Combination of liquid chromatography/tandem mass spectrometry and gas chromatography/mass spectrometry for the detection of 21 anabolic steroid residues in bovine urine

For the detection of anabolic steroid residues in bovine urine, a highly sensitive liquid chromatographic/electrospray ionization tandem mass spectrometric (LC/ESI-MS/MS) method was developed using both positive and negative ionization. For four compounds the ESI mode was not sensitive enough and gas chromatographic/mass spectrometric GC/MS detection was therefore still necessary as a complementary method. The sample clean-up consisted of solid-phase extraction (SPE) on a C(18) column followed by enzymatic hydrolysis and a second solid-phase extraction on a combination of a C(18) and a NH(2) column. After this last SPE clean-up, the eluate was split into two equal aliquots. One aliquot was further purified and after derivatization used for GC/MS analysis. The other aliquot was analyzed with LC/MS/MS in both ESI+ and ESI- modes. The method was validated according to the European Commission Decision 2002/657/EC. Decision limits (CCalpha) were between 0.16 and 1 ng ml(-1) for the compounds detected with the LC/MS/MS method. The developed method is used in routine analysis in our laboratory.     

Fast analysis of 19 anabolic steroids in bovine tissues by high performance liquid chromatography with tandem mass spectrometry

For the detection of 19 steroid hormones in bovine muscle, a fast and sensitive liquid chromatography with electrospray ionization tandem mass spectrometry method was developed using both positive and negative ionization mode. Chromatographic separation on Poroshell 120‐EC C18 column was achieved in less than 10 min using isocratic elution of mobile phase of acetonitrile/methanol/water. The compounds were extracted from muscle tissue using ethyl acetate and quick, easy, cheap, effective, rugged, and safe technique. The purification of the obtained extract was performed by dispersive solid‐phase extraction with sorbents C18, primary secondary amine and magnesium sulphate. The method was validated in accordance with the Commission Decision 2002/657/EC. For all steroids tested good recoveries were obtained (from 51.2 to 121.4%) in the concentration range from decision limits until 5 µg/kg. The values of decision limits and the detection capabilities for individual compounds were in the range 0.10–0.48 and 0.17–0.95 µg/kg, respectively. The method was characterized by satisfactory linearity for most compounds (correlation coefficients  > 0.99) and the reproducibility was lower than 35%. The elaborated procedure has met the criteria for confirmatory methods and is currently used in the official control of hormones.     

Quantitation of 17alpha-ethinylestradiol in aquatic samples using liquid-liquid phase extraction, dansyl derivatization, and liquid chromatography/positive electrospray tandem mass spectrometry

A rapid and sensitive method for the analysis of 17alpha-ethinylestradiol (EE2) in environmental aqueous samples has been developed. Aquatic samples were extracted using liquid-liquid extraction, and organic phase extracts were concentrated and derivatized with dansyl chloride. Analysis was performed using high-performance liquid chromatography with positive electrospray ionization and tandem mass spectrometry (HPLC/ESI-MS/MS). Deuterated 17alpha-ethinylestradiol was used as internal standard and was added to samples before extraction. A limit of quantitation of 1 ng/L was obtained using a 25 mL aqueous sample. The average recovery of EE2 spiked into a 25 mL tapwater sample was 100%. This highly sensitive quantitation method is useful for measuring low levels of EE2 in aqueous environmental samples.     

Determination of chloramphenicol residues in bee pollen by liquid chromatography/tandem mass spectrometry

A novel liquid chromatographic/tandem mass spectrometric (LC/MS/MS) method was developed for the trace residue determination of chloramphenicol (CAP) in bee pollen. CAP was extracted from bee pollen with a mixture of methanol and 1% metaphosphoric acid solution, followed by a 2-stage solid-phase extraction enrichment and cleanup. The first stage involved a polymeric cartridge, and the second stage involved an alumina neutral cartridge. The LC separation was performed on a C18 column with 10 mM ammonium formate-acetonitrile (7 + 3) as the mobile phase and MS detection with negative-ion electrospray ionization. CAP-d5 was used as the internal standard. The method was validated according to Commission Decision 2002/657/EC. The calibration curves were linear between 0.1 and 5.0 ng/mL, and overall recoveries ranged from 98 to 113%. Decision limits (CCalpha) ranged from 0.05 to 0.07 microg/kg, and detection capabilities (CCbeta) ranged from 0.08 to 0.12 microg/kg. The developed method was applied to 11 samples.     

