液相色谱-电喷雾串联质谱法同时检测鸡组织中5种抗病毒类药物的残留量

建立了鸡组织中抗病毒类药物多残留检测的液相色谱-电喷雾串联质谱法(LC-ESI-MS/MS)。采用三氯乙酸-乙腈溶液提取鸡组织中的金刚烷胺、金刚乙胺、美金刚、咪喹莫特和吗啉胍,离心过滤后经强阳离子交换柱(SCX)净化,色谱柱Xamide(100 mm×2.1 mm, 5 μm)分离,多反应监测(MRM)正离子扫描方式进行质谱检测。结果表明,鸡组织与鸡肝中5种药物的检出限为0.06～0.30 μg/kg,定量限为0.2～1.0 μg/kg。当5种药物的添加水平为0.2～10.0 μg/kg时,在鸡肉中的平均回收率为72.3%～94.2%,相对标准偏差(RSD)(n=6)为3.5%～11.3%;在鸡肝中的平均回收率为70.8%～92.7%, RSD(n=6)为5.3%～12.6%。该方法选择性好,抗干扰能力强,可作为鸡肉和鸡肝中抗病毒药物残留检测的确证方法。     

A rapid determination of propiverine and its N-oxide metabolite in human plasma by high performance liquid chromatography-electrospray ionization tandem mass spectrometry

A simple, fast and sensitive high-performance liquid chromatography (HPLC)-electrospray ionization (ESI) tandem mass spectrometric method (LC-MS/MS) has been developed for determination of propiverine and propiverine N-oxide metabolite in human plasma using oxybutynin as internal standard. Instead of extracting propiverine from plasma using organic solvents, which should be separated from the aqueous phase and evaporated before injecting the sample into the chromatograph, plasma sample containing propiverine and N-oxide was directly injected after precipitating proteins with acetonitrile. Numerous compounds in the plasma did not interfere with the highly specific multiple reaction monitoring in tandem mass spectrometric detection following C₈ reversed-phase chromatographic separation under conditions that eluted propiverine, N-oxide and oxybutynin within 2 min (0.1% formic acid in water/acetonitrile, 25:75, v/v). The LC-MS/MS method and an alternative LC-MS method, using methyl-t-butyl ether extraction and selected ion monitoring, were validated over 1-250 ng ml⁻¹ of propiverine and 2 to 500 ng ml⁻¹ of N-oxide, and successfully applied in a pharmacokinetic study. The lower limit of quantitation was 1 ng ml⁻¹ for propiverine and 2 ng ml⁻¹ for N-oxide in both methods.     

超高效液相色谱-电喷雾串联四极杆质谱法同时测定配合饲料中29种兽药

建立了同时测定配合饲料中喹诺酮类、磺胺类、大环内酯类和硝基呋喃类共计29种兽药的超高效液相色谱-电喷雾串联四极杆质谱(UPLC-ESI-MS/MS)检测方法。饲料样品用甲醇-乙腈(1:1, v/v)混合溶液提取,提取液经Oasis HLB固相萃取柱净化,采用UPLC-ESI-MS/MS检测。以甲醇和含0.1%(v/v)甲酸的水溶液作为流动相,进行梯度洗脱,用C18色谱柱分离,正离子模式扫描,多反应监测模式检测。29种兽药在0.01～5.0 mg/L范围内线性关系良好,相关系数(r)均大于0.99;在复合预混饲料、全价配合饲料、浓缩饲料中4个添加水平下29种兽药的平均回收率在61.2%～94.3%范围内,相对标准偏差(RSD)为2.2%～15.0%;方法的检出限(以信噪比大于10计)为0.01 mg/kg或0.05 mg/kg。该方法简便、快速、准确,重现性好,灵敏度高,适用于配合饲料中多种兽药的同时检测。     

高效液相色谱-电感耦合等离子体质谱测定血浆中铂类抗癌药物

建立了高效液相色谱-电感耦合等离子体质谱快速测定血浆中顺铂、卡铂、奥沙利铂的方法。实验表明顺铂和奥沙利铂在纯水和血浆中不稳定,奥沙利铂在0.9%的NaCl中不稳定,因此采集的样品需尽快分析。提出了可通过测定顺铂、奥沙利铂色谱保留时间和卡铂的标准曲线来间接测定血浆中不稳定的顺铂和奥沙利铂含量的简化方法。方法检出限以铂计为0.04 ng/mL,顺铂、卡铂、奥沙利铂线性回归曲线的回归系数r均大于0.9995。方法的加标回收率在84%～102%之间,相对标准偏差在0.9%～6.4%之间。     

