气相色谱-质谱结合准确质量测定快速鉴定合成大麻素

采用气相色谱-质谱（GC-MS）结合准确质量测定方法,利用保留指数、特征碎片丰度以及高分辨准确质量数据信息,快速鉴定样品中的合成大麻素组分。建立了JWH-018、JWH-073等12种合成大麻素标准物质的分析信息数据库,检测限达到了2.5~30 ng/mL,并成功利用该方法对未知样品进行了检测,确认样品中含有“JWH-081和JWH-018”合成大麻素组分。结果表明方法可以满足合成大麻素快速鉴定的要求。     

高效液相色谱-三重四极杆质谱法同时测定新型“香料”毒品中的10种合成大麻素

为测定新型"香料"毒品中常见的合成大麻素成分,研究开发了高效液相色谱-三重四极杆质谱联用分析方法。采用安捷伦Poroshell 120 EC-C18(3.0 mm×50 mm,2.7μm)色谱柱,以高纯水-甲醇作流动相进行梯度洗脱,柱温30℃,流速0.3 m L/min。采用电喷雾电离-正离子(ESI+)、负离子(ESI-)分段检测模式,并对合成大麻素的质谱特征和离子碎裂规律进行研究。结果表明,采用该方法可以实现对常见10种合成大麻素的定性和定量分析,正、负离子模式下检测的目标物分别在1~100,10~1 000 ng/m L范围内呈良好线性,日内相对标准偏差(RSD)均不大于3.2%,日间RSD均不大于6.3%。经加标回收率测定和实际样本检验,该方法快速、准确、灵敏、可靠,适用于新型"香料"毒品中常见合成大麻素成分的定性定量检测。     

超高效液相色谱法同时测定电子烟油中的5种吲哚/吲唑酰胺类合成大麻素北大核心CSCD

合成大麻素是目前世界上滥用最多的新精神活性物质之一,其结构多变,更新迅速,目前已发展至新型第八代吲哚/吲唑酰胺类。近年来与吲哚/吲唑酰胺类合成大麻素相关的案件逐渐增多,在实际案件中对缴获物中合成大麻素的定量分析需求随之增多,但相应的检验鉴定技术仍处于发展阶段。本研究针对电子烟油中5种常见的吲哚/吲唑酰胺类合成大麻素,建立了超高效液相色谱法对其同时进行定量分析测定。实验对流动相的种类、洗脱梯度、柱温、检测波长等色谱条件进行了优化,再结合外标法定量,实现了对5种合成大麻素的定量分析。样品用甲醇提取,在Waters ACQUITY UPLC CSH C18(100 mm×2.1 mm,1.7μm)色谱柱上进行分离,柱温35℃,流速0.3 mL/min,进样量1μL,乙腈和超纯水作为流动相进行梯度洗脱,检测波长为290 nm和302 nm。结果表明,采用该方法,5种合成大麻素可在10 min内完全分离,在1~100 mg/L范围内线性关系良好,相关系数(r^(2))均可以达到0.9999,检出限为0.2 mg/L,定量限为0.6 mg/L,满足实际样品分析需求。采用1、10、100 mg/L 3个水平的5种合成大麻素混合标准溶液进行精密度试验,日内精密度(n=6)均小于1.5%,日间精密度(n=6)均小于2.2%。以空白电子烟油为基质样品,在2、10、50 mg/L 3个加标水平下进行加标回收试验,各待测物的平均加标回收率为95.5%~101.9%,相对标准偏差(RSD,n=6)为0.2%~1.5%,准确度为-4.5%~1.9%。本方法具有准确、快速、灵敏、分离效果好等优点,适用于电子烟油中5种吲哚/吲唑酰胺类合成大麻素的定量测定,可满足相关鉴定工作的要求,也可为具有相似结构的合成大麻素的液相色谱定量分析提供参考。     

