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Review Hyphenation in Sample Preparation: Advancement From The Micro To The Nano World
Review Hyphenation in Sample Preparation: Advancement From The Micro To The Nano World
hyphenated separation. Kato et al. [15] also described an loop. The sol-gel titania – PDMS coated capillaries were
on-line solid-phase extraction method, coupled with used for on-line extraction and HPLC analysis of poly-
HPLC – MS-MS for the determination of 16 phthalate cyclic aromatic hydrocarbons, ketones, alkylbenzenes,
metabolites in human urine. The method employed a and a wide range of other less volatile or thermally labile
conventional analytical column for the chromato- compounds [21] that are not amenable to GC separation.
graphic separations of these analytes and the mobile Hashi et al. [22] described the determination of polycyclic
phase used was A (0.1% acetic acid in water) and B (0.1% aromatic hydrocarbons (PAH) in the atmospheric partic-
acetic acid in acetonitrile) at a flow rate of 0.35 mL/min. ulates by using on-line enrichment coupled with fast
The limits of detection ranged from 0.11 to 0.90 ng/mL. A high-performance liquid chromatography with fluores-
similar set-up was described by Kuklenyik et al. [16] for cence detection. The limits of detection of PAH were in
the extraction and measurement of perfluorinated the range of 0.02 – 0.23 ng/mL with recoveries between
organic acids and amine in human serum and milk. A 12- 87 and 12% for spiked atmospheric particulate sample.
lL volume of the reconstituted serum or milk extract The mobile phase used was acetonitrile – water (72:28,
was auto-injected on to HPLC at a 300-lL/min flow rate v/v) at a flow rate of 1.0 mL/min. Altun et al. [23] developed
with 20 mM ammonium acetate (pH 4) in water and and validated a method for local anesthetics in human
methanol as mobile phase. The HPLC gradient program plasma through on-line MEPS by using a cation-
(14 min) was started at 60% methanol in the mobile exchanger with a flow rate of 0.20 mL/min.
phase followed by an increase in organic content to 80% Abdel-Rehim [24] developed and validated a new sensi-
in 0.5 min, which was kept for 9 min. Later on, the tive, selective, and accurate on-line micro-extraction in
mobile phase organic content was decreased in 0.5 min packed syringe (MEPS) technique hyphenated with HPLC
to 60% methanol, where it was kept for 3 min to equili- for the determination of lidocaine, prilocaine, ropiva-
brate the column. Furthermore, the same group [17] caine, and mepivacaine in human plasma. The extrac-
described an automated on-line hyphenation of SPE with tion recoveries were in the range of 60 – 90%. Veuthey et
HPLC – MS for the extraction and measurement of isofla- al. [25] described on-line solid phase extraction to achieve
vones and lignans in urine. The mobile phases used were nano analysis of drugs in biological samples. In this
10 mM ammonium acetate (pH 6.5) and methanol – ace- hyphenation technique, the single column performs two
tonitrile (50:50 v/v) at a flow rate of 0.8 mL/min, respec- functions, i. e. extraction and separation. The column was
tively. The detection limits were in the range of 0.2 – connected to a detection system via a switching valve.
0.7 ng/mL. These authors described these hyphenations The sample was directly injected on to the extraction
as an innovation in separation science. support with and after the extraction, the valve was
Koster et al. [18] reported the analysis of lidocaine in switched, and analytes were transferred to the detector
urine by an on-line SPME – LC method. A polydimethylsi- with the eluting mobile phase followed by extraction
loxane (PDMS) coated fiber was directly immersed into support re-equilibration. According to the authors, the
buffered urine with optimized contact time, pH, ionic method was simple and several applications have been
strength, and temperature. The extraction yields were published for the direct analysis of biofluids. Quintana et
22% in about 45 min with a reproducibility of a 5% al. [26] described an automated on-line hyphenation of
expressed as relative standard deviation. The detection SPE – HPLC incorporating multi syringe flow injection
limits were 25 – 1000 ng/mL. Volmer et al. [19] studied the analysis (MSFIA), bead injection, and lab-on-valve (BI –
eleven corticosteroid and two steroid conjugations in a LOV) prior to HPLC. The potential of the novel MSFI – BI –
urine sample by SPME – LC – MS. Several SPME optimiza- LOV hyphenation for on-line handling of complex envi-
tion factors such as polarity of fibres, extraction time ronmental and biological samples prior to reversed-
and effect of ionic strength, were investigated, and their phase chromatographic separations was assessed for the
impact on the SPME/LC/MS technique was studied. The expeditious determination of five acidic pharmaceutical
method was sensitive with detection limits between 4 residues viz. ketoprofen, naproxen, bezafibrate, diclofe-
and 300 ng/mL and precision between 4.9 and 11.1% nac, and ibuprofen along with one metabolite, i. e. sali-
RSD. Kim et al. [20]. developed sol-gel titania-based coat- cylic acid, in surface water, urban wastewater, and urine.
ing capillary micro extraction (CME) coupled with HPLC The column used was an Xterra RP-18 (3.96150 mm)
for the extraction and analyses of polycyclic aromatic with the mobile phases A: MeOH – water (20:80, v/v) and
hydrocarbons, ketones, and alkylbenzenes at high pH. To B: MeOH – water (95/5, v/v), both containing 0.1% (v/v) for-
perform CME – HPLC, a so-gel TiO2 – PDMS capillary was mic acid at flow rates of 1.0 mL/min. The detection limit
installed in an HPLC injection port as an external sam- was 0.02 – 0.67 ng/mL.