超高效液相色谱-电喷雾串联质谱法测定生活饮用水中49种药物及5种代谢物

建立了超高效液相色谱-电喷雾串联质谱(UPLC-ESI MS/MS)分析生活饮用水中54种药物的方法。采用HLB固相萃取柱对水样中的目标化合物进行富集净化,以5 mL甲醇洗脱;洗脱液用氮气吹至近干,用0.4 mL 0.1%甲酸水溶液定容,上机分析;ACQUITY UPLC^TMBEH C₁₈柱用作色谱分离,以0.1%甲酸水溶液-甲醇为流动相进行梯度洗脱;多反应监测(MRM)模式进行检测;目标药物使用基质外标法定量。54种药物在自备井水、市政末梢水和地表水中的加标回收率分别为58.7%~104.4%、53.1%~109.5%和50.7%~118.8%,相对标准偏差(n=6)分别为0.3%~12.8%、1.0%~15.5%和0.4%~19.3%;方法定量限为0.002~5.000 ng/L。将建立的方法应用于北京部分自备井水、市政末梢水和地表水样品的分析,结果在自备井水样中检出26种药物。     

液相色谱-电喷雾离子阱串联质谱同时分析烟草中的20种游离氨基酸

建立了液相色谱-电喷雾离子阱串联质谱(LC-ESI-IT-MS/MS)同时分析烟草中20种游离氨基酸的方法。烟草样品经萃取后过滤直接进样,无需进行衍生和固相萃取等其他前处理步骤。液相色谱采用HyPURITY C18反相色谱柱(200 mm×2.1 mm, 5 μm),采用1%(体积分数,下同)乙腈水溶液(含0.1%九氟戊酸)和90%乙腈水溶液(含0.1%九氟戊酸)为流动相进行梯度洗脱。结果表明,20种氨基酸的检出限(LOD)为0.01～0.05 μmol/L (S/N=3),线性相关系数均大于0.9977,峰面积测定的相对标准偏差(RSD)为0.78%～4.93%。该方法分析效率、灵敏度和选择性高,已成功应用于多种烟草样品中氨基酸的分析测定。     

Quantification of zolpidem in human plasma by liquid chromatography-electrospray ionization tandem mass spectrometry

A simple and robust method for quantification of zolpidem in human plasma has been established using liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI MS/MS). Es-citalopram was used as an internal standard. Zolpidem and internal standard in plasma sample were extracted using solid-phase extraction cartridges (Oasis HLB, 1 cm3/30 mg). The samples were injected into a C8 reversed-phase column and the mobile phase used was acetonitrile-ammonium acetate (pH 4.6; 10 mm) (80:20, v/v) at a flow rate of 0.7 mL/min. Using MS/MS in the selected reaction-monitoring (SRM) mode, zolpidem and Es-citalopram were detected without any interference from human plasma matrix. Zolpidem produced a protonated precursor ion ([M+H]+) at m/z 308.1 and a corresponding product ion at m/z 235.1. The internal standard produced a protonated precursor ion ([M+H]+) at m/z 325.1 and a corresponding product ion at m/z 262.1. Detection of zolpidem in human plasma by the LC-ESI MS/MS method was accurate and precise with a quantification limit of 2.5 ng/mL. The proposed method was validated in the linear range 2.5-300 ng/mL. Reproducibility, recovery and stability of the method were evaluated. The method has been successfully applied to bioequivalence studies of zolpidem.     

Analysis of primary aromatic amines in the mainstream waterpipe smoke using liquid chromatography-electrospray ionization tandem mass spectrometry

In recent years waterpipe smoking has spread worldwide and emerged as global health issue. Yet only little is known on the composition of waterpipe smoke. Here, we present a study on the identification and quantification of primary aromatic amines (PAAs) in this complex environmental matrix. Smoking of the waterpipe was conducted with a smoking machine and particulate matter was collected on glass fiber pads. We developed a fast, simple and specific liquid chromatography-electrospray ionization tandem mass spectrometry (LC-MS/MS) approach to simultaneously detect 31 different PAAs in this matrix. The detection limits comprised a range of 0.45-4.50 ng per smoking session, represented by 2-aminobiphenyl and 3,4,5-trichloroaniline, respectively. Intra- and inter-day precision were determined and proved excellent. We detected 31.3 ± 2.2 ng aniline and 28.0 ± 1.6 ng 4,4'-oxydianiline in the smoke of one waterpipe session. The water in the bowl exerted a small but considerable filter effect on PAAs. The method worked-out showed excellent sensitivity and specificity and is thus highly suited for the determination of PAAs in mainstream waterpipe smoke.     

