血中水合氯醛及其代谢物的GC-MS/MS和HPLC-MS/MS检测

Determination of chloral hydrate and its metabolites in blood samples by GC-MS/MS and HPLC-MS/MS

目的建立血液中水合氯醛及其代谢物的色谱串联质谱检测方法,用于法医案件的检验。方法血液检材经乙酸乙酯萃取后气质联用选择离子扫描(SIM)检测水合氯醛和三氯乙醇,经乙腈沉淀蛋白后液质联用负离子模式下多反应监测(MRM)检测三氯乙酸。结果水合氯醛及其代谢物的线性范围分别是100ng/mL~15μg/mL、500ng/mL~20μg/mL和500ng/mL~15μg/mL,线性关系良好,R2均大于0.998,最低检出限分别为15ng/mL、130ng/mL和30ng/mL,最低定量限分别为50ng/mL、500ng/mL和100ng/mL;日内精密度分别在2.26%~7.31%、3.09%~7.23%和2.79%~5.37%;日间精密度分别在4.14%~7.03%、2.18%~4.43%和2.75%~4.96%;提取回收率分别为92.72%~106.30%、94.22%~103.70%和89.05%~104.50%。并对水合氯醛相关案件进行检测,心血中水合氯醛浓度为411.34ng/mL,三氯乙醇浓度为9.49μg/mL,三氯乙酸浓度为8.32μg/mL。结论本文建立的水合氯醛及其代谢物的检测方法准确简单快速,可应用于水合氯醛中毒案件的法医学鉴定。

Objective To establish a chromatography-tandem mass spectrometry method for the determination of chloral hydrate and its metabolites in blood for forensic cases. Methods Chloral hydrate and trichloroethanol were detected by mass spectrometry with selective ion scanning(SIM) after ethyl acetate extraction, and trichloroacetic acid was detected by liquid mass spectrometry with multiple reaction monitoring(MRM) under negative ion mode after acetonitrile precipitation. Results The calibration curves for chloral hydrate, trichloroethanol and trichloroacetic acidshowed good linear relationships(R2 > 0.998) in the concentration range of 100 ng/mL~15 μg/mL, 500 ng/mL~20 μg/mL and 500 ng/mL~15 μg/m L, respectively. The lowest limits of detection were 15 ng/mL, 130 ng/mL and 30 ng/mL, respectively. The lowest limits of quantification were 50 ng/mL, 500 ng/mL and 100 ng/mL, respectively. The intra-day precisions were 2.26%~7.31%, 3.09%~7.23% and 2.79%~5.37%, respectively. The daily precisions were 4.14%~7.03%, 2.18%~4.43% and 2.75%~4.96% respectively. The extraction recoveries w e r e 9 2. 7 2 % ~ 1 0 6. 3 0 %, 9 4. 2 2 % ~ 1 0 3. 7 0 % a n d 89.05%~104.50%, respectively. A case related to chloral hydrate was analyzed, and the concentrations of chloral hydrate, trichloroethanol and trichloroacetic acid in blood were 411.34 ng/m L, 9.49 μg/m L and 8.32 μg/m L respectively. Conclusion The method established in this paper is accurate, simple and rapid, and can be used for forensic identification of chloral hydrate poisoning.