Kerrigan S, Brooks D E
Department of Chemistry, University of British Columbia, Vancouver, Canada.
J Immunol Methods. 1999 Apr 22;224(1-2):11-8. doi: 10.1016/s0022-1759(98)00194-x.
Polyclonal antibodies raised against the hallucinogenic drug, lysergic acid diethylamide (LSD), were used to detect and extract drug from whole blood samples. An indirect ELISA was used to detect as little as 1 pg of total drug in 25 microl blood. The limit of detection of the immunoassay, calculated from the mean - 3 SD was 39 pg/ml. The analytical recovery of LSD (2.5-0.2 ng/ml) from whole blood was 102-113%. Within-run CVs for LSD spiked in blood at 1.25, 0.16 and 0.04 ng/ml were 5.6, 3.1, and 8.9%, respectively (n = 4). There was an overall decrease in precision when whole blood was used in place of urine, due to the increased complexity of the matrix. However, using this technique LSD was calibrated in blood in the sub-ng/ml region of forensic interest. Immunoaffinity extraction was used to isolate LSD from blood and urine samples. The affinity support was prepared by covalently attaching anti-LSD antibodies to Protein A-coated agarose beads. No pre-treatment of the sample was required other than the addition of neutral buffer. Sub-ng/ml concentrations of LSD were routinely extracted from blood and urine samples with greater than 80% recovery of drug. This technique, which could be used to extract LSD from blood and urine samples prior to confirmatory drug analysis, could be completed in about 10 min.
用针对致幻药物麦角酸二乙酰胺(LSD)产生的多克隆抗体从全血样本中检测和提取药物。采用间接酶联免疫吸附测定法(ELISA)检测25微升血液中低至1皮克的总药物量。根据平均值减去3倍标准差计算,该免疫测定法的检测限为39皮克/毫升。从全血中回收LSD(2.5 - 0.2纳克/毫升)的分析回收率为102% - 113%。添加到血液中浓度为1.25、0.16和0.04纳克/毫升的LSD在批内变异系数(CV)分别为5.6%、3.1%和8.9%(n = 4)。由于基质复杂性增加,使用全血代替尿液时精密度总体下降。然而,采用该技术可在法医学关注的亚纳克/毫升范围内对血液中的LSD进行校准。采用免疫亲和提取法从血液和尿液样本中分离LSD。通过将抗LSD抗体共价连接到蛋白A包被的琼脂糖珠上来制备亲和支持物。除添加中性缓冲液外,无需对样本进行预处理。能从血液和尿液样本中常规提取亚纳克/毫升浓度的LSD,药物回收率大于80%。该技术可在确证性药物分析前用于从血液和尿液样本中提取LSD,大约10分钟即可完成。