Multi-residue confirmatory method for the determination of twelve coccidiostats in chicken liver using liquid chromatography tandem mass spectrometry

The confirmatory LC-MS/MS method for the determination of residues for twelve coccidiostats including ionophore antibiotics (lasalocid, maduramycin, monensin, narasin, salinomycin, semduramycin) and chemical coccidiostats (clazuril, decoquinate, diclazuril, halofuginone, nicarbazin and robenidine) in poultry liver has been developed. The sample preparation was based on extraction with acetonitrile, defatting with Alumina columns and clean-up on Oasis HLB spe. The separation of analytes was performed on PhenylHexyl column with a gradient of acetonitrile, methanol and the ammonium formate pH 4.0. For all analytes, at least 2 diagnostic fragmentation ions were monitored. The validation, performed according to the CD 2002/657/EC, proved the suitability of the method for the confirmatory analysis of the coccidiostats. For lasalocid, however, low reproducibility was observed and the proper quantification could not be performed with this method.     

同位素稀释液相色谱-串联质谱法准确测定猪肉中的氯霉素

建立了同位素稀释质谱测定猪肉中氯霉素残留的方法,对影响溶剂提取、萃取净化、液相色谱-质谱检测方法准确度的因素进行了量化与优化。猪肉样品经液氮冷冻粉碎,加入氯霉素-D5同位素内标和0.1 mol/L醋酸钠缓冲溶液(pH 5),用乙酸乙酯提取4次,正己烷脱脂,HLB固相萃取小柱净化后采用负离子多反应监测模式下的液相色谱-质谱技术检测。方法的线性范围为0.01～0.5 ng/g,相关系数r2大于0.999;检出限和定量限分别为0.004 ng/g和0.02 ng/g;在0.02 ng/g和1.0 ng/g加标水平的回收率(n=3)分别为95.2%～109.1%和99.7%～102.5%;日内和日间相对标准偏差均小于2%。研究了氯霉素及氯霉素-D5的基体效应,测得不同流动相和稀释溶剂组成时基体效应因子k的范围在0.950～1.015之间。研究结果对同位素稀释质谱痕量残留检测结果准确度的控制、方法的建立与验证具有参考意义。     

Determination of drugs of abuse in bovine dentin using liquid chromatography–electrospray ionization tandem mass spectrometry

Drugs deposited in human teeth are well preserved; the spectrum of toxicological investigations may therefore be supplemented by an analysis method for drugs in teeth. A liquid chromatography–electrospray ionization tandem mass spectrometry assay for the detection and quantification of basic drugs of abuse in bovine dentin samples was developed and validated. The drugs and metabolites amphetamine, methamphetamine, methylenedioxymethylamphetamine, methylenedioxyethylamphetamine, codeine, morphine, cocaine and benzoylecgonine were extracted from 50 mg ground dentin powder by ultrasonication for 60 min in methanol 3 times. The extracts were analyzed on a triple‐quadrupole mass‐spectrometer in multiple reaction monitoring mode. The method was validated and proved to be accurate, precise, selective, specific and stable with good linearity within the calibration range and a lower limit of quantification of 10 to 20 pg/mg. To artificially load bovine dentin samples with drugs, the natural process of de‐ and remineralization in the oral cavity was mimicked by a pH‐cycling experiment. The artificially drug‐loaded dentin samples showed drug concentrations of 20 to 80 pg/mg. The method can be applied in further in vitro experiments as well as in post‐mortem cases, especially where limited sample tissue is available. Copyright © 2014 John Wiley & Sons, Ltd.     

超高效液相色谱-串联质谱法同时测定调味料中的17种防腐剂和抗氧化剂

建立了超高效液相色谱-串联质谱(UPLC-MS/MS)同时测定调味料中11种防腐剂和6种抗氧化剂的定性、定量分析方法。低脂肪和中等脂肪含量的调味料样品用饱和NaCl水溶液(用磷酸调pH为2～3)稀释混合均匀,然后用乙腈提取,正己烷液-液萃取净化(中等脂肪含量的样品提取液还需经C8固相萃取柱进一步的净化处理);脂肪含量高的样品先用正己烷稀释,再用饱和NaCl水溶液(用磷酸调pH为2～3)溶解样品,然后用乙腈提取,提取液进一步经C8固相萃取柱净化处理。提取液经C18反相色谱柱(150 mm×2.1 mm, 1.7μm)分离,流动相为20 mmol/L乙酸铵水溶液和乙腈,梯度洗脱,以电喷雾离子源负离子多反应监测(MRM)模式进行MS/MS检测。17种分析物在线性范围内具有较好的线性关系,相关系数r≥0.9955,其定量限(LOQ,信噪比为10)为0.05～5 mg/kg;空白样品中的添加回收率为79.7%～118%,精密度(以相对标准偏差计)为0.57%～13.1%。该方法适用于调味品中防腐剂和抗氧化剂的检测。     

