液相色谱-串联质谱同时分析尿液中的大麻酚类和△^9-四氢大麻酸

目的建立LC/MS-MS同时检测尿液中Δ9-四氢大麻酚(THC)、大麻酚(CBN)、大麻二酚(CBD)和大麻主要代谢物Δ9-四氢大麻酸(THC-COOH)的方法.方法屎液样本经碱水解,加入氘代四氢大麻酸Δ9-d9-THC-COOH)内标,经V(正己烷)V(乙酸乙酯)=91提取,吹干,以100μL乙腈定容,利用LC/MS-MS方法进行分析.结果THC-COOH、CBN、THC和CBD的最低检测出质量浓度为0.2、0.4、1.0和2.0ng/mL;在阳性尿液中检出THC-COOH成分,质量浓度为335.9 ng/mL.结论所建立的方法简便快速、灵敏度高、专属性强,可满足检测尿液中THC、CBN、CBD以及大麻主要代谢物THC-COOH的要求.     

GC／MS同时分析头发中大麻酚类和△^9-四氢大麻酸

目的建立同时分析毛发中THC、CBN、CBD和大麻主要代谢物THC-COOH的方法.方法头发检材去污处理后,加入内标氯灭酸,经NaOH水解和正己烷:乙酸乙酯(9:1)提取,吹干后衍生化,利用GC/MS-SIM方法分析.结果THC、CBN和THC-COOH的最低检出限分别为0.01、0.05和0.01ng/mg.10例阳性头发中均检出THC成分,THC浓度范围为0J 1～8.84ng/mg,有3例未检出THC-COOH,检出者的量亦低于定量下限.结论同时分析头发中的大麻酚类和△9-四氢大麻酚是可行的,头发中THC-COOH浓度明显都低于THC浓度.     

反相HPLC法同时测定大麻植物中的三种有效成分

目的建立同时测定大麻植物中四氢大麻酚(tetrahydrocannabinol,THC)、大麻二酚(cannabidiol,CBD)和大麻酚(cannabinol,CBN)三种有效成分的高效液相色谱(HPLC)法。方法采用通用C_(18)色谱柱,以乙腈-磷酸盐缓冲液(0.015 mol/L KH_2PO_4)为流动相,流速为1.0 m L/min,检测波长为220 nm,同时收集波长190~400nm的紫外光谱图,并以此光谱图及保留时间作为定性依据。结果所建方法能良好地分离THC、CBD和CBN,三种成分在0.4~40μg/m L范围内线性关系良好(R~2≥0.999 3),回收率大于87%,最低检出限分别为1.8、2.0和1.3 ng,日内精密度与日间精密度均小于5%。结论反相HPLC法简便、快速、准确,适用于大麻植物中THC、CBD和CBN的定性定量检测。     

大麻毒品的检验方法

本文用改良的薄层色谱和气相色谱法对走私贩毒的大麻毒品进行定性、定量分析,两种方法均能完全分离出大麻的3种主要成分:大麻酚(CBN)、四氢大麻酚(THC)及大麻二酚(CBD),灵敏度:薄层法0.15μg,气相色谱法10ng     

新疆麻烟活性成分气相色谱法测定

本文报告了气相色谱法测定新疆麻烟中活性成分Δ~9-四氢大麻酚(Δ~9-THC)、大麻二酚(CBD)、大麻酚(CBN)含量的方法,以24烷为内标,采用3％OV-17玻璃柱。实验所得平均回收率为98.0～103.0％。CV％不大于5％。方法简便、准确。     

大麻中主要成份的色谱予柱净化和HPLC测定

<正> 大麻成份极其复杂,多达近百种。法庭科学主要是对大麻中的 CBD(大麻二酚)、CBN(大麻酚)和△~9—THC(四氢大麻酚)三种有效成份进行定性及定量测定。高效液相测定大麻中的主要成份是将有机溶剂提取液未经净化,直接进高效液相色谱仪,样品分析时间长。本实验将SEP—PAK ALUMINAN 小滤柱应用于大麻的     

胶体金标记检测大麻单克隆抗体免疫试剂盒研制

目的 建立准确、快速、简便的检测尿液中大麻的胶体金免疫层析技术(ICT)。方法 采用柠檬酸三钠还原法制备胶体金颗粒,标记抗主要代谢物四氢大麻酚-9-羧酸,THC与GC/MS检测方法相比,用ICT法的216份尿样,其检测限为50 ng/ml,灵敏度为96.67％,准确性为98.61％。结论 ICT法检测尿液中THC,其特异性强,对确定大麻的存在具有广泛的应用价值。     

