International Journal of Mass Spectrometry 488 (2023) 117040

Contents lists available at ScienceDirect

International Journal of Mass Spectrometry

journal homepage: www.elsevier.com/locate/ijms

A fast, accurate and comprehensive LC-MS/MS method validation for

the sensitive quantification of water-soluble vitamins in walnut,
almond, hazelnut and pistachio fruits
Mustafa Abdullah Yilmaz a, b, *, Oguz Cakir b, Ismail Yener a
a
Department of Analytical Chemistry, Faculty of Pharmacy, Dicle University, 21280, Diyarbakir, Turkey
b
Dicle University Science and Technology Application and Research Center, 21280, Diyarbakir, Turkey

a r t i c l e i n f o a b s t r a c t

Article history: Evaluating nutritional values of food products such as nuts has been of great importance nowadays. In
Received 30 September 2022 the current study, development and validation of a robust and fast LC-MS/MS method was carried out for
Received in revised form simultaneous determination of pantothenic acid (B5), riboflavin (B2), biotin (B7), thiamine (B1), cyano-
8 March 2023
cobalamine (B12), nicotinamide (B3), nicotinic acid (B3), pyridoxine (B6) and folic acid (B9) in walnut,
Accepted 10 March 2023
Available online 16 March 2023
almond, hazelnut and pistachio samples from Turkey. Isotope labeled vitamin standards were used to
compensate the sensitivity losses and increase the degree of precision. The validation parameters of the
developed analytical method were accuracy (recovery), linearity, limit of quantification (LOQ), intra-day
Keywords:
LC-MS/MS
and inter-day precision (repeatability), relative standard uncertainty (U% at 95% confidence level (k ¼ 2)),
Method validation and limit of detection (LOD). Chromatographic parameters were optimized to have optimum separation
Water soluble vitamins of nine water soluble vitamins (WSVs) and MS detection was performed using a tandem mass spec-
Isotope labeled internal standard trometer. Linearity of the established LCeMS/MS method was in the range of 25e1000 ng/mL for the
Nuts studied vitamins. The coefficients of determination for the vitamins were between 0.9973 and 0.9999.
LOD/LOQ values of the analytes were 10.48/12.54, 13.08/19.59, 5.77/7.70, 12.48/17.71, 12.38/14.90, 6.55/
10.08, 8.08/9.81, 13.03/16.38, 13.08/19.59 for B1, B2, nicotinamide (B3), nicotinic acid (B3), B5, B6, B7, B9 and
B12, respectively.
© 2023 Elsevier B.V. All rights reserved.

1. Introduction into fat-soluble and water-soluble groups. Water-soluble vitamins

Tree nuts, especially walnut (Juglans regia L.), almond (Prunus
dulcis L.), hazelnut (Corylus avellana L.) and pistachio (Pistacia vera
L.) are recommended as part of a healthy diet owing to their
beneficial nutritional value containing proteins, vitamins, fatty
acids, and phytochemicals. Studies of recent date favor the rela-
tionship between nut consumption and a reduced risk for diabetes,
cholesterol, postprandial glycemia, insulinemia, obesity, cardio-
vascular diseases and cancer [1e7]. Moreover, regular nut con-
sumption is associated with an adverse risk for the metabolic
syndrome and obesity. A daily intake of 25 g is recommended to
merge nuts in a healthy diet [8].
Based on their physical properties, vitamins could be divided
* Corresponding author. Department of Analytical Chemistry, Faculty of Phar-
macy, Dicle University, 21280, Diyarbakir, Turkey.
E-mail address: mustafaabdullahyilmaz@gmail.com (M.A. Yilmaz).
contain the B-complex with reference to all known water-soluble
vitamins saving vitamin C [10,11]. Each of the B-complex vitamins
functions biologically either as effective elements in the regulation
of biological functions in the human body within normal limits or
as micronutrients necessary for human health [9]. Deficit or surfeit
of certain vitamins might indicate or cause certain illnesses. To
illustrate, the deficiency of vitamins B1 might cause biochemical
brain lesions called Wernicke's encephalopathy and vitamin B6 and
B12 deficiencies may cause anemia [12,13].
Considering the crucial role of vitamins and the nutritional
importance of nuts, investigating the vitamin content of nuts is of
great importance. Vitamins in food and pharmaceutical formula-
tions have been studied using several analytical techniques such as
liquid chromatography with fluorescence, UV-VIS and mass spec-
trometric detection, voltammetry, micellar electrokinetic chroma-
tography, charge coupled device (CCD) photodetector, sequential
injection chromatography and capillary zone electrophoresis
[14e25]. Being a golden standard, LC-MS/MS is a stable and

