Determination of fentanyl, metabolite and analogs in urine by GC/MS

A rapid and sensitive method for the simultaneous determination of alfentanyl, sufentanyl and fentanyl (and its major metabolite norfentanyl) in urine was developed and validated. The method involved a liquid–liquid extraction in alkaline conditions, derivatization with pentafluoropropionic anhydride to improve the sensitivity for norfentanyl and subsequent analysis in GC/MS. The LODs are 0.08 ng ml^?1 for all substances (0.04 ng ml^?1 for alfentanyl). Intra‐ and inter‐day precision coefficient of variation was always below 15%; mean relative error (accuracy) was always below 15%. The method was linear for all analytes, with quadratic regression of calibration curves always higher than 0.99. The method was applied to real samples of subjects who had received therapeutic doses of fentanyl, showing its suitability for the determination of low levels of these substances. The method was also applied to a subject whose death was attributed to fentanyl overdose. Copyright © 2010 John Wiley & Sons, Ltd.     

GC/MS分析牛尿中瘦肉精残留

目的:建立牛尿中"瘦肉精"的GC/MS分析方法,为保障食品安全提供有效的分析手段。方法:调节样品的pH为5.2,用乙酸乙酯-异丙醇提取,蒸干后用乙酸铵溶解,经固相萃取小柱净化,氮气吹干后经双三甲基硅基三氟乙酰胺(BSTFA)衍生,GC/MS分析。结果:以牛尿为基质,"瘦肉精"在20.0～160.0μg·L-1添加浓度范围内线性关系良好,r=0.9980,其回收率在65.6%～99.0%之间,检出限为0.3μg·L-1。结论:本方法专属性好,准确,杂质少,适用于牛尿中"瘦肉精"的检测。     

家兔血清、尿中甲基苯丙胺及其代谢产物苯丙胺的气相色谱/质谱分析

目的:建立血清、尿中甲基苯丙胺(MA)及其主要活性代谢产物苯丙胺(AP)的气相色谱/质谱(GC/MS)定性定量分析方法,探讨样品稳定性和家兔血、尿中AP和MA的比值关系。方法:样品中加入内标物丙基解痉素(SKF525A)后碱化,乙醚萃取,三氟醋酸酐衍生化,GC/MS法分析甲基苯丙胺和苯丙胺。结果:血清和尿中甲基苯丙胺与苯丙胺的线性范围分别为0.01~5.0mg.L-1和0.1~50.0mg.L-1;最低检出限为0.005mg.L-1(S/N≥3);提取回收率均大于74.3%;方法回收率为95.50%~103.16%;日内及日间RSD均小于10%。18h内苯丙胺与甲基苯丙胺在血、尿中比值分别为0.16~1.5和0.021~0.079。血清样品在室温和冷冻条件下存放24h甲基苯丙胺、苯丙胺的相对误差为5.3%~6.1%。结论:甲基苯丙胺中毒血清样品在室温和冷冻条件下稳定,建立的GC/MS分析方法同时测定血清和尿液中甲基苯丙胺及其主要代谢产物苯丙胺,方法简便、灵敏、重复性好,适用于甲基苯丙胺中毒与滥用案例的快速鉴定。     

LC/MS/MS与GC/MS法分析鉴定小鼠尿中沙美特罗的代谢产物

目的:鉴定沙美特罗在小鼠尿中的主要代谢产物.方法:ig给药后,收集小鼠尿液,经固相提取,葡萄糖醛酸酶水解,进行LC/MS/MS分析和硅烷化后进行GC/MS分析同时分离鉴定沙美特罗代谢产物.结果和结论:在给药后尿样中发现沙美特罗原型和4种代谢产物M1～M4,其结构推测为19-羟基沙美特罗(M1)、2-羰基沙美特罗(M2)、19-羰基沙美特罗(M3)和19-羟基-8-甲氧基沙美特罗(M4).     

Solid-phase extraction and GC/MS confirmation of barbiturates from human urine.