Highly sensitive determination of haloperidol in human serum by capillary gas chromatography using a surface ionization detector

A method has been developed for highly sensitive determination of haloperidol in human serum involving a simple extraction procedure followed by gas chromatographic separation. Target components were separated from the extracting solvents with a Van den Berg type solventless sample injector before introduction Into a DB-1 capillary separation column. A surface ionization detector (SID), which has highly selective sensitivity for Substituted amines, was employed for quantitation using bromperidol as an internal standard. Chloroform proved to be the best extracting solvent, yielding a quantitative detection limit of 5 ng/ml (S/N = 2). Comparison of the response to target compounds obtained by the SID, FTD (flame thermionic detector), and FID (flame ionization detector) showed the SID to be superior.     

Determination of ochratoxin A at parts-per-trillion levels in cereal products by immunoaffinity column cleanup and high-performance liquid chromatography/mass spectrometry

Ochratoxin A (OTA) is a toxic and potentially carcinogenic fungal toxin found in a variety of food commodities. A new sensitive method has been developed to quantify OTA in cereal products by reversed-phase liquid chromatography (LC) with mass spectrometric (MS) detection. Ochratoxin B was used as the internal standard. OTA was extracted from cereal products with acetonitrile-water, and the extract was diluted with a buffer; the diluted extract was cleaned up on an immunoaffinity column before LC/MS analysis. Two multiple-reaction monitoring transitions were used, one for quantification of OTA and one for confirmation of identity. The method was shown to be highly sensitive, with a low decision limit (CCalpha) of 0.012 microg/kg and a detection capability (CCbeta) of 0.021 microg/kg. Within-laboratory repeatability coefficient of variation values were 7.1, 3.7, and 3.1%, and the corresponding recoveries were 104, 106, and 103% for rice samples fortified with OTA at 0.05, 0.10, and 0.15 microg/kg, respectively. Method validation was performed according to the criteria of European Commission Decision 2002/657/EC. All criteria as presented in the Commission Decision were fulfilled. This method is the first fully validated method using immunoaffinity chromatography for cleanup and MS for detection in the analysis of cereals for OTA. The method was also successfully applied to cereal-derived products. The analytical results for determination of the OTA content of cereal products commercially available in Hong Kong are also reported.     

Multiresidue analysis of beta-agonists in pork by coupling polymer monolith microextraction to electrospray quadrupole time-of-flight mass spectrometry

A novel method of polymer monolith microextraction (PMME) using poly(methacrylic acid-co-ethylene glycol dimethacrylate) monolith combined with electrospray ionization quadrupole time-of-flight mass spectrometry (ESI-QTOF MS) was developed for the rapid and sensitive determination of beta-agonists in pork samples. The conditions of PMME were optimized for the improvement of extraction efficiency and reduction of the matrix interferences from pork. Under the optimal condition, the eluate solution allowed direct analysis by mass spectrometry. In the positive ion mode and in the multiple reaction monitoring (MRM) mode, the limits of detection (LODs) for beta-agonists were found to be 0.08 ng/g (clenbuterol, CLB), 0.18 ng/g (salbutamol, SBTM) and 0.26 ng/g (terbutaline, TBTL) in pork, respectively, with good inter- and intra-day precisions (2-10% for CLB, 11-23% for SBTM and 4-16% for TBTL). The proposed PMME/ESI-QTOF MS method was successfully applied to the determination of beta-agonist residues in thirteen real samples, and the positive samples were confirmed according to the identification points (IPs) system defined by Commission Decision 2002/657/EC. To investigate the matrix effect, the proposed method was compared with PMME-HPLC/ESI-QTOF MS and the slight decrease in sensitivity of PMME/ESI-QTOF MS was ascribed to the inter-analyte ion suppression.     