https://doi.org/10.1016/j.ijms.2023.117040
1387-3806/© 2023 Elsevier B.V. All rights reserved.
M.A. Yilmaz, O. Cakir and I. Yener International Journal of Mass Spectrometry 488 (2023) 117040

efficient analytical method, having advantages of the ability to be
applied to various complex matrices, extreme sensitivity, and
unique specificity. Several studies have been conducted mentioning
the analysis of secondary metabolites in plant species [26e30].
There are several studies involving the LC-MS analysis of fat- and
water-soluble vitamins together. However, this might not be a good
idea as the extraction conditions required for water soluble vita-
mins are incompatible with fat soluble vitamins [31,32].
In this study a fast, accurate, comprehensive and robust LC-MS/
MS method was developed and validated for simultaneous
determination of nine water soluble vitamins B1, B2, B3 (nicotin-
amide), B3 (nicotinic acid), B5, B6, B7 (biotin), B9 and B12 in walnut,
almond, hazelnut and pistachio samples from Turkey.
2. Materials and methods
2.1. Chemicals
Reference standards of B group water soluble vitamins (B1:
thiamine hydrochloride (99.7%), B2: riboflavin (98.9%), B3:

Fig. 1. Molecular structures of the water soluble vitamin standards used in the study.

2
M.A. Yilmaz, O. Cakir and I. Yener International Journal of Mass Spectrometry 488 (2023) 117040

nicotinamide (100%), B3: nicotinic acid (99.5%), B5: pantothenic acid stored in their shells at room temperature prior to analysis in order
calcium salt (97.7%), B6: pyridoxine hydrochloride (99.9%), B7: to determine their water-soluble vitamins in the fruits.
biotin (99.3%), folic acid (91.5%), cyanocobalamine (95.7%)) were
obtained from Dr. Ehrenstorfer (Augsburg, Germany) (Fig. 1).
2.3. Preparation of vitamin standard solutions
Isotope labeled internal standards (13C4-thiamine hydrochloride
(98% assay and 99% isotopic purity), 13C4,15N2-riboflavin (97% assay
The stock solutions of WSV standards with 250 mg/L (B1, B2, B3
and 98% isotopic purity), 2H4-nicotinamide (98% assay and 98%
(nicotinic acid), B3 (nicotinamide), B5, B6, B7 and B12) were prepared
isotopic purity) and 2H4-nicotinic acid (98% assay and 98% isotopic
individually in water. Folic acid (B9) was prepared in 0.01 M NH4OH
purity) were purchased from Sigma Aldrich (Germany). 13C3,15N-
solution. 1000 mg/L Stock solutions of each isotope labeled internal
pantothenic acid calcium salt (98% assay, 13C3, 99%, 15N 98%), 13C4-
standards (13C4eB1, 13C15 2 2
4 N2eB2, H4-nicotinamide (B3), H4-nico-
pyridoxine hydrochloride (95% assay and 98% isotopic purity), 13C5- 13 15 13 13 13
tinic acid (B3), C3, NeB5, C4eB6, C5eB7, C5eB9) were pre-
biotin (95% assay and 98% isotopic purity) and 13C5-folic acid (95%
pared from their purchased forms accordingly. A mixture of the
assay and 99% isotopic purity)) were purchased from Cambridge
studied vitamin standards and a mixture of internal standards were
Isotope Laboratories (Hungary). Acetic acid, ammonium acetate,
freshly prepared every day. Calibration point standards of the
hexane and HPLC grade methanol were purchased from Merck
studied vitamins were prepared from these secondary daily stock
(Darmstadt, Germany). Ultrapure water was obtained from Sarto-
solutions by diluting with ultrapure water. Internal standard solu-
rious (Goettingen, Germany) Arium Pro Ultrapure Water System.
tion mixture was added to each calibration points, so that the
concentration of each internal standard would be 100 mg/L. Table 1
shows the names, retention times, mother ions, fragment ions and
2.2. Plant material
ionization modes for the vitamin standards utilized in the method.
Ten walnut genotypes were selected from Mardin (Derik dis-
trict) province in the south-east part of Turkey. Two almond (Fer- 2.4. Extract preparation from nuts for LC-MS/MS study
raduel and Ferragnes), two walnut (Franquette and Chandler), and
two pistachio standard varieties (Uzun and Siirt) were collected The air-dried walnut, almond, hazelnut, and pistachio fruits
from commercial fields of Sanliurfa province (south-east Turkey). were separated from their shells. Then, nut samples were
Two hazelnut standard varieties (Acı and Foşa) were collected from powdered and the fruit samples (0.2 g) were placed in a centrifuge
Giresun province (north-east Turkey). Nut samples were dried and tube (15 mL). Afterwards, 50 mL of internal standard mixture was