新型靛红腙类合成大麻素质谱裂解规律研究

研究了新型靛红腙类合成大麻素在电子轰击(EI)和电喷雾(ESI)电离模式下的质谱裂解规律,并建立了可疑物中该类合成大麻素的鉴定方法。采用气相色谱-质谱联用(GC-MS)和液相色谱-高分辨质谱联用(LC-Q-Orbitrap/MS)技术,对5种新型靛红腙类合成大麻素(MDA-19 (BZO-HEXOXIZID),5C-MDA-19 (Pentyl MDA-19,BZO-POXIZID),CHM-MDA-19 (BZO-CHMOXIZID),4en-pentyl MDA-19(BZO-4en-POXIZID),5F-MDA-19 (5F-BZO-POXIZID))的主要碎片离子和碎裂过程进行分析,并对获得的质谱图进行解析,推测该类合成大麻素的EI-MS及ESI-MSⁿ碎裂规律。EI-MS可获得比ESI-MSⁿ更多的碎片离子用于该类合成大麻素的结构推断。碎片离子6,7和8对应的质荷比(m/z)118 (C₈H₈N⁺),132(C₈H₆NO<...     

LTQ-Orbitrap组合式高分辨质谱法快速筛查毛发中7种毒品及代谢物

采用超高压液相色谱-二维线性离子阱结合静电场轨道阱组合式高分辨质谱联用技术(Accela U HPLC/LTQ Orbitrap XL),建立了人毛发中吗啡、O~6-单乙酰吗啡、可待因、乙酰可待因、氯胺酮、去甲氯胺酮和美沙酮毒品及代谢物快速筛查方法。取毛发样品经表层清洗后冷冻研磨粉碎,置于硼酸盐缓冲液(pH 9.2)中超声90 min,离心取上清液,用Oasis HLB柱固相萃取制备。通过静电场轨道阱全扫描得到毒品及其代谢物的精确相对分子质量,同时进行7种毒品及其代谢物的快速筛查。高分辨率质谱可有效去除毛发基质干扰,毒品及其代谢物筛查检出限在0.001～0.02 ng/mg,在0.05～50 ng/mg范围内存在良好线性关系(r>0.9975);本方法平均加标回收率为76.1%～109.6%;日内及日间精密度RSD≤14.9%。本方法灵敏度高,样品制备简便,适用于常见毒品的快速筛查。     

固相萃取/气相色谱-质谱法测定燃香烟气中9种大麻素类新精神活性物质

建立了同时测定燃香烟气中9种大麻素类新精神活性物质的气相色谱-质谱（GC-MS）分析方法。样品在燃香烟气收集装置中经溶剂提取,浓缩离心后,采用HLB固相萃取小柱净化,毛细管色谱柱Agilent HP-5MS毛细管柱（30 m×0.25 mm×0.25μm）分离,使用电子轰击电离（EI）源检测,SIM模式进行质谱监测,内标法对9种大麻素类化合物进行定量。结果表明,9种大麻素类化合物在27 min内完成分离分析,并在0.01～4.0 mg/L范围内具有良好的线性关系,相关系数（r2）均不小于0.999 5,方法的检出限（LOD,S/N=3）和定量下限（LOQ,S/N=10）分别为0.003～0.06 mg/L和0.01～0.2 mg/L。在阴性样品中进行1、2和10倍定量下限的加标回收实验,9种大麻素类化合物的回收率为74.8%～114%,相对标准偏差（RSD,n=6）为1.6%～9.1%。采用该方法对10种市售的燃香样品进行测定,均未检出大麻素类化合物。建立的方法操作简便,可快速对燃香烟气中大麻素类化合物进行定性和定量分析,且能满足发烟量大燃香的检测需求。该方法的建立为我国燃香中大麻素类...     

