Inc.
Woodbridge, ON, Canada
Saba Hariri and Feng Qin
PerkinElmer, Inc.
Woodbridge, ON, Canada
Quantitation of Pesticide
and Mycotoxin Residues
in Cannabis as Defined Introduction
by AZ, MI, OK, CO, OR, NV, Over half of states in the U.S. have legalized
the use of recreational or medical cannabis
and WA State Recreational and like other agricultural commodities,
pesticides are used in cannabis cultivation to
Cannabis Regulations protect cannabis plants from pests and improve
growth yield. After a pesticide is applied to a
cannabis or hemp crop, trace amounts of pesticide residues may stay on the flower. Pesticides
intended for use on cannabis must be tested for the safety of these residues. Since cannabis
is regarded as a Schedule 1 Controlled Substance by the U.S.A federal government, there are
no federal regulations for pesticide analysis in cannabis. Currently individual states in the US
have developed their own regulations outlining the list of pesticides to monitor and acceptable
maximum residue limits or action limits for each pesticide. Oregon (OR) was the first state* in
the U.S. to come up with comprehensive guidelines for pesticide residues analysis in cannabis
and set regulatory limits for 59 pesticides in cannabis1. Arizona has implemented regulations
for 56 out of 59 pesticides on the Oregon list and added an extra herbicide (pendimethalin) to its
list2. Similar to Arizona, a number of other states - Michigan, Oklahoma, Colorado, Washington
and Nevada have enacted regulations with action limits for a list of pesticides that are
a subset of Oregon's state list of pesticides with few additional pesticides3-4. In addition to
pesticides, different states requires cannabis products to be tested for 5 mycotoxins. Mycotoxins,
including 4 aflatoxins and ochratoxin A, are naturally occurring toxins produced by certain
types of mold and fungi. Mycotoxin contamination can occur during cultivation or storage of
cannabis and pose a serious health risk to consumers. As a result, testing for the levels of
pesticide and mycotoxins in cannabis is important to ensure quality control and the health of
consumers, particularly those who may already have compromised health.
For purposes of this study, Arizona (AZ), Colorado (CO), Michigan (MI), Nevada (NV), Oklahoma (OK),
Oregon (OR ), and Washington (WA) are collectively referred to herein as the “states.
76
Quantitation of Pesticide and Mycotoxin Residues in Cannabis as Defined by AZ, MI, OK, CO, OR, NV, and WA State Recreational Cannabis Regulations
Conventionally, multiresidue pesticide analysis is performed using Experimental