A highly selective and sensitive procedure has been developed for isolating and identifying barbiturates in human urine. With a new disposable bonded silica gel solid-phase extraction (SPE) column and hexobarbital as an internal standard (IS), amobarbital, butabarbital, pentobarbital, phenobarbital, secobarbital, and methaqualone were selectively isolated from endogenous urine components. Capillary gas chromatography/ion trap mass spectrometry (GC/MS) analysis of the extracts generated a full mass spectrum for the detection, identification, and quantitation of barbiturates. Linear quantitative response curves for the drugs have been generated over a concentration range of 20-500 ng/mL. Overall extraction efficiencies for drugs averaged greater than 90%, and the quantitative response curves exhibited correlation coefficients of 0.996 to 0.999.     

Evaluation of full-scanning GC/ion trap MS analysis of NIDA drugs-of-abuse urine testing in urine.

Analytical methods developed for the Finnigan MAT ITS40 gas chromatograph-ion trap mass spectrometer (GC/MS) were evaluated for the confirmation of drugs-of-abuse in urine. The specific drugs evaluated are those listed by the National Institute on Drug Abuse (NIDA): 11-nor-9-carboxy-delta 9-tetrahydrocannabinol (9-carboxy-THC), benzoylecgonine (BE), codeine and morphine, phencyclidine (PCP), amphetamine, and methamphetamine. Drugs were extracted from urine using solid-phase columns, separated by capillary gas chromatography, and analyzed by ion trap mass spectrometry following electron impact ionization. All drugs except PCP were derivatized prior to analysis. The full scan limits of detection (LOD), quantitation (LOQ), and linearity were 2.5, 5.0, and 1000 ng/mL, respectively, for 9-carboxy-THC; 37, 75, and 5000 ng/mL for BE; 50, 100, and 2500 ng/mL for the opiates; 0.25, 0.50, and 500 ng/mL for PCP; and 50, 100, and 5000 ng/mL for the amphetamines. The limits of detection (LOD) and limits of quantitation (LOQ) met the minimum criteria for the signal-to-noise (S/N) ratio and spectral match criteria for drug identification. Absolute LODs and LOQs (in ng/mL) for the ITS40 based on single ion monitoring of blank urines were: 0.8 and 2.0 for 9-carboxy-THC; 8.9 and 25 for BE; 3.3 and 9.6 for codeine; 6.2 and 16.7 for morphine; 0.25 and 0.32 for PCP; 0.7 and 2.0 for amphetamine; and 2.4 and 5.7, for methamphetamine, respectively. The coefficient of variation ranged from 5 to 10%, and analytical recoveries were in the range of 90-114%. The ion trap mass spectrometer permits full scan identification of drugs while maintaining analytical LOQ that are below NIDA guidelines, and has equivalent or better detection limits to quadrupole analyzers for high sensitivity applications.     

GC/MS confirmatory method for etorphine in horse urine

A highly sensitive procedure for GC/MS determination of etorphine in horse urine is described. This assay provides both specificity and reliability and is particularly well suited for the confirmation of radioimmunoassay screening procedures usually used for etorphine. After solvent extraction and purifications, the etorphine is characterized as a pentafluoroacetic derivative (PFAA) by using mass fragmentography. The detection limit is 0.1 ng/mL in urine; the coefficient of variation of the estimations is 10.9%. The procedure has been validated after on-field administration of 5 to 90 micrograms of etorphine to five thoroughbred horses (10 to 180 ng/kg).     

Detection of diuretics in horse urine by GC/MS.

The use of diuretics in horses subject to doping control is prohibited. Thus, a sensitive screening procedure is required to identify the chemically different diuretics. We communicate here a method to detect three commonly employed acidic diuretics: bumetanide, ethacrynic acid, and furosemide. A liquid-liquid extraction on Extrelut 3 was performed at weak acidic and basic conditions using ethyl acetate as organic solvent. For analysis by GC, the diuretics were methylated on-column in the presence of MSTFA/TMAH, avoiding the commonly employed highly toxic derivatizing agent methyl iodide. For identification of diuretics, we used a mass selective detector operating in the SIM (selected ion monitoring) mode. Confirmation analysis may be obtained with a full scan run. Recoveries for the individual drugs ranged from 31 to 48% at the 100-ng/mL level for 3 mL urine, using calibration curves of drug standards with linearity from 2.5 to 20 ng injected. The limit of detection amounts to 40 ng/mL for the three diuretics. The method permits rapid and sensitive detection of diuretics in horse urine and is recommended for doping control.     

