CHAPTER: 2
Review of Literature
64
2.1. Methods used for the analysis of drugs
Analysis of drugs in pharmaceutical products and biological samples is growing in
importance, both in the development of more selective and effective drugs and in
understanding their therapeutic and toxic effects. Knowledge of drug levels in body fluids,
such as serum and urine, allows the optimization of pharmacotherapy and provides a basis
for studies of patient compliance, bioavailability, pharmacokinetics and the influences of
co-medications. The quantitative and qualitative analysis of drugs and their metabolites has
been applied extensively in pharmacokinetic studies because pharmacokinetic variables
such as time to reach maximum plasma concentration, clearance and bioavailability have
to be known for a new drug to be approved. In addition, therapeutic drug monitoring
(TDM) is used to improve drug therapy. In contrast drugs of abuse, illicit drugs,
intoxicating drugs and poisons are analyzed in clinical and forensic toxicology. The
screening of drugs of abuse in body fluids is also important for identifying and treating
users of these drugs and for monitoring drug addicts following withdrawal from therapy.
Also, these analytes are often present at low concentrations in biological samples. Drug
analyses have been performed using various analytical instruments under many
circumstances including clinical control for diagnosis and treatment of diseases, doping
control, forensic analysis and toxicology.
An important place is occupied by chromatographic methods based on high-
performance liquid chromatography (HPLC), thin layer chromatography (TLC), and gas
chromatography (GC)) for the determination of drugs for therapeutic drug monitoring in
biological samples and as organic pollutants in environment. Unification of the equipment
65
used necessitates preparation of a very accurate and detailed description of conditions for
carrying out the analysis. Other meaningful methods are ultraviolet-visible (UV-vis) &
infrared (IR) spectrophotometry, atomic absorptive spectrometry (AAS), nuclear magnetic
resonance (NMR), mass spectrometry (MS) or spectrofluorimetry, capillary electrophoresis
(CE), capillary zone electrophoresis (CZE), micellar electrokinetic capillary
chromatography (MEKC)) and voltamperometric methods. Any determination of organic
pollutants in environment requires either a direct analysis using these analytical
instruments or a prior preconcentration step followed by analysis. Various methods are
available for the estimation of the pharmaceuticals using chromatographic methods like
thin layer chromatography [1-10], high performance liquid chromatography with different
detectors and derivatization [11-40], gas chromatography with different detectors and
derivatization [41-78], capillary electrophoresis [79-94], ion chromatography [95-104],
ultra-violet spectrophotometry [105-114], flow injection analysis [115-119], voltammetry
and polarography [120, 121] and fluorimetric methods [122-124]. The detection methods
combined with above separation methods including ultra-violet, mass spectra, fluorescent
light, refractive index (RI) and electrochemical detection are also described.
The most widely used approaches in quality control of pharmaceuticals and
pharmacokinetic studies of drugs are HPLC and GC separations, which have become a
powerful and important technology in various other fields as well. These methods are
efficient and versatile, currently available and increasingly used analytical techniques for
qualitative and quantitative analysis of endogenous and exogenous substances in biological
samples. In case of drug formulations, since the purified molecule is routinely used in
drugs assays and it is critical that preparations being tested be devoid of antimicrobial
66
components introduced during purification. In this regard, HPLC techniques can provide a
valuable tool for generating highly pure preparations for characterizing the antimicrobial
activities. Also HPLC with its ability to analyze both volatile and non-volatile compounds,
to determine ultra trace to preparative to process scale separations, may be employed in
clinical laboratories.
Sample preparation is necessary to isolate the desired components from complex
matrices, because most analytical instruments cannot handle the matrix directly. Recent
trends in sample preparation include various forms of solid-phase extraction (SPE) [125-
138] , solid-phase microextraction (SPME) [139-142], stir-bar sorptive extraction (SBSE)
[143-148], membrane extraction [149-154], liquid-phase microextraction (LPME) [155-
159], supercritical fluid extraction (SFE) [160-163], pressurized liquid extraction (PLE)
[164-168], matrix solid-phase dispersion (MSPD) [169-172], dispersive solid-phase
extraction (DSPE) [173-176], ultrasonic assisted extraction (USAE) [177-179],
microwave-assisted solvent extraction (MASE) [180-185], etc. Some of these methods
often employ large volumes of hazardous organic solvents; others are time-consuming
and/or expensive. Most of these methods require collection of the samples and their
transportation to the laboratory for further processing. Incorrect sample handling during
collection, transportation, and preservation may result in significant variability in the
results.
Solid-phase extraction (SPE) is today the most commonly used sample preparation
method. SPE is used to extract, concentrate and clean-up compounds of interest from a
sample matrix using a solid support. Here, the analytes are adsorbed on the packing bed
and this is followed by the elution or thermal desorption for recovery. Compared to SPE or
67
liquid-liquid extraction (LLE), microextraction in packed syringe (MEPS) reduces the
sample preparation time and organic solvent consumption. MEPS is a new technique for
miniaturised solid-phase extraction that can be connected online to GC or LC without any
modifications [118-122]. MEPS can be fully automated; the sample processing, extraction
and injection steps are performed online using the same syringe. Compared to solid-phase
micro extraction, MEPS reduces both sample preparation time (Approx. 1 min) and sample
volume (10-1000 µL) and a much higher recovery (>50%) can be obtained.
2.2. Application of pre-concentration techniques with HPLC/GC-MS to
antidepressants
Antidepressant drugs are widely used for the treatment of depression and these
drugs are frequently encountered in emergency toxicology screening, drug-abuse testing
and forensic medical examinations [186]. Various methods for determination of
antidepressant drugs have been reported including HPLC, GC, GC-MS and HPLC-MS.
LLE, SPE, column switching approach and, more recently, SPME and SBSE have been
adopted for that purpose. Two noradrenergic and specific serotonergic antidepressants
mirtazapine and mianserine has been determined and separated simultaneously by using
simple TLC-densitometry method and validated for their determination in commercially
available tablets [187]. A sensitive HPLC method has been described for the simultaneous
determination of eleven cyclic antidepressants in human biological samples [188]. An
isocratic reversed-phase HPLC method with UV detection has been devised and optimized
to quantify antidepressants in human serum [189]. In another approach, a restricted access
material alkyl-diol-silica (RAM-ADS) has been used to prepare a highly biocompatible
68
SPME capillary for the automated and direct in-tube extraction of several benzodiazepines
from human serum [190]. A novel RAM-SBSE bar has been developed for the direct
extraction and desorption of caffeine and three of its metabolites in biological samples
[191]. LC methods with online sample clean-up and column switching are advantageous,
since they allow automated analysis after preparation of serum or plasma [192-195]. A
selective and reproducible in-tube SPME-LC-UV method has been reported for
simultaneous determination of few antidepressants in human plasma [196]. A few
antidepressants and metabolites have been analyzed and separated by an isocratic HPLC
method with column switching and ultraviolet detection in human serum [197]. Boron-
doped diamond (BDD) electrodes for the electrochemical detection of six tricyclic
antidepressant drugs have been examined [198]. A sensitive method using sample
preparation technique MEPS with LC-UV has been reported for the determination of new
generation antidepressants in human plasma samples [199].