tandem quadrupole mass spectrometry (MS/MS) combined with
Materials and Reagents
either liquid chromatography (LC) or gas chromatography (GC)
or both. High performance liquid chromatography-tandem mass A PerkinElmer One Pesticide420TM ISO17034 CRM Reagent Kit
spectrometry (LC/MS/MS) has emerged as the method of choice was used for preparation of pesticide and mycotoxins standards
for pesticide and mycotoxin analysis because it offers superior at different concentrations for collecting validation data with an
selectivity, sensitivity, ruggedness, and does not require extensive LC/MS/MS method. To compensate for matrix effects observed
sample preparation before analysis for a wide panel of analytes. in complex cannabis samples, 30 internal standards were
Although gas chromatography-mass spectrometry (GC/MS/MS) used from this kit to improve method recovery and accuracy
methods have been developed for pesticide analysis in cannabis of quantitation.
samples, they are only applicable to a smaller subset number of
analytes. Compounds such as abamectin, a high molecular weight Pesticides Hardware/Software
compound, are not amenable to analysis by GC/MS/MS because Chromatographic separation was conducted on a PerkinElmer®
they are heat labile and degrade in either the GC injection port or LC/MS/MS® LX50 UHPLC system, while detection was achieved
the column at high temperature. A robust and single instrument using a PerkinElmer Q-Sight® 420 MS/MS detector with a dual
platform (LC/MS/MS) method is ideal for simultaneous analysis of ionization ESI and APCI source, which operate independently
pesticide and mycotoxins in cannabis labs. with two separate inlets. All instrument control, data acquisition
and data processing was performed using the Simplicity 3Q™
Numerous reports for pesticide analysis in cannabis have been
software platform.
published, most of which have certain workflow limitations 5-7.
Most of these studies use time-consuming, and costly, sample Sample Preparation Method
preparation methods (eg. QuEChERS with dSPE or SPE) with
Below is the step by step sample preparation procedure with
poor sample preparation recoveries for some of the pesticides,
10-fold dilution:
and require the use of both LC/MS/MS and GC/MS/MS based
instruments for analysis of all pesticides. This can increase cost, • Take approximately 5 grams of cannabis flower as a
complexity, and turnaround time of analysis since they require representative of each sample batch and grind it finely using
the use of two instruments and time consuming and expensive a grinder (e.g. OMNI Bead Ruptor 96).
sample preparation methods. Moreover, GC/MS/MS methods
• Measure 1 gram of sample and place it into 50 mL centrifuge tube.
are not as robust as LC/MS/MS methods for pesticide analysis
in complex cannabis matrices since they require extensive • Add 5 mL of LC/MS grade acetonitrile to the tube and cap it.
sample preparation to prevent GC injection port contamination • Place the tube on multi-tube vortex mixer and allow it to
from complex matrices. The aim of this study is to demonstrate vortex for 10 minutes.
the performance of a novel LC/MS/MS method with a dual ESI
• Centrifuge extract in tube for 10 minutes at 3000 rpm.
and APCI source and simple solvent extraction to meet the
regulations of the states for 64 pesticides and 5 mycotoxins in • Filter the solvent into a 5 mL glass amber vial using 0.22
place as of June 2021 in a complex cannabis flower matrix. micron nylon syringe-filter and cap it.
• Label the bottle with the sample ID.
• Transfer 0.5 mL of extracted sample into a 2 mL HPLC vial and
dilute it with 0.49 mL of LC/MS grade acetonitrile and mix it.
Spike 10 µL of internal standard solution from a PerkinElmer
One Pesticide420TM ISO17034 CRM Reagent Kit.
LC Method and MS source conditions were measured with an APCI source. Figures 1 and 2 show
MRM chromatograms with excellent signal to noise ratio for a
The LC method and MS source parameters are shown in Table 1.
representative set of challenging pesticides (daminozide,
MGK-264, abamectin and acequinocyl) and mycotoxins
LC Conditions (aflatoxin-B1 and ochratoxin-A) spiked at low levels in range of
LC Column PerkinElmer Quasar SPP Pesticides (4.6 × 100 mm, 2.7 µm) 0.005-0.025 µg/g (5-25ppb) in the cannabis flower, respectively
using the LC/MS/MS method with an ESI source. Figure 3 shows
An 18 min. (this time includes both analysis time and
column equilibration time) LC/MS/MS method with MRM chromatograms for two pesticides (methyl parathion and
optimized gradient using an ESI source was used for PCNB) analyzed at low levels of 0.010 µg/g (10 ppb) in the
Mobile Phase separation and analysis of 61 out of 64 pesticides and cannabis flower using the LC/MS/MS method with an APCI
Gradient 5 mycotoxins residues at low levels of a cannabis matrix.
source. For determining the limits of quantitation, cannabis
A fast 6 min. LC/MS/MS method with short gradient,
optimum mobile phase composition and an APCI source extracts were fortified with different low levels of pesticides and
was used for the measurement of remaining 3 pesticides. mycotoxins. The limits of quantification (LOQs) and response
Column Oven reproducibility at LOQ level for each of the pesticides and the
30 ºC
Temperature mycotoxins in cannabis extract are summarized in Tables
Auto sampler 2 and 3. The LOQs were determined by measuring signal to
20 ºC
Temperature noise ratio for the cannabis flower matrix spiked standards.
Injection 3.0 µL for the LC/MS/MS method with an ESI source. LOQ is the lowest concentration for spiked cannabis matrix
Volume 10 µL for the LC/MS/MS method with an APCI source. at which S/N = 10 or higher is measured for quantifier ion,
MS Source Conditions for ESI Source and APCI Source response RSD (n=7) is less than 20% and ion ratio for qualifier
ESI Voltage
to quantifier ion was within 30% of the reference ion ratio
+5500 V obtained for qualifier to quantifier ion using solvent standards.
(Positive)
ESI Voltage If the reference ion ratio for qualifier to quantifier ion was
-4200 V less than 20%, then we used the criteria that ion ratio at LOQ
(Negative)
APCI Corona level in cannabis matrix was within 50% of reference ion ratio
-5 µA obtained using solvent standards. As demonstrated in Tables 2
Discharge
Drying Gas 150 arbitrary units and 3, the LOQs in this study are well below the action limit for
the states by a factor of roughly 2 to 400 for all of 64 pesticides
Nebulizer Gas 350 arbitrary units
and 5 mycotoxins listed. The response RSD for each pesticide and
Source mycotoxin at its LOQ level in the cannabis matrix was less than
315 ºC
Temperature
20%. This demonstrates that the method is more than adequately
HSID
200 ºC sensitive and reproducible for pesticides and mycotoxins analysis
Temperature
in cannabis at the regulatory limit specified by the states.
Detection mode Time-managed MRM™
Table 1. LC Method and MS Source Conditions. Sample Preparation Recovery with Solvent Extraction
The Quick, Easy, Cheap, Effective, Rugged and Safe (QuEChERS)

