Determination of Tetrahydrocannabinol and Δ9-Tetrahydrocannabinic Acid in Urine by Liquid Chromatography-Tandem Mass Spectrometry with Solid Phase Extraction
摘 要
提出了固相萃取-液相色谱-串联质谱法测定尿液中四氢大麻酚(THC)和Δ9-四氢大麻酸(THC-COOH)含量的方法。取尿液样品1 mL,加入1 mol·L-1 NaOH溶液1 mL,于60 ℃加热15 min后过活化好的MAX混合阴离子固相萃取柱。用1 mL 2%(体积分数)氨水溶液固定目标物,用1 mL 80%(体积分数)甲醇溶液进行淋洗,最后用1 mL含5%(体积分数)甲酸的甲醇溶液进行洗脱,收集洗脱液,按照仪器工作条件进行测定。以Agilent Poroshell HPH-C18色谱柱(50 mm×4.6 mm,4 μm)为固定相,以不同体积比的5 mmol·L-1乙酸铵溶液-甲醇混合液为流动相进行梯度洗脱;分离后的THC和THC-COOH经电喷雾离子源负离子模式扫描,多反应监测模式检测。结果表明:THC和THC-COOH的质量浓度在一定范围内与对应的峰面积呈线性关系,检出限(3S/N)分别为0.003,0.05 μg·L-1;按照标准加入法对空白样品进行加标回收试验,回收率为81.2%~99.0%,日内精密度(n=5)为1.1%~6.0%,日间精密度(n=5)为2.2%~6.4%;方法用于一份疑似吸食大麻人员的尿液样品分析,检出THC和THC-COOH,检出量分别为5.8 μg·L-1和56.6 μg·L-1。
Abstract
A method for the determination of tetrahydrocannabinol (THC) and Δ9-tetrahydrocannabinic acid (THC-COOH) in urine by liquid chromatography-tandem mass spectrometry with solid phase extraction was proposed. The urine sample (1 mL) was taken, and 1 mL of 1 mol·L-1 NaOH solution was added. The mixed solution was heated at 60 ℃ for 15 min, and then passed through activated MAX mixed anionic solid phase extraction column. Using 1 mL of 2% (volume fraction) ammonia solution to fix the targets, with 1 mL of 80% (volume fraction) methanol solution for rinsing, and 1 mL of methanol solution containing 5% (volume fraction) formic acid for elution, the collected eluent was determined according to the working conditions of the instrument. Gradient elution was performed using Agilent Poroshell HPH-C18 column (50 mm×4.6 mm, 4 μm) as the stationary phase, and the mixed solutions composed of 5 mmol·L-1 ammonium solution and methanol with different volume ratios as the mobile phase. The separated THC and THC-COOH were scanned by electrospray ion source with negative ion mode, and detected by multi-reaction monitoring mode. It was shown that linear relationships between the mass concentrations and corresponding peak areas of THC and THC-COOH were kept in definite ranges, with detection limits (3S/N) of 0.003 μg·L-1 and 0.05 μg·L-1, respectively. Test for recovery was made on the blank sample by standard addition method, giving results in the range of 81.2%-99.0%, with precisions of intra-day (n=5) and inter-day (n=5) were in the ranges of 1.1%-6.0% and 2.2%-6.4%, respectively. The method was used to analyze one urine sample of a suspected cannabis user, in which THC and THC-COOH were detected, with detection amounts of 5.8 μg·L-1 and 56.6 μg·L-1, respectively.
中图分类号 O657.63 DOI 10.11973/lhjy-hx202310008
所属栏目 工作简报
基金项目 贵州省科技厅科技支撑项目(黔科合支撑[2021]一般529);贵州省哲学社会科学规划课题(20GZYB47)
收稿日期 2022/1/28
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联系人作者王燕(66515076@qq.com)
备注赵志东,副教授,主要研究方向为毒物毒品与微量物证分析
引用该论文: ZHAO Zhidong,YU Liyu,MENG Jiao,WANG Yan. Determination of Tetrahydrocannabinol and Δ9-Tetrahydrocannabinic Acid in Urine by Liquid Chromatography-Tandem Mass Spectrometry with Solid Phase Extraction[J]. Physical Testing and Chemical Analysis part B:Chemical Analysis, 2023, 59(10): 1156~1161
赵志东,余丽宇,孟娇,王燕. 固相萃取-液相色谱-串联质谱法测定尿液中四氢大麻酚和Δ9-四氢大麻酸的含量[J]. 理化检验-化学分册, 2023, 59(10): 1156~1161
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【2】张晓京,陆峰,马丽娟.用气相色谱法分析大麻籽油中的Δ9-四氢大麻酚[J].宁夏大学学报(自然科学版), 2004,25(3):253-254.
【3】向平,沈敏,沈保华,等.GC/MS同时分析头发中大麻酚类和Δ9-四氢大麻酸[J].法医学杂志, 2002,18(4):216-219.
【4】杨志勇,王占良,张建丽,等.气相色谱-质谱联用快速筛查尿样中麻醉剂类药物[J].质谱学报, 2012,33(3):161-167.
【5】JAMERSON M H, WELTON R M, MORRIS-KUKOSKI C L, et al. Rapid quantification of urinary 11-nor-Δ9-tetrahydrocannabinol-9-carboxylic acid using fast gas chromatography-mass spectrometry[J]. Journal of Analytical Toxicology, 2005,29(7):664-668.
【6】MOORE C, RANA S, COULTER C, et al. Application of two-dimensional gas chromatography with electron capture chemical ionization mass spectrometry to the detection of 11-nor-Δ9-tetrahydrocannabinol-9-carboxylic acid (THC-COOH) in hair[J]. Journal of Analytical Toxicology, 2006,30(3):171-177.
【7】MARALIKOVA B, WEINMANN W. Simultaneous determination of Δ9-tetrahydrocannabinol, 11-hydroxy-Δ9-tetrahydrocannabinol and 11-nor-9-carboxy-Δ9-tetrahydrocannabinol in human plasma by high-performance liquid chromatography/tandem mass spectrometry[J]. Journal of Mass Spectrometry, 2004,39(5):526-531.
【8】卓先义,赵晖,向平.液相色谱-串联质谱同时分析尿液中的大麻酚类和Δ9-四氢大麻酸[J].中国司法鉴定, 2008(1):21-25.
【9】WEINMANN W, GOERNER M, VOGT S, et al. Fast confirmation of 11-nor-9-carboxy-Δ9-tetrahydrocannabinol (THC-COOH) in urine by LC/MS/MS using negative atmospheric-pressure chemical ionisation (APCI)[J]. Forensic Science International, 2001,121(1/2):103-107.
【10】GRAUWILER S B, SCHOLER A, DREWE J. Development of a LC/MS/MS method for the analysis of cannabinoids in human EDTA-plasma and urine after small doses of Cannabis sativa extracts[J]. Journal of Chromatography B, 2007,850(1/2):515-522.
【11】FERNANDEZ M M R, BOECK G D, WOOD M, et al. Simultaneous analysis of THC and its metabolites in blood using liquid chromatography-tandem mass spectrometry[J]. Journal of Chromatography B, 2008,875(2):465-470.
【12】CONCHEIRO M, LEE D Y, LENDOIRO E, et al. Simultaneous quantification of Δ9-tetrahydrocannabinol, 11-nor-9-carboxy-tetrahydrocannabinol, cannabidiol and cannabinol in oral fluid by microflow-liquid chromatography-high resolution mass spectrometry[J]. Journal of Chromatography A, 2013,1297:123-130.
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