大鼠血液外泌体中外源性γ-羟基丁酸的检测  

Detection of Exogenous γ-Hydroxybutyric Acid in Rat Blood Exosomes

作　　者：郜峥翔 罗奇志 张亮[2] 裴茂清[2] 王慧君 岳霞 GAO Zheng-xiang;LUO Qi-zhi;ZHANG Liang;PEI Mao-qing;WANG Hui-jun;YUE Xia(School of Forensic Medicine,Southern Medical University,Guangzhou 510080,China;Criminal Tech-nology Center,Guangdong Provincial Public Security Department,Guangzhou 510050,China)

机构地区：[1]南方医科大学法医学院,广东广州510080 [2]广东省公安厅刑事技术中心,广东广州510050

出　　处：《法医学杂志》2022年第2期212-216,共5页Journal of Forensic Medicine

基　　金：国家自然科学基金面上项目(81871528)。

摘　　要：目的建立基于外泌体的超高效液相色谱-质谱法(ultra-high performance liquid chromatographymass spectrometry,UPLC-MS)定量检测大鼠血液中的γ-羟基丁酸(gamma-hydroxybutyrate,GHB),寻找区分内源性GHB和外源性GHB的方法。方法将成年雄性SD大鼠分为给药1 h、5 h、10 h组与对照组,给药组单次腹腔注射GHB前体物质γ-丁内酯(gamma-butyrolactone,GBL)后,分别于注射后1 h、5 h、10 h腹主动脉采血5 mL,对照组给予等量生理盐水后1 h采血5 mL。其中0.5 mL血液经前处理后直接进行UPLC-MS检测,剩余血液经差速超速离心提取外泌体后进行检测。结果对照组大鼠血液中GHB质量浓度为(87.36±33.48)ng/mL,给药1 h、5 h和10 h组大鼠血液中GHB质量浓度分别为(110400.00±1766.35)、(1479.00±687.01)和(133.60±12.17)ng/mL。外泌体检测结果显示,对照组及给药10 h组均未检测到GHB峰值信号,给药1 h和5 h组GHB质量浓度分别为(91.47±33.44)和(49.43±7.05)ng/mL。结论通过提取大鼠血液外泌体后经UPLC-MS法检测GHB,证实外源性GHB可在血液外泌体中检出,而内源性GHB则不能检出,提示该方法可能作为判断是否有外源性药物摄入的依据。Objective To find a method to distinguish exogenous gamma-hydroxybutyrate(GHB)from endogenous GHB by establishing ultra-high performance liquid chromatography-mass spectrometry(UPLC-MS)based on exosome for quantitative detection of GHB in the rat blood.Methods Adult male SD rats were divided into 1 h,5 h,10 h administration group and control group.After 1 h,5 h and 10 h of single precursor of GHB gamma-butyrolactone(GBL)intraperitoneal injection in administration groups,5 mL blood was collected from the abdominal aorta.Meanwhile,the control group was given a same dose of normal saline,and 5 mL blood was collected at 1 h.Among the 5 mL blood,0.5 mL was directly detected by HPLC-MS after pretreatment,and exosomes were extracted from the remaining blood by differential centrifugation and detected.Results The concentration of GHB in the control group was(87.36±33.48)ng/mL,and the concentration with administration at 1 h,5 h and 10 h was(110400.00±1766.35)ng/mL,(1479.00±687.01)ng/mL and(133.60±12.17)ng/mL,respectively.The results of exosome detection showed that no peak GHB signal was detected in the control group and the 10 h administration group,and the concentrations of GHB at 1 h and 5 h administration groups were(91.47±33.44)ng/mL and(49.43±7.05)ng/mL,respectively.Conclusion GHB was detected in blood exosome by UPLC-MS,which indicated that exogenous GHB could be detected in plasma exosomes,while endogenous GHB could not be detected,suggesting that this method may be used as a basis to determine whether there is exogenous drug intake.

关 键 词：法医病理学 外泌体 Γ-羟基丁酸 Γ-丁内酯 超高效液相色谱-质谱法 血液 大鼠 

分 类 号：DF795.1[医药卫生—法医学]