Results and Discussion extraction is the most widely used sample preparation method
Detectability and Reproducibility for extraction of low levels of contaminants such as pesticides
from fruit and vegetable matrices with higher water content8.
The LC/MS/MS method with dual ESI and APCI source
The method includes extraction of a broad range of pesticides
analyzes each state’s list of regulated pesticides and
and also the removal of sugars, organic acids and other
mycotoxins in cannabis flower with two separate injections
compounds commonly found in fruits and vegetables9-12. It
using the same instrument platform and total run time of 24
is not a suitable method for very polar pesticides, such as
min. A diverter valve enables automated fast switching of
daminozide, which is included in certain states’ cannabis
mobile phase eluent to APCI source from ESI source and vice
regulations, including Arizona’s. Since daminozide is too polar
versa to facilitate the fast analysis using this method without
to be extracted efficiently with QuEChERS, it remains in the
requiring time consuming step of switching APCI and ESI ion
aqueous phase and does not partition into the organic solvent
sources in LC/MS/MS systems with a single source. With this
during salting out step. The recovery of Daminozide from a
LC/MS/MS method, 61 out of 64 pesticides and 5 mycotoxins,
cannabis matrix with QuEChERS extraction has been reported
listed in the states cannabis regulations, were analyzed using
to be less than 10%5. Different groups have tried to develop
an ESI source and remaining three pesticides (chlorfenapyr,
an advanced QuEChERS method with a d-SPE step which
methyl parathion and pentachloronitrobenzene (PCNB) )
utilizes PSA ( primary secondary amine ) or other adsorbents
to remove the matrix from cannabis extract. However, the cannabis flower matrix and solvent standard. The calculation
addition of the d-SPE step to the QuEChERS method not was performed by taking the difference between the signal
only makes this method more laborious and expensive, but of the analyte in the post spiked cannabis flower extract and
also leads to low recoveries of a few compounds such as clean solvent, and dividing it by the signal in clean solvent.
spinosad, spirotetramat, spioroxamine, and ochratoxin A, Figure 4 shows these calculated matrix effects as a function of
among others. This is a result of these compounds binding the analyte’s retention time on the column using the LC/MS/MS
to the PSA adsorbent in the d-SPE step, and resulting in poor method with an ESI source. Among the 66 analytes (61 pesticides
recoveries. Recently Moulins et al. developed three different and 5 mycotoxins) measured with the ESI source, most analytes
complex sample preparation methods using SPE ( solid phase showed signal suppression effects. Since a cannabis flower matrix
extraction) with C18 columns and d-SPE with Enhanced Matrix is very hydrophobic, significant matrix ion suppression effects of <
Removal – Lipid (EMR-Lipid) sorbents13. A number of analytes -20% were observed mainly at higher elution time for analytes. For
such as imazalil, spirotetramat, and pyridaben, among others, those analytes measured with the APCI source, ion suppression
showed very poor recoveries with their intricate, expensive effects greater than 20% were only observed for 1 out of 3
and time consuming sample preparation methods. Due to low analytes. A solution containing 30 internal standards from the
recoveries of some of pesticides using sample preparation PerkinElmer One Pesticide420 TM ISO17034 CRM Reagent Kit
methods such as QuEChERS, d-SPE and SPE to extract was spiked in both solvent standards and cannabis samples
pesticides and mycotoxins from a cannabis matrix, we used to improve the accuracy of quantitative analysis as well as the
a simple acetonitrile based solvent extraction method to get overall recovery by compensating for the matrix ion suppression
good recovery with high throughput. To confirm this method, effects and inaccuracies in sample injection in LC. The overall
fortified cannabis flower samples were used to determine recovery (accuracy) of the LC/MS/MS method takes into
pesticides and mycotoxin recovery. The cannabis flower account both the recovery from sample preparation as well as
samples were tested to confirm the absence of pesticides ion suppression effects and is given by following equation:
before they were spiked. Five cannabis flower samples were
spiked at 2 levels (low and high) of 64 pesticides (0.1 and 1 µg/g)
and 5 mycotoxins (0.01 and 0.1 µg/g) standard. The absolute Signal in pre-spiked extract Signal in post-spiked extract
Over All Recovery (%) = * * 100
sample preparation recoveries or extraction efficiency of all of 64 Signal in post-spiked extract Signal in solvent standard
pesticides and five mycotoxins at two different levels were within