Some tricyclic antidepressants and neuroleptics in their quaternary mixtures have
been simultaneously determined by a reversed-phase HPLC method with UV detection at
252 nm [200]. Santos neto et al. studied the application of a system that joins the known
advantages of capillary LC with those of column-switching using restricted access material
to the analysis of fluoxetine in plasma samples [201]. SPME coupled with HPLC-DAD has
been described for the analysis of heterocyclic aromatic amines [202]. A new and simple
analytical methodology for the simultaneous determination of twenty antidepressant drugs
in human plasma sample has been reported. The method was based on the LC-MS with
sonic spray ionization (SSI) technique [203]. Other recent approach based on in-tube
SPME-LC-MS has been developed for the analysis of ten antidepressants in urine and
69
plasma [204]. A SPME combined with LC-ESI-MS/MS to determine trace levels
amphetamine (AM) and methamphetamine (MA) in serum has been investigated [206].
Fourteen antidepressants and their metabolites has been separated and analyzed by fully
automated on-line SPE-LC-MS/MS method for the direct analysis in plasma [207]. A LC-
MS/MS method for the simultaneous determination of seventeen antipsychotic drugs in
human postmortem brain tissue has been developed. Sample preparation was performed
using hybrid SPE-precipitation technology for the removal of endogenous protein and
phospholipid interferences [208]. Tricyclic antidepressant drugs have been analyzed by a
fully-automated turbulent-flow LC-MS/MS method in serum [209]. A simple capillary gas
chromatography (CGC) procedure for the analysis of three active ingredients (fluoxetine,
fluvoxamine and clomipramine) in their respective pharmaceutical formulations has been
reported [210]. A few methods based on SBSE in combination with thermal desorption on-
line coupled to CGC-/MS to the analysis of pharmaceutical drug compounds and
metabolites and organic solutes in urine and blood are reported [212, 213].
2.3. Application of pre-concentration techniques with HPLC/GC-MS to
antiepileptics
Several methods have been reported for the determination of one or more
antiepileptic drugs in biological fluids for therapeutic drug monitoring (TDM) or for
toxicology purposes. There are various HPLC methods for the simultaneous determination.
A newly developed HPTLC method for quantitative determination of LTG, ZNS and LVT
in human plasma, in comparison to HPLC and LC-MS/MS methods, has been reported
[214]. PRM and its three major metabolites have been analyzed in rat urine by HPLC using
70
SPE [215]. A method based on SBSE-HPLC-UV for therapeutic drug monitoring of CBZ,
CBZE, PTN and PHB in plasma samples and compared with a LLE-HPLC-UV method
[216]. Contin et al. proposed a very simple and fast method for the simultaneous
determination of the new generation antiepileptic drugs LTG, OXC and main active
metabolite monohydroxycarbamazepine and FLM in plasma of patients with epilepsy
using HPLC with spectrophotometric detection [217]. Oxcarbazepine and its main
metabolites have been simultaneously determined by a method based on HPLC with UV
detection in combination with SPE for sample pretreatment in human plasma [218].
A simple and fast method for the determination of the new generation antiepileptic
drug LEV in plasma of patients with epilepsy using HPLC with UV detection has been
developed and validated [219]. The newer antiepileptic drugs RFN, ZNS, LTG, OXC and
FBM in plasma of patients with epilepsy using HPLC-UV has been reported [220]. The
separation and simultaneous estimation of the antiepileptic drugs LTG, PHB, CBZ and
PTN has been proposed by reversed-phase HPLC in human serum using a simple single-
step extraction procedure [221]. CBZ has been analyzed and estimated by an HPLC-UV
method in both solution form and rabbit plasma [222]. An interesting study has been
carried out by Thomas et al. to determine the potential impurities of eslicarbazepine
acetate. The impurities were identified by HPLC coupled with ESI and IT/MS/MS [223].
A simple method for the simultaneous determination of seven antiepileptic drugs in serum
by HPLC-DAD has been developed [224]. After SPE, separation is achieved on a C18
analytical column using isocratic elution with a mixture of acetonitrile, methanol and
phosphate buffer at 45◦C. In another analytical method, the simultaneous determination of
seven non-steroidal anti-inflammatory drugs and the anticonvulsant carbamazepine has
been examined in river and wastewater. The method involved pre-concentration and clean-
up by SPME followed by analysis with HPLC-DAD [225].
71
A rapid and reliable HPLC-DAD has reported for the simultaneous determination
of the oxcarbazepine and its metabolites in plasma and saliva from psychiatric and
neurological patients [226]. In another approach, HPLC-PDA has been developed for the
simultaneous determination of six antiepileptic drugs and two metabolites in human
plasma [227]. A simple and reliable method has been developed for the simultaneous
determination of seven non steroidal anti-inflammatory drugs and the anticonvulsant CBZ.
The method involved preconcentration and clean-up by SPME followed by HPLC-DAD
analysis [228]. In another recent application, a simple and sensitive high-performance
liquid chromatographic method for determination of gabapentin in human serum using
LLE and 9-fluorenylmethyl chloroformate (FMOC-Cl) as pre-column labeling agent has
been developed [229].
A HPLC-FD method for the simultaneous determination of the three antiepileptic
drugs in human plasma has been presented [230]. A HPLC-ELSD (evaporative light
scattering detector) method has been studied for simultaneous separation and quantitation
of four commonly used AEDs [231]. A specific and sensitive LC-MS method for the
simultaneous determination of CBZ and eight metabolites in human plasma is also
reported [232]. A restricted access media-molecularly imprinted polymer (RAM-MIP) for
cyclobarbital has been developed for selective extraction of antiepileptics in river water
samples. The RAM-MIP for cyclobarbital showed molecular recognition abilities for PHB,
AMB and PTN as well as cyclobarbital. The analysis was performed by column-switching
HPLC-MS/MS [233]. OXC and its pharmacologically active dihydro metabolite have been
determined by a HPLC-MS method. The method was successfully applied to several
authentic plasma samples from patients treated or intoxicated with OXC [234]. The CBZ
and its five main metabolites have been analyzed in aqueous samples using SPE followed
72
by LC-ES-MS/MS analysis [235]. In another different approach, a simple HPLC-MS has
been developed for the determination of PTN in human plasma. The sample preparation
involves a simple procedure based on liquid-liquid extraction [236]. A simple and accurate
method based on a LC-ESI-MS/MS for the simultaneous determination of PCM, NAP,
IBP, ETD, DCL, LTG, CBZ, PTN, PHB, CYB, AMB and CAF has been developed in
human live and post-mortem whole blood [237].
A sensitive LC-MS/MS method has been developed for the simultaneous
quantification of ten antiepileptic drugs in human plasma as a tool for drug monitoring
[238]. OXC, 10-hydroxycarbazepine (MHD) and trans-diol-carbazepine (DHD), in human
serum, have been analyzed by using LC-MS/MS. Serum drugs were extracted by C8 solid-
phase cartridges [239]. Valproic acid has been examined by using sensitive and high
throughput LC-MS/MS detection with SPE as clean up procedure in human plasma [240].
AM, CAF, PTN, RNT, and THP has been determined simultaneously by an LC-MS/MS
assay in small volume human plasma specimens for pharmacokinetic evaluations in
neonates [241]. In a recent GC-MS method, the detection of pharmaceutical residues in
various waters applying SPE has been developed [242]. A method for a range of acidic
pharmaceuticals, CBZ, and endocrine disrupting compounds has been reported in soils
with final analysis by GC-MS [243]. Another method based upon GC/MS separation has
been reported for the simultaneous determination of thirteen pharmaceuticals and five
wastewater-derived contaminants by SPE and derivatization with N,O-(bistrimethylsilyl)-
trifluoroacetamide (BSTFA). The method was applied to the analysis of raw and treated
sewage samples obtained from a wastewater treatment plant [244]. ETSX, LTG, CBZ and
CBZE have been determined after solute extraction followed by analysis using CE [245].