Chemically enhanced liquid chromatography/tandem mass spectrometry determination of glutamic acid in the diffusion medium of retinal cells

A rapid, reproducible and highly sensitive method, based on liquid chromatography mass spectrometry, was developed for the determination of the excitatory amino acid glutamic acid released in the diffusion medium of control, ischemic and mutant cells from retinas. Signal intensity of glutamic acid was enhanced by dansyl chloride derivatization giving rise to a detection limit in the order of pmol/mL. Further, in HPLC-ESI-MS detection an MS-friendly dansyl group to glutamic acid enhanced both ionization efficiency in the ESI source and collision-activated dissociation in the collision cell. The sample processing procedure included liquid-liquid extraction, derivatization with dansyl chloride and a final cation-exchange extraction to generate clean extracts for LC/MS/MS analysis. This approach has been validated as sensitive, linear (20-300 ng/mL), accurate and precise for the differential quantification of glutamic acid in the diffusion medium of retina cells. This is the first report of using chemical derivatization to enhance MS/MS detection of the glutamic acid released in the diffusion medium of wild-type and mutant retina cells, under ischemic conditions.     

Determination of a series of quinolones in pig plasma using solid-phase extraction and liquid chromatography coupled with mass spectrometric detection Application to pharmacokinetic studies

A simple and sensitive liquid chromatography (LC) method was developed for the simultaneous determination of eight quinolones in pig plasma samples. The following two methods of detection were used: ultraviolet (UV) and mass spectrometry with electrospray ionization (ESI/MS). Sample preparation consisted of solid-phase extraction (SPE) on Strata X cartridges prior to the analysis by LC/UV or LC/ESI/MS. The recovery, linearity, limit of detection (LOD) and limit of quantification (LOQ), precision and accuracy of the method were evaluated using spiked pig plasma samples. The suitability of the method for pharmacokinetic studies was evaluated by determining the concentrations of enrofloxacin (ENR) and ciprofloxacin (CIP) also in pig plasma, after administration of 200mg of enrofloxacin per kilogram of fodder during 5 consecutive days.     

High operationally stable sol-gel diglycidyloxycalix[4]arene fiber for solid-phase microextraction of propranolol in human urine

A simple, sensitive, and accurate method for the determination of propranolol in human urine has been developed based on solid-phase microextraction (SPME) followed by GC-flame ionization detection (FID). The sol-gel 5,11,17,23-tetra-tert-butyl-25,27-dihydroxy-26,28-diglycidyloxycalix[4]arene/hydroxy-terminated silicone oil (diglycidyloxy-C[4]/OH-TSO) fiber was prepared to accommodate to the harsh extraction conditions. It possesses excellent alkali-proof ability and retains its extraction characteristics intact even after treatment with highly alkaline (4 mol/L) NaOH solution. Direct chemical bonding of the coating to the fiber surface provides it with excellent solvent resistance and the introduction of calixarene enhances its thermal stability. The newly developed sol-gel calixarene coating was effectively used for the extraction of propranolol in human urine. No interference with the determination of propranolol was observed from the urine components. Standard curves were linear in the range 50-5000 microg/L for headspace-SPME (HS-SPME) and 25-25000 microg/L for direct-SPME (Dir-SPME) with correlation coefficients better than 0.9999. The detection limit was 0.275 microg/L for HS-SPME and 0.193 microg/L for Dir-SPME. The method was validated using standard addition methodology and recovery values were between 91.4 and 117% for both the sampling modes with the RSDs less than 6% at different concentration levels in the linear ranges. The results obtained by both the sampling modes were feasible, and no significant differences between them regarding accuracy, precision, and detection limits were seen.     