an acceptable range of 80-120% with RSD less than 20% for five
cannabis flower samples. The sample preparation recovery was The first ratio in the above equation represents sample
calculated by taking the ratio of signal of analyte in pre-spiked to preparation recovery and the second ratio accounts for
post-spiked extract and multiplying this number by 100. ion suppression/enhancement effects. After canceling the
common term (signal in post spiked extract) in numerator
and denominator of above equation, the overall recovery
Signal in prespiked Extract * 100
Sample Preparation Recovery (%) =
calculation can be simplified by taking the percentage ratio of
Signal in post spiked extract
signal in pre-spiked extract to solvent standard. The following
example would explain how the addition of internal standards
would improve overall recovery by compensating for matrix
Internal Standards to Compensate for Sample Matrix Effects effects. Figure 5a shows the overlay of acequinocyl’s signal
and Improve Overall Recovery or Accuracy in a clean standard solvent and an extract of cannabis flower
spiked with acequinocyl before and after extraction. The
A cannabis flower matrix contains a number of highly
recovery from the sample preparation for acequinocyl is
concentrated plant components such as cannabinoids and
calculated by taking the percentage ratio of the signal of analyte
terpenes which are co-extracted in extract and can cause severe
in pre-spiked and post spiked extract, and it was calculated
matrix effects such as ion suppression or enhancement for a
to be 90%.The overall recovery is calculated by taking the
number of pesticides. Moreover, quantification of pesticide and
percentage ratio of the signal in pre-spiked extract to the signal
mycotoxins residues in different cannabis products is difficult
in solvent standard. Figure 5a clearly demonstrates that the
due to the great disparity in high concentration levels of naturally
overall recovery of acequinocyl without internal standard is about
occurring cannabinoids as well as terpene content and other
55% due to significant ion suppression effects caused by the
compounds, which can would lead to different sample matrix
matrix. Acequinocyl-d25, a deuterated analog of acequinocyl,
effects in different cannabis products. The ion suppression/
was added in fixed amounts to both the solvent standard and
enhancement effects were determined by checking signal for
spiked pesticides and mycotoxins at a fixed concentration in a
the cannabis flower matrix to compensate for ion suppression interference and achieve LOQ of 0.01 µg/g for propiconazole
effects. The internal standard has a similar chemical structure in the cannabis matrix. Figure 7 shows the signal overlay
to the analyte and its elution and ionization efficiency (or ion of blank cannabis matrix and propiconazole spiked at level
suppression effect) is therefore very similar to the analyte in of 0.025 µg/g in cannabis using MRM transitions with and
both the solvent standard and the sample matrix, meaning that without matrix interference. This figure demonstrates that
response ratios of the analyte to the internal standard in the optimum propiconazole MRM transition helped in achieving
solvent standard and cannabis sample would also be similar. lower detection limits due to minimal matrix interference
According to experimental results shown in Figures 5b and 5c, use from cannabis. For some of pesticides and mycotoxins, it is
of an internal standard significantly improved the overall recovery not possible to determine alternative MRM transitions with
of acequinocyl – calculated based on ratio of response ratio (RR) good signal and less matrix interference. In this case, the LC
of acequinocyl to its internal standard in pre-spiked cannabis matrix gradient can be optimized to separate the analyte and matrix
(RR=1.91) and solvent standard (RR=1.89) from 55% to 102% due interfering compound in time to reduce matrix interference.
to the correction of matrix effects. Figure 6 shows a comparison For example, Figure 8 a shows co-elution of pendimethalin
of the overall recoveries of 61 pesticides and 5 mycotoxins at a level of 0.01 µg/g in a cannabis flower matrix and matrix
analyzed using an ESI source with and without internal standard. interfering compound with generic LC gradient. After optimizing
Similarly, the overall recoveries for 3 pesticides analyzed using LC gradient, Figure 8b demonstrates base line separation for
an APCI source were in the range of 80-120% with RSD less than signal of pendimethalin at a level of 0.01 µg/g in cannabis
20%. Finally, the overall recoveries in the range of 80-120% with flower matrix and matrix interfering compound with optimum
RSD less than 20% were achieved for all 64 pesticides (at level of LC gradient. Similarly, we used optimized MRM transitions
0.1 and 1 ug/g) and 5 mycotoxins (at level of 0.01 and 0.1 ug/g) and LC gradient for a number of pesticides and mycotoxins
with the addition of 30 internal standards to the cannabis flower such as acequinocyl, prallethrin spiromesifen, pyrethrins and
matrix the for LC/MS/MS method with an ESI and APCI source. For aflatoxin-B1 to reduce matrix interference.
one of pesticides (cyfluthrin), the overall recovery values were not
determined at low spiked value of 0.1 µg/g since it was below their Analysis of Pesticides Normally Analyzed by GC/MS/MS/
LOQ value. Using LC/MS/MS with an APCI Source
LC/MS/MS with Optimum MRM Transitions and LC Gradient Some of pesticides (chlorfenapyr, methyl parathion and
to Minimize Matrix Interference PCNB) in cannabis, regulated by different states in USA, are
traditionally analyzed traditionally using GC/MS/MS with an EI
As stated, cannabis is a challenging matrix to test, and this is source since these pesticides have low proton affinity, which
compounded by the low concentration level of the pesticides. results in very low ionization efficiency with the ESI source in
Cannabis is a complex plant comprised of at least 554 different positive ion mode. An APCI ion source is much better suited
compounds including over 113 phytocannabinoids and 120 for ionization of very hydrophobic and halogenated analytes,
