Centre Hospitalier Universitaire d'Angers, Laboratoire de Pharmacologie-Toxicologie, France.
Service de Médecine Légale et Pénitentiaire, Centre Hospitalo-Universitaire, Angers, France.
J Forensic Leg Med. 2024 Oct;107:102764. doi: 10.1016/j.jflm.2024.102764. Epub 2024 Sep 27.
Thanatological biochemistry has gained prominence in determining causes of death, especially when suspected