Development of a pressurised liquid extraction and liquid chromatography with electrospray ionization-tandem mass spectrometry method for the determination of domoic acid in shellfish

Amnesic shellfish poisoning (ASP) is a potentially lethal human toxic syndrome which is caused by domoic acid (DA) that originates in marine phytoplankton belonging to the Pseudonitzschia genus. A confirmatory and sensitive procedure has been developed and validated for the determination of DA in shellfish. The proposed method includes pressurised liquid extraction (PLE) with methanol/acetone (9:1), florisil clean-up purification inside the PLE extraction cell and detection by liquid chromatography (LC) coupled to electrospray ionization in positive mode tandem mass spectrometry (ESI-MS-MS). Comparison of ionization sources (ESI, atmospheric pressure ionization (APCI) atmospheric pressure photoionization (APPI) and combined APCI/APPI) were carried out in order to improve the analytical signal. The main parameters affecting the performance of the different ionization sources and PLE parameters were previously optimised using statistical design of experiments (DOE). Linear calibrations were obtained using mussel tissue extracts 0.05-5 microg DA/ml (R2>0.999). The limits of detection (LOD) and quantitation (LOQ) of the method were 0.2 and 0.5 microg/g respectively and recoveries ranged from 81 to 95%. This method was successfully applied to determine DA levels in 46 shellfish samples collected from Valencian (Spain) supermarkets, showing high sample throughput.     

Influence of matrix on suitability of four methods for organochlorine pesticide analysis in waters

The monitoring of organochlorine pesticides has raised a great concern in the last years due to their toxicity (some of them are carcinogenic and endocrine disruptor compounds) and persistence. European Directive 2008/105/EC establishes very restrictive levels for organochlorine pesticides in surface waters. Therefore, simple, fast, highly sensitive and low cost analytical methods are required to detect and quantify these pollutants in water. In the present work, four procedures for extraction and determination are proposed and compared for the analysis of 28 organochlorine pesticides in tap, surface and sea waters. The suitability of each method of analysis was evaluated for each kind of water. The extraction methods proposed were: two solid-phase extraction methods using C₁₈ laminar disk and Oasis HLB cartridges, a solid-phase microextraction procedure using a polydimethylsiloxane/divinylbenzene (PDMS/DVB) fibre, and a micro liquid–liquid extraction procedure using ethyl acetate as solvent. Determination of pesticides was performed by large volume on-column injector-gas chromatography-electron capture detection (LVOCI-GC-ECD), splitless-GC-ECD and GC-MS (mass spectrometry). All methods present a good sensitivity with method detection limits lower than 10?ng?L^?1, good accuracy with recoveries between 75 and 120% (with some exceptions) and good precision (relative standard deviations <15%), according to the Commission Decision 2002/657/EC criteria. The advantages and disadvantages of each method are discussed in terms of the green chemistry principles, the figures of merit and the matrix effect. This work tries to be a useful guidance for routine and control analysis laboratories.     

Determination of pesticide residues in red wines with microporous membrane liquid–liquid extraction and gas chromatography

A sample pretreatment method based on microporous membrane liquid-liquid extraction (MMLLE) was developed for the subsequent gas chromatographic determination of pesticides in wine. MMLLE provided efficient and selective extraction with enrichment factors in the range 3-13. The gas chromatographic separation was carried out using on-column injection and flame ionization detection. The method was linear, repeatable and sensitive. The limits of quantification were better than 0.006 mg/L for all the analytes except for iprodione (0.37 mg/L). The method was applied to the determination of pesticides in several red wines of different origin.     