terpenes. Other components of the plant include hydrocarbons, and therefore it was used to determine the detection limits
nitrogenous compounds, carbohydrates, flavonoids, fatty acids, of these non-polar analytes in cannabis matrices. Using a
non-cannabinoid phenols, phytosterols, vitamin K, carotene fast six-minute LC/MS/MS method, with an APCI source
and xanthophylls as pigments, and various simple alcohols, in negative on mode, developed earlier by our group15-16, the
aldehydes, ketones, carboxylic acids, esters and lactones14. LOQs of pentachloronitrobenzene, methyl parathion and
Some of these constituents like cannabinoids and terpenes chlorfenapyr in cannabis flower matrix were in the range of
are present in the plant at percentage level. These chemicals 0.010-0.025 µg/g, well below the different state action limits
can get co-extracted with trace level of pesticides and can in cannabis. Although, both methyl parathion and chlorfenapyr
cause matrix interference if they have same retention time can be measured using an ESI source in positive ion mode,
and MRM transition as pesticide. To improve the selectivity the use of APCI ion source in negative ion mode for both these
of pesticides analysis in cannabis flower, it is necessary analytes improved their sensitivity by a factor of 5-10. Earlier,
to have multiple transitions for few compounds in order to publications have previously claimed that the analysis of these
find a transition that does not have matrix interference. For three pesticides (chlorfenapyr, methyl parathion and PCNB) can
example, propiconazole can be ionized easily as a protonated only be done with a GC/MS/MS to meet the regulatory limits in
molecular ion in a standard, and this MRM transition in figure cannabis by different states. Our data demonstrates clearly that
7a, based on monoisotopic mass ion in the cannabis matrix, these three pesticides can be measured with good sensitivity
showed poor LOQ of 0.1 µg/g due to matrix interference from and selectivity to meet state regulations for different states
coextracted compounds isobaric to this pesticide in cannabis using LC/MS/MS with an ESI source.
flower matrix. Therefore, as shown in figure 7b, MRM transition
based on M+2 isotope mass was determined to reduce matrix
Conclusions References
This study describes a novel LC/MS/MS method with dual 1. Exhibit A, Table 3. Pesticide analytes and their action levels.
ESI and APCI source for analysis of different pesticides and Oregon Administrative Rules 333-007-0400; Oregon/gov/oha,
mycotoxins residues regulated by seven different states in effective 5/31/2017.
cannabis samples as of June 2021. A number of internal
2. Arizona state regulations for pesticide and mycotoxin
standards were added to the cannabis matrix to get overall
analysis in cannabis accessed from https://www.azdhs.
recovery in the range of 80-120% and thereby improving the
gov/documents/licensing/medical-marijuana/az-medical-
accuracy of quantitation by compensating for ion suppression
marijuana-rules.pdf.
effects. Both LC gradient and MS method parameters were
optimized to minimize matrix interference from complex 3. Title 310. Oklahoma state department of health
cannabis samples. This method allowed identification and Chapter 681. Medical Marijuana regulations accessed from
quantification of all 64 pesticides and five mycotoxins at low https://oklahoma.gov/content/dam/ok/en/omma/docs/osdh
levels (0.001 to 0.25 µg/g), which is well below the actions current rules.pdf.
limits set by the states. The ability to screen and quantitate all
4. Michigan Marijuana regulatory agency sampling and testing
64 pesticides, including the very hydrophobic and chlorinated
technical guidance for marijuana products, https://www.
compounds normally analyzed on a GC/MS/MS system, and
michigan.gov/documents/mra/Sampling_and_Testing-_
the five mycotoxins, makes this method a novel way to screen
Technical_Guidance_for_Marijuana_Products_694124_7.pdf.
and quantitate pesticides and mycotoxins in cannabis with a
single instrument LC/MS/MS platform. 5. K. K. Stenerson and G. Oden, Cann. Sci. & Tech., 1(1), 48-53
(2018).
Highlights of this application
6. J. Kowlaski, J. H. Dahl, A. Rigdon, J. Cochran, D. Laine and G.
• Analysis of all seven states (regulated pesticides and Fagras, LCGC, 35(5) 8-22 (2017).
mycotoxins) by a LC/MS/MS with ESI and APCI source
7. X. Wang, D. Mackowsky, J. Searfoss and M. Telepchak,
• A method, with LOQs below the states action limits, that is LCGC, 34(10), 20-27 (2016).
reliable and reproducible
8. M. Anastassiades, S. J. Lehotay, D. Stajnbaher and F.J.
• Most straightforward sample preparation procedure to
Schenk, J. AOAC Int., 86(2), 412-431 (2003).
achieve short sample turn around times
9. S. W. C. Chung and B.T. P. Chan, J. Chromatogr. A, 1217,
• PerkinElmer One Pesticide420 ISO17034 CRM Reagent Kit can
4815-4824 (2010).
reduce time for procurement and preparation of pesticides,
mycotoxins and internal standards. 10. S.C. Cunha, S. J. Lehotay, K. Mastovska, J. O. Fernandes, M.
Beatriz and P. P. Oliveria, J. Sep. Sci., 30(4), 620-626 (2007).
• The internal standards (available from PerkinElmer One
Pesticide420TM ISO17034 CRM Reagent Kit) are added to 11. Y. Sapozhinikova, J. Agric. Food Chem., 62, 3684-3689 (2014).
compensate for ion suppression effects to improve overall
12. J. Wang and W. Cheung, J. AOAC Int. 99(2), 539-557 (2016).
recovery and accuracy of quantitation.
• Optimized MS and LC methods to minimize matrix 13. J. R. Moulins, M. Blais, K. Montsion, J. Tully, W. Mohan,
interference from complex cannabis flower matrix M. Gagnon, T. McRitchie, K. Kwong, N. Snider and D. R.
Blais, J. AOAC Int.,101(6),1948-1960 (2018).
• Dual source, ESI/APCI, removes the need for a GC/MS/MS by
using a LC/MS/MS with an APCI source for analysis of PCNB, 14. S. N. Atapattu and K. R. D. Johnson, J. Chromatogr. A, 1612
Methyl parathion and Chlorfenapyr. (2020) 460656.
• Complete application includes analysis of mycotoxins 15. A. Dalmia, E. Cudjoe, T. Astill, J. Jalali, J.P. Weisenseel,
and pesticides with one instrument (sample preparation, F. Qin, M. Murphy, and T. Ruthenberg, Cannabis Science
chromatography, and mass spectrometry) and Technology 1(3), 38-50 (2018).
16. A. Dalmia, C. Johnson, S. Hariri, J. Jalali, E. Cudjoe,