A validated SPE-LC-MS/MS assay for Eplerenone and its hydrolyzed metabolite in human urine

An automated LC-MS/MS assay was validated to quantitate the first selective aldosterone blocker Eplerenone (I) and its hydrolyzed metabolite (II) in human urine. After the addition of the stable isotope labeled internal standards, human urine samples were extracted on a C(18) solid phase extraction (SPE) cartridge using a Zymark RapidTrace automation system. The extraction eluates were diluted with 20 mM ammonium acetate aqueous solution and directly injected onto the LC-MS/MS system. The chromatographic separation was performed on a reverse phase Zorbax XDB-C(8) HPLC column (2.1 x 50 mm, 5 microm) with a mobile phase of acetonitrile:water (40:60, v/v) containing 10 mM ammonium acetate (pH 7.4). I and II were ionized using positive and negative ionization mass spectrometry, respectively, to achieve the best sensitivity. The ionization polarity was switched during the run at approximately 2.5 min after the injection. Multiple reaction monitoring (MRM) with a tandem mass spectrometer was used to detect the analytes. The precursor to product ion transitions of m/z 415-->163 and m/z 431-->337 were used to measure I and II, respectively. The assay exhibited a linear dynamic range of 50-10000 ng/ml of urine for both of I and II. The lower limit of quantitation (LLOQ) was 50 ng/ml for I and II. Acceptable precision and accuracy were obtained for concentrations over the standard curve range. Sample analysis time for each injection was 5 min; a throughput of 100 human urine standards and samples per run was achieved.     

GC/MS analysis of propylated barbiturates

A simple and reproducible method for the analysis of barbiturates by GC/MS after derivatization with dimethylformamide dipropyl acetal is reported. The method is readily adapted to screening, confirmation, and quantitation.     

Confirmation of marijuana, cocaine, morphine, codeine, amphetamine, methamphetamine, phencyclidine by GC/MS in urine following immunoassay screening

Rapid, reliable, sensitive, qualitative, and quantitative methods using small urine volumes (0.2-0.5 mL) were developed primarily for confirmation of marijuana, cocaine, benzoylecgonine, ecgonine methyl ester, morphine, codeine, amphetamine, methamphetamine, and phencyclidine. Using capillary gas chromatography/mass spectrometry (GC/MS) and selected ion monitoring (SIM), mass spectra were obtained for each analyte. Samples were prepared by hydrolysis where applicable, organic solvent extraction, and derivatization where necessary. Confirmation was achieved by comparing abundance of major ions and retention time of the total ion current (TIC) of an analyte with those of the appropriate analytical standard. Quantitation was achieved and calibration curves derived by obtaining the molecular ion ratios of that analyte/internal standard (IS) over a concentration range of 10-300 ng/mL (0.16-4.0 ng total injected into GC/MS). The overall extraction efficiency for these analytes ranged from 53% to 96%. Statistically significant cut-off values (p less than 0.01) were obtained for each analyte. The slope, y-intercept, and coefficient of determination (r2) were calculated for each analyte. All of the GC/MS methods were extensively tested against urine samples determined positive or negative by immunoassay (IA) and are now used in our laboratory.     

Methamphetamine in antemortem blood and urine by radioimmunoassay and GC/MS

Methamphetamine abuse is increasing and methamphetamine is second only to alcohol as a positive finding in cases submitted to the San Diego Sheriff's Crime Laboratory. In general, whole blood specimens are submitted more often than urine. A modified version of a commercially available radioimmunoassay, Coat-A-Count (CAC) Methamphetamine, was investigated as a screen for methamphetamine in whole blood and urine. The assay was modified by using 100 microL of sample, making up standards in whole beef blood, extending the incubation time to 2 h or overnight, and using a cutoff reference of 50 ng/mL methamphetamine. The detection limit for the CAC Methamphetamine kit was 20 ng/mL methamphetamine in whole blood. The CAC Methamphetamine results were compared to Abuscreen Amphetamine High Specificity results and to gas chromatography/mass spectrometry (GC/MS) quantitation of amphetamine and methamphetamine for 157 positive and 48 negative blood specimens. With the CAC Methamphetamine assay there were 2 false negatives detected, both less than the 50 ng/mL cutoff level. There were 12 (6%) false positives with the CAC Methamphetamine assay and 29 (14%) false positives with the Abuscreen Amphetamine assay. Of the positive samples, 95% contained only methamphetamine, with an average concentration of 308 ng/mL, range 25-2030 ng/mL.     

