毛发中7种常见合成大麻素类新精神活性物质的分析及应用

目的建立毛发中常见合成大麻素类新精神活性物质的超高效液相色谱-串联质谱(ultra-highperformance liquid chromatography-tandem mass spectrometry,UPLC-MS/MS)测定方法。方法将20 mg毛发加入1m L内标甲醇溶液,经冷冻研磨、超声提取后,提取物经ACQUITY UPLC HSS T3柱(100mm×2.1mm,1.8μm)分离,流动相A为含20 mmol/L乙酸铵、0.1%甲酸、5%乙腈的水溶液,流动相B为乙腈,采用电喷雾离子源正离子模式,在多反应监测模式下采集数据。结果毛发中7种合成大麻素类物质在各自线性范围内线性关系良好(r0.99),检出限为0.5～2 pg/mg,定量限为1～5 pg/mg,日内、日间精密度为0.1%～12.6%,日内、日间准确度为89.2%～110.7%,提取回收率为52.3%～93.3%,基质效应为19.1%～95.2%。结论建立的测定方法样品制备简便、灵敏度高,适用于法医学鉴定实践毛发中常见合成大麻素类新精神活性物质的定性定量分析。     

QuEChERS结合UPLC-MS/MS检测血液中3种色胺类新精神活性物质

目的建立一种QuEChERS结合超高效液相色谱-串联质谱法(ultra-high performance liquid chro-matography-tandem mass spectrometry,UPLC-MS/MS)快速筛查和检验人体血液中3种色胺类新精神活性物质——N,N-二烯丙基-5-甲氧基色胺(5-Me O-DALT)、N-甲基-N-异丙基-5-甲氧基色胺(5-Me O-Mi PT)、N,N-二异丙基-5-甲氧基色胺(5-Me O-Di PT)的方法。方法分别考察了萃取剂种类、盐析剂种类及用量、吸附剂用量等条件对3种色胺类物质检验结果的影响。血液样品经Qu ECh ERS方法处理后采用UPLC-MS/MS法进行检测。结果人体血液中5-Me O-DALT、5-Me O-Mi PT、5-Me O-Di PT分别在0.5～100、0.5～100、0.2～100 ng/m L范围内具有良好的线性关系,相关系数均大于0.99,检出限为0.1～0.2 ng/m L,回收率范围在84.86%～94.57%,日内及日间精密度良好。结论该方法简单、快速、易于操作、回收率高,适用于血液中色胺类物质的定性定量分析,能够为公安机关处理相关案件提供借鉴和参考。     

赛洛新和赛洛西宾分析方法研究进展

致幻剂是一类低剂量就使人意识、知觉和视觉异常,引起幻觉和情绪障碍的精神活性物质。赛洛新和赛洛西宾是一类具有致幻作用的色胺类化合物,存在于裸盖伞属和斑褶菇属的致幻蘑菇中。当前滥用及娱乐性使用含此类物质的致幻蘑菇的现象日趋严重,由此引起的危害公共安全案(事)件不断增多。因此,对于致幻蘑菇滥用的鉴定是当前司法鉴定领域的重要需求和国际热点研究问题之一。通过对赛洛新和赛洛西宾的药理毒理、体内过程、检材处理方法和分析检测方法进行了综述,以期为司法鉴定实践提供参考。     

苯环利定类新型致幻剂的分析方法研究进展

苯环利定是一种人工合成的新精神活性物质，具有强烈的致幻作用，吸食后暴力倾向增加，该物质在受到管制后大量结构类似物被合成，严重威胁公众健康和社会安定。苯环利定类新型致幻剂滥用引起了国际社会的广泛关注，对其药理毒理及分析方法的研究成为法医毒物分析领域的热点。结合近几年的研究，本文综述了苯环利定类新型致幻剂的理化性质、药理毒理作用、体内过程及检测方法，以期为苯环利定类新型致幻剂的监测管控提供依据。     

