SPE—GC／MS、GC／NPD法检测血液中苯丙胺类毒品

目的利用GC／MS、GC／NPD与固相萃取（SPE）技术相结合,建立血液中苯丙胺类毒品的定性定量分析方法。方法采用Bond—ElutCerti{y固相柱、甲醇淋洗、二氯甲烷／异丙醇／氨水（78／20／2）洗脱固相萃取分离提取,比较了不同PH体系、稀释状态、洗脱溶剂对提取回收率的影响,建立血液中苯丙胺类毒品的GC／MS、GC／NPD定性定量分析方法。结果以GC／NPD分析AM、MA、MDA和MDMA浓度在15ng／mL-2000ng／mL、10ng／mL～1600ng／mL、20ng／mL-3000ng／ml、20ng／mL-3000ng／mL范围内线性关系良好,AM、MA、MDA和MDMA的检测限分别为10ng／mL、8ng／mL、15ng／mL、15ng／mL,方法平均回收率大于85％,标准偏差小于5％,GC／MS-Scan检测限分别为40ng／mL、32．0ng／mL、60．0ng／mL、60．0ng／mL。结论此方法可满足苯丙胺类毒品滥用者的血液定性定量分析。     

GC/MS、GC/NPD法检测血液中氯胺酮

目的利用GC/MS、GC/NPD与固相萃取（SPE）技术相结合,建立血液中氯胺酮的定性定量分析方法。方法选择4-苯基丁胺为内标,采用Bond-Elut Certify固相柱萃取、二氯甲烷：异丙醇：氨水（78∶20∶2,v/v/v）洗脱的固相萃取分离技术,比较不同pH体系、洗脱溶剂对回收率的影响,建立血液中氯胺酮的GC/MS、GC/NPD定性定量分析方法。结果以GC/NPD分析氯胺酮在6.0～5000ng/mL范围内线性关系良好,GC/MS-Scan定性检测限为20.0ng/mL。方法平均回收率达96.9%,标准偏差小于5%。结论此方法可满足氯胺酮毒品滥用者血液定性定量分析。     

应用分子印迹固相萃取法提取血液中大隆和溴敌隆

目的采用分子印迹固相萃取,UPLC/MS分析,检验血液中大隆和溴敌隆。方法以大隆为模板分子,丙烯酰胺为单体,二甲基亚砜为有机溶剂,按照模板分子与单体摩尔配比关系为1∶4合成大隆分子印迹聚合物,作为固相萃取剂萃取血液中大隆和溴敌隆,以2×1mL去离子水、1mL 30%甲醇去离子水洗涤杂质,2×1mL 1%甲酸/甲醇洗脱,提取血中目标物后进行UPLC/MS分析检测。结果溴敌隆和大隆回收率均在90%以上,在50～500ng/mL浓度范围内线性关系良好r,2溴敌隆为0.998 4,大隆为0.991,检测限（S/N=5/1）低于10ng/mL。结论本方法回收率高,基质效应降低,杂质干扰少,可用于血液中低浓度大隆和溴敌隆的检验。     

固相萃取/气相色谱法检测全血中的扎来普隆

目的建立全血中扎来普隆的固相萃取/气相色谱检测方法。方法采用Oasis HLB固相萃取柱,5%甲醇水淋洗,二氯甲烷洗脱,GC/NPD定量检验,GC/MS定性检验,并对检测条件进行考察。结果扎来普隆与血中杂质分离良好,平均萃取回收率达89%以上,最低检出限20ng/mL,50～2000ng/mL内线性良好,相关系数R2=0.9945,日内RSD为3.1%～4.2%,日间RSD 5.8%～6.7%。结论该方法操作简便快速,提取回收率高,重现性好,提取物杂质少,可用于实际案件的检验。     