大麻遗传标记的法医学应用研究进展

大麻是大麻科大麻属一年生雌雄异株的草本植物,其内含有具有强烈成瘾性和麻醉性的四氢大麻酚(THC).大麻价格低廉、获取方便、且受到一些国家和地区合法化的影响,目前已成为滥用最广泛的毒品之一.因此,大麻植株的鉴定对于打击毒品犯罪、维护社会稳定具有重要意义.近年来,基于DNA遗传标记的大麻鉴定为案件侦破提供了新的技术手段,针...     

电喷雾和大气压化学法对大麻酚类物质离子化的比较

目的比较电喷雾电离(ESI)和大气压化学电离(APCI)两种模式对大麻酚类物质的离子化效果。方法采用UFLC-(ESI/APCI)MS分析方法,分别考察使用ESI和APCI时雾化电压、雾化气流量、干燥气流量、加热块温度、解离管温度等参数变化对大麻酚类物质的影响规律,确定最优条件参数组合,并比较ESI和APCI对大麻酚类物质的离子化效果。结果对于0.5μg/mL大麻二酚、大麻酚和四氢大麻酚标准品,ESI峰高分别为215 006、143 051、216 944,信噪比41.74、49.88、42.12,峰面积日内标准偏差(RSD)<3.96%;APCI峰高分别为140 238、226 505、247 753,信噪比78.37、131.03、138.46,峰面积日内标准偏差(RSD)<11.98%。结论检测大麻样品时,ESI为首选,基质复杂时可以使用APCI作为ESI的补充手段。     

涉及新型合成大麻素ADB-BUTINACA的疑似性侵案1例

正近年来,新型合成大麻素已成为毒品检测报告中最常见的新精神活性物质之一[1],有研究表明其毒性远大于四氢大麻酚,滥用会导致心肌梗死、中风、急性肾损伤、癫痫及各种并发症[2-5],生产者和使用者为了回避法律管控是合成大麻素种类不断出新的重要原因[6, 7]。本案例中检出的合成大麻素N-(1-氨甲酰基-2,2-二甲基丙基)-1-丁基吲唑-3-甲酰胺(ADB-BUTINACA)是一种大麻受体的完全激动剂,     

Validated method for the simultaneous determination of Delta9-THC and Delta9-THC-COOH in oral fluid, urine and whole blood using solid-phase extraction and liquid chromatography-mass spectrometry with electrospray ionization

A fully validated, sensitive and specific method for the extraction and quantification of Delta(9)-tetrahydrocannabinol (THC) and 11-nor-9-carboxy-Delta(9)-THC (THC-COOH) and for the detection of 11-hydroxy-Delta(9)-THC (11-OH THC) in oral fluid, urine and whole blood is presented. Solid-phase extraction and liquid chromatography-mass spectrometry (LC-MS) technique were used, with electrospray ionization. Three ions were monitored for THC and THC-COOH and two for 11-OH THC. The compounds were quantified by selected ion recording of m/z 315.31, 329.18 and 343.16 for THC, 11-OH THC and THC-COOH, respectively, and m/z 318.27 and 346.26 for the deuterated internal standards, THC-d(3) and THC-COOH-d(3), respectively. The method proved to be precise for THC and THC-COOH both in terms of intra-day and inter-day analysis, with intra-day coefficients of variation (CV) less than 6.3, 6.6 and 6.5% for THC in saliva, urine and blood, respectively, and 6.8 and 7.7% for THC-COOH in urine and blood, respectively. Day-to-day CVs were less than 3.5, 4.9 and 11.3% for THC in saliva, urine and blood, respectively, and 6.2 and 6.4% for THC-COOH in urine and blood, respectively. Limits of detection (LOD) were 2 ng/mL for THC in oral fluid and 0.5 ng/mL for THC and THC-COOH and 20 ng/mL for 11-OH THC, in urine and blood. Calibration curves showed a linear relationship for THC and THC-COOH in all samples (r(2)>0.999) within the range investigated. The procedure presented here has high specificity, selectivity and sensitivity. It can be regarded as an alternative method to GC-MS for the confirmation of positive immunoassay test results, and can be used as a suitable analytical tool for the quantification of THC and THC-COOH in oral fluid, urine and/or blood samples.     