J. Kingstad and F. Qin, Current Trends in Mass. Spec., 18(3),
22-29 (2020).
Note: The following table includes some non-polar compounds typically analyzed using a GC/MS/MS in the past, which are marked with an asterisk (*) and a dagger (†).
The non-polar compounds analyzed using a LC/MS/MS with an ESI source are indicated using an asterisk (*). The non-polar compounds analyzed using LC/MS/MS with
an APCI source are indicated by a dagger (†). The rest of compounds marked with no asterisk or dagger were determined using LC/MS/MS with an ESI source.
LC‐MS/MS % CV OR AZ MI WA OK CO NV
Pesticide
(µg/g) (n=7) (µg/g) (µg/g) (µg/g) (µg/g) (µg/g) (µg/g) (µg/g)
Abamectin 0.025 8.7 0.5 0.5 0.5 0.5 0.5 0.07 0.2
Acephate 0.010 9.5 0.4 0.4 0.4 0.4 NA NA NA
Acequinocyl 0.005 5.1 2 2 2 2 NA NA 4
Acetamiprid 0.005 6.4 0.2 0.2 0.2 0.2 NA NA NA
Aldicarb 0.005 8.4 0.4 0.4 0.4 0.4 NA NA NA
Azoxystrobin 0.005 5.7 0.2 0.2 0.2 0.2 0.2 0.02 NA
Bifenazate 0.005 9.5 0.2 0.2 0.2 0.2 0.2 0.2 0.4
Bifenthrin 0.005 4.3 0.2 0.2 0.2 0.2 NA NA 0.1
Boscalid 0.005 9.1 0.4 0.4 0.4 0.4 NA NA NA
Carbaryl 0.005 8.9 0.2 0.2 0.2 0.2 NA NA NA
Carbofuran 0.010 8.2 0.2 0.2 0.2 0.2 NA NA NA
Chlorantraniliprole 0.005 8.1 0.2 0.2 0.2 0.2 NA NA NA
Chlorfenpyr † 0.025 14.4 1 1 1 1 NA NA NA
Chlorpyrifos 0.010 9.5 0.2 0.2 0.2 0.2 NA NA NA
Clofentezine 0.005 9.8 0.2 0.2 0.2 0.2 NA NA NA
Cyfluthrin* 0.25 6.7 1 1 1 1 NA NA 2
Cypermethrin* 0.050 6.3 1 1 1 1 NA NA 1
Daminozide 0.010 8.6 1 1 1 1 NA NA 0.8
Dimethomorph 0.005 8.6 NA NA NA NA NA NA 2
DDVP (Dichlorvos) 0.010 9.2 0.1 0.1 0.1 0.1 NA NA NA
Diazinon 0.010 7.6 0.2 0.2 0.2 0.2 NA NA NA
Dimethoate 0.010 7.8 0.2 0.2 0.2 0.2 NA NA NA
Ethoprophos 0.010 9.7 0.2 0.2 0.2 0.2 NA NA NA
Etofenprox 0.005 6.8 0.4 0.4 0.4 0.4 NA NA NA
Etoxazole 0.005 7.9 0.2 0.2 0.2 0.2 0.2 0.01 0.4
Fenoxycarb 0.005 9 0.2 0.2 0.2 0.2 NA NA NA
Fenpyroximate 0.005 5.8 0.4 0.4 0.4 0.4 NA NA NA
Fenhexamid 0.005 9.9 NA NA NA NA NA NA 1
Fipronil 0.010 7.8 0.4 0.4 0.4 0.4 NA NA NA
Flonicamid 0.005 8.1 1 1 1 1 NA NA 1
Fludioxonil 0.010 11.4 0.4 0.4 0.4 0.4 NA NA 0.5
Hexythiazox 0.005 5.6 1 1 1 1 NA NA NA
Imazalil 0.010 8.5 0.2 0.2 0.2 0.2 0.2 0.04 NA
Imidacloprid 0.005 8.1 0.4 0.4 0.4 0.4 0.4 0.02 0.5
Kresoxim‐methyl 0.010 9.4 0.4 0.4 0.4 0.4 NA NA NA
Malathion 0.005 9.7 0.2 0.2 0.2 0.2 0.2 0.05 NA
LC‐MS/MS % CV OR AZ MI WA OK CO NV
Pesticide
(µg/g) (n=7) (µg/g) (µg/g) (µg/g) (µg/g) (µg/g) (µg/g) (µg/g)
Metalaxyl 0.005 8.2 0.2 0.2 0.2 0.2 NA NA NA
Methiocarb 0.005 9.4 0.2 0.2 0.2 0.2 NA NA NA
Methomyl 0.010 12.3 0.4 0.4 0.4 0.4 NA NA NA
Methyl parathion† 0.010 6.2 0.2 NA 0.2 0.2 NA NA NA
MGK-264* 0.005 7.2 0.2 NA 0.2 0.2 NA NA NA
Myclobutanil 0.005 8.2 0.2 0.2 0.2 0.2 0.2 0.04 0.4
Naled* 0.005 18 0.5 0.5 0.5 0.5 NA NA NA
Oxamyl 0.005 8.5 1 1 1 1 NA NA NA
Paclobutrazol 0.005 8.1 0.4 0.4 0.4 0.4 NA NA 0.4
PCNB† 0.010 10 NA NA NA NA NA NA 0.8
Permethrins* 0.025 6.4 0.2 0.2 0.2 0.2 0.2 0.04 NA
Pendimethalin* 0.010 7.3 NA 0.1 NA NA NA NA NA
Phosmet 0.005 7.6 0.2 0.2 0.2 0.2 NA NA NA
Piperonyl Butoxide 0.005 8.9 2 NA NA 2 NA NA 3
Prallethrin* 0.010 9.5 0.2 0.2 0.2 0.2 NA NA NA
Propiconazole 0.010 9.9 0.4 0.4 0.4 0.4 NA NA NA
Propoxur 0.005 8.2 0.2 0.2 0.2 0.2 NA NA NA
Pyrethrins* 0.010 8.3 1 1 1 1 NA NA 2
Pyridaben 0.005 5.7 0.2 0.2 0.2 0.2 NA NA NA
Spinetoram 0.010 9.1 NA NA NA NA NA NA 1
Spinosad 0.010 9.6 0.2 0.2 0.2 0.2 0.2 0.06 1
Spiromesifen 0.010 7.8 0.2 0.2 0.2 0.2 0.2 0.03 NA
Spirotetramat 0.005 8.5 0.2 0.2 0.2 0.2 0.2 0.02 1
Spiroxamine 0.005 8.2 0.4 0.4 0.4 0.4 NA NA NA
Tebuconazole 0.005 7.5 0.4 0.4 0.4 0.4 0.2 0.01 NA
Table 2. The limits of quantitation and response reproducibility for pesticides in cannabis flower at LOQ level using LC/MS/MS method.
This table also displays list of pesticides and their action limits for cannabis flower in the seven states.
Action Limit for

