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通过荧光偏振免疫分析法监测大麻素的尿排泄:大麻素与肌酐比值研究

Monitoring urinary excretion of cannabinoids by fluorescence-polarization immunoassay: a cannabinoid-to-creatinine ratio study.

作者信息

Fraser Albert D, Worth David

机构信息

Department of Pathology and Laboratory Medicine, Queen Elizabeth II Health Sciences Center, Halifax, Nova Scotia, Canada.

出版信息

Ther Drug Monit. 2002 Dec;24(6):746-50. doi: 10.1097/00007691-200212000-00011.

Abstract

Drug testing in substance abuse treatment programs is focused on urine analysis of parent drugs and major metabolites. Huestis reported that serial monitoring of the major urinary cannabinoid metabolite (delta9-THC-COOH)-to-creatinine ratios in paired urine specimens (collected at least 24 hours apart) could differentiate new marijuana or hashish use from residual cannabinoid metabolite excretion in urine after previous drug use. Subjects with a history of chronic marijuana use were screened for cannabinoids in urine over several months by an enzyme immunoassay (EMIT) with a cut-off value of 50 ng/mL. Presumptive positive specimens were confirmed by gas chromatography-mass spectrometry (GC-MS) for delta9-THC-COOH with a cut-off value of 15 ng/mL. The objective of this study was to determine whether a semiquantitative cannabinoids immunoassay (corrected for creatinine concentration) could differentiate new marijuana use from residual cannabinoid excretion in chronic users of marijuana or hashish compared with GC-MS. The criterion for new marijuana use was a cannabinoid-to-creatinine ratio > or =0.5 (dividing the immunoassay quantitative result to creatinine ratio of specimen 2 by the specimen 1 ratio, specimen 3 by the specimen 2 ratio, etc.). Urine specimens were analyzed by fluorescence-polarization immunoassay (FPIA) on an Abbott TDxFLx analyzer after analysis by GC-MS. In 90 urine specimens (group A) with delta9-THC-COOH values determined by GC-MS, the mean delta9-THC-COOH concentration was 44.4 ng/mL (range, 16-100), and the mean FPIA total cannabinoids value was 91.7 ng/mL (range, 21-204 ng/mL) with a correlation coefficient of 0.993 (group A). In 111 specimens (group B), the mean delta9-THC-COOH concentration was 361 ng/mL (range, 101-960 ng/mL). The mean FPIA value was 657 ng/mL (range, 211-1,270 ng/mL), and the correlation coefficient of the B series was 0.975. Percent cross-reactivity for delta9-THC-COOH standards prepared in drug-free urine by FPIA was 82% at 25 ng/mL, 45% at 50 ng/mL, and 50% at 100 ng/mL. Overall, there was 89% agreement (132 of 148 specimens) between FPIA and GC-MS. In 16 of 148 specimens, however, the FPIA and GC-MS paired urine data did not agree. The sensitivity of the FPIA assay was 95.3%, and the specificity was 44.4%. The authors conclude that FPIA cannabinoid analysis should be further evaluated as an alternative to GC-MS quantitation to help distinguish new marijuana use from residual marijuana metabolite excretion in clinical drug treatment programs.

摘要

药物滥用治疗项目中的药物检测主要集中在对母体药物及其主要代谢物的尿液分析上。休斯蒂斯报告称,对配对尿液样本(采集时间间隔至少24小时)中主要尿大麻素代谢物(δ9-四氢大麻酚-COOH)与肌酐的比率进行连续监测,能够区分新的大麻或哈希什使用情况与先前使用药物后尿液中残留大麻素代谢物的排泄情况。对有慢性大麻使用史的受试者,通过酶免疫测定法(EMIT)对尿液中的大麻素进行了数月筛查,临界值为50纳克/毫升。推定阳性样本通过气相色谱-质谱联用仪(GC-MS)进行确认,检测δ9-四氢大麻酚-COOH,临界值为15纳克/毫升。本研究的目的是确定一种半定量大麻素免疫测定法(校正肌酐浓度后)与GC-MS相比,能否区分大麻或哈希什慢性使用者中新的大麻使用情况与残留大麻素排泄情况。新的大麻使用标准是大麻素与肌酐的比率≥0.5(将样本2的免疫测定定量结果与肌酐的比率除以样本1的比率,样本3的比率除以样本2的比率等)。尿液样本在通过GC-MS分析后,在雅培TDxFLx分析仪上采用荧光偏振免疫测定法(FPIA)进行分析。在90份通过GC-MS测定δ9-四氢大麻酚-COOH值的尿液样本(A组)中,δ9-四氢大麻酚-COOH的平均浓度为44.4纳克/毫升(范围为16 - 100),FPIA总大麻素的平均值得为91.7纳克/毫升(范围为21 - 204纳克/毫升),相关系数为0.993(A组)。在111份样本(B组)中,δ9-四氢大麻酚-COOH的平均浓度为361纳克/毫升(范围为101 - 960纳克/毫升)。FPIA的平均值得为657纳克/毫升(范围为211 - 1270纳克/毫升),B组系列的相关系数为0.975。通过FPIA在无药物尿液中制备的δ9-四氢大麻酚-COOH标准品的交叉反应率在25纳克/毫升时为82%,在50纳克/毫升时为45%,在100纳克/毫升时为50%。总体而言,FPIA与GC-MS之间的一致性为89%(148份样本中的132份)。然而,在148份样本中的16份中,FPIA和GC-MS配对尿液数据不一致。FPIA检测的灵敏度为95.3%,特异性为44.4%。作者得出结论,FPIA大麻素分析作为GC-MS定量的替代方法应进一步评估,以帮助在临床药物治疗项目中区分新的大麻使用情况与残留大麻代谢物排泄情况。

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