Extraction and simultaneous determination of metoprolol and α-hydroxy metoprolol in latent fingerprints by liquid chromatography tandem mass spectrometry

Abstract

A simple extraction and sensitive simultaneous analysis method for metoprolol and its metabolite alpha-hydroxy metoprolol in latent fingerprint using liquid chromatography-mass spectrometry was developed. The extraction procedure was optimized as scrubbing using cotton swabs for 30 times followed by ultrasonic assistance in 30?°C methanol for 5?min with power of 2000 W. Drug analysis was performed using the mixture of methanol and 0.1% formic acid solution (pH 3.5, 30:70, v/v) as the mobile phase under positive electrospray ionization condition. The linear range obtained was 1.0–500.0?ng/mL for MET, and α-MET, and the limit of detection and the limit of quantification were 0.3?ng/cotton swab and 1.0?ng/cotton swab, respectively. The method showed very slight matrix effect and good recovery to the analytes in the fingermarks. Evaluation of extraction substrates and developing methods showed that the method developed is best used for impermeable substrates and that the commercially development powders have very slight influence on the qualitative detection of metoprolol in fingermarks. Finally, the drug users distinguishing test proved that the method developed for pharmaceuticals could be applied in the forensic analysis for circling out the pool of suspects in the criminal investigation based on their drug use history.     

高效液相色谱-串联质谱法测定食品中硝磺草酮

1 引 言 硝磺草酮(Mesotrione),化学名2-(4-甲磺酰基2硝基苯甲酰基)环己烷1,3二酮,又名甲基磺草酮、硝磺酮,是瑞士先正达公司发明的玉米田芽前和苗后广谱选择性除草剂,因具有低毒性、高活性、对环境友好等特点[1,2],是近几年广泛使用的除草剂.但最近研究发现,硝磺草酮具有高效的起始活性和残留活性,长期食用含有硝磺草酮残留的食物会对人畜产生致癌作用,或引起胎儿畸形[3,4].欧盟和世界贸易组织对浆果、亚麻籽、越橘、栗草料等物质中硝磺草酮的限量为0.05 mg/kg.而美国、加拿大等国对芦笋、草杆、草料等物质中硝磺草酮的限量为0.01 mg/kg[10-13].     

固相萃取-高效液相色谱-串联质谱法同时测定养殖海水中23种抗生素

建立了固相萃取-高效液相色谱-串联质谱(SPE-HPLC-MS/MS)同时检测水中6类23种抗生素的分析方法。水样用固相萃取柱富集净化,通过对比水样在不同上样pH、洗脱液用量等条件下的回收率,优化了前处理方法。采用0.1%(v/v)甲酸-1 g/L甲酸铵水溶液和甲醇-乙腈(1:1, v/v)体系作为流动相,经过梯度洗脱进行分离,在HPLC-MS/MS多反应监测模式下进行定性定量分析。结果显示,23种抗生素的方法检出限(MDL)范围为0.1~2.9 ng/L,加标回收率为47.3%~132.6%。采用该方法对东营海水养殖区5个养殖池水样进行了检测,除青霉素类之外的各类抗生素均有检出,其中磺胺增效剂甲氧苄氨嘧啶的检出率达100%,氯霉素类抗生素氟甲砜霉素检出的最高质量浓度达到261.0 ng/L。结果表明,所建立的方法高效、灵敏、可靠,可用于海水中多种抗生素的分析。     

液相色谱-串联质谱法测定小型家用电器塑料部件中全氟辛酸

建立了高效液相色谱-串联质谱(HPLC-MS/MS)结合加速溶剂萃取测定小型家用电器塑料部件中全氟辛酸(PFOA)的分析方法。样品冷冻粉碎后采用甲醇作溶剂进行快速溶剂萃取,萃取液经C18固相萃取柱富集净化后,以C18柱为分离柱,以甲醇-水为流动相,梯度洗脱,电喷雾负离子模式下多反应监测(MRM))模式检测。PFOA在0.5～100.0μg/L范围内线性关系良好(r2=0.998 9),回收率为93%～107%,相对标准偏差为2.9%～7.6%,检出限(S/N=3)为0.5μg/kg。该方法操作简便、灵敏度高,适用于小型家用电器塑料部件中全氟辛酸残留分析。     