Table 1
Parent ions, transition ions and ionization modes for each vitamin and the corresponding internal standard.13C4,15N2-riboflavin was used as internal standard for both
riboflavin and cyanocobalamine.

# Vitamin standards R.T. Mother ion (m/z) Fragment ions Ionization mode

1 Thiamine (B1) 0.72 265.15 144.00e81.10 Pozitive

13
C4-thiamine 0.72 269.20 122.05e81.10 Pozitive
2 Pyridoxine (B6) 1.09 170.15 77.25e152.10 Pozitive
13
C4-pyridoxine 1.09 174.10 138.15e156.15 Pozitive
3 Nicotinic Acid (B3) 1.18 124.15 78.20e53.20 Pozitive
2
H4-nicotinic acid 1.18 128.15 84.20e109.15 Pozitive
4 Nicotinamide (B3) 1.63 123.10 80.05e96.10 Pozitive
2
H4-nicotinamide 1.63 127.10 84.15e100.10 Pozitive
5 Pantothenic Acid (B5) 2.27 220.10 98.00e183.90 Pozitive
13
C3,15N-pantothenic acid 2.27 224.15 94.15e206.05 Pozitive
6 Folic Acid (B9) 5.69 442.10 176.05e120.05 Pozitive
13
C5-folic acid 5.69 447.05 176.10e295.10 Pozitive
7 Cyanocobalamine (B12) 6.36 678.40 358.90e456.80 Pozitive
8 Biotin (B7) 6.52 245.20 97.15e123.20 Pozitive
13
C5-biotin 6.52 250.20 97.10e126.10 Pozitive
9 Riboflavin(B2) 6.70 377.10 172.20e198.00 Pozitive
13
C4,15N2-riboflavin 6.70 383.10 175.20e249.15 Pozitive

(m/z: mass to charge ratio, R.T.: Retention time).

Table 2
Analytical method validation parameters which belong to the developed LC-MS/MS method.

# Vitamin standards R.T. Equation r2 R.S.D.% Linearity Range (mg/L) LOD/LOQ (mg/L) Recovery (%) U

Interday Intraday Interday Intraday

1 Thiamine (B1) 0.72 y ¼ 14386.7  -643916 0.9999 0.0010 0.0010 25e1000 10.48/12.54 0.9852 1.0395 0.0174
2 Pyridoxine (B6) 1.09 y ¼ 19786.7  -30761 0.9988 0.0028 0.0027 25e1000 6.55/10.08 1.0082 1.0399 0.0177
3 Nicotinic Acid (B3) 1.18 y ¼ 1792.8  -14418 0.9989 0.0018 0.0015 25e1000 12.48/17.71 1.0494 1.0963 0.0225
4 Nicotinamide (B3) 1.63 y ¼ 7487.1  -246166 0.9999 0.0006 0.0011 25e1000 5.77/7.70 1.0853 1.0671 0.0307
5 Pantothenic Acid (B5) 2.27 y ¼ 841.4  þ2756 0.9973 0.0010 0.0008 25e1000 12.38/14.90 1.1517 1.1151 0.0326
6 Folic Acid (B9) 5.69 y ¼ 388.2  þ5977 0.9999 0.0425 0.0443 25e1000 13.03/16.38 0.9242 0.9313 0.0942
7 Cyanocobalamine (B12) 6.36 y ¼ 851.2  þ9370 0.9999 0.0404 0.0513 25e1000 13.14/17.65 0.9604 0.9635 0.1163
8 Biotin (B7) 6.52 y ¼ 765.8  þ6754 0.9998 0.0362 0.0485 25e1000 8.09/9.81 0.9782 0.9845 0.0748
9 Riboflavin(B2) 6.70 y ¼ 409.9  -7105 0.9999 0.0007 0.0012 25e1000 13.08/19.59 1.1219 1.0880 0.0148

(r2: Coefficient of determination, LOD/LOQ (mg/L): Limit of detection/quantification, RT: Retention time, RSD: Relative standard deviation, U (%): percent relative uncertainty at
95% confidence level (k ¼ 2)).