73
2.4. Application of pre-concentration techniques with HPLC/GC-MS to
fluoroquinolones (FQs)
Several efforts have been made during the last few years to develop analytical
methods suitable for monitoring of fluoroquinolone residues in foodstuffs [249].
Appropriate methods for the determination of these antibiotics in milk [252, 253], eggs
[255, 256], poultry [257, 259-263] and fish [269, 270] have been recently reported. Turiel
et al. analyzed several quinolones in soil samples [246]. The method was based on the
extraction of these analytes by an USAE in small columns and their subsequent
quantification by HPLC using UV detection. A HPLC method with UV detection for seven
quinolones in plasma and amniotic fluid has been presented [247]. Another method has
been developed based on HPLC-UV for the determination of four quinolones in urine,
ground water, chicken muscle, hospital wastewater and pharmaceutical samples using C18
and reverse phase amide columns [248]. The separation and analysis of several quinolones
and fluoroquinolones has been proposed in baby-food samples. The method involves
isolation of these analytes by USAE procedure followed by a SPE sample clean-up step
and final determination of the analytes by HPLC using UV detection [249]. A different
approach based on UA-DLLME coupled with LC-UV for the determination of four
fluoroquinolones in pharmaceutical wastewater has been developed [250]. An interesting
study has been carried out for the simultaneous analysis of the fluoroquinolones in bovine
serum. In this method, HPIAC column containing covalently bound anti-sarafloxacin
antibodies was used to capture the fluoroquinolones while allowing the remainder of the
serum components to elute to waste [251]. A HPLC method with DAD for the
determination of seven tetracyclines in milk has been developed [252]. Ten quinolones has
74
been examined with HPLC followed by a simple SPE cleanup procedure in cow’s milk
[253].
A HPLC-FD after MI-SPE sample pretreatment has been reported for simultaneous
analysis of few fluoroquinolones in environmental water samples [254]. In another
method, the simultaneous determination of seven quinolones in egg samples of laying hens
by HPLC-FD has been carried out [255]. A method based on PLE and HPLC-FD has been
developed for the simultaneous determination of three fluoroquinolones in table eggs
[256]. Norfloxacin and ofloxacin from chicken breast muscles has been examined using
HPLC-FD with SFE as a sample preparation [257]. The traces of the most common
veterinary fluoroquinolones marbofloxacin and enrofloxacin used as antibacterial agents
have been determined in cattle and swine farms in natural waters. Quantitative analysis
was done by HPLC-FD with SPE [258]. A sample cleanup procedure combining molecular
imprinting and matrix solid-phase dispersion (MI-MSPD) for the simultaneous isolation of
ofloxacin, pefloxacin, norflorxacin, ciprofloxacin, and enrofloxacin in chicken eggs and
swine tissues followed by HPLC-FD has been reported [259]. Enrofloxacin and its active
metabolite ciprofloxacin have been identified simultaneously by a HPLC method in
chicken muscle [260]. A cloud point extraction process to extract two fluoroquinolone
antimicrobial agents, ofloxacin and gatifloxacin, from aqueous media has been described
[261]. LC-UV, LC-MS and LC-MS/MS have been used for the simultaneous quantification
of quinolones antibiotics in turkey and chicken muscles [262, 263]. Simultaneous
determination of the structurally different antibiotics from environmental and biological
monitoring using HPLC-UV, single mass and tandem mass spectrometry has been
performed and compared [264].
Three widely used fluoroquinolones have been determined by HPLC coupled to
pneumatically assisted ESI-MS in human urine. The determination of FQs in honey sample
75
based on the combination of SRSE with HPLC-ESI-MS has been proposed [266]. A LC-
MS/MS has been utilized for the quantification of the quinolone residues in poultry muscle
and eggs [267]. More recently, a new method in which MIP material was packed as
sorbent in a device for MEPS combined with LC-MS/MS for the analysis of selected FQs
drugs in municipal wastewater samples has also been developed [268]. Two CE-MS
methods for the simultaneous determination of twelve antibacterial residues in fish and
livestock, and five quinolone residues in chicken and fish have been reported [269, 270].
2.5. Application of pre-concentration techniques with HPLC/GC-MS to
N-acyl homoserine lactones (AHLs)
A method for the determination of N-acyl homoserine lactones in the form of their
hydrolysis products has been presented. Real samples were analyzed by CZE-MS after
alkaline lactonolysis and extraction by mixed-mode anion-exchange SPE [271]. AHLs in
lung tissues of mice infected with Pseudomonas aeruginosa has been detected [272].
AHLs produced by sequential Pseudomonas aeruginosa isolated from chronically infected
patients with cystic fibrosis have been measured by thin-layer chromatography [273]. In
another method, AHL production in Gram-negative psychrotrophic bacteria has been
detected in raw milk [274]. A total of 84.9% of the bacteria were identified as AHL
producers eliciting a diversity of responses in the AHL-monitor systems. These results
demonstrate that AHL-production is common among psychrotrophic bacteria isolated from
milk and indicate that quorum sensing may play an important role in the spoilage of this
product. The isolation of AHL-degrading Shewanella sp. strain MIB015 from the intestinal
microflora of Plecoglossus altivelis (the ayu fish).17) MIB015 interrupted quorum-sensing
and exoprotase production in Aeromonas sp. by degrading AHL has been reported [275]. A
76
method based on SPE followed by UPLC for the determination of five derivatives of
AHLs has been proposed. In order to demonstrate the applicability of the method,
supernatants with the known AHL producer Burkholderia cepacia LA3 grown in different
media were investigated [276]. Another method based on fast and MS compatible UPLC-
DAD for the rapid quantitative determination of AHLs and their corresponding hydrolysis
products has been optimized and was successfully applied to a bacterial culture supernatant
real sample containing AHLs [277].
The direct evidence for the presence of AHLs in CF sputum were established.
AHLs were detected in sputum from patients colonised by P. aeruginosa or B. cepacia but
not Staphylococcus aureus. Furthermore, using HPLC-MS and thin layer chromatography,
the presence of N-hexanoylhomoserine lactone and N-(3-oxododecanoyl) homoserine
lactone respectively in sputum samples from patients colonised by P. aeruginosa was
confirmed [278]. A method using reversed-phase HPLC coupled with positive-ion ESI and
ion trap mass spectrometry for the identification and quantification of AHLs in crude cell-
free supernatants of bacterial cultures has been described. The selectivity was based on the
MS-MS fragment ions of the molecular [M+H]+
ions and on their relative intensities and
was successfully applied to Vibrio vulnificus, a marine bacterium [279]. The production of
AHLs by bacteria associated with marine sponges has been identified [280].