Sample Number Mycotoxin LC‐MS/MS (ppb) %CV (n=7)
7 States in USA (ppb)
1 Ochratoxin A 5 8.7 20
2 Aflatoxin B1 1 9.9 NA
3 Aflatoxin B2 2.5 9.1 NA
4 Aflatoxin G1 1 12 NA
5 Aflatoxin G2 2.5 11.4 NA
6 Aflatoxin sum (B1+B2+G1+G2) 7 NA 20
Table 3. LOQs and response reproducibility for five mycotoxins at LOQ level with a LC/MS/MS in cannabis flower. The table also displays action limits for mycotoxins in
cannabis for the seven states.
A B
S/N = 33 S/N = 30
C D
S/N = 18 S/N = 36
Figure 1: MRM chromatogram of a representative set of pesticides:(a) daminozide, (b) abamectin, (c) acequinocyl, and (d) MGK-264 spiked at level of 0.01, 0.025, 0.005 and
0.005 µg/g, respectively in a cannabis matrix using a LC/MS/MS method with an ESI Source.
A B
S/N = 25 S/N = 51
Figure 2: MRM chromatogram of a representative set of mycotoxins:(a) ochratoxin-A, and (b) mycotoxin-B1 spiked at level of 0.005 µg/g (5 ppb) in cannabis matrix using a
LC/MS/MS method with an ESI Source.
84
A A
S/N = 27
B B
RR = 1.91
S/N = 18
C
Figure 3: MRM chromatogram of representative set of pesticides:(a) methyl
parathion, and (b) PCNB spiked at level of 0.010 µg/g in cannabis matrix using LC/ RR = 1.88
MS/MS method with an APCI Source.
Figure 5: (a) Overlay of the response of acequinocyl in solvent (Red) and cannabis
flower matrix spiked with acequinocyl before (Green) and after (Blue) extraction
without any internal standard. (b) Overlay of the response of acequinocyl (Green)
and acequinocyl-d25 internal standard (Red) in pre-spiked cannabis flower matrix
with a response ratio (RR) of 1.91 for analyte to internal standard. (c) Overlay of
response of acequinocyl (Green) and acequinocyl-d25 internal standard (Red) in
solvent with a response ratio (RR) of 1.88 for analyte to internal standard.
Figure 4: Sample matrix effects in cannabis flower matrix for 66 analytes (61 pesticides
and 5 mycotoxins) analyzed using a LC/MS/MS method with an ESI source.
85
Figure 6: The overall recovery (accuracy) % for analysis of 66 analytes (61 pesticides and 5 mycotoxins) calculated using with and without using an internal standard for a
LC/MS/MS method with an ESI source.
A B
Propiconazole
Isomers
Propiconazole
Isomers
Figure 7: (a) Overlay of the response of blank cannabis matrix (Red) and propiconazole (Green) spiked at level of 0.025 µg/g in cannabis matrix with MRM transition showing
matrix interference and (b) overlay of the response of blank cannabis matrix (Red) and propiconazole (Green) spiked at level of 0.025 µg/g in cannabis matrix with MRM
transition showing minimized matrix interference.
86
A B
Matrix Pendimethalin
Pendimethalin
Matrix
Figure 8: (a) MRM chromatogram for pendimethalin at a level of 0.010 µg/g in cannabis matrix with generic LC gradient and (b) MRM chromatogram for
pendimethalin at a level of 0.010 µg/g in cannabis matrix with optimum LC gradient.
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ANALYTICAL