UPLC-MS/MS法同时测定尿液中的6种苯丙胺类毒品

建立了固相萃取-超高效液相色谱-质谱法(SPE-UPLC-MS/MS)联用技术定量测定尿液中的6种苯丙胺类毒品。样品经Oasis HLB柱提取、纯化后,采用电喷雾离子源电离(ESI)、正离子多反应监测(MRM)模式质谱进行定性和定量分析。6种苯丙胺类毒品分别在0.1~10 ng/m L,0.2~20 ng/m L和0.5~50 ng/m L范围线性良好,相关系数不低于0.9993,提取回收率高于85%,RSD小于10.0%。方法可用于尿液中痕量苯丙胺类毒品的分析。     

超高效液相色谱串联质谱法检验全血中三种合成大麻素

建立了同时测定全血中合成大麻素JWH-018,JWH-250和AM-2201超高效液相色谱-三重四极杆质谱(UPLC-TQ/MS)快速检验方法。分别考察了沉淀蛋白法,改良后的Qu ECh ERS方法和Oasis HLB固相萃取3种前处理方法,以回收率为考察标准,比较了3种前处理方法的优缺点。沉淀蛋白法使用乙腈沉淀血液中的蛋白质,振荡、离心、过膜后直接进样;改良后的Qu ECh ERS方法采用无水M g SO4平衡水相;固相萃取选用HLB柱,对p H、淋洗液、缓冲液等条件进行优化,选用p H 9的硼砂-硼酸(4:1,V/V)缓冲液,甲醇-水(5:95,V/V)溶液为淋洗液,0.1%甲酸-乙腈(1:4,V/V)为洗脱液。选用ZORBAX Eclipse Plus C18色谱柱,以A相0.1%甲酸水和B相0.1%甲酸-乙腈作为流动相,进行梯度洗脱。采用液相色谱-串联质谱仪的电喷雾电离,正离子模式扫描,多反应监测(MRM)模式检测合成大麻素JWH-018,JWH-250及AM-2201。结果表明,3种合成大麻素在其各自的浓度范围内线性关系良好(R20.999);3个添加水平(5,50,100 ng/m L)下,固相萃取方法回收率在70.1%~98.5%之间,改良后的Qu ECh ERS方法回收率在80.3%~108.6%之间,沉淀蛋白法回收率在62.0%~92.3%之间,3种合成大麻素检出限(S/N=3)在0.01~0.05 ng/m L范围内,定量限(S/N=10)在0.05~0.1 ng/m L范围内。     

QuEChERS/高效液相色谱-四极杆-飞行时间质谱法测定巧克力中的18种合成大麻素

建立了检测巧克力中18种合成大麻素的QuEChERS/高效液相色谱-四极杆-飞行时间质谱方法。通过优化提取溶剂种类、提取条件和净化条件，确定200.0 mg巧克力采用1 mL甲醇超声提取10 min，取上层清液加入0.05 g C₁₈和0.05 g PSA净化后，以乙腈和0.1%甲酸水溶液为流动相进行梯度洗脱，采用ZORBAX Eclipse Plus C₁₈(3.0 mm×100 mm,1.8μm)色谱柱进行色谱分离，利用四极杆-飞行时间质谱检测。18种合成大麻素在11 min内实现了分离，5F-EMB-PICA与5F-MDMB-PICA同分异构体通过色谱保留时间和二级质谱碎片实现分辨；5F-EMB-PINACA与5F-ADB同分异构体通过二级质谱碎片实现分辨。18种合成大麻素在1～200μg/L范围内呈良好线性关系，相关系数均不小于0.997 0，检出限为0.02～0.20μg/L，定量下限为0.07～0.66μg/L，在50、100、150μg/kg加标水平下样品的回收率为86.2%～104%，仪器的相对标准偏差(RSD)为0.040%...     

合成大麻素类毒品情报分析：掺杂成分的监测

合成大麻素毒品蔓延趋势严峻,缴获毒品中的掺杂成分既包括生产过程中引入的掺杂物（如前驱体、中间体以及溶剂等）,又包括流通过程中引入的掺杂物（如稀释剂、食品添加剂等）。掺杂成分能体现出特异性的制贩毒路径,可以精细刻画作案手法。本文针对2021年收到的16份疑似合成大麻素类毒品样品,基于其中2种主要毒品成分MDMB-4en-PINACA和ADB-BUTINACA,进行掺杂成分的预判。使用气相色谱-质谱联用法,根据总离子流图确定9种掺杂成分,基于吲唑酰鎓离子（MW=145）确定17种掺杂成分;通过系统分析16份毒品样品的掺杂物,总结出一系列合成大麻素类毒品情报,为相关禁毒工作提供技术支撑。     