A method involving direct separation by GC with EI-MS to determine some AHLs has
been employed and simultaneous separation and characterization of AHLs were possible
without prior derivatization. The method was applied for the analysis of AHLs in
Burkholderia cepacia (strains JA-7 and LA-10) extracts [281]. The occurrence of AHLs in
extracts of some Gram-negative bacteria by GC-MS has been determined. Crude cell-free
77
supernatants of bacterial cultures of Aeromonas hydrophila, Aeromonas salmonicida,
Pseudomonas aeruginosa, Pseudomonas fluorescens, Yersinia enterocolitica and Serratia
liquefaciens were screened for AHL production in selected ion monitoring mode using the
prominent fragment at m/z 143 [282]. The extracts of mucopurulent respiratory secretions
from thirteen cystic fibrosis patients infected with P. aeruginosa and/or strains of the B.
cepacia complex has been studied using reverse-phase HPLC and analyzed for the
presence of AHLs using a traI-lux CDABE-based reporter that responds to AHLs with acyl
chains ranging between 4 and 12 carbons [283]. Using this assay system, a broad range of
AHLs were detected and identified despite being present at low concentrations in limited
sample volumes. N-(3-oxo-dodecanoyl)-L-homoserine lactone, N-(3-oxo-decanoyl)-L-
homoserine lactone and N-octanoyl-L-homoserine lactone (OHL) were the AHLs most
frequently identified. OHL and N-decanoyl-L-homoserine lactone were detected in
nanomolar concentrations compared to picomolar amounts of the 3-oxo-derivatives of the
AHLs identified [283]. A comparison tabulation of data on HPLC and GC-MS methods
has been given in Table 2.1.
78
Table 2.1 Survey of HPLC and GC-MS methods for quantitative determination of antiepileptics, antidepressants and
quinolones and their applications
Analytes Matrix
analyzed
Sample Preparation
Method
Analytical Technique
Used
LOD
(S/N=3)
LOQ
(S/N=10)
Recover
y (%)
Reference
AMITRI,
AMOXI,
CLOMI,
DESI, IMI,
MAPRO,
MAIN, NRT,
etc.
Biological
Samples
LLE HPLC-UV ((column 1
C8 RP columns:
a) TSK gel Super octyl
(100 mm×4.6 mm i.d.,
particle size 2 µm),
b) Hypersil MOS-C8cle
(100 mm ×4.6 mm i.d.,
particle size 5 µm),
Yokogawasize
-- 0.5
µg/mL
94-103 [188]
AMITRI,
NRT, IMI,
CLOMI,
NORCLM,
TRIMI,
MAPRO,
DESI, DOXE,
NORDX, etc.
Human serum SPE
3-ml 3M-Empore
disk cartridges
HPLC-UV
Nucleosil 100-Protect 1
(250 mm×4.6 mm i.d.,
particle size 5 µm),
Macherey and Nagel
-- -- 75-99 [189]
Benzodiazepi
ne
Human Serum SPME
(RAM-ADS)
HPLC-UV
LiChrospher 100 RP-18
(15.0 cm×4.0 mm i.d.,
particle size 5 µm),
Merck
22-29
ng/mL
74-98
ng/mL
>90 [190]
Caffeine and Rat Plasma SBSE HPLC-UV 25 ng/mL -- >50 [191]
79
metabolites (RAM-ADS), ODS Hypersil (60 mm
×4.6 mm, particle size 5
µm), Thermo Hypersil-
Keystone
ARP Human Serum RAM Column
Switching (10
mm× 4 mm i.d.,
particle size 20
µm)
HPLC-UV
LiChrospher CN column
(250 mm × 4.6 mm i.d.
particle size 5 µm), MZ-
Analysentechnik
-- <50 µg/L 94.7-
111.5
[192]
QUE, CLZ,
PRZ, OLZ
and
metabolites
Human Blood Column Switching
Silica C8 material,
particle size 20 µm
HPLC-UV
(ODS Hypersil C18
material (250 mm×4.6
mm i.d., particle size 5
µm), MZ-
Analysentechnik
-- 10-50
ng/mL
87-123 [193]
FLUVO and
its metabolites
Plasma LLE and Column
Switching
Hydrophilic meta
acrylate polymer
column, (35
mm×4.6 mm i.d.),
particle size 10 µm
HPLC-UV
C18 STR ODS-II column
(150 mm×4.6 mm
i.d., particle size 5 µm),
Shinwa Chemical
Industry
-- 0.9-1.2
ng/mL
96-100 [195]
MIRTA, CIT,
PARO,
DULO,
FLUVO and
SRT
Human Plasma In tube SPME
Fused-silica
capillary (80
cm×250 µm i.d.)
coated with the
OV-1701 phase
HPLC-UV
LiChrospher 60 RP-
select B (C18) column
(250 mm×4 mm, particle
size 5 µm), Merck
5-20
ng/mL
20-50
ng/mL
95-102 [196]
CLOMI, Serum Column Switching HPLC-UV -- -- 95-108 [197]
80
AMITRI,
ARP, CLZ,
DESI, FLU,
IMI, TRIMI,
DELO, NRT,
etc.
Perfect-Bond, (10
mm×4 mm),
particle size 20
µm, MZ-
Analysentechnik
LiChrospher 60 RP
Select (125 mm×4 mm
i.d., particle size 5 µm),
MZ-Analysentechnik
IMI, DESI,
CLOMI,
AMITRI,
NRT, DOXE
Human Plasma Centrifugation HPLC-UV
C18 column Inertsil ODS-
3 (150 mm×4.6 mm i.d.,
particle size 5 µm)
0.5-1080
nM
90.89-
92.83
[198]
SRT, MIRTA,
FLU, CIT,
PARO
Human Plasma MEPS
C8 and strong
cationic exchange
sorbent (2 mg) in
250 µL syringe
HPLC-UV
RP 18 LichroCART
(125 mm×4 mm i.d.,
particle size 5µm), Merck
-- 10-25
ng/mL
84-97 [199]
AMITRI, IMI,
CHLOR,
THIO
Pharmaceutical
Formulations
Dilution HPLC-UV
(Lichrospher100
RP-18 (250 mm×4 mm
i.d., particle size 5 µm)
with a guard column
(4 mm×4 mm, particle
size 5 µm), Merck
0.332-
0.451
µg/mL
1.5-1.69
µg/mL
98.4-
101.9
[200]
FLU Human Plasma RAM-BSA-C18
column
(50 mm×520 µm)
Capillary LC-UV (C18
analytical column (100
mm×520 µm)
-- 20 ng/mL [201]
Heterocyclic
aromatic
amine
Food Samples SPME
CW-TPR (50 µm),
CW-DVB (65 µm),
PDMS-DVB (60
HPLC-DAD
TSK-Gel ODS-80TM
column (150 mm×4.6mm
i.d., particle size 5 µm),
0.1-14
ng/mL
-- 17.8-
74.9
[202]
81
µm), PA (85 µm) Tosoh Biosep
IMI, AMITRI,
DESI,
CLOMI,
TRIMI,
MIAN,
FLUVO,
PARO, etc.
Human Plasma SPE
Oasis1 HLB
cartridge column (1
mL)
LC/MS
Inertsil
C8 (150 mm×2.0 mm,
particle size 5 µm)
0.03-0.63
µg/ mL
0.10-1.0
µg/mL
69- 102 [203]
TRZ, CLZ,
CIT, DOXE,
PARO,
FLUVO, IMI,
AMITRI,
FLU, SRT,
CLOMI, etc.