头发中合成大麻素5F-MDMB-PICA检测

目的一种新型合成大麻素成分5F-MDMB-PICA被非法添加至电子烟油中,其滥用对吸食者自身健康和公共安全产生了极大的危害,因此亟待建立一种5F-MDMB-PICA的检测方法并进行评价。方法采用UPLC-MS/MS方法定性和定量检测电子烟成瘾者头发中的合成大麻素成分5F-MDMB-PICA,称取约20mg头发,加1mL含内标THC-D3(0.4ng/mL)甲醇,经研磨、超声、离心、过滤后,采用Waters Acquity UPLC HSS T3柱(100mm×2.1mm,1.8μm)分离,流动相A为20mmol/L乙酸铵、乙腈和水,流动相B为乙腈。结果头发中5F-MDMB-PICA在1～200pg/mg的浓度范围内线性良好(r0.99),检测限为0.5pg/mg,定量下限为1pg/mg,日内、日间精密度均小于6.6%,日内、日间准确度为96.8%～105.0%,提取回收率为72.5%～93.3%。结论本文方法灵敏度高、样品制备简便,已成功应用于实际案例的电子烟成瘾者头发中合成大麻素成分5F-MDMB-PICA的定性和定量检测。     

新型微流体液相色谱(micro-LC)串联四极杆质谱检测头发中5种苯二氮卓类药物

目的建立了新型微流体液相色谱(micro-LC)串联四级杆质谱检测头发中5种苯二氮卓类药物(三唑仑,α-羟基咪达唑仑,咪达唑仑,α-羟基阿普唑仑,普拉西泮)。方法采用液-液萃取法(LLE)提取毛发样品,微流体液相色谱串联四级杆质谱检测。结果在回归系数大于0.99的情况下,目标分析物的良好线性。精密度和萃取效率分别在2.7%~17.1%和66.5%~120.5%之间。每个分析物的检出限和定量限分别为0.001~0.08 pg·mg^-1和0.0125~0.25pg·mg^-1。结论本方法与传统的液相色谱串联质谱法(LC/MS-MS)相比灵敏度出高2-10倍。本研究展示了新型微流体分离系统串联质谱方法的实用性,并为法医毒理学领域关于头发样本中痕量药物检测提供了一种新的检测手段。     

离子色谱法用于新精神活性物质盐型测定

目的建立新精神活性物质(new psychoactive substance,NPS)盐型测定的离子色谱分析方法。方法建立离子色谱法分析NPS样品中6种有机酸根离子(乙酸根、酒石酸根、马来酸根、草酸根、富马酸根、柠檬酸根)和5种无机阴离子(氟离子、氯离子、硝酸根离子、硫酸根离子、磷酸根离子)的定性和定量分析方法。采用该方法对222份缴获NPS样品(103份含合成大麻素类,81份含卡西酮类,44份含苯乙胺类,12份含色胺类,7份含苯环利啶类,6份含哌嗪类,2份含氨基茚满类,26份含芬太尼类,43份含其他类)中的盐型进行分析。结果各阴离子在相应的线性范围内线性良好,相关系数(r)均大于0.999,检出限为0.01～0.05 mg/L,定量限为0.1～0.5 mg/L。合成大麻素类样品中,除(4-苄基哌嗪-1-基)[1-(5-氟戊基)-1H-吲哚-3-基]甲酮(5F-BEPIRAPIM)为盐酸盐外,其余102份均为碱型;81份卡西酮类、44份苯乙胺类、7份苯环利啶类、2份氨基茚满类样品的盐型均为盐酸盐;色胺类样品的盐型包括碱型、盐酸盐、富马酸盐、草酸盐4种;哌嗪类样品的盐型包括盐酸盐和碱型2种;芬太尼类和其他类样品的盐型包括碱型、盐酸盐、柠檬酸盐3种。结论采用离子色谱法对NPS样品的盐型进行分析,简单、准确、高效,使得NPS定性和定量结论更加科学、严谨。     