A preliminary investigation on the distribution of cannabinoids in man

An LC/MS/MS procedure to determine THC along with its major metabolites 11-OH-THC, THC-COOH and its glucuronide as well as the cannabinoids CBD and CBN was applied to 5 post mortem cases to study their distribution into some less commonly studied matrices. Analytes were determined in fluids and tissue homogenates following protein precipitation and liquid-liquid extraction. Gall bladder fluid exhibited maximum concentrations of all analytes except THC, which was detectable in high concentrations in muscle tissue along with CBD. THC was also present in lung specimens, whereas its concentration in liver samples was low or not detectable at all. Liver und kidney specimens contained appreciable amounts of THC-COOglu. Findings from bile support extensive enterohepatic recirculation of the glucuronide. Muscle tissue seems an interesting specimen to detect multiple cannabis use, and brain may serve as an alternative specimen for blood; nevertheless, the present findings should be substantiated by further investigations.     

Weather-induced changes in cannabinoid content of hair

Authentic hair samples from Cannabis users and a drug free hair sample which was separately spiked with tetrahydrocannabinol (THC), cannabidiol (CBD) or cannabinol (CBN) were exposed outside as well as to natural sunlight at prevailing and elevated humidity in quartz glass tubes during 8 weeks. In addition, authentic and spiked hair samples were exposed to xenon arc radiation in a light exposure cabinet for 24 hours. Stability of THC, CBD and CBN in authentic samples differed from that of the spiked hair. The radiation experiment revealed that CBN could not be measured in hair which had been spiked with THC. Under all conditions chosen the concentrations of THC, CBD and CBN decreased. At high humidity the concentrations declined more rapidly. In both authentic and spiked samples THC was most unstable compared to CBD and CBN. Therefore, in hair analysis determination of CBD and CBN seems promising to detect Cannabis exposure even under unfavorable conditions.     

Validated method for the simultaneous determination of Δ-THC and Δ-THC-COOH in oral fluid, urine and whole blood using solid-phase extraction and liquid chromatography–mass spectrometry with electrospray ionization

A fully validated, sensitive and specific method for the extraction and quantification of Δ⁹-tetrahydrocannabinol (THC) and 11-nor-9-carboxy-Δ⁹-THC (THC-COOH) and for the detection of 11-hydroxy-Δ⁹-THC (11-OH THC) in oral fluid, urine and whole blood is presented. Solid-phase extraction and liquid chromatography–mass spectrometry (LC–MS) technique were used, with electrospray ionization. Three ions were monitored for THC and THC-COOH and two for 11-OH THC. The compounds were quantified by selected ion recording of m/z 315.31, 329.18 and 343.16 for THC, 11-OH THC and THC-COOH, respectively, and m/z 318.27 and 346.26 for the deuterated internal standards, THC-d₃ and THC-COOH-d₃, respectively. The method proved to be precise for THC and THC-COOH both in terms of intra-day and inter-day analysis, with intra-day coefficients of variation (CV) less than 6.3, 6.6 and 6.5% for THC in saliva, urine and blood, respectively, and 6.8 and 7.7% for THC-COOH in urine and blood, respectively. Day-to-day CVs were less than 3.5, 4.9 and 11.3% for THC in saliva, urine and blood, respectively, and 6.2 and 6.4% for THC-COOH in urine and blood, respectively. Limits of detection (LOD) were 2 ng/mL for THC in oral fluid and 0.5 ng/mL for THC and THC-COOH and 20 ng/mL for 11-OH THC, in urine and blood. Calibration curves showed a linear relationship for THC and THC-COOH in all samples (r² > 0.999) within the range investigated.The procedure presented here has high specificity, selectivity and sensitivity. It can be regarded as an alternative method to GC–MS for the confirmation of positive immunoassay test results, and can be used as a suitable analytical tool for the quantification of THC and THC-COOH in oral fluid, urine and/or blood samples.     