3
M.A. Yilmaz, O. Cakir and I. Yener
added and the samples were defatted by hexane (10 mL), filtered
and left to dry. Dried nut samples were put into another centrifuge
tube (15 mL). Later, acetic acid solution (5 mL, 0.1 M) was added to
the tube and the mixture was vigorously shaken (5 min). The
mixture in the centrifuge tube was kept in the ultrasonic bath
(15 min). Afterwards, the centrifuge tube was filled up to 15 mL
with 2 M ammonium acetate solution. The mixture was shaken for
5 min. After that, the sample solution was centrifuged at 4400 rpm
for 10 min. Lastly, 2 mL of the supernatant was taken slowly and
filtered via syringe filter (0,2 mm) and stored in the freezer (-20  C)
till the LC-MS/MS analysis.
2.5. Instrumentation and analytical conditions
The chromatograph and the mass spectrometer instrument
conditions and the details of the analytical method were given in
the supplementary information file.
3. Results
3.1. LC-MS/MS analytical method validation studies
In this study a comprehensive and robust LC-MS/MS method was
optimized and validated for the quantification of nine water soluble
vitamins (B1, B2, B3 (nicotinic acid), B3 (nicotinamide), B5, B6, B7, B9
and B12) in ten walnut genotypes, two walnut (Chandler and Fran-
quette), two almond (Ferraduel and Ferragnes), two hazelnut (Acı
and Foşa) and two pistachio standard varieties. Standard solutions,
spiked and non-spiked samples were used to evaluate the perfor-
mance characteristics of the analytical method. In this context, the
developed method was validated in terms of limits of quantification
and detection (LOQ/LOD), intra-day and inter-day precision
(repeatability), accuracy (recovery), relative standard uncertainty (U
% at 95 % confidence level (k ¼ 2)), and linearity (Table 2).
The linearity was examined using external standard calibration
curve with six concentration levels for each analyte, and each
concentration level was assayed in triplicate. The developed
method demonstrated to be linear for all vitamin compounds, be-
tween the ranges of tested concentrations (25e1000 mg/L) during
the validation of the method with r20.9973. The coefficients of
determination (r2) and the equations of the calibration curves were
demonstrated in Table 2.
Standard addition procedure was used to perform precision and
accuracy of the developed analytical method. Three levels of spike
concentrations (25 ppb for low concentration, 100 ppb for medium
concentration and 500 ppb for high concentration) were chosen for
Fig. 2. TIC chromatogram of B group vitamin standards at 500 ppb concentrations (1: Thiamine, 2: Pyridoxine, 3: Nicotinic acid, 4: Nicotinamide, 5: Pantothenic acid, 6: Folic acid, 7:
Cyanocobalamine, 8: Biotin, 9: Riboflavin) analysed by LC-MS/MS.
4
International Journal of Mass Spectrometry 488 (2023) 117040
each analyte. The three chosen levels of concentrations were be-
tween the lowest (25 ppb) and highest (1000 ppb) points of the
calibration points. To evaluate the intra-day variability, six repli-
cates of the spiked samples were measured within a single day,
while to examine the inter-day assay, spiked samples were assessed
in triplicate per day for three consecutive days. The accuracy was
investigated using the recovery data calculated by the percentage
ratio of observed concentration to the nominal concentration of the
spiked sample. The precision was evaluated via the intermediate
precision (inter-day) and repeatability (intra-day).
In order to determine the LOQ and LOD values of the studied
vitamins in the LC-MS/MS, 10 identical analyte mixture solutions
were prepared at the minimum concentration signaled by the
vitamin standards and injected to the LC-MS/MS. Following equa-
tions were used to calculate the LOQ and LOD values (1 and 2):
LOQ ¼ Mean þ 10  Standard Deviation (1)
LOD ¼ Mean þ 3  Standard Deviation (2)
The detailed data on LOQ and LOD studies were given in sup-
plementary information file (Table S1). The standard uncertainties
of the vitamin analytes were assessed by the performed precision
(repeatability) and accuracy (recovery) studies in accordance with
the Eurachem/CITAC Guide [33].
4. Discussion
4.1. Method validation
Simultaneous determination of multiple water soluble vitamins
in nuts has analytically important challenges both in terms of
extraction and separation. The fact that B group water soluble vi-
tamins have different functional groups and polarities entails an
appropriate extraction procedure followed with a sensitive detec-
tion method like LC-MS/MS. Although there were several studies
covering the detection of some water soluble vitamins in different
nut samples by HPLC coupled to UVeVis, DAD, fluorescence or mass
spectrometric detector, these methods were limited and might not
show wide applicability or comprehensiveness [33e39].
These detection methods either have sensitive to the matrix
effects or limited specificity as they change the relative response
between standards and samples. In this study, these drawbacks
were compensated by using isotope labeled analogs of the studied
B group vitamins with tandem mass spectrometric detection. The
developed LC-MS/MS method to quantify nine water soluble group
B vitamins was fully validated. The method demonstrated to be
M.A. Yilmaz, O. Cakir and I. Yener International Journal of Mass Spectrometry 488 (2023) 117040

