血尿肝中巴比妥类药物硅藻土提取紫外差示导数光谱测定

Determination of Barbiturates in Blood ,Urine and Liver by Solid-phase Extraction with Celite and UV Differential Derivative Spectrophotometry

目的建立一种操作简单、快速、去除杂质能力强、提取率高的硅藻土提取血、尿、肝中巴比妥类药物的方法.方法尿液不经稀释、血液经稀释过硅藻土柱,乙醚洗脱;肝匀浆用6%高氯酸沉淀蛋白,上清液过硅藻土柱,二氯甲烷洗脱药物.洗脱液挥干用0.45mol/L氢氧化钠溶液溶解,将溶液等分为两份,分别用等量的0.45mol/L氢氧化钠溶液和0.6mol/L硼酸-氯化钾溶液稀释,得到pH10和pH14水溶液,以pH10溶液为参比,测定pH14溶液的紫外二阶导数光谱进行药物检测.结果该法提取率血98.6%～100.3%,尿99.7%～103.2%,肝78.4%～102%,检出限均低于1.0μg/g(ml),变异系数小于2.9%,线性范围0.5～5.0μg/ml.结论硅藻土提取血、尿、肝中巴比妥类药物、紫外差示导数光谱法进行测定,适合作为法医毒物常规检验方法.

Objective To establish a simple and rapid method for the determination of barbiturates in blood, urine and liver by solid -phase extraction with celite and UV differential derivative spectrophotometry. Methods Blood diluted with water or urine or supernant of the homogenized liver deproteined with 6% perchloric acid were added to celite column and the elutions were effected with ether for blood and urine, or dichloromethane for liver. The eluations were evaporated on water bath at 45℃ or 50℃. The dried residues were redissolved with 0.45moL/L NaOH solution respectively. The reconstituted solution was divided two parts and diluted with equal volume of 0.45moL/L NaOH solution and 0.6moL/L KC1 - H3BO3 buffer, and pH20 and pH14 solutions were obtained respectively, The 2nd derivative spectrum of pH14 solution was measured with pill0 solution as reference. Results Limit of detection for barbiturates is below 1.0μg/g（ml）. The recoveries of barbiturates were almost 100% for blood and urine and between 78.4%～102% for liver. The relative standard deviation of recoveries was less than 2.9%. The linear range was between 0.5～ 5.0μg/ml. Conclusion The method can be used for determination of barbiturates in blood, urine and liver in forensic practice.