超高效液相色谱-串联质谱法测定人尿液中甲氨蝶呤浓度的不确定度评定

目的：评价超高效液相色谱-串联质谱（UHPLC-MS/MS）测定人尿液中甲氨蝶呤浓度的不确定度。方法：通过考察测定方法,分析实验过程中的不确定度来源,包括对照品称量、标准溶液配制和样品制备、回收率及基质效应、校正曲线拟合、仪器的精密性、实验温度和对照品纯度等,然后逐一评定并计算合成不确定度和扩展不确定度。结果：置信概率P为95%时,人尿液中甲氨蝶呤低浓度0.06 μmol·L^-1和高浓度3 μmol·L^-1质控样品的扩展不确定度分别为0.006 5 μmol·L^-1和0.20 μmol·L^-1。结论：根据不确定度的大小及来源,改良测定方法,提高分析准确性。UHPLC-MS/MS法测定人尿液中甲氨蝶呤浓度的不确定度在低浓度时主要由曲线拟合、生物样品配制和标准液配制、回收率及基质效应引入;高浓度的不确定度主要由生物样品配制和标准液配制、回收率引入。     

GC/MS在苯丙胺类药物分析中的应用

苯丙胺类药物的滥用已经成为世界严重的社会问题之一。在过去的几年中,大量的文章报道了血、尿及毛发中苯丙胺类药物的检测和鉴定。通过使用不同的检测器,最低检测浓度已经达到了ng·ml-1。这里我们主要介绍GC/MS在苯丙胺类药物分析中的应用以及还存在的一些不足之处。用于确定血、尿及毛发中苯丙胺类药物的大多数GC/MS程序,基本上都遵循下列原则:样品在经过液液提取(LLE)或固相提取(SPE)之后,随之进行衍生化,然后目标分析物在熔融的石英毛细管柱上被分离,并以选择离子检测(SIM)模式进行定量分析与检测,而且在大多数情况下,选择的内…     

Confirmation and quantitation of cocaine, benzoylecgonine, ecgonine methyl ester in human urine by GC/MS

A rapid, sensitive, reliable quantitative GC/MS method using 0.2 mL of urine was developed for the confirmation of cocaine use. After a simple organic solvent extraction and derivatization with pentafluoropropionic anhydride, cocaine, benzoylecgonine, and ecgonine methyl ester were identified by GC/MS through the retention time for the total ion current and selected ion monitoring (SIM) for each analyte. Quantitation was achieved by obtaining the calibration curves for the molecular ion ratios of the analyte/ketamine (IS) over a range of 12.5-250 ng/mL (0.1-2 ng total). The extraction efficiency for these analytes ranged from 70 to 82%. The sensitivity limit of detection for each analyte was 12.5 ng/mL (0.1 ng) at p less than 0.01. Intra- and interday precision for these analytes ranged between 14.7 and 29.5% CV. This method is in routine use in our laboratory for the GC/MS confirmation of enzyme immunoassay cocaine-positive urine samples.     

GC/MS and EMIT analyses for delta 9-tetrahydrocannabinol metabolites in plasma and urine of human subjects

Ten male subjects smoked placebo marijuana cigarettes spiked with delta 9-tetrahydrocannabinol (THC) at 150 micrograms/kg body weight. Plasma and urine samples were collected for 22 hr after smoking. Total cross-reacting cannabinoids were measured by immunoassay (EMIT) and THC and major metabolites were identified and quantitated by gas chromatographic/mass spectrometric (GC/MS) procedures. In plasma, THC concentration provided the best indication of recent (less than 6 hr) smoking. In enzyme-hydrolyzed urine, 8 beta, 11-dihydroxy-THC at high concentration was identified in the earliest voidings, falling rapidly to the limit of detection before 22 hr in most subjects. Levels above 15 to 20 ng/mL were indicative of use within the previous 4 to 6 hr.     