在线固相萃取-超高效液相色谱-串联质谱法同时测定血液样品中12种卡西酮类毒品

目的 建立一种同时测定血液样品中12种卡西酮类毒品的在线固相萃取-超高效液相色谱-串联质谱法。方法 血液样品用乙腈沉淀蛋白，经离心、稀释、过滤后上样，采用PLRP-S在线固相萃取柱（2.1mm×12.5mm,15～20μm）富集纯化，Poroshell 120 EC-C18色谱柱（3.0mm×150mm,2.7μm）进行分离，在线固相萃取柱以乙腈-5%（体积分数）甲醇作为流动相进行流速1.0 mL/min的梯度洗脱，色谱柱以5 mmol/L乙酸铵缓冲液[含0.1%（体积分数）甲酸]-乙腈作为流动相进行流速0.4m L/min的梯度洗脱。离子源为电喷雾离子源，采用多反应监测模式进行测定。结果 12种卡西酮类毒品线性关系良好，相关系数均大于0.998，方法检出限为0.1～0.5ng/mL，定量限为0.3～1.5ng/mL。12种卡西酮类毒品在3个不同质量浓度条件下的回收率为70.9%～108%，日内精密度和日间精密度分别为1.5%～8.9%、5.1%～44.5%(n=6)。结论 该方法操作简单方便、样品需求量少、灵敏度高、检出限低，可用于血液样品中卡西酮类毒品的测定。     

固相萃取-气相色谱法检测全血中佐匹克隆

目的采用固相萃取-气相色谱法检测全血中佐匹克隆。方法采用Oasis HLB固相萃取柱对样品进行前处理,去离子水、0.5%氨水-甲醇/水((V/V 40∶60)溶液先后淋洗,二氯甲烷/异丙醇(V/V 75∶25)洗脱后进行GC/NPD检测。结果血液中佐匹克隆在50～5 000ng/mL范围内线性良好(R2=0.998 8),平均萃取回收率为96.9%,检出限为30ng/mL,日内RSD为2.1%～5.7%,日间RSD为3.3%～6.2%,结论固相萃取-气相色谱检测法灵敏度高,重现性好,可在血液中佐匹克隆的检测中选用。     

毛细管固相微萃取/UPLC-MS/MS测定血浆中毒品成分

目的建立毛细管固相微萃取UPLC-MS/MS测定血浆中毒品的方法,并用于实际样品分析。方法取含毒品成分的血浆样品1mL,加入2mL硼砂缓冲液后吸入注射器中,联接内壁上固定有DB-1701毛细管柱的不锈钢针头,流动注射泵以300μL/min流速使样品全部排出,1mL纯水冲洗,50μL乙腈洗脱2min,UPLC-MS/MS梯度洗脱-正离子-多反应监测测定洗脱液毒品成分,并对28例案件样品进行分析。结果血浆中毒品回收率为64.5%~82.1%,RSD10%,LOD40 ng/mL,28份样品中检出19例。结论采用毛细管固相微萃取(in-tube SPME),方法快速、简捷、环保,与UPLC-MS/MS技术结合可应用于血浆中毒品的快速筛查。     

固相萃取技术与GC／MS、GC／NPD结合检验血中毒鼠强

利用GDX-403固相萃取技术对血样直接提取,以咖啡因为内标,对毒鼠强进行GC/MS、GC/NPD分析,毒鼠强回收率65%～80%.并利用GC/NPD高灵敏度,使GC/MS和GC/NPD在检测分析中互补.本方法操作简便、快捷,适合检案要求.     