高分辨质谱联合核磁共振波谱技术鉴定色胺类新精神活性物质4-OH-MET和4-AcO-DMT

目的利用高分辨质谱和核磁共振波谱技术检测涉毒案件中涉及的未列管的色胺类新精神活性物质。方法将实际案例中缴获的白色和褐色粉末经提取后使用气相色谱-四极杆飞行时间质谱(gas chro-matography-quadrupole time-of-flight mass spectrometry,GC-QTOF-MS)、超高效液相色谱-线性离子阱-四极杆-轨道阱质谱(ultra-high performance liquid chromatography-linear ion trap quadrupole-orbitrap massspectrometry,UPLC-LTQ-Orbitrap MS)和核磁共振氢谱(1H-nuclear magnetic resonance spectroscopy,1H-NMR)等方法进行分析。结果白色粉末经GC-QTOF-MS检测,组分的主要特征碎片离子峰有m/z 218.1410(分子离子峰)、72.080 6(基峰)等;经UPLC-LTQ-Orbitrap MS检测,质子化分子离子为m/z 219.149 4,碰撞诱导解离模式下主要的二级质谱离子有m/z 160.076 3、72.080 8。褐色粉末经GC-QTOF-MS检测,组分的主要特征碎片离子峰有m/z 246.135 7(分子离子峰)、58.065 1(基峰)等;经UPLC-LTQ-Orbitrap MS检测,质子化分子离子为m/z 247.145 0,碰撞诱导解离模式下主要的二级质谱离子有m/z 202.087 1、160.076 3、134.060 5。经NIST 17谱库检索和1H-NMR共同确认白色粉末和褐色粉末分别含有色胺类新精神活性物质N-甲基-N-乙基-4-羟基色胺(4-OH-MET)和N,N-二甲基-4-乙酰氧基色胺(4-Ac O-DMT)。结论 GC-QTOF-MS、UPLC-LTQ-Orbitrap MS和1H-NMR多种方法联合应用可对未知的新精神活性物质进行鉴定。     

气相色谱-质谱法检验新型毒品“G点液”

<正>5-甲氧基-N,N-二异丙基色胺(5-MethoxyN,N-Diisopropyltryptamine,5-MeO-DIPT)是一种新型精神活性物质,俗称"火狐狸(Foxy)"、"媚药"。此种物质属于色胺类化合物,其分子式为C17H26N2O,分子量274。2004年美国、日本等地陆续将5-MeO-DIPT列为管制药品,2015年我国将5-MeO-DIPT列为管制精神类药品。5-MeO-DIPT多以粉末形式存在,然而近来有不法分子在互联网上     

5-MeO-MiPT对斑马鱼毒性及生物转化途径研究

目的 为评价色胺类新精神活性物质N,N-甲基异丙基-5-甲氧基色胺(N-methyl,N-isopropyl-5-methoxytryptamine,5-MeO-MiPT)的毒性效应及生物转化途径.方法 以斑马鱼为模式生物,进行自发活动行为测试及镜像反射行为模型测试,分析5-MeO-MiPT对斑马鱼的毒性效应;并通过超...     

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.     

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.     

Hair analysis for fenfluramine and norfenfluramine as biomarkers for N-nitrosofenfluramine ingestion

In this paper, a high performance liquid chromatographic method with fluorescence detection (HPLC-FL) for the determination of fenfluramine (Fen) and norfenfluramine (Norf) in human hair as biomarker metabolites of N-nitrosofenfluramine (N-Fen) is described. Washed and cut hair segments were extracted by ultrasonication for 1h at room temperature in methanol. The extract was evaporated and applied for derivatization with the fluorescent reagent 4-(4,5-diphenyl-1H-imidazol-2-yl)benzoyl chloride (DIB-Cl). An HPLC-FL analysis was performed using an ODS column with mobile phase composition of acetonitrile and water (65:35, v/v) and monitored at 430 nm (excitation 325 nm). The method was sensitive with detection limits of 36 and 16 pg/mg hair for Fen and Norf, respectively. The linearity was assessed in the range 0.036-144 ng/mg for Fen and 0.016-127 ng/mg for Norf with correlation coefficients larger than 0.999. The method was successfully used for the segmental determination of Fen and Norf in hair samples obtained from hospitalized patients diagnosed with hepatotoxicity and suspected to ingest N-Fen. Both Fen and Norf could be detected in these patients' hair samples in the ranges 43-1389 pg/mg for Fen and 18-680 pg/mg for Norf and the results showed that the patients might ingest N-Fen for a period of not less than 5 months. As well, the method was applied for the determination of Fen and Norf in rats that possess pigmented and non-pigmented hair after an intraperitoneal administration of Fen. Both compounds were determined in black as well as in white hair.     

Windows of detection of zolpidem in urine and hair: application to two drug facilitated sexual assaults

A LC-MS/MS method for the detection of zolpidem in hair was developed to detect this drug after a single dose in possible drug facilitated sexual assaults. To determine the window of detection of zolpidem in both urine and hair, three volunteers received a 10 mg dose. Urine specimens were collected each 12 h for 144 h. Hair was sampled 3-5 weeks after exposure. Hair and urine extracts were separated on a Xterra MS C18 column using a gradient of acetonitrile and formate buffer. For each compound, detection was related to two daughter ions. Zolpidem was detected for up to 60 h in urine with peak concentrations obtained at 12 h. A single exposure to zolpidem was detected in hair at concentrations ranging from 1.8 to 9.8 pg/mg. Hair analysis was applied to two possible criminal cases. In the first case, zolpidem tested positive in the corresponding hair segment at 4.4 pg/mg. In the other case, zolpidem was detected in all the segments analyzed, demonstrating likely previous drug use in addition to recent exposure associated with a positive blood result.     