Highly sensitive method for the determination of omeprazole in human plasma by liquid chromatography-electrospray ionization tandem mass spectrometry: application to a clinical pharmacokinetic study

A highly sensitive, rapid assay method has been developed and validated for the estimation of omeprazole (OPZ) in human plasma with liquid chromatography coupled to tandem mass spectrometry with electrospray ionization in the positive-ion mode. The assay procedure involves alkalinization of plasma followed by simple liquid-liquid extraction of OPZ and lansoprazole (internal standard, IS) from human plasma with acetonitrile. Chromatographic separation was achieved with 0.01 M ammonium acetate:acetonitrile (40:60, v/v) at a flow rate of 0.25 mL/min on an Inertsil ODS 3 column with a total run time 2.5 min. The MS/MS ion transitions monitored were 346.1 --> 198.1 for OPZ and 370.1 --> 252.1 for IS. Method validation and clinical sample analysis were performed as per FDA guidelines and the results met the acceptance criteria. The lower limit of quantitation achieved was 0.05 ng/mL and the linearity was observed from 0.05 to 10.0 ng/mL. The intra-day and inter-day precisions were in the ranges 2.09-8.56 and 5.29-8.19%, respectively. This novel method has been applied to a pharmacokinetic study of OPZ in humans.     

Bacterial response to eukaryotic cells. Analysis of differentially expressed proteins using nano liquid chromatography-electrospray ionization tandem mass spectrometry

Host-bacteria interactions have mostly been investigated with regard to the host response or to activities of pathogenic bacteria. In contrast, we aim to identify reactions of non-pathogenic bacteria that result from their contact with host cells of the gastrointestinal tract. In a proteomic approach, the response of non-pathogenic human Escherichia coli bacteria on gut epithelial cells (rat IEC-6) was investigated in an in vitro co-culture model. For this purpose, a sensitive analytical procedure was developed based on the identification of two-dimensional polyacrylamide gel electrophoresis separated proteins by online nanoLC-electrospray ionization MS/MS using a quadrupole time-of-flight tandem mass spectrometer for accurate mass determination. We demonstrate here the efficiency of this technique by the identification of a total of 43 differentially expressed proteins, out of which 25 were up-regulated and 18 were down-regulated. They represent a wide range of molecular weight and different metabolic and physiological functions.     

超高效液相色谱-串联质谱法快速测定镇咳祛痰药中阿片类物质的含量

建立了镇咳祛痰药中吗啡、可待因、海洛因、蒂巴因、罂粟碱、那可汀6种阿片类物质含量的LC-MS/MS快速测定方法.药品经超声浸取,甲醇稀释过滤后经Waters C18柱分离,以乙腈和10 mmol/L乙酸铵(含0.1％甲酸)溶液进行梯度洗脱,采用电喷雾正离子化(ESI+)、多反应监测模式进行测定.6种阿片类物质在相应的线...     

超高效液相色谱-电喷雾串联质谱法同时测定地表水中28种皮质激素

建立了同时测定地表水中28种皮质激素的超高效液相色谱-电喷雾串联质谱（UHPLC-ESI-MS/MS）的分析方法。水样经HLB柱固相萃取、C₈反相色谱柱分离,在动态多反应监测（DMRM）模式下采用电喷雾离子化正、负离子模式（ESI±）进行信号采集与测定,内标法定量。结果发现,28种皮质激素在1.0~100 μg/L范围内具有良好的线性关系（R²>0.99）,方法检出限为0.21~0.48 ng/L,方法定量限为0.32~0.72 ng/L。在5.0、10、50 ng/L的基质加标水平下,28种目标物的平均回收率为68.8%~108.7%,相对标准偏差为0.1%~8.1%。该方法灵敏、准确、可靠,将广泛应用于环境中糖/盐皮质激素的痕量监测及其行为、风险研究。     

Development and validation of a liquid chromatography–tandem mass spectrometry analytical method for the therapeutic drug monitoring of eight novel anticancer drugs