HS-SPME-GC/MS法检测尿液及毛发中苯丙胺类毒品

目的采用顶空固相微萃取（HS-SPME）、GC/MS分析方法,对生物样品中苯丙胺（AM）、甲基苯丙胺（MAM）、3,4-亚甲二氧基苯丙胺（MDA）和3,4-亚甲二氧基甲基苯丙胺（MDMA）4种苯丙胺类毒品进行定性定量分析。方法在碱性和饱和盐处理状态下,采用100μm聚二甲基硅氧烷（PDMS）萃取纤维,于顶空瓶中进行生物样品AM、MAM、MDA、MDMA 4种毒品萃取,以2-甲基苯乙胺为内标,经气-质联用选择离子检测（GC/MS/SIM）模式进行定性定量分析。对HS-SPME条件优化,对方法的精密度、准确度和检出限进行测定。结果 AM、MAM、MDA、MDMA 4种毒品尿液中的最低检出限为5ng/mL,毛发中的最低检出限为0.5ng/mg。尿液中线性关系范围为0.05μg/mL~5μg/mL,r〉0.991,回收率为82%~108%,RSD为2.6%~6.1%（n=5）;毛发中线性关系范围为5ng/mg~500ng/mg,r〉0.992,回收率为80%~113%,RSD（%）为1.4%~6.8%（n=5）。结论 HS-SPME-GC/MS各项定量参数符合分析要求。该方法简单、灵活、经济、快速、无溶剂,适用于生物检材中该类毒品的分析。     

血清中盐酸曲马多的膜式固相萃取及GC／MS／SIM测定

目的建立血清中盐酸曲马多的膜式固相萃取(SPE)气-质联用分析方法。方法1mL血清用2mL0.1mol/L磷酸盐缓冲液(pH6)稀释后,用含有C18和强酸型强阳离子交换基团的SPECC18AR/MP3固相萃取柱萃取,洗脱液为含2%氨水的乙酸乙酯;选择SKF525作为内标物,用GC/MS/SIM定量测定。结果在加标量为0.1、0.2和0.5μg/mL的血清样品中,盐酸曲马多的回收率分别为98.9%、92.5%和84.8%,5次测定的RSD分别为3.2%、8.7%和10.9%;线性范围为0.1～4μg/mL,多项式回归相关系数r2=0.9939;检出限为21ng/mL;同一根萃取柱,连续使用5次,没有出现堵塞和污染,回收率及RSD未见下降。结论本方法适合于法医毒物分析。     

Quantitative determination of amphetamines, cocaine, and opiates in human hair by gas chromatography/mass spectrometry

Hair of young subjects (N = 36) suspected for drug abuse was analysed for morphine, codeine, heroin, 6-acetylmorphine, cocaine, methadone, amphetamine, methamphetamine, 3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxymethamphetamine (MDMA), and 3,4-methylenedioxyethylamphetamine (MDEA). The analysis of morphine, codeine, heroin, 6-acetylmorphine, cocaine, and methadone in hair included incubation in methanol, solid-phase extraction, derivatisation by the mixture of propionic acid anhydride and pyridine, and gas chromatography/mass spectrometry (GC/MS). For amphetamine, methamphetamine, MDA, MDMA, and MDEA analysis, hair samples were incubated in 1M sodium hydroxide, extracted with ethyl acetate, derivatised with heptafluorobutyric acid anhydride (HFBA), and assayed by GC/MS. The methods were reproducible (R.S.D. = 5.0-16.1%), accurate (85.1-100.6%), and sensitive (LoD = 0.05-0.30ng/mg). The applied methods confirmed consumption of heroin in 18 subjects based on positive 6-acetylmorphine. Among these 18 heroin consumers, methadone was found in four, MDMA in two, and cocaine in two subjects. Cocaine only was present in two, methadone only in two, methamphetamine only in two, and MDMA only in seven of the 36 subjects. In two out of nine coloured and bleached hair samples, no drug was found. Despite the small number of subjects, this study has been able to indicate the trend in drug abuse among young people in Croatia.     

Detection of phentermine in hair samples from drug suspects

Phentermine (PT) has been widely used as an anti-obesity drug. This drug has to be used with caution due to its close resemblance with amphetamines in its structure and toxicity profile. Recently, PT is in distribution by illegal modes and is found to be available through sources such as the internet, thus their misuse and/or abuse is threatening to be a serious social issue. In the present study, 32 cases of drug suspects were observed for PT abuse, detected using hair samples for drug analysis. PT and other amphetamines, such as methamphetamine (MA), amphetamine (AP), 3,4-methylenedioxyamphetamine (MDMA) and 3,4-methylenedioxyamphetamine (MDA), were extracted using 1% HCl in methanol for 20 h at 38°C. The extracts were derivatized with trifluoroacetic anhydride (TFAA) and analyzed using gas chromatography/mass spectrometry (GC/MS). Among the 32 cases of PT abuse, MA and its main metabolite, AP were identified in seven cases and MDMA and its main metabolite, MDA were detected in two other cases.     

