LC-MS/MS法研究酒精对人体血液中内源性GHB的影响

Researches on the Effect of Alcohol on Endogenous GHB in Human Blood by LC-MS/MS Method

目的运用LC-MS/MS法研究酒精对人体血液中内源性GHB含量的影响。方法建立血液中γ-羟基丁酸(GHB)、γ-丁内酯(GBL)和1,4-丁二醇(1,4-BD)的LC-MS/MS法,以22名健康受试者为研究对象,模拟真实案例饮酒,测定研究对象饮酒前、饮酒7 h后血液中内源性GHB的含量,比较饮酒前后血液中内源性GHB的含量。结果GHB在0.1~50μg/mL浓度范围、GBL和1,4-BD在0.15~50μg/mL浓度范围内,线性关系良好(R2>0.999),GHB、GBL和1,4-BD的检出限分别为0.015μg/mL,0.025μg/mL和0.01μg/mL,定量限分别为0.1μg/mL,0.15μg/mL和0.15μg/mL。3种目标物的基质效应在18.2%~20.1%之间,回收率为81.6%~92.3%,日内和日间精密度RSD均小于10%,准确度为-5.8%~7.0%。饮酒7 h后血液中内源性GHB浓度范围为0.71~3.81μg/mL,相比饮酒前GHB含量升高,差异有统计学意义(P<0.05)。结论饮酒7 h后人体血液中内源性GHB含量增高,但浓度低于4μg/mL,为真实案例涉及的内外源性GHB的判定提供数据支持,丰富了GHB的法医毒物学数据。

Objective To study the effect of alcohol on endogenous GHB content in human blood.Methods LC-MS/MS inspection method was established in the blood forγ-Hydroxybutyric acid(GHB),γ-butyrolactone(GBL)and 1,4-butanediol(1,4-BD).22 healthy subjects were selected as the research subjects to simulate the real cases of drinking alcohol.The levels of GHB in their blood were detected before and after drinking to compare the endogenous GHB levels in the blood before and after drinking.Results GHB in the concentration range of 0.1~50μg/mL had a good linear relationship and there was a good linear relationship between GBL and 1,4-BD in the concentration range of 0.15~50μg/mL(R2>0.999).The limits of detection of GHB,GBL and 1,4-BD were 0.015μg/mL,0.025μg/mL and 0.01μg/mL,respectively,and the limits of GHB,GBL and 1,4-BD quantification were 0.1μg/mL,0.15μg/mL and 0.15μg/mL.The matrix effects of the three compounds ranged from 18.2%to 20.1%,the recoveries were 81.6%to 92.3%,the intra-day and inter-day precision of RSD were less than 10%,and the accuracy was-5.8%-7.0%.The content of GHB in the blood after 7 hours of drinking were higher than that before drinking alcohol(P<0.05),and the concentration range was 0.71-3.81μg/mL.Conclusion The content of endogenous GHB in human blood has increased after 7 hours of drinking alcohol,but the concentration was lower than 4μg/mL,which can provide the data support for the assessment of endogenous and exogenous GHB involved in real cases,enriching the forensic toxicology data of GHB.