To support therapeutic drug monitoring of patients with cancer, a fast and accurate method for simultaneous quantification of the registered anticancer drugs afatinib, axitinib, ceritinib, crizotinib, dabrafenib, enzalutamide, regorafenib and trametinib in human plasma using liquid chromatography tandem mass spectrometry was developed and validated. Human plasma samples were collected from treated patients and stored at −20°C. Analytes and internal standards (stable isotopically labeled analytes) were extracted with acetonitrile. An equal amount of 10 mm NH₄CO₃ was added to the supernatant to yield the final extract. A 2 μL aliquot of this extract was injected onto a C₁₈‐column, gradient elution was applied and triple‐quadrupole mass spectrometry in positive‐ion mode was used for detection. All results were within the acceptance criteria of the latest US Food and Drug Administration guidance and European Medicines Agency guidelines on method validation, except for the carry‐over of ceritinib and crizotinib. These were corrected for by the injection order of samples. Additional stability tests were carried out for axitinib and dabrafenib in relation to their reported photostability. In conclusion, the described method to simultaneously quantify the eight selected anticancer drugs in human plasma was successfully validated and applied for therapeutic drug monitoring in cancer patients treated with these drugs.     

高效液相色谱-电喷雾串联质谱法检测环境水样中22种抗生素类药物

建立了高效液相色谱-电喷雾串联质谱(HPLC-ESI MS/MS)分析环境水样中22种抗生素类药物的方法。采用HLB固相萃取柱对环境水样中的目标化合物进行富集、净化,然后以6 mL氨水-甲醇(5:95, v/v)溶液洗脱。收集的洗脱液经氮气吹干至1 mL,然后进行HPLC-ESI MS/MS分离分析。色谱流动相A相为甲醇-乙腈(1:1, v/v), B相为0.3%(体积分数)甲酸水溶液(含0.1%(体积分数)甲酸铵,pH 2.9);色谱柱为XTerra MS C18柱。质谱检测采用正离子扫描,多反应监测模式。分别以自来水和污水作为基质,22种抗生素类药物的加标平均回收率分别为54.9%～130%和57.4%～138%,相对标准偏差(n=3)分别为2.85%～28.6%和2.02%～23.2%;方法的检出限为0.05～0.5 ng/L。将建立的方法应用于北京市高碑店湖和小清河水样的分析,结果表明在两个水样中均有部分抗生素类药物检出。     

Simultaneous analysis of oral anticancer drugs for renal cell carcinoma in human plasma using liquid chromatography/electrospray ionization tandem mass spectrometry

An analytical method using high‐performance liquid chromatography/electrospray ionization tandem mass spectrometry has been developed and validated for simultaneous measurement of four tyrosine kinase inhibitors used for renal cell carcinoma and their metabolites in human plasma. Despite their similar structures, it is difficult to measure plasma levels of these compounds simultaneously using optimal MS parameters for each compound because a quantitative range exceeding 50,000‐fold is required. To overcome this problem, we used a linear range shift technique using in‐source collision‐induced dissociation. Linearity ranges of sorafenib, sorafenib N‐oxide, sunitinib, N‐desethyl sunitinib, axitinib and pazopanib were 100–10,000, 10–1,000, 1–100, 1–100, 1–100 and 500–50,000 ng/mL, respectively. The intra‐ and inter‐day precision and accuracy were high, and coefficients of variation and relative error were <10.3% and within ±11.8%, respectively. The matrix effects of all analytes ranged from 87.7 to 114.8%. Extraction recoveries and overall recoveries showed small extraction loss (<15.0%) for all analytes. Moreover, all cancer patient samples used in this study were successfully quantified and fell within the linear range of measurement. Therefore, this novel analytical system using in‐source collision‐induced dissociation has sufficient performance to measure plasma concentrations of these four tyrosine kinase inhibitors and their metabolites for therapeutic drug monitoring.     

液相色谱-电喷雾串联质谱法检测鸡组织中5种聚醚类药物残留

建立了鸡组织中聚醚类药物多残留检测的高效液相色谱-电喷雾串联质谱方法。采用甲醇提取鸡组织中的拉沙洛菌素、盐霉素、莫能菌素、甲基盐霉素和马杜霉素,经硅胶柱净化,以乙腈(含0.1%甲酸)-0.1%甲酸水溶液(体积比为97:3)为流动相,Symmetry Shield RP18作为色谱分析柱,多反应监测(MRM)正离子扫描方式进行质谱检测。当5种聚醚类药物的添加水平为鸡肉0.1～1500 μg/kg、鸡肝0.2～4500 μg/kg时,平均回收率为71.6%～99.1%,日内测定的相对标准偏差(RSD)(n=5)为3.2%～10.7%,日间RSD(n=3)为4.6%～14.7%。2种鸡组织中5种聚醚类药物的定量限为0.1～1.0 μg/kg。该方法的灵敏度、准确度和精密度均符合兽药残留分析技术的要求,适用于鸡肉和鸡肝中5种聚醚类药物的多残留检测。