Evaluation of ephedrine, pseudoephedrine and phenylpropanolamine concentrations in human urine samples and a comparison of the specificity of DRI amphetamines and Abuscreen online (KIMS) amphetamines screening immunoassays

The purpose of this study was to evaluate the ability of two amphetamine class screening reagents to exclude ephedrine (EPH), pseudoephedrine (PSEPH), and phenylpropanolamine (PPA) from falsely producing positive immunoassay screening results. The study also sought to characterize the prevalence and concentration distributions of EPH, PSEPH, and PPA in samples that produced positive amphetamine screening results. Approximately 27,400 randomly collected human urine samples from Navy and Marine Corps members were simultaneously screened for amphetamines using the DRI and Abuscreen online immunoassays at a cutoff concentration of 500 ng/mL. All samples that screened positive were confirmed for amphetamine (AMP), methamphetamine (MTH), 3,4-Methylenedioxyamphetamine (MDA), 3,4-Methylenedioxymethamphetamine (MDMA), EPH, PSEPH, and PPA by gas chromatography/mass spectrometry (GC/MS). The DRI AMP immunoassay identified 1,104 presumptive amphetamine positive samples, of which only 1.99% confirmed positive for the presence of AMP, MTH, MDA, or MDMA. In contrast, the online AMP reagent identified 317 presumptive amphetamine positives with a confirmation rate for AMP, MTH, MDA, or MDMA of 7.94%. The presence of EPH, PSEPH, or PPA was confirmed in 833 of the 1,104 samples that failed to confirm positive for AMP, MTH, MDA, or MDMA; all of the 833 samples contained PSEPH. When compared to the entire screened sample set, PSEPH was present in approximately 3%, EPH in 0.9%, and PPA in 0.8% of the samples. The results indicate that cross reactivities for EPH, PSEPH, and PPA are greater than reported by the manufacturer of these reagents. The distribution of concentrations indicates that very large concentrations of EPH, PSEPH, and PPA are common.     

Fast LC-MS/MS method for the determination of amphetamine, methamphetamine, MDA, MDMA, MDEA, MBDB and PMA in urine

A fast method was designed for the simultaneous determination of amphetamine (A), methamphetamine (MA), PMA, MDA, MDMA, MDEA and MBDB in urine. The drugs were analysed by LC (ESI)-MS/MS, after a simple liquid-liquid extraction in the presence of the deuterated analogues. Reverse phase separation on an Atlantis dC18 Intelligent Speed column was achieved in less than 4 min under gradient conditions, and the total run time was 8 min. The method was fully validated, including linearity (1-1000 ng/mL for A, MDMA, MDEA and MBDB; 2-1000 ng/mL for MDA and PMA; 1-200 ng/mL for MA; r2>0.99 for all compounds), recovery (>80%), within-day and between-day precision and accuracy (CV and MRE<12.7% for intermediate level and ULOQ, and <17.2% for LLOQ), limit of detection (0.2 ng/mL for MDMA, MDEA and MBDB; 0.5 ng/mL for A, MA and PMA; 1 ng/mL for MDA) and quantitation (1 ng/mL for A, MA, MDMA, MDEA and MBDB; 2 ng/mL for MDA and PMA) and relative ion intensities. No matrix effect was observed. The procedure proved to be sensitive, specific and rapid, and was applied to real forensic cases.     