pling loop. HPLC conditions were ODS column Clarkson et al. [27] described hyphenation of solid-
(25064.6 mm) with acetonitrile – water (80:20v/v) as phase extraction with liquid chromatography and
mobile phase. The target analytes were extracted on-line nuclear magnetic resonance: application for identifica-
by passing the aqueous sample through this sampling tion of flavonol glycosides (kaempferol 3-O-(6-O-a-L-rham-
Biological matrices
Trazodone Plasma SPE – GC – FID 3 lg/L [118]
Benzodiazepines Plasma SPE – GC – NPD 5 – 25 lg/L [119]
Benzodiazepines Plasma ISP – CGC 0.5 – 2 lg/L [120]
Biogenic amines Cells and culture medium Dialysis – RPLC – fluorescence – 10 fmol/lL [121]
ED
Ciprofloxacin Biological samples Dialysis – RPLC – fluorescence 0.1 nM [122]
Amino acids Beverages and feed stuff Dialysis – RPLC – fluorescence 1 – 278 lg/L [123]
Fluoroquinolones Fortified tissue Dialysis – RPLC – fluorescence 2.5 – 5 ng/g [124]
Sterols Oils and fats LLE – GC – FID 0.04 – 0.08 ng/100 g [125]
Methylenedioxylated amphet- Plasma and serum samples Dialysis – RPLC – fluorescence 10 lL [126]
amines
Clozapine & N-desmethylcloza- Human plasma Dialysis – RPLC – UV 0.050 – 0.055 lmol/L [10]
pine
Verapamil & Norverapamil Human plasma Dialysis – RPLC – fluorescence 5 lg/L [127]
Local anesthetics Plasma SLM – GC 1 lg/L [128]
Ciprofloxacin Biological samples Dialysis – RPLC – fluorescence 0.1 nM [122]
Phenytoin, Carbamazepine & Plasma Dialysis – RPLC – UV 0.1 – 0.8 lg/L [129]
Phenobarbitone
Tramadol Human plasma RAM – MIP – RPLC – fluoresence a10 ng/mL [130]
Arsenic species Urine Dialysis – IC – HGAAS 1.0 – 2.18 lg/L [131]
Ropivacaine & metabolite Plasma Dialysis – lRPLC – MS 0.1 nM [132]
Ropivacaine Urine SLM – IPLC – UV 2 – 18 nM [133]
Phenols Plasma SLM – LC – biosensor 50 lg/L [134]
Meropenem Rat bile Dialysis – lRPLC – UV 0.1 mg/L [135]
Bambuterol Plasma SLM – lRPLC – UV 80 nM [136]
Atrazine mercapturate Urine SPE – RPLC 0.0 5ng/mL [137]
& planar PCBs Biological samples SFE – NPLC – UV a0.3 – 7.4 ng/g [138]
Blood, milk, tissue
N-Methylcarbamates Food PHWE – PRLC – fluorescence 1 lg/mL [139]
Piritramide Human plasma and urine SPE – LC/MS/MS 0.05 ng/mL [140]
Cyproterone acetate Human plasma RAM – RPLC – MS/MS 300 pg/mL [141]
Sugars and organic acids Foods and beverages Dialysis – RPLC – RI 0.6 – 0.41 lg/g [142]
Fluconazole Blood and dermal rat micro- Dialysis – RPLC – UV 0.10 mg/L [143]
dialysates
Bismuth, cadmium & lead Urine SPE – GF – AAS 0.002 – 0.013 ng/mL [144]
Cadmium Biological RM SPE – ICP-AES 0.05 ng/mL [145]
Environmental matrices
Endocrine disruptors Water SPE – GC – MS 0.1 – 20 ng/L [146]
HCH & ethers Water LLE – GC – ECD or FID 20 – 480 pg/L [147]
Organic pollutants Water SPE – GC – ECD 4.1 – 6.3 ng/L [148]
Phenols Water SPE – GC – FID 1 – 27 ng/L [149]
Phenols Water SPE – GC – FID 0.3 – 2 lg/L [150]
OPPs Water SPE – GC – AED 2 – 5 ng/L [151, 152]
Micro contaminants Water SPE – GC – FTIR 100 – 1000 ng/L [153]
Micro contaminants Water SPE – GC – MS 0.2 – 20 ng/L [154, 155]
Pesticides Water LLE – GC – AED 1 – 5 lg/L [156]
Pesticides Water SPE – GC – MS 2 – 20 ng/L [157]
Pesticides Water SPE – GC – MS a1 lg/L [158]
Pesticides Water SPE – RPLC – MS – MS 0.4 – 13 ng/L [159]
Endocrine disruptors Water SPE – GC – MS 0.1 – 20 ng/L [142]
Triazines & OPPs Wastewater SPE – GC – NPD or MS 15 – 25 ng/L (NPD) [160]
1.5 ng/L (MS)
Alkylthio-s-triazineherbicides River water SLM – RPLC – UV 0.03 lg/L [161]
Vinclozolin Water MMLLE – NPLC – UV 1 lg/L [162]
Cationic surfactants Aqueous samples MMLLE – NPLC – UV 0.7 – 5 lg/L [163]
Drugs Water SFE – RPLC – UV – MS 200 ppb [164]
Wine aroma compounds Wine SFE – GC – FID 0.8 – 3.4 lg [165]
Organic compounds Aerosol particles SFE – NPLC – GC – MS 0.02 – 0.04 ng/m3 [166]
Organic acids Aerosol particles SFE – NPLC – GC – MS 0.4 ng/m3 [167]
Table 1. Continued
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