Human Urine
and Plasma
SPME
Hybrid silica
monolith with
cyanoethyl
functional groups)
LC-MS
(Inertsil
C8 (150 mm×2.0 mm,
particle size 5 µm) with a
C18 guard column),
Shimadzu
0.06-2.95
ng/mL
-- 75.2-113 [204]
Antidepressan
t and
neuroleptic
drugs
Serum LLE LC-EI-MS -- 1.2-54
nmol/L
[205]
AM and MA Human serum SPME
(7 µm, 100 µm)
(PDMS), 60 µm,
65 µm
(PDMS/DVB), 50
µm (CW/TPR) and
75 µm
(CAR/PDMS)
LC-ESI-MS/MS
Supelco Discovery C18
(15 cm×3.0 mm, particle
size 5 µm), Supelco
0.04-0.3
µg/L
0.13- 0.9
µg/L
95-96 [206]
AMITRI, Plasma SPE LC-MS/MS -- 10 µg/L >99 [207]
82
NRT, IMI,
DESI, FLU,
PARO,
FLUVO,
SRT, etc.
SPE MCX
cartridge
(cation-exchange
mode)
Gemini C18 guard
column (4 mm×2.0 mm,
particle size 5 µm),
Phenomenex
SLP, TRP,
ASLP, PLP,
RSP, QUE,
CLZ, ARP,
THIO, etc.
Human brain
tissue
SPE
1 mL Hybrid SPE-
PPT, Supelco
(Sigma-Aldrich)
LC-MS/MS
ZORBAX Eclipse Plus
C8 Narrow Bore (150
mm×2.1 mm, particle
size 5 µm), and the guard
column, Agilent
-- 2-80 ng/g [208]
AMITRI,
DESI, IMI,
NRT
Serum SPE
Cyclone-P online
SPE column
(0.5×50 mm)
LC-MS/MS
Hypersil Gold C18 (50
mm×3 mm, particle size
of 5 μm), Thermo Fisher
Scientific
< 3
ng/mL
< 20
ng/mL
97-114 [209]
FLU,
FLUVO,
CLOMI
Pharmaceutical
formulations
Homogenizations
and Centrifugation
CGC-F.I.D.
HP-5 (5% phenyl
methylsilicone, 15
m×0.25 mm i.d., 0.25 µm
film thickness), Hewlett-
Packard
10.1- 105
µg/L
33-300
µg/L
98-102 [210]
SSRI’s
Antidepressan
t
Urine LLE GC-MS 100
ng/mL
-- [211]
MTD, EUG,
DDA, DZP,
TMZ, BZP,
NDZP, etc.
Urine SBSE
Twister Gerstel,
coated with
25 μL PDMS
CGC-MS
HP-5MS column (30
m×0.25 mm i.d., 0.25 μm
df), Agilent Technologies
1 µg/L 5 µg/L 32-52 [212]
83
EUG, LNL,
NMT, MNT,
CRV, GRN,
FRN, etc.
Urine and
Blood
SBSE
Twister Gerstel,
coated with
25 μL PDMS
CGC-MS
HP-5MS column (30
m×0.25 mm i.d., 0.25 μm
df), Agilent Technologies
0.3 µg/L 1 µg/L [213]
LTG, ZNS,
LEV
Human Plasma Centrifugation HPTLC
Silica gel 60F254 (10
cm×10 cm, 250 µm
thicknesses), Merck
1.3-2.25
µg/mL
3.69-6.85
µg/mL
98-104 [214]
PRM and its
metabolites
Rat Urine SPE Bond Elut
Certify LRC
columns containing
C8 sorbent and a
SCX
HPLC-UV
Nucleosil 100-5 µm, C18,
(250 mm×4.6 mm )
Macherey-Nagel
0.5
µg/mL
1.5- 2
µg/mL
59-100 [215]
CBZ, CBZE,
PTN, PHB
Plasma SBSE 10 mm
long glass-
encapsulated
magnetic stir bar,
externally coated
with 0.5 mm thick
22µg of PDMS,
HPLC-UV
LiChrospher 100 RP-18
column (125 mm×4 mm,
particle size 5 µm),
Merck
-- 0.08-
0.125
µg/mL
72-86 [216]
LTG, OXC,
FLM
Plasma Centrifugation HPLC-UV
Synergi 4 µm Hydro-RP
column, (150 mm×4 mm
i.d.), Phenomenex
0.25-2.5
µg/mL
0.5-5
µg/mL
100.2-
104.4
[217]
OXC and its
metabolites
Plasma SPE
Oasis HLB
cartridges (30 mg,
1 ml), Waters
HPLC-UV
Varian Microsorb MV
Rainin RP column (C18,
150 mm×4.6 mm i.d.,
particle size 5 µm) with a
5 ng/mL 15 ng/mL 94.7-
98.8
[218]
84
Varian C18 precolumn
(30 mm×4.6 mm i.d.,
particle size 5 µm).
LEV Plasma Deproteinization
and centrifugation
HPLC-UV
Synergi Hydro-RP
column, (150 mm×4.6
mm i.d., particle size 4
µm), Phenomenex
-- 2 µg/mL <90 [219]
RFN, ZNS,
LTG, OXC,
FLM
Plasma Centrifugation HPLC-UV
Synergi Hydro-RP
column, (150 mm×4.6
mm i.d., particle size 4
µm), Phenomenex
-- 2 µg/mL 97-103 [220]
LTG, PHB,
CBZ, PTN
Human Serum Centrifugation HPLC-UV
NOVA PAK C18
Hypersil ODS stainless
steel column (250
mm×4.6 mm. particle
size 5 µm) with guard
cartrige Hypersil ODS
(7.5 mm×4.6 mm,
particle size 5 µm), Flexit
Jour Pvt. Ltd.
-- 0.2
µg/mL
95-102 [221]
CBZ Rabbit Plasma Centrifugation HPLC-UV
C18 µ-Bondapak, (150
mm×4.6 mm i.d., particle
size 10 µm), Waters
-- 0.5
µg/mL
98.37-
100.45
[222]
ECBZA Water Dissolution HPLC-UV
RP-8, (250 mm×4.6 mm,
0.020
µg/mL
0.060
µg/mL
93.55-
103.28
[223]
85
particle size 5 µm),
Waters
PTN, PHB,
CBZ, VPA,
ESM, LTG,
OXC, ZNS
Serum SPE
Oasis HLB
disposable
extraction columns
(30 mg, 1 ml),
Waters
HPLC-DAD
(15 cm×0.46 cm) packed
with Alltima C18, Alltech
Nederland
0.009-
0.039
mg/L
0.014-
0.065
mg/L
98-103 [224]
CBZ, NPR,
DCL, KTP,
PIR, INDO,
DFL
River and
wastewater
SPME
Fused-silica fiber
coated with
(PDMS-DVB
60-µm film
thickness),
Supelco.
HPLC-DAD
Discovery RP-Amide
C16 column, (150
mm×4.6 mm, particle
size 5 µm), Supelco.