Deposition of JWH-018, JWH-073 and their metabolites in hair and effect of hair pigmentation

Analysis of drugs in hair is often used as a routine method to obtain detailed information about drug ingestion. However, few studies have been conducted on deposition of synthetic cannabinoids and metabolites in hair. The first purpose of this study was to establish and validate an analytical method for detection of JWH-018, JWH-073, and their metabolites in hair, by use of UHPLC–MS–MS, for forensic application. The second purpose was to investigate the distribution of synthetic cannabinoids metabolites in hair and the effect of hair pigmentation, by use of an animal model. For this, JWH-073 was chosen as a representative synthetic cannabinoid. Finally, the developed method was applied to hair samples from 18 individuals suspected of synthetic cannabinoids use. JWH-018, JWH-073, and their metabolites were extracted from hair with methanol. The extract was then filtered and analyzed by UHPLC–MS–MS with an electrospray ion source in positive-ionization mode. Validation proved the method was selective, sensitive, accurate, and precise, with acceptable linearity within the calibration ranges. No significant variations were observed when different sources of both human and rat hair were used. The animal study demonstrated that JWH-073 N-COOH M was the major metabolite of JWH-073 in rat hair, and hair pigmentation did not have a significant effect on incorporation of JWH-073 and its metabolites into hair. In the analysis of 18 authentic hair samples, only JWH-018, JWH-018 N-5-OH M, and JWH-073 were detected, with wide variation in concentrations.     

Simultaneous analysis of synthetic cannabinoids in the materials seized during drug trafficking using GC-MS

A rapid and simple gas chromatography–mass spectrometry (GC-MS) method was developed and validated to identify and quantify synthetic cannabinoids in the materials seized during drug trafficking. Accuracy and reproducibility of the method were improved by using deuterated JWH-018 and JWH-073 as internal standards. Validation results of the GC-MS method showed that it was suitable for simultaneous qualitative and quantitative analyses of synthetic cannabinoids, and we analyzed synthetic cannabinoids in seized materials using the validated GC-MS method. As a result of the analysis, ten species of synthetic cannabinoids were identified in dried leaves (n?=?40), bulk powders (n?=?6), and tablets (n?=?14) seized in Korea during 2009–2012, as a single ingredient or as a mixture with other active co-ingredients. JWH-018 and JWH-073 were the most frequently identified compounds in the seized materials. Synthetic cannabinoids in the dried leaves showed broad concentration ranges, which may cause unexpected toxicity to abusers. The bulk powders were considered as raw materials used to prepare legal highs, and they contained single ingredient of JWH-073, JWH-019, or JWH-250 with the purity over 70 %. In contrast, JWH-018 and JWH-073 contents in the tablets were 7.1–13.8 and 3.0–10.2 mg/g, respectively. Relatively low contents in the tablets suggest that the synthetic cannabinoids may have been added to the tablets as supplements to other active co-ingredients.     

LC-QTOF-MS as a superior strategy to immunoassay for the comprehensive analysis of synthetic cannabinoids in urine

The objective of this study was to compare the performance of an immunoassay screening for synthetic cannabinoids with a newly developed confirmation method using liquid chromatography quadrupole time-of-flight mass spectrometry. The screening included metabolites from JWH-018, JWH-073, and AM-2201. The confirmation included metabolites from AM-2201, JWH-018, JWH-019, JWH-073, JWH-081, JWH-122, JWH-210, JWH-250, JWH-398, MAM-2201, RCS-4, and UR-144. The immunoassay was tested and found to have no cross-reactivity with UR-144 metabolites but considerable cross-reactivity with MAM-2201 and JWH-122 metabolites. Sensitivity and specificity for the immunoassay were evaluated with 87 authentic urine samples and found to be 87 % and 82 %, respectively. With a cutoff at 2 ng/ml, the confirmation showed 80 positive findings in 38 cases. The most common finding was JWH-122 5-OH-pentyl, followed by JWH-018 5-OH-pentyl. There were 9 findings of UR-144 metabolites and 3 of JWH-073 metabolites. In summary, the immunoassay performed well, presenting both high sensitivity and specificity for the synthetic cannabinoids present in the urine samples tested. The rapid exchange of one cannabinoid for another may pose problems for immunoassays as well as for confirmation methods. However, we consider time-of-flight mass spectrometry to be superior since new metabolites can be quickly included and identified.

