Journal of Forensic Medicine ›› 2022, Vol. 38 ›› Issue (2): 212-216.DOI: 10.12116/j.issn.1004-5619.2021.410116

• Original Articles • Previous Articles     Next Articles

Detection of Exogenous γ-Hydroxybutyric Acid in Rat Blood Exosomes

Zheng-xiang GAO1(), Qi-zhi LUO1, Liang ZHANG2, Mao-qing PEI2, Hui-jun WANG1, Xia YUE1()   

  1. 1.School of Forensic Medicine, Southern Medical University, Guangzhou 510080, China
    2.Criminal Technology Center, Guangdong Provincial Public Security Department, Guangzhou 510050, China
  • Received:2021-01-26 Online:2022-04-25 Published:2022-04-28
  • Contact: Xia YUE

Abstract: 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 (110 400.00±1 766.35) ng/mL, (1 479.00±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.

Key words: forensic pathology, exosome, gamma-hydroxybutyrate (GHB), gamma-butyrolactone (GBL), ultra-high performance liquid chromatography-mass spectrometry (UPLC-MS), blood, rats

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