Table 3
Amounts of group B vitamins in 10 walnut genotypes and standard walnut (Chandler and Franquette), almond (Ferraduel and Ferragnes), hazelnut (Acı and Foşa) and pistachio
(Uzun and Siirt) varieties (mg/L).

Sample B1 B2 B3 (Nac) B3 (Nam) B5 B6 B7 B9 B12

MDE2 2.78 1.35 9.57 1.98 6.08 4.84 0.052 ND ND

MDE 18 3.58 1.76 8.95 1.26 5.20 5.32 0.053 ND ND
MDE 24 3.42 1.67 8.99 1.86 5.26 4.61 0.050 ND ND
MDE 32 2.56 1.23 7.29 1.76 5.34 5.16 0.054 ND ND
MDE 39 3.37 1.65 9.06 1.85 5.66 4.87 0.062 ND ND
MDE 42 4.00 1.98 8.79 1.31 5.16 5.13 0.063 ND ND
MDE 45 2.43 1.16 7.77 2.09 5.24 5.29 0.050 ND ND
MDE 50 2.82 1.36 8.80 1.77 5.62 5.35 0.066 ND ND
MDE 56 3.26 1.59 8.24 1.27 5.08 5.05 0.052 ND ND
MDE 67 2.40 1.15 8.23 1.15 5.84 5.23 0.050 ND ND
Chandler 3.61 1.77 9.49 1.99 5.99 4.70 0.064 ND ND
Franquette 2.97 1.44 8.94 1.07 5.07 5.14 0.056 ND ND
Ferraduel 1.87 11.26 29.63 3.42 4.25 1.57 0.136 ND ND
Ferragnes 2.23 10.83 31.85 3.88 4.89 1.39 0.143 ND ND
Acı 6.45 1.25 16.87 3.05 10.62 6.26 0.156 ND ND
Foşa 6.98 1.07 15.72 2.64 9.58 5.89 0.162 ND ND
Uzun 8.45 1.65 11.63 2.41 5.86 18.09 0.212 ND ND
Siirt 7.65 1.76 10.89 2.57 5.31 16.78 0.197 ND ND

Note: Nac: nicotinic acid, Nam: nicotinamide, ND: Not detected.

Fig. 3. TIC chromatogram of the Chandler walnut variety analysed by LC-MS/MS.