Analysis of ketamine and norketamine in hair samples using molecularly imprinted solid-phase extraction (MISPE) and liquid chromatography–tandem mass spectrometry (LC-MS/MS)

An anti-ketamine molecularly imprinted polymer (MIP) was synthesized and used as the sorbent in a solid-phase extraction protocol to isolate ketamine and norketamine from human hair extracts prior to LC-MS/MS analysis. Under optimised conditions, the MIP was capable of selectively rebinding ketamine, a licensed anaesthetic that is widely misused as a recreational drug, with an apparent binding capacity of 0.13 μg ketamine per mg polymer. The limit of detection (LOD) and lower limit of quantification (LLOQ) for both ketamine and norketamine were 0.1 ng/mg hair and 0.2 ng/mg hair, respectively, when 10 mg hair were analysed. The method was linear from 0.1 to 10 ng/mg hair, with correlation coefficients (R ²) of better than 0.99 for both ketamine and norketamine. Recoveries from hair samples spiked with ketamine and norketamine at a concentration of 50 ng/mg were 86% and 88%, respectively. The method showed good intra- and interday precisions (<5%) for both analytes. Minimal matrix effects were observed during the LC-MS/MS analysis of ketamine (ion suppression −6.8%) and norketamine (ion enhancement +0.2%). Results for forensic case samples demonstrated that the method successfully detected ketamine and norketamine concentrations in hair samples with analyte concentrations ranging from 0.2 to 5.7 ng/mg and from 0.1 to 1.2 ng/mg, respectively.     

Identification of a novel cannabimimetic phenylacetylindole, cannabipiperidiethanone, as a designer drug in a herbal product and its affinity for cannabinoid CB₁ and CB₂ receptors

A new cannabimimetic phenylacetylindole (cannabipiperidiethanone, 1) has been found as an adulterant in a herbal product which contains two other known synthetic cannabinoids, JWH-122 and JWH-081, and which is distributed illegally in Japan. The identification was based on analyses using GC-MS, LC-MS, high-resolution MS and NMR. Accurate mass spectrum measurement showed the protonated molecular ion peak of 1 at m/z 377.2233 [M+H]? and the molecular formula of 1 was C??H??N?O?. Both mass and NMR spectrometric data revealed that 1 was 2-(2-methoxyphenyl)-1-{1-[(1-methylpiperidin-2-yl)methyl]-1H-indol-3-yl}ethanone. Compound 1 has a mixed structure of known cannabimimetic compounds: JWH-250 and AM-2233. Namely, the moiety of phenylacetyl indole and N-methylpiperidin-2-yl-methyl correspond to the structure of JWH-250 and AM-2233, respectively. However, no synthetic, chemical or biological information about 1 has been reported. A binding assay of compound 1 to cannabinoid receptors revealed that 1 has affinity for the CB? and CB? (IC??=591, 968 nM, respectively) receptors, and shows 2.3- and 9.4-fold lower affinities than those of JWH-250. This is the first report to identify cannabimimetic compound (1) as a designer drug and to show its binding affinity to cannabinoid receptors.     