-- 5-20
µg/L
72-125 [225]
OXC and its
metabolites
Plasma MEPS
4mg C18 material,
inserted into a 250
µL gas-tight
syringe, SGE
Analytical Science
HPLC-DAD
Gemini C18 reversed-
phase column (150
mm×4.6 mm i.d., particle
size 5 µm) equipped with
a C18 cartridge (4 mm×3
mm i.d., particle size 5
µm precolumn),
Phenomenex
0.015-
0.037
µg/mL
0.050-
0.125
µg/mL
86.5-
96.8
[226]
OXC, CBZ,
LTG, PRM,
PTN, PHB
Plasma SPE
Oasis HLB
cartridges (30 mg,
1 ml), Waters
HPLC-DAD
Spherisorb RP column
(C18 150 mm× 4.0 mm,
i.d. particle size 4.5 µm),
25-100
ng/mL
70-300
ng/mL
87-103 [227]
86
Varian
NPR, KTP,
DCL, PIR,
INDO, DFL
River Water SPME
Fused-silica fiber
coated with
(PDMS-DVB 60
µm film thickness),
Supelco
HPLC-DAD
Discovery RP-Amide
C16 column, (150
mm×4.6 mm, particle
size 5 µm), Supelco
0.5-3
µg/L
1-4 µg/L 71.6-
122.8
[228]
GBP Human Serum LLE/
Derivatization
HPLC-FD
Shimpack CLC-C18 (150
mm×4.6 mm i.d., particle
size 5 µm) with Shim-
pack G-C18 guard column
(10 mm×4.0mm i.d.,
particle size 5 µm),
Shimadzu
-- 0.03
µg/mL
90 [229]
GBP, VGB,
TPR
Human Plasma SPE
Oasis MCX
cartridges (30 mg,
1mL), Waters
HPLC-F
Synergy Hydro-RP
column, (150 mm×4.6
mm i.d., particle size 4
µm), Phenomenex
0.1-0.3
µg/mL
0.2-1
µg/mL
92-98 [230]
CBZ, PCT,
PRM, VPA
Water Dissolution HPLC-ELS
Hibar pre-packed column
RT 250-4, Lichrosorb
RP-8 (250 mm × 4 mm,
particle size 5µm), Merck
0.01-0.1
µg/mL
0.09-0.51
µg/mL
[231]
CBZ, OXC
and
metabolites
Human Plasma Centrifugation LC-MS Zorbax eclipse
XD8 C8 column, (150
mm×4.6 mm, i.d.,
particle size 4 µm)
-- 0.02-0.5
mg/L
80-105 [232]
87
PTN, PHB,
AMB, CYB
River Water RAM-MIP
(4-vinylpyridine
and ethylene glycol
dimethacrylate)
LC-MS
Cosmosil 5 C18 MS-II
(150 mm×2.0 mm i.d.)
and Cosmosil 5 C18-MS-
II guard column
(10 mm×4.6 mm i.d.),
Nacalai Tesque
0.5-5
ng/L
2-15
ng/L
96.5-113 [233]
OXC and its
metabolites
Plasma Centrifugation and
filtration
LC-MS
Merck LiChroCART
Column, (125 mm×2 mm
i.d.) with a LiChroCART
10-2 Superspher 60 RP
Select B guard column,
Merck
0.01
mg/L
0.1 mg/l 60-86 [234]
CBZ and its
metabolites
Aqueous
Samples
SPE
Oasis HLB
cartridges, (500
mg, 6 mL)Waters
LC-EI-MS
Genesis C8 column (150
mm×2.1 mm i.d., particle
size 3µm), Jones
Chromatography
0.8-4.8
pg
-- 83.6-
103.5
[235]
PTN Plasma Derivatization and
centrifugation
LC-EI-MS
(Hypersil Hypurity C18,
50 mm×4.6 mm, particle
size 5μm)
-- 101.2
ng/mL
78.33 [236]
Acidic and
neutral drugs
Human Blood Centrifugation and
filtration
LC-ESI-MS/MS
Synergi Polar-RP column
(150 mm×2.0 mm i.d.,
particle size 4 μm)
connected to a Polar-RP
Security Guard pre-
18-470
µg/L
60-1600
µg/L
92-101 [237]
88
column cartridge (2.0
mm i.d.×4 µm),
Phenomenex
GBP, VPA,
LEV, LTG,
CBZE, CBZ,
OXC, ZNS,
TPR, PTN
Human Plasma Protein
Precipitation with
CAN
LC-MS/MS Luna
C18 column (100 mm×2.0
mm, particle size 3 μm),
Phenomenex,
-- 1 µg/mL 85-114.5 [238]
OXC and its
metabolites
Human Serum SPE
C8 sorbent
LC-MS/MS
Symmetry C18, (100
mm×2.1 mm i.d, particle
size 3.5 μm), Waters
3.9-7.8
ng/mL
95.6-104 [239]
VPA Human Plasma SPE
Oasis HLB
cartridges, Waters
LC-MS/MS
Betabasic C8 column,
(100 mm×4.6 mm i.d.,
particle size 5 μm),
Thermo Electron
-- 2 µg/mL 74.47-
99.73
[240]
AMP, CAF,
PTN, RNT,
TPL
Human Plasma Protein
Precipitation with
methanol
LC-MS/MS
Luna C18 column (150
mm×3 mm, particle size
3 μm), Phenomenex
-- 12.2-48.8
ng/mL
90.2-
113.3
[241]
KTP, IBP,
CLF, NPR,
GEM, FPF,
etc.
Water Samples SPE
RP-C18 material (1
g) (BAKERBOND
Polar Plusm,
Mallinckrodt-
Baker
GC-MS
HP5MS, (30 m×0.25 mm
i.d., 0.25 μm film
thickness), Agilent
Technologies
1-10 ng/L 1-40
ng/L
70-110 [242]
CBZ, EDs,
etc.
Soil PLE
RP Oasis HLB
GC-MS
HP5-MS Fused silica
0.025-2.5
ng/g
-- 54-118 [243]
89
cartridge, (200 mg) capillary column (30
m×0.25 mm, 0.25μm
film thickness)
CBZ and
other
pharmaceutica
l compounds
Wastewater SPE
Oasis HLB
cartridges (200 mg,
N vinylpyrrolidone
and divinylbenzene
mixture, particle
size 30 μm,
Waters,
C18-U-MCAX (200
mg), Supelco,
Oasis MCX (200
mg) Waters,
Strata X, and
StrataXC (both 500
mg), Phenomenex
GC-MS
DB-5MS column, (30
m×0.25 mm
i.d., 0.25 μm phase
thickness), Agilent
1-30 ng/L 3-90
ng/L
21-111 [244]
CBZ, CBZE,
LTG, ESM
Plasma LLE CE Fused-silica
capillaries (360 mm×50
μm i.d.), Polymicro
Technologies
0.40-15
µM
-- 94.76-
108
[245]
CINO, OXO,
NAL, FLQ
Soil USAE
Glass columns (10
cm×2 cm i.d.),
Scharlab
HPLC-UV
Atlantis C18 column (150
mm×3.0 mm, particle
size 3 μm), Waters
0.05-0.08
µg/g
0.15-0,25
µg/g
90.2-
104.3
[246]
LEVO, ENO,
MOXI,
LOME, OXO,
Plasma and
Amniotic
flui.d.
SPE
Strata X (30 mg,1
mL), Phenomonex
HPLC-UV
a) Zorbax Eclipse XDB-
C8 column (150 mm×4.6
0.005-
0.01
µg/mL
0.020-
0.035
µg/mL
95-98.6 [247]
90
MARB,
PEFLO
mm i.d.) connected to a
Kromasil C8 column (20
mm×4.5 mm i.d.)
b) Zorbax Eclipse XDB-
C18 column (150 mm×4.6
mm i.d.) in connection to
a Phenomenex C18
column (4 mm×3.0 mm
i.d.).