Cannabinoids determination in oral fluid by SPME–GC/MS and UHPLC–MS/MS and its application on suspected drivers

The confirmation of Δ9-tetrahydrocannabinol (THC) in oral fluid (OF) is an important issue for assessing Driving Under the Influence of Drugs (DUID). The aim of this research was to develop a highly sensitive method with minimal sample pre-treatment suitable for the analysis of small OF volumes (100 μL) for the confirmation of cannabinoids in DUID cases. Two methods were compared for the confirmation of THC in residual OF samples, obtained from a preliminary on-site screening with commercial devices. An ultra high performance LC–MS (UHPLC–MS/MS) method and an SPME–GC/MS method were hence developed. 100 μL of the residual mixture OF/preservative buffer or neat OF was simply added to 10 μL of THC-D3 (1 μg/mL) and submitted to the two different analyses: A — direct injection of 10 μL in UHPLC–MS/MS in positive electrospray ionisation (ESI) mode and B — sampling for 30 min with SPME (100 μm polydimethylsiloxane or PDMS fibre) and direct injection by desorption of the fibre in the GC injection port.The lowest limit of detection (LLOD) of THC was 2 ng/mL in UHPLC–MS/MS and 0.5 ng/mL in SPME–GC/MS. In addition, cannabidiol (CBD) and cannabinol (CBN) could be detected in GC/MS equipment at 2 ng/mL, whilst in UHPLC–MS/MS the LLOD was 20 ng/mL.Both methods were applied to 70 samples coming from roadside tests. By SPME–GC/MS analysis, THC was confirmed in 42 samples, whilst CBD was detected in 21 of them, along with CBN in 14 samples. THC concentrations ranged from traces below the lowest limit of quantification or LLOQ (2 ng/mL) up to 690 ng/mL.     

Characterization of Algerian-Seized Hashish Over Eight Years (2011–2018). Part II: Chemical Categorization

In Algeria, large quantities of hashish are seized every year. This study aimed to investigate the total content of major cannabinoids in the illicit seized hashish in Algeria over an 8-year period (2011–2018) in order to establish the chemical profile of North African hashish. A total of 3265 hashish samples were analyzed using a validated high-performance liquid chromatography–diode array detection (HPLC-DAD) method, allowing the simultaneous quantification of both the acidic and the neutral forms of Δ9-tetrahydrocannabinol (THC), cannabidiol (CBD), and cannabinol (CBN). The results revealed a slight upward trend in the mean THC content, from 7.0% in 2011 to 9.4% in 2018, with an overall mean value of 8.4%. The overall means of CBD and CBN content were 3.5% and 0.8%, respectively. The number of high-potency hashish samples gradually increased to reach 6% in 2018. Two distinct hashish chemotypes were identified: the highly populated chemotype II, corresponding to the traditional medium-potency hashish ([THC + CBN]/CBD ~ 2.16), and chemotype I, containing hashish samples of relatively high THC levels and low levels of CBD (ratio ~ 4.90). Both chemotypes I and II were characterized in the ternary plot, and the proportions (THC:CBD:CBN) were about 85%:13%:2% and 60%:35%:5%, respectively.     

A survey of the potency of Japanese illicit cannabis in fiscal year 2010

In recent years, increased 'cannabis potency', or Δ(9)-tetrahydrocannabinol (THC) content in cannabis products, has been reported in many countries. A survey of Japanese illicit cannabis was conducted from April 2010 to March 2011. In Japan, all cannabis evidence is transferred to the Minister of Health, Labour and Welfare after criminal trials. The evidence was observed at Narcotics Control Department offices in major 11 cities. The total number of cannabis samples observed was 9072, of which 6376 were marijuana. The marijuana seizures were further classified, and it was found that in terms of the number of samples, 65.2% of them were seedless buds, and by weight 73.0% of them were seedless buds. Seedless buds were supposed to be 'sinsemilla', a potent class of marijuana. THC, cannabinol (CBN) and cannabidiol (CBD) in marijuana seizures exceeding 1g were quantified. The number of samples analyzed was 1115. Many of them were shown to contain CBN, an oxidative product from THC. This was a sign of long-term storage of the cannabis and of the degradation of THC. Relatively fresh cannabis, defined by a CBN/THC ratio of less than or equal to 0.1, was chosen for analysis. Fresh seedless buds (335 samples) contained an average of 11.2% and a maximum of 22.6% THC. These values are comparable to those of 'high potency cannabis' as defined in previous studies. Thus, this study shows that highly potent cannabis products are distributed in Japan as in other countries.     