linear for all analytes (25e1000 mg/L), between the ranges of
studied concentrations during the validation of the analytical
method with r20.9973. The intraday and interday RSD values of
the analytes were smaller than 0.0513 and 0.0425, respectively.
Moreover, LOD values of the vitamin standards were between 5.77
and 13.14 while the interday and intraday recoveries values of the
analytes were between 0.9242 and 1.1517 (Table 2, Fig. 2).
4.2. Quantitative evaluation of WSVs in nut samples by LC-MS/MS
The developed analytical method was applied to determine the
amounts of WSVs in ten walnut genotypes, two walnut (Chandler
and Franquette), two almond (Ferraduel and Ferragnes), two
hazelnut (Acı and Foşa) and two pistachio standard varieties. Ac-
cording to the quantitative analyses, certain B group vitamins
appeared to be in high amounts in the studied nut samples. To
begin with, all of the commercial cultivars and studied genotypes of
nut samples contained considerable amounts of vitamins B1, B2, B3
(nicotinic acid), B3 (nicotinamide), B5 and B6, while vitamins B9, B12
were absent in all of the studied species and varieties (Table 3).
Fig. 3 shows the LC-MS/MS TIC chromatogram of Chandler standard
walnut variety. Figs. S1eS17 shows the LC-MS/MS TIC chromato-
grams of other studied nut samples in the supplementary infor-
mation file.
5
Pistachio varieties (8.45 and 7.65 mg/kg dry fruit in Uzun and
Siirt, respectively) was the richest in terms of thiamine that serves
as a coenzyme facilitating the breakdown of amino acids and car-
bohydrates. Thiamine contents of hazelnut, walnut and almond
varieties followed pistachio samples in amount. On the other hand,
almond varieties (11.26 and 10.83 mg/kg dry fruit in Ferraduel and
Ferragnes, respectively) remarkable amounts of riboflavin (almost
ten times the other nut varieties). Nicotinic acid and nicotinamide
contents (different forms of vitamin B3) of almond (29.63 and
31.85 mg/kg dry fruit in Ferraduel and Ferragnes, respectively) and
hazelnut (16.87 and 15.72 mg/kg dry fruit in Acı and Foça, respec-
tively) varieties were also notably high.
Hazelnut (10.62 and 9.58 mg/kg dry fruit in Acı and Foça,
respectively) varieties were the richest nut samples in terms of
pantothenic acid (B5). Additionally, Pistachio (18.09 and 16.78 mg/
kg dry fruit in Uzun and Siirt, respectively) varieties were quite rich
interms of pyridoxine. Being necessary for the production of
glucose in the body and thought to support the growth of the hair
and nails, biotin content of pistachio (0.212 and 0.197 mg/kg dry
fruit in Uzun and Siirt, respectively) samples were noteworthy.
Several recent studies reported the water soluble vitamin con-
tents of nutsamples. California-grown raw almond varieties
detected to have different amounts of riboflavin (between 5.8 and
22.7 mg/kg) and niacin (14.0e50.2 mg/kg) [35]. Moreover, in
M.A. Yilmaz, O. Cakir and I. Yener
another study water soluble vitamin contents of Turkish Tombul
hazelnut variety detected by fluorometric and microbiological
methods were 0.42 ± 0.03, 0.10 ± 0.01, 0.63 ± 0.04, 1.94 ± 0.15,
1.12 ± 0.07 and 0.08 ± 0.01 mg/100 g in thiamin, riboflavin, pyri-
doxine, niacin, pantothenic acid and biotin, respectively [40]. In
addition, in some Tunisian pistachio varieties vitamin B1 content
varied from 0 to 1 mg/g in the varieties Meknessy 1, Kermizi,
Mateur-2, Mateur-3 and Mumtez, and was 0.019 mg/g in Lybie
rouge. B2 vitamin content varied from 0.001 mg/g in the variety
Mateur-3 to 0.07 mg/g in Lybie rouge, and vitamin B6 content
varied from 0. 0016 mg/g in Mateur-3 to 0.0164 mg/g in Red Aleppo
[41].
5. Conclusion
In conclusion, in this study a fast, comprehensive and robust LC-
MS/MS method was developed and validated to simultaneously
qualify and quantify water soluble vitamins B1, B2, B3 (nicotin-
amide), B3 (nicotinic acid), B5, B6, B9 and B12 in ten walnut ge-
notypes, two walnut (Chandler and Franquette), two almond
(Ferraduel and Ferragnes), two hazelnut (Acı and Foşa) and two
pistachio standard varieties. Isotope labeled vitamin standards
were used to compensate for the sensitivity losses and increase the
degree of precision. The results of the current study demonstrates
that almond, walnut, hazelnut and pistachio fruits can provide
valuable amounts of water soluble vitamins. The data confirm that
studied nut varieties are quite rich sources of significant vitamins
that would be very beneficial to human health. Furthermore, the
developed and comprehensively validated LC-MS/MS method is
applicable not only to nuts but also to a wide range of foods and
other natural products.
Author statement
Mustafa Abdullah YILMAZ: Data curation, Methodology, Vali-
dation, Writing original draft. Oguz Cakir: Conceptualization, Re-
sources, Supervision. Ismail Yener: Formal analysis, Visualization,
Writing review & editing.
Declaration of competing interest
The authors declare that they have no known competing
financial interests or personal relationships that could have
appeared to influence the work reported in this paper.
Data availability
The data that has been used is confidential.
Acknowledgements
Authors are thankful to DUBTAM (Dicle University Science and
Technology Application and Research Center) for allowing the us-
age of its instrumental facilities and they are grateful to Prof.Dr.
Mikdat Şimşek for supplying the nut samples.
Appendix A. Supplementary data
Supplementary data to this article can be found online at
https://doi.org/10.1016/j.ijms.2023.117040.
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