OFLO,
LOME,
CINO, NAL
Urine, Water
samples,
Chicken
muscles
Filtration and
centrifugation
HPLC-UV
C18 RP analytical column
Acclaim 120, (250
mm×4.6 mm i.d., particle
size 5 μm) Dionex
Supelco RP-amide
column, (150 mm×4.6
mm i.d., particle size 5
μm), Ascentis
0.55-1.41
ng/mL
1.67-4.27
ng/mL
>90 [248]
ENRO, ENO,
CINO, CIP,
NOR, DANO,
NAL, FLQ,
OXO
Baby food USAE/MI-SPE
(MIP-SAX, 150
mg)
HPLC-UV
Atlantis C18 HPLC
column (150 mm×3.9
mm, particle size 3 μm)
coupled to an Atlantis
C18 guard column (20
mm×3.9 mm, particle
size 3 μm), Waters
0.03-0.11
µg/g
0.10-0.35
µg/g
>85 [249]
OFLO, NOR,
ENRO,
LOME
Pharmaceutical
wastewater
UA-DLLME
HPLC-UV
Zorbax Eclipse XDB-C18
column (150 mm×4.6
0.14-0.81
µg/L
-- 82.7-
110.9
[250]
91
mm, particle size 5 μm),
Agilent Company
CIP, ENRO,
SARA,
DIFLO
Bovine Serum
PEEK cartridges
(2.1 × 30 mm) with
POROSE media
containing protein
G (PE Biosystems)
HPIAC
Inertsil phenyl column
(150 mm×4.6 mm,
particle size 5 μm),
Alltech
0.18-0.47
ng/mL
1 ng/mL 95-100.8 [251]
MNC, OTC,
MTC, DMC,
CTC, DC
Milk SPE (Abselut
Nexus), Varian,
(Discovery,
Supelco), and
(Lichrolut), Merck.
HPLC-DAD
Inertsil ODS-3 analytical
column, (250 mm×4
mm2
, particle size 5 μm)
3-6
ng/mL
10-20
ng/mL
93.8-
103.7
[252]
ENO, OFLO,
NOR, DANO,
CIP, ENRO,
NAL, FLU
Milk SPE
LiChrolut RP-18
(200 mg, 3 mL)
cartridges, Merck
HPLC-PDA Perfect Sil
Target ODS-3 analytical
column (250 mm×4 mm2,
particle size 5 μm), MZ-
Analysentechnik
1.5-6.8
ng/µL
-- 75-92 [253]
ENRO, CIP,
NOR, LOME,
DANO,
SARA, FLQ,
OXO, AMX,
etc.
Aqueous
Samples
MI-SPE
The template
enrofloxacin
(183.4 mg, 0.5
mmol), functional
monomer 1 (187.1
mg, 0.5 mmol),
methacrylamide
(85 mg, 1 mmol),
EDMA (3.8 mL,
20 mmol) and the
HPLC-FD Aqua C18
column (polar
endcapped; 250 mm×4.6
mm i.d., particle size 5
μm) protected by an
RP18 guard column (4.0
mm×3.0 mm i.d., particle
size 5 μm), Phenomenex
0.01-0.30
µg/L
-- [254]
92
free radical
initiator ABDV
(42.4 mg, 1%
(w/w) total
monomers)
dissolved in ACN
(5.6 mL)
ENRO,DIFL
O, DANO,
CIP, FLQ,
OXO, SARA
Eggs Protein
Precipitation
HPLC-FD
Waters Symmetry C18
column, (150 mm×3.0
mm), Waters
4-12 ng/g 99.2-
100.6
[255]
ENRO,
CIPRO
Eggs PLE
HPLC-FD
AQUA C18 column,
(polar endcapped, 250
mm×4.6 mm, particle
size 5 µm) protected by a
RP18 guard column (4.0
mm×3.0 mm, particle
size 5 µm), Phenomenex
LC-MS
Synergi MAX-RP
column, (150 mm×2 mm
i.d., particle size 4 µm),
Phenomenex
17-20
ng/g
30-41
ng/g
67-90 [256]
NOR, OFLO Chicken
muscles
SFE
10 ml stainless
SFE vessel (150
mm×10mm o.d.)
HPLC-F
Novapak C18 stainless
column (300 mm × 3.9
mm i.d., particle size 4
-- 2.5
ng/mL
70-87 [257]
93
µm), Waters
MARB,
ENRO
Surface water SPE
(Envi-18 and SDB-
XC, Strata-XC,
MM1, WAX-HLB)
cartriges
HPLC-FD
Hypersil C18 column,
(250 mm×4.6 mm,
particle size 5 µm),
Varian
0.7-2.2
ng/L
2-6 ng/L 90-116 [258]
OFLO,
PEFLO,
NOR, CIP,
ENRO
Chicken eggs
and Swine
tissue
MI-MSPD
MAA, TRIM, and
AIBN sorbent
HPLC-FD
ODS C18 stationary
phase VP-ODS, (150 mm
× 4.6-mm i.d., particle
size 5 µm), Shimadzu
0.05-0.09
ng/g
-- 85.7-
104.6
[259]
ENRO, CIP Chicken
muscles
MI-SPE
EDMA and AIBN
as polymerization
mixture
HPLC-FD
ODS C18 stationary phase
VP-ODS, (150 mm × 4.6
mm i.d., particle size 5
µm), Shimadzu
0.07-0.09
ng/g
-- 77.8-
94.6
[260]
OFLO,GATI CPE
FD F-4500 recording
spectrofluorometer with a
xenon lamp, Hitachi Ltd.
0.04-0.06
ng/mL
-- 96.5-
99.39
[261]
CIP, DANO,
ENRO,
DIFLO, FLQ
Turkey muscles SPE
ENV+ Isolute
cartridges
LC-UV Zorbax Eclipse
XDB-C8 column (150
mm×4.6 mm i.d., particle
size 5 µm), Agilent
Technologies and using a
pre-column Kromasil C8
(20 mm×4.5 mm),
Aplicaciones Analíticas.
LC-MS
4-10
µg/kg
0.4-2
13-33
µg/kg
2-6 µg/kg
70-87
72-85
[262]
94
LC-MS/MS
µg/kg
0.05-0.1
µg/kg
0.2-0.5
µg/kg
73-85
NOR, CIP,
SARA,
DIFLO,
ENRO,
DANO, OXO,
FLM
Chicken
muscles
LLE/SPE
ENV+ cartridges
(200 mg, 3 mL)
LC-UV
LC-MS Zorbax Eclipse
XDB-C8 (150 mm×4.6
mm i.d., particle size 5
µm), Agilent
Technologies
5-20
µg/kg
0.15-0.50
µg/kg
15-60
µg/kg
0.50-1.50
µg/kg
70-85
70-85
[263]
CIP, OFLO,
CEFA, CEFT,
CEFU,
CHLOR
Urine and wipe
samples
SPE
Bakerbond C18
cartridges, (1000
mg, 6 mL), Baker
HPLC-UV
Nucleosil 100-5 C18 HD
(250 mm×3 mm i.d.),
Macherey-Nagel
HPLC-MS
Nucleodur 100-5 C18 EC
(125 mm×3 mm i.d.),
Macherey-Nagel
HPLC-MS/MS
Nucleodur 100-5 C18 EC
column (125 mm×2 mm
i.d.), Macherey-Nagel
30-75
µg/L
0.4-70 µg
/L
0.05-0.3
µg/L
-- >70 [264]
NOR, CIP,
OFLO, ENRO
Human Urine SPE
3M-Empore MPC
Extraction
cartridges, Supelco
HPLC-ESI-MS
Kromasil C8 column,
(250 mm×4.6 mm i.d.),
Teknokroma
13-21
µg/L
44-58
µg/L
46-62 [265]
PEFLO,
DANO, CIP,
Honey SRSE
Monolithic
HPLC-ESI-MS
Agilent Eclipse-XDB-C18
0.06-0.14
ng/g
0.21-0.48
ng/g
70.3-
122.6
[266]
95
DIFLO polymer AMPS,
OCMA, EDMA,
DMF, PEG and
AIBN.