Solid-phase microextraction of N-nitrosamines

A method was developed for the extraction of seven N-nitrosamine compounds from water by solid-phase microextraction (SPME). The method developed requires a total analysis time of only 1.25 h for both extraction and detection (versus 3-20 h for other isolation techniques). Three gas chromatography (GC) detection systems were tested with the SPME method, nitrogen chemiluminesence detection (NCD), nitrogen-phosphorus detection (NPD) and chemical ionization mass spectrometry (CI-MS), with method detection limits (MDLs) found in the ng/L range. This method was used to analyze wastewater samples and showed excellent selectivity of extraction. The detection limits of this method for N-nitrosodimethylamine (NDMA) range from 30 to 890 ng/L as a function of detector type. The excellent selectivity of SPME in addition to the fast analysis time would make this method ideal for general surveys, wastewater analysis and laboratory studies (e.g. degradation kinetics or formation potential).     

Sensitive analysis of anti-HIV drugs, efavirenz, lopinavir and ritonavir, in human hair by liquid chromatography coupled with tandem mass spectrometry

A highly sensitive and selective method using liquid chromatography coupled with tandem mass spectrometry (LC/MS/MS) was developed and validated for the measurement of three antiretroviral agents, efavirenz, lopinavir and ritonavir, in human hair. Hair samples from adherent HIV-infected patients on antiretroviral therapies were cut into about 1 mm length segments and drugs were extracted by first shaking the samples with methanol in a 37 degrees C water bath overnight (>14 h), followed by methyl tert-butyl ether/ethyl acetate (1:1) extraction under weak alkaline conditions. The extracted lopinavir and ritonavir were separated by reversed-phase chromatography and detected by tandem mass spectrometry in electrospray positive ionization mode with multiple reaction monitoring (MRM), while efavirenz was monitored in negative ionization MRM mode. This method was validated from 0.01 to 4.0 ng/mg hair for ritonavir and 0.05-20 ng/mg hair for lopinavir and efavirenz by using 2 mg of a human hair sample. The interday and intraday assay precision (coefficients of variation, CV) for spiked quality control (QC) samples at low, medium and high concentrations were within 15% and accuracy ranged from 89% to 110%. Assay reproducibility was also demonstrated by analysis of incurred hair QC samples (CV <14%). No significant matrix ionization suppression was observed. This developed method allowed for the monitoring of these target medications in the hair samples of HIV-infected women on antiretroviral therapy in an observational study using small amounts of hair.     

Analysis of polar pesticides in water and wine samples by automated in-tube solid-phase microextraction coupled with high-performance liquid chromatography-mass spectrometry

A simple and sensitive method for the determination of polar pesticides in water and wine samples was developed by coupling automated in-tube solid-phase microextraction (SPME) to high-performance liquid chromatography-electrospray ionization mass spectrometry (HPLC-ESI-MS). To achieve optimum performance, the conditions for both the in-tube SPME and the ESI-MS detection were investigated. In-tube SPME conditions were optimized by selecting the appropriate extraction parameters, especially the stationary phases used for SPME. For the compounds studied, a custom-made polypyrrole (PPY)-coated capillary showed superior extraction efficiency as compared to several commercial capillaries tested, and therefore, it was selected for in-tube SPME. The influence of the ethanol content on the performance of in-tube SPME was also investigated. It was found that the amount of pesticides extracted decreased with the increase of ethanol content in the solutions. The ESI-MS detection conditions were optimized as follows: nebulizer gas, N2 (30 p.s.i.; 1 p.s.i.=6894.76 Pa); drying gas, N2 (10 l/min, 350 degrees C); capillary voltage, 4500 V; ionization mode, positive; mass scan range, 50-350 amu; fragmentor voltage, variable depending on the ions selected. Due to the high extraction efficiency of the PPY coating and the high sensitive mass detection, the detection limits (S/N = 3) of this method for the compounds studied are in the range of 0.01 to 1.2 ng/ml, which are more than one order of magnitude lower than those of the previous in-tube SPME-HPLC-UV method. A linear relationship was obtained for each analyte in the concentration range of 0.5 to 200 ng/ml with MS detection. This method was applied to the analysis of phenylurea and carbamate pesticides in spiked water and wine samples.