Optimized conditions for simultaneous determination of opiates, cocaine and benzoylecgonine in hair samples by GC--MS

The present paper describes a qualitative and quantitative method for the simultaneous detection of opiates, cocaine and benzoylecgonine from human hair samples. Every step of the analytical procedure was studied to find the optimized conditions. Nine different incubation systems were examined. The influence of different pH values of samples on the isolation of analytes from the incubation media by Bond Elut cartridges and the stability of the compounds of interest in the different incubation media and conditions were investigated. The extracting power of different incubation media was studied as well. The phosphate buffer 0.1 N at pH 5 was chosen as the extraction medium in an optimized procedure for simultaneous determination of opiates, cocaine and benzoylecgonine in hair samples. The method developed was validated. Recoveries were 90% for morphine (M), 81% for 6-monoacetylmorphine (6-AM), 90% for codeine (CD), 86% for cocaine (C) and 90% for benzoylecgonine (BE). Relative standard deviation for inter-day precision was better than 12%. The limits of detection resulted as 0.05 ng/mg for M and C, as 0.08 for 6-AM and as 0.2 ng/mg for BE. Forty hair samples collected from drug abusers admitted to centers for detoxification treatment were analyzed obtaining 23 positive results for opiates and/or cocaine. Twelve hair specimens longer than 10 cm were analyzed following a sectional approach. In the six positive cases, it was interesting to find that the 6-AM/M ratio generally decreased for each sample from the proximal segment to the distal segments. Moreover, the 6-AM/M ratio was generally lower than 1 in the intermediate and distal segments.     

Ethyl Glucuronide Concentration in Hair of Detainees: A Preliminary Study

Through the measurement of ethyl glucuronide in hair (hETG), it is possible to assess chronic alcohol abuse over time. In this paper, we present a study on hETG in Italian prison inmates. Analyses were performed by LC-MS according to a previously published method. Results were evaluated using the cut-offs established by the Society of Hair Testing. Positives samples (ETG > 30 pg/mg) accounted for 6% of all subjects, with concentrations ranging from 42 pg/mg up to 270 pg/mg, abstinent subjects (ETG < 7 pg/mg) accounted for 88%, and moderate alcohol consumption (7 < ETG < 30 pg/mg) for 6% of the subjects. No females displayed ETG values above 30 pg/mg. Among positive samples, only two subjects did not declare heavy alcohol consumption and were found strongly positive at 210 and 270 pg/mg. To the best of our knowledge, this represents the first study on ETG hair concentration on prison inmates.     

Hair analysis and detectability of single dose administration of androgenic steroid esters

Detection of anabolic steroids in hair samples has been possible only in fatal cases or in cases of high-continuous dosages. In order to verify the possibility of detecting an acute administration, a sensitive and specific assay has been developed for the simultaneous determination of testosterone, nandrolone and some of their esters in hair. The analytes were extracted from finely cut hair with methanol-trifluoroacetic acid overnight. After the incubation, the mixture was evaporated to dryness, redissolved and extracted with hexane. The dried organic layer was silanised and analysed by GC-MS and GC-MS-MS. A sensitivity of at least 20 pg injected was obtained for all the analytes. In guinea pigs treated with a single intramuscular dose of 10 mg/kg nandrolone decanoate, neither nandrolone decanoate nor nandrolone were found in hair collected after 13 days, while both compounds were clearly detectable after four repeated doses (each dose every 3-4 days) of 20 mg/kg nandrolone decanoate. Neither nandrolone decanoate nor nandrolone could be detected in hair from a male healthy volunteer 1 month after treatment with 50 mg nandrolone decanoate, while his urine still tested highly positive for the main nandrolone metabolite (> 100 ng/ml). Testosterone esters could not be detected in hair of healthy subjects collected respectively 3, 2 and 1 month after a single intramuscular administration of 250 mg testosterone enanthate (five subjects), a single intramuscular coadministration of 25 mg testosterone propionate plus 110 mg testosterone enanthate (one subject), or a single oral administration of 120 mg testosterone undecanoate (three subjects). Otherwise, hair analysis revealed an increase of testosterone concentration corresponding to the period of treatment. Analysis of blood and urine samples confirmed the absorption of those compounds. At the sensitivity achieved by the present method, no detection of nandrolone, nandrolone decanoate nor testosteron esters in hair seems to be obvious after a single dose administration.     