Hair analysis for delta(9)-THC,delta(9)-THC-COOH,CBN and CBD,by GC/MS-EI. Comparison with GC/MS-NCI for delta(9)-THC-COOH

A sensitive analytical method was developed for quantitative analysis of delta(9)-tetrahydrocannabinol (delta(9)-THC), 11-nor-delta(9)-tetrahydrocannabinol-carboxylic acid (delta(9)-THC-COOH), cannabinol (CBN) and cannabidiol (CBD) in human hair. The identification of delta(9)-THC-COOH in hair would document Cannabis use more effectively than the detection of parent drug (delta(9)-THC) which might have come from environmental exposure. Ketamine was added to hair samples as internal standard for CBN and CBD. Ketoprofen was added to hair samples as internal standard for the other compounds. Samples were hydrolyzed with beta-glucuronidase/arylsulfatase for 2h at 40 degrees C. After cooling, samples were extracted with a liquid-liquid extraction procedure (with chloroform/isopropyl alcohol, after alkalinization, and n-hexane/ethyl acetate, after acidification), which was developed in our laboratory. The extracts were analysed before and after derivatization with pentafluoropropionic anhydride (PFPA) and pentafluoropropanol (PFPOH) using a Hewlett Packard gas chromatographer/mass spectrometer detector, in electron impact mode (GC/MS-EI). Derivatized delta(9)-THC-COOH was also analysed using a Hewlett Packard gas chromatographer/mass spectrometer detector, in negative ion chemical ionization mode (GC/MS-NCI) using methane as the reagent gas. Responses were linear ranging from 0.10 to 5.00 ng/mg hair for delta(9)-THC and CBN, 0.10-10.00 ng/mg hair for CBD, 0.01-5.00 ng/mg for delta(9)-THC-COOH (r(2)>0.99). The intra-assay precisions ranged from <0.01 to 12.40%. Extraction recoveries ranged from 80.9 to 104.0% for delta(9)-THC, 85.9-100.0% for delta(9)-THC-COOH, 76.7-95.8% for CBN and 71.0-94.0% for CBD. The analytical method was applied to 87 human hair samples, obtained from individuals who testified in court of having committed drug related crimes. Quantification of delta(9)-THC-COOH using GC/MS-NCI was found to be more convenient than GC/MS-EI. The latter may give rise to false negatives due to the detection limit.     

First systematic evaluation of the potency of Cannabis sativa plants grown in Albania

Cannabis products (marijuana, hashish, cannabis oil) are the most frequently abused illegal substances worldwide. Delta-9-tetrahydrocannabinol (THC) is the main psychoactive component of Cannabis sativa plant, whereas cannabidiol (CBD) and cannabinol (CBN) are other major but no psychoactive constituents. Many studies have already been carried out on these compounds and chemical research was encouraged due to the legal implications concerning the misuse of marijuana. The aim of this study was to determine THC, CBD and CBN in a significant number of cannabis samples of Albanian origin, where cannabis is the most frequently used drug of abuse, in order to evaluate and classify them according to their cannabinoid composition. A GC-MS method was used, in order to assay cannabinoid content of hemp samples harvested at different maturation degree levels during the summer months and grown in different areas of Albania. This method can also be used for the determination of plant phenotype, the evaluation of psychoactive potency and the control of material quality. The highest cannabinoid concentrations were found in the flowers of cannabis. The THC concentrations in different locations of Albania ranged from 1.07 to 12.13%. The influence of environmental conditions on cannabinoid content is discussed. The cannabinoid content of cannabis plants were used for their profiling, and it was used for their classification, according to their geographical origin. The determined concentrations justify the fact that Albania is an area where cannabis is extensively cultivated for illegal purposes.     

Simultaneous quantitation of delta-9-tetrahydrocannabinol (THC) and 11-nor-9-carboxy-delta-9-tetrahydrocannabinol (THC-COOH) in serum by GC/MS using deuterated internal standards and its application to a smoking study and forensic cases.

A new procedure for the simultaneous detection of delta-9-tetrahydrocannabinol (THC) and its major metabolite, 11-nor-9-carboxy-delta-9-tetrahydrocannabinol (THC-COOH) in serum has been evaluated. The method combines rapid, efficient, solid-phase extraction and simple derivatization by methylation. Analysis and quantitation is performed by gas chromatography/mass spectrometry (GC/MS) using deuterated cannabinoids as internal standards (IS). Reproducibility and sensitivity of the method are good. The procedure is applied to serum specimens collected from a smoking study with 24 volunteers and 212 forensic cases. Results are interpreted based upon the current knowledge about THC metabolism and pharmacokinetics.