Hair analysis by GC/MS/MS to verify abuse of drugs

Because of its peculiar characteristics, hair analysis provides a way of obtaining information that cannot be acquired by other commonly used forensic medical analyses, such as blood or urine analysis. In the keratin matrix many xenobiotics are incorporated permanently, in contrast to the situation with blood or urine where they are generally only detectable for a few hours or days. Therefore hair analysis should be the method of choice in the clinical and forensic toxicology field when the assessment of repeated or chronic exposure to a drug is required, e.g. in the case of criminal responsibility, revocation/restoration of a driving licence or in workplace testing. Some factors that can affect the concentrations of drugs in hair, such as passive contamination, age, ethnicity and cosmetic treatment, must be considered. Analytical methodology is also very important: GC/MS/MS has proved to be a highly sensitive and specific technique for the detection of very low concentrations of such drugs in hair. In this study five cases of the application of hair analyses using this technique for the determination of abused drugs (opiates, cocaine, amphetamine, anabolic steroids) are described.     

液相色谱-电喷雾离子阱质谱法研究人尿中1,5-二咖啡酰奎宁酸的代谢产物

目的 研究健康受试者口服1,5-二咖啡酰奎宁酸后尿液中的代谢产物.方法 健康受试者每人口服1,5-二咖啡酰奎宁酸600 mg,收集0-24 h的尿样,经C_(18)小柱固相萃取纯化后,用液相色谱-电喷雾离子阱质谱联用技术对人尿中的代谢产物进行分析鉴定.结果 在人尿中发现了1,5-二咖啡酰奎宁酸的甲基化、葡萄糖醛酸化及甲基-葡萄糖醛酸化代谢产物共28个.其中,有2个代谢产物结构经标准品对照得到确证.结论 甲基化、葡萄糖醛酸化和异构化反应是1,5-二咖啡酰奎宁酸在人体内的3种重要代谢途径.     

LC-MS/MS 法测定人尿液中氯法拉滨的浓度

目的建立高效液相色谱-串联质谱（LC-MS/MS）方法测定人尿样中氯法拉滨的浓度。方法采用AB SCIEX QTRAP 5500串联质谱仪及Agilent1200高效液相色谱仪进行检测。尿样经甲基叔丁基醚提取处理,以克拉屈滨为内标。色谱柱为ThermoC18柱（150mm×4.6mm,5μm）,流动相为乙腈—4mM乙酸铵（含0.3%的甲酸）（250∶3,v/v）;流速为0.5mL·min-1。氯法拉滨和克拉屈滨的MRM扫描离子通道m/z分别为304.2→170.0,286.1→170.0。进样量：10μL。结果氯法拉滨和克拉屈滨分离良好,保留时间分别为3.77min,3.88min。氯法拉滨在2.5~500ng·mL-1范围内线性关系良好（r=0.9995）,日内、日间RSD均低于6.39%,准确度（RE）均低于10.17%。结论本法样品预处理简便快捷,检测结果专属性强,灵敏度好,准确度高,适用于氯法拉滨药代动力学的研究。     

毛细管气相色谱法测定人尿中右美沙芬及其代谢物

目的：为研究细胞色素Ｐ４５０同工酶ＣＹＰ２Ｄ６在人群中的代谢多态性。本文建立了人尿中右美沙芬（ＤＭ）及其Ｏ－去甲基代谢物３－羟基－Ｎ－甲基－吗喃（ＤＴ）的气相色谱分析方法。方法：尿样经提取后,以ＨＰ－１毛细管柱作为分离柱,ＦＩＤ为检测器,正二十烷为内标进行气相色谱分析。结果：在此实验条件下,右美沙芬、内标物和代谢物的保留时间分别为８．５、９．９和１１．２ｍｉｎ;尿样中ＤＭ在０．１０－２．０μｇ／ｍ