LC/MS Pesticides Application Notes Compendium

Tests on Other Food Matrices

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Conclusion sample extraction utilized in this study demonstrated good recovery

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Analysis of Target Pesticide Residues in Introduction

Mobile Phase: Solvent A: 5 mM ammonium formate in water

6 20 90 10 0.3 Table 2. Optimized compound-dependent MS parameters for tested pesticides

Atrazine 216.1 174.1 15 132.0 20

Azoxystrobin 404.1 372.1 18 344.1 34

Source Temp: 325 °C

Nebulizer gas: 350 Pyrimethanil 200.0 107.0 33 82.0 32

Detection Mode: MRM Mode Thiamethoxam 292.0 211.0 18 181.1 28

Figure 1. MRM chromatogram of pesticides at 10 ng/mL in neat solution.

Table 3. Summary of Pesticides found in berry samples.

Pesticide Concentration (µg/kg)

The simple/routine sample preparation approach used in this

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Direct Analysis of Glyphosate

Gradient Conditions 2.00 20 80

Spiked Level (50 μg/kg) Spiked Level (1 mg/kg)

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Analysis of 213 Pesticide

# Pesticide Residues RT (min)

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GuangZhou Sugarcane Industry Research Institute

Mobile Phase Solvent A: Water

Column Brownlee SPP C18, 2.1x100 mm, 2.7μm

Table 2. MS/MS parameters. Results

Table 3. MS source parameters.

Source Voltage – 4500 V

Dyring Gas Setting 100

Nebulizer Gas Setting 180

Source Temperature 450 °C

Fipronill Fipronil Sulphone

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Florida Testing pesticide analysis in cannabis, individual

Below is the step by step sample preparation procedure with

Acceptable Limit/ Acceptable Limit/ Acceptable Limit/

Table 2: Continued ...

Acceptable Limit/ Acceptable Limit/ Acceptable Limit/

Acceptable Limit/ Acceptable Limit/ Acceptable Limit/

Signal in prespiked Extract * 100

Internal Standards to Compensate for Sample Matrix Effects

Figure 5 shows a comparison of the overall recoveries of 61

2. Arizona state regulations for pesticide and mycotoxin

3. Chapter 5. Testing Laboratories Section 5313 Residual

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“No Dilute” Just Shoot:

Analysis in Wine preparation is usually labor intensive and requires trained

Standards and Samples

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Analysis of near 400

In the following work, a simple solvent extraction approach Analytical Protocols

Solvent calibrants with stable isotope labelled internal standards

Matrix effects were determined by comparing slopes between

Table 1. LC/MS/MS Instrumental Conditions for ESI and APCI Methods

Further Matrix Effects Investigation

Following the promising method validation results in black tea,

LOQs Compared with Pesticide Maximum Regulatory

Blank tea sample

tea consistently between locals. The most comprehensive