Simultaneous analysis of six amphetamines and analogues in hair, blood and urine by LC-ESI-MS/MS. Application to the determination of MDMA after low ecstasy intake

A rapid and sensitive method using LC-MS/MS triple stage quadrupole for the determination of traces of amphetamine (AP), methamphetamine (MA), 3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxymethamphetamine (MDMA, "ecstasy"), 3,4-methylenedioxyethamphetamine (MDEA), and N-methyl-1-(3,4-methylenedioxyphenyl)-2-butanamine (MBDB) in hair, blood and urine has been developed and validated. Chromatography was carried out on an Uptisphere ODB C(18) 5 microm, 2.1 mm x 150 mm column (Interchim, France) with a gradient of acetonitrile and formate 2 mM pH 3.0 buffer. Urine and blood were extracted with Toxitube A (Varian, France). Segmented scalp hair was treated by incubation 15 min at 80 degrees C in NaOH 1M before liquid-liquid extraction with hexane/ethyl acetate (2/1, v/v). The limits of quantification (LOQ) in blood and urine were at 0.1 ng/mL for all analytes. In hair, LOQ was <5 pg/mg for MA, MDMA, MDEA and MBDB, at 14.7 pg/mg for AP and 15.7 pg/mg for MDA. Calibration curves were linear in the range 0.1-50 ng/mL in blood and urine; in the range 5-500 pg/mg for MA, MDMA, MDEA and MBDB, and 20-500 pg/mg for AP and MDA. Inter-day precisions were <13% for all analytes in all matrices. Accuracy was <20% in blood and urine at 1 and 50 ng/mL and <10% in hair at 20 and 250 pg/mg. This method was applied to the determination of MDMA in a forensic case of single administration of ecstasy to a 16-year-old female without her knowledge during a party. She suffered from hyperactivity, sweating and agitation. A first sample of urine was collected a few hours after (T+12h) and tested positive to amphetamines by immunoassay by a clinical laboratory. Blood and urine were sampled for forensic purposes at day 8 (D+8) and scalp hair at day 60 (D+60). No MDMA was detected in blood, but urine and hair were tested positive, respectively at 0.42 ng/mL and at 22 pg/mg in hair only in the segment corresponding to the period of the offence, while no MDA was detectable. This method allows the detection of MDMA up to 8 days in urine after single intake.     

GC法检测血液和尿液中甲基苯丙胺和咖啡因

目的建立同时测定血、尿中甲基苯丙胺和咖啡因含量的方法。方法应用GC／NPD技术,以4-苯基丁胺为内标,直接碱化,用氯仿提取,三氟乙酸酐衍生化,8CB熔融石英毛细管柱（30m×0．25mm×0．25μm）分析。结果生物样品中甲基苯丙胺与咖啡因在0．012—7．5μg/mL浓度范围内线性关系良好,检测限（S／N=3）依次为1．2ng／mL,0．6ng／mL（血）;1．6ng／mL,0．8ng／mL（尿）。苯丙胺在0．017—10．0μg／mL浓度范围内线性关系良好,检测限为1．6mg／mL（血）,3．2ng／mL（尿）。所有样本回收率均大于85％。结论本方法准确、灵敏,适用于血、尿中甲基苯丙胺及其代谢物苯丙胺的三氟乙酸酐衍生化物和咖啡因的同时检测,为判定滥用毒品种类、追查毒品来源以及研究生物体内甲基苯丙胺和咖啡因的交互影响提供了检测手段。     

Rapid and simple method for direct determination of several amphetamines in seized tablets by GC--FID

A simple and rapid method for direct simultaneous determination of amphetamine, methamphetamine, 3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxymethamphetamine (MDMA), 3,4-methylenedioxy-N-ethylamphetamine (MDEA) and N-methyl-1-(3,4-methylenedioxyphenyl)-2-butanamine (MBDB) in seized tablets was developed using gas chromatography with flame ionization detection. Separation of all six underivatized amphetamines, including diphenylamine as internal standard, was performed in about 6 min, using SPB-50 capillary column. Amphetamine and methamphetamine eluted with negligible tailing while the other amphetamines had highly symmetrical peaks. Sensitivity per component on-column was in the nanogram range, and reproducibility from 2.6 to 6.6% at low concentration (2.4 microg/mL) and from 1.2 to 2.6% at high (70 microg/mL) concentration. The method has a wide linear range, from Limit of detection (LOD) to almost 200 microg/mL, thus allowing analysis of different samples across a wide range of possible concentrations of amphetamines. This simple, fast and precise method using gas chromatography--flame ionization detector (GC--FID), in conjunction with other methods (TLC, IR, HPLC), can be used for identification of amphetamines and direct determination in seized tablets, especially in laboratories with heavy workload.     