column (150 mm×4.6
mm i.d., particle size 5
µm)
ENRO,
DIFLO,
DANO, FLU,
OXO
Poultry
muscles and
eggs
Homogenization
and centrifugation
LC-MS/MS
Waters Symmetry C18
column (150 mm× 3.0
mm), Waters
1-10
µg/Kg
2-20
µg/Kg
-- [267]
CIP, NOR,
OFLO, FLQ
Wastewater MI-MEPS
Polymerization
solution, CIP,
MAA, EGDMA,
AIBN and MeOH,
(100 µL gas-tight
syringe with 4mg
polymer)
LC-MS/MS
Hypersil Gold PFP
column (30 mm×2.1 mm,
particle size 5 µm),
Thermo Scientific
0.5-8.1
ng/L
-- 87-115 [268]
Sulfonamides
and
quinolones
Fish and
Livestock
SPE
ODS (MFE-Pak
C18) particle
diameter in the
range of 45-55 µm
and pore diameter
60 Å), Análisis
Vínicos
CE-MS
Fused silica capillary, 75
cm total length, 50 cm
thermostated, 25 cm at
room temperature,
75 mm i.d., and 375 mm
o.d., Supelco
1-10
µg/kg
15-30
µg/kg
78-97 [269]
DANO, FLQ,
OFLO,
ENRO,
PPMA
Chicken
muscles and
Fish
SPE
ODS (MFE-Pak
C18) particle
diameter in the
range of 45-55 µm
CE-MS
Fused silica capillary, 75
cm total length, 50 cm
thermostated, 25 cm at
room temperature, 75
20 ng/g -- 62-99 [270]
96
and pore diameter
60 Å), Análisis
Vínicos
mm i.d., and 375 mm
o.d., Supelco
AHLs Culture
supernatant of
B. cepacia
SPE
Oasis MAX
cartridges, Waters
CZE-MS
Fused-silica capillaries
(50 cm length, 75 mm
i.d., 360 mm o.d.),
Polymicro Technologies
0.01
µg/mL
0.05
µg/mL
95-105 [271]
AHLs Mice lung
tissue
-- TLC
C18 RP TLC plates,
aluminium sheets RP- 18
F254g (20 cm×20 cm),
Merck Chrom line
-- -- -- [272]
AHLs P. aeruginosa -- TLC C18 reversed-phase
TLC plates, Merck
-- -- -- [273]
AHLs Milk Agitation TLC
-- -- -- [274]
AHLs Shewanella sp. -- HPLC
Crestpak C18T-5 C18
reverse phase column,
Jasco
-- -- -- [275]
AHLs (C4-
C14)
Barley sees SPE
(Bond Elut LRC
C18-OH, Mega
Bond Elut C18,
Bond Elut PPL,
Bond Elut PRS and
Bond Elut SCX),
UPLC
(100 mm×2.1 mm i.d.,
particle size 1.7 µm),
filled with BEH C18
packing material,
0.4-10
µM
3.2-6.6
µM
94-97 [276]
97
Varian
(Bakerbond C18,
Octadecyl polar
plus, Bakerbond
phenyl, Bakerbond
Silica Gel,
Bakerbond
Florisil, Bakerbond
Diol, Bakerbond
WP CBX,
Bakerbond
Cation Exchange),
Baker
(Strata-X Cation
Exchange), Pheno-
menex (Oasis
MAX), Waters
(Chromabond HR-
P), Macherey and
(Adsorbex NH2),
Merck
AHLs B. cepacia -- UPLC-MS Acquity
BEH C18 column, (100
mm×2.1 mm, particle
size 1.7 µm), Waters
Corporation
0.11-1.64
mg/L
1.03-4.90
mg/L
-- [277]
AHLs Sputum Dilution and
centrifugation
TLC/LC-MS
Kromasil KR100-5C8
column (250 mm×8 mm),
-- -- -- [278]
98
Hichrom
AHLs E. coli Centrifugation HPLC-MS/MS
C18 RP column Hypersil
ODS, (250 mm×4.6 mm,
particle size 5 µm)
0.28-93
pM
-- 63-116 [279]
AHLs Marine sponges -- -- -- -- -- [280]
AHLs B. cepacia -- GC-MS HP-5 MS
capillary column, (30
m×250 mm i.d., 0.25 µm
film thickness) coated
with 5% Ph Me siloxane
-- -- -- [281]
AHLs Y. enterocolitia Centrifugation GC-MS
HP-5 MS capillary
column, (30 m×250 mm
i.d., 0.25 µm film
thickness) coated with
5% Ph Me siloxane.
3.2-6.2
µM
-- -- [282]
AHLs Respiratory
secretion
Centrifugation -- 0.02 µM-
250 nM
-- -- [283]
99
2.6. Conclusions
Sample preparation is a process required for the transformation of a sample to make
it amenable for chemical analysis or to improve the analysis. This is necessary when a
given sample cannot be directly analyzed or when direct analysis generates poor results.
Typical problems with analyses are interferences and low sensitivity. Sample preparation
is usually needed to eliminate interferences and to increase sensitivity.
SPE has evolved rapidly as a major sample pretreatment technique with a wide
application area. There is a continuously growing interest in this technique from various
fields. Application of a SPE technique makes sample preparation very simple, rapid and
accurate. SPE is chiefly used to prepare liquid samples and extracts of semi-volatile or
nonvolatile analytes but may also be used for solids pre-extracted into solvents. SPE has
been widely adopted for preparing samples in the analysis of pharmaceuticals and drugs of
abuse in biological matrices. The choice of sorbent is the key factor in SPE because this
can control parameters such as selectivity, affinity and capacity. This choice depends
strongly on the analytes and their physic-chemical properties, which should define the
interactions with the chosen sorbent. However, results also depend on the kind of sample
matrix and interactions with both the sorbent and the analyte. SPE sorbents range from
chemically bonded silica of the C8 and C18 organic groups, grafitized carbon, ion-exchange
materials up to polymeric materials, mixed-mode sorbents, immuno sorbents, molecularly
imprinted polymers as well as restricted access materials and recently developed monolith
sorbents. MIPs are capable of molecular recognition and are stable enough for long-term
storage, easy to prepare and inexpensive. Thus, they may be considered to be a new
artificial affinity media. Different modes of MIP based SPE have been demonstrated
100
including various modes of off-line and on-line SPE for pre-concentration or pre-treatment
of analytes and for conventional SPE where the MIP is packed into columns or cartridges.
MEPS relates the versatility of the new tool to provide an avenue for improved
sample preparation to aid the speed, sensitivity and selectivity options provided by HPLC
and GC. Although, MEPS is in its infancy; potential exists for this technique to be applied
on a larger scale since it provides an opportunity for easy, efficient and cleaner sample
preparation. It could fit in well with the available tools in both the qualitative and
quantitative aspects of separation science. The technique may lead to newer innovations
since it provides flexibility in different parameters including type of adsorbent materials,
loading environment, sample load size, etc. It is anticipated that the participation of
researchers should further aid in refining and defining the optimal use of MEPS with LC or
GC strategy including the control of the matrix effects.
101
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