Determination of ethyl glucuronide in human hair by SPE and LC-MS/MS

A method for the sensitive and selective determination of ethyl glucuronide (EtG) in hair has been developed using solid-phase extraction (SPE) and liquid chromatography-tandem mass spectrometry (LC-MS/MS). Washed and cut hair segments were extracted by ultrasonication (3h, 50 degrees C) and the extracts were cleaned-up with aminopropyl SPE columns. LC-MS/MS analysis was performed using a polar-endcapped phenyl-hexyl-RP-phase with negative mode electrospray ionisation (ESI) using a triple quadrupole mass spectrometer (Sciex API 365) with a turboionspray source and post-column addition of acetonitrile for enhanced sensitivity. The MS/MS transitions monitored were m/z 221 -->75 for EtG and 226 -->75 for D(5)-EtG as an internal standard. The method was selective and sensitive, with a detection limit of 51 pg/mg hair at a signal-to-noise ratio of 3:1. The mean recovery was 96%, with an intra- and inter-day precision of less than 11.7% at a concentration of 200 pg/mg. The linearity was assessed in the range of 25-2000 pg/mg hair, with a correlation coefficient of 0.997. The method was successfully applied to 97 human hair samples which were taken at autopsies from persons with known alcoholism or were obtained from alcoholics who were hospitalized for ethanol withdrawal, from social drinkers and from children having not consumed any alcohol. Although, approximately two-third of the alcoholics showed EtG concentrations in hair of higher than 51 pg/mg (up to >4000 pg/mg), in one-third the EtG concentration was below the detection limit. However, only in one of five hair samples of "social drinkers", the EtG concentration was above the detection limit (51 pg/mg). No EtG has been detected in the hair of children. These investigations demonstrate that heavy alcohol consumption may be but not necessarily has to be detectable by EtG analysis in hair.     

Target screening and confirmation of 35 licit and illicit drugs and metabolites in hair by LC-MSMS

A liquid chromatography-tandem mass spectrometry (LC-MSMS) target screening in 50mg hair was developed and fully validated for 35 analytes (Δ9-tetrahidrocannabinol (THC), morphine, 6-acetylmorphine, codeine, methadone, fentanyl, amphetamine, methamphetamine, 3,4-methylenedioxyamphetamine, 3,4-methylenedioxymethamphetamine, benzoylecgonine, cocaine, lysergic acid diethylamide, ketamine, scopolamine, alprazolam, bromazepam, clonazepam, diazepam, flunitrazepam, 7-aminoflunitrazepam, lorazepam, lormetazepam, nordiazepam, oxazepam, tetrazepam, triazolam, zolpidem, zopiclone, amitriptyline, citalopram, clomipramine, fluoxetine, paroxetine and venlafaxine). Hair decontamination was performed with dichloromethane, and incubation in 2 mL of acetonitrile at 50°C overnight. Extraction procedure was performed in 2 steps, first liquid-liquid extraction, hexane:ethyl acetate (55:45, v:v) at pH 9, followed by solid-phase extraction (Strata-X cartridges). Chromatographic separation was performed in AtlantisT3 (2.1 mm × 100 mm, 3 μm) column, acetonitrile and ammonium formate pH 3 as mobile phase, and 32 min total run time. One transition per analyte was monitored in MRM mode. To confirm a positive result, a second injection monitoring 2 transitions was performed. The method was specific (no endogenous interferences, n=9); LOD was 0.2-50 pg/mg and LOQ 0.5-100 pg/mg; linearity ranged from 0.5-100 to 2000-20,000 pg/mg; imprecision <15%; analytical recovery 85-115%; extraction efficiency 4.1-85.6%; and process efficiency 2.5-207.7%; 27 analytes showed ion suppression (up to -86.2%), 4 ion enhancement (up to 647.1%), and 4 no matrix effect; compounds showed good stability 24-48 h in autosampler. The method was applied to 17 forensic cases. In conclusion, a sensitive and specific target screening of 35 analytes in 50mg hair, including drugs of abuse (THC, cocaine, opiates, amphetamines) and medicines (benzodiazepines, antidepressants) was developed and validated, achieving lower cut-offs than Society of Hair Testing recommendations.