Development and validation of a liquid chromatography-tandem mass spectrometry method for the quantitation of synthetic cannabinoids of the aminoalkylindole type and methanandamide in serum and its application to forensic samples

After the discovery of synthetic cannabimimetic substances in 'Spice'-like herbal mixtures marketed as 'incense' or 'plant fertilizer' the active compounds have been declared as controlled substances in several European countries. As expected, a monitoring of new herbal mixtures which continue to appear on the market revealed that shortly after control measures have been taken by legal authorities, other compounds were added to existing mixtures and to new products. Several compounds of the aminoalkylindole type have been detected so far in herbal mixtures but still their consumption cannot be detected by commonly used drug-screening procedures, encouraging drug users to substitute cannabis with those products. There is a increasing demand on the part of police authorities, hospitals and psychiatrists for detection and quantification of synthetic cannabinoids in biological samples originating from psychiatric inpatients, emergency units or assessment of fitness to drive. Therefore, a liquid chromatography-tandem mass spectrometry method after liquid-liquid extraction for the quantitation of JWH-015, JWH-018, JWH-073, JWH-081, JWH 200, JWH-250, WIN 55,212-2 and methanandamide and the detection of JWH-019 and JWH-020 in human serum has been developed and fully validated according to guidelines for forensic toxicological analyses. The method was successfully applied to 101 serum samples from 80 subjects provided by hospitals, detoxification and therapy centers, forensic psychiatric centers and police authorities. Fifty-seven samples or 56.4% were found positive for at least one aminoalkylindole. JWH-019, JWH-020, JWH-200, WIN 55,212-2 and methanandamide were not detected in any of the analyzed samples.     

Metabolism of JWH-015, JWH-098, JWH-251, and JWH-307 in silico and in vitro: a pilot study for the detection of unknown synthetic cannabinoids metabolites

This pilot study was performed to study the main metabolic reactions of four synthetic cannabinoids: JWH-015, JWH-098, JWH-251, and JWH-307 in order to setup a screening method for the detection of main metabolites in biological fluids. In silico prediction of main metabolic reactions was performed using MetaSite^? software. To evaluate the agreement between software prediction and experimental reactions, we performed in vitro experiments on the same JWHs using rat liver slices. The obtained samples were analyzed by liquid chromatography-quadrupole time-of-flight and the identification of metabolites was executed using Mass-MetaSite^? software that automatically assigned the metabolite structures to the peaks detected based on their accurate masses and fragmentation. A comparison between the experimental findings and the in silico metabolism prediction using MetaSite^? software showed a good accordance between experimental and in silico data. Thus, the use of in silico metabolism prediction might represent a useful tool for the forensic and clinical toxicologist to identify possible main biomarkers for synthetic cannabinoids in biological fluids, especially urine, following their administration.

Analysis of 30 synthetic cannabinoids in serum by liquid chromatography‐electrospray ionization tandem mass spectrometry after liquid‐liquid extraction

The analysis of synthetic cannabinoids in human matrices is of particular importance in the fields of forensic and clinical toxicology since cannabis users partly shift to the consumption of ‘herbal mixtures’ as a legal alternative to cannabis products in order to circumvent drug testing. However, comprehensive methods covering the majority of synthetic cannabinoids already identified on the drug market are still lacking. In this article, we present a fully validated method for the analysis of 30 synthetic cannabinoids in human serum utilizing liquid‐liquid extraction and liquid chromatography‐electrospray ionization tandem mass spectrometry. The method proved to be suitable for the quantification of 27 substances. The limits of detection ranged from 0.01 to 2.0 ng/mL, whereas the lower limits of quantification were in the range from 0.1 to 2.0 ng/mL. The presented method was successfully applied to 833 authentic serum samples during routine analysis between August 2011 and January 2012. A total of 227 (27%) samples was tested positive for at least one of the following synthetic cannabinoids: JWH‐018, JWH‐019, JWH‐073, JWH‐081, JWH‐122, JWH‐200, JWH‐203, JWH‐210, JWH‐307, AM‐2201 and RCS‐4. The most prevalent compounds in positive samples were JWH‐210 (80%), JWH‐122 (63%) as well as AM‐2201 (29%). Median serum concentrations were all below 1.0 ng/mL. These findings demonstrate a significant shift of the market of synthetic cannabinoids towards substances featuring a higher CB₁ binding affinity and clearly emphasize that the analysis of synthetic cannabinoids in serum or blood samples requires highly sensitive analytical methods covering a wide spectrum of substances. Copyright © 2012 John Wiley & Sons, Ltd.