尿中苯丙胺、甲基苯丙胺、MDA和MDMA的固相微萃取和GC/MS/SIM测定

目的研究固相微萃取(SPME)用于尿中苯丙胺(AMP)、甲基苯丙胺(MET)、3,4-亚甲二氧基苯丙胺(MDA)和3,4-亚甲二氧基甲基苯丙胺(MDMA)的提取。方法样品调节至碱性和用盐饱和后用顶空SPME,内标为MET-d5。萃取纤维为100μm聚二甲基硅氧烷(PDMS)。用气质联用选择离子检测(GC/MS/SIM)。结果0.2μg/ml加标尿样,AMP、MET、MDA和MDMA的富集倍数分别为22,60,13和47。检出限(S/N=3)为0.4～9.5ng/ml。线性范围为0.05～1μg/ml。0.2、0.5和1.0μg/ml加标尿样,相对回收率77.9%～112.4%,变异系数2.7%～18.0%(n=5)。用该方法分析5个案件样品,和常规液液萃取结果接近。结论顶空SPME法用于尿中AMP、MET、MDA和MDMA等化合物的分析,无需有机溶剂,富集效率高,提取-富集-进样一体化,简单方便实用。     

Simultaneous analysis of six amphetamines and analogues in hair, blood and urine by LC-ESI-MS/MS: Application to the determination of MDMA after low Ecstasy intake

A rapid and sensitive method using LC-MS/MS triple stage quadrupole for the determination of traces of amphetamine (AP), methamphetamine (MA), 3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxymethamphetamine (MDMA, “ecstasy”), 3,4-methylenedioxyethamphetamine (MDEA), and N-methyl-1-(3,4-methylenedioxyphenyl)-2-butanamine (MBDB) in hair, blood and urine has been developed and validated. Chromatography was carried out on an Uptisphere ODB C₁₈ 5 μm, 2.1 mm × 150 mm column (Interchim, France) with a gradient of acetonitrile and formate 2 mM pH 3.0 buffer. Urine and blood were extracted with Toxitube A^® (Varian, France). Segmented scalp hair was treated by incubation 15 min at 80 °C in NaOH 1 M before liquid–liquid extraction with hexane/ethyl acetate (2/1, v/v). The limits of quantification (LOQ) in blood and urine were at 0.1 ng/mL for all analytes. In hair, LOQ was <5 pg/mg for MA, MDMA, MDEA and MBDB, at 14.7 pg/mg for AP and 15.7 pg/mg for MDA. Calibration curves were linear in the range 0.1–50 ng/mL in blood and urine; in the range 5–500 pg/mg for MA, MDMA, MDEA and MBDB, and 20–500 pg/mg for AP and MDA. Inter-day precisions were <13% for all analytes in all matrices. Accuracy was <20% in blood and urine at 1 and 50 ng/mL and <10% in hair at 20 and 250 pg/mg. This method was applied to the determination of MDMA in a forensic case of single administration of ecstasy to a 16-year-old female without her knowledge during a party. She suffered from hyperactivity, sweating and agitation. A first sample of urine was collected a few hours after (T + 12 h) and tested positive to amphetamines by immunoassay by a clinical laboratory. Blood and urine were sampled for forensic purposes at day 8 (D + 8) and scalp hair at day 60 (D + 60). No MDMA was detected in blood, but urine and hair were tested positive, respectively at 0.42 ng/mL and at 22 pg/mg in hair only in the segment corresponding to the period of the offence, while no MDA was detectable. This method allows the detection of MDMA up to 8 days in urine after single intake.