Fleming Steven W, Dasgupta Amitava, Garg Uttam
Department of Pathology and Laboratory Medicine, Children's Mercy Hospitals and Clinics, Kansas City, MO, USA.
Methods Mol Biol. 2010;603:145-56. doi: 10.1007/978-1-60761-459-3_14.
Cocaine, a stimulant, is a commonly abused drug. Cocaine and its metabolites are measured in various biological specimens for clinical and forensic purposes. Urine or plasma or serum is spiked with deuterated internal standards cocaine-d3, benzoylecgonine-d3, ecgonine methyl ester-d3, and cocaethylene-d3 and buffered with phosphate buffer. The drugs in the sample are extracted by cation-exchange solid phase extraction. The drugs from the solid phase cartridge are eluted and the eluent is dried under the stream of nitrogen. The residue is incubated with pentafluoropropionic acid anhydride and pentafluoropropanol to form pentafluoropropionyl derivatives of ecgonine methyl ester and benzoylecgonine. Cocaine and cocaethylene are refractory to derivatization. The extract is dried, reconstituted in ethyl acetate, and injected into gas chromatography mass-spectrometry analyzer. Quantitation of the drugs in the samples is made, using selected ion monitoring, from a 3-point calibration curve.
可卡因是一种兴奋剂,是一种常见的滥用药物。为了临床和法医目的,在各种生物样本中检测可卡因及其代谢物。尿液、血浆或血清中加入氘代内标物可卡因 - d3、苯甲酰芽子碱 - d3、芽子碱甲酯 - d3和可卡乙碱 - d3,并用磷酸盐缓冲液缓冲。样品中的药物通过阳离子交换固相萃取法提取。从固相柱中洗脱药物,洗脱液在氮气流下干燥。残渣与五氟丙酸酐和五氟丙醇一起孵育,形成芽子碱甲酯和苯甲酰芽子碱的五氟丙酰衍生物。可卡因和可卡乙碱难以衍生化。提取物干燥后,用乙酸乙酯复溶,然后注入气相色谱 - 质谱分析仪。使用选择离子监测,根据三点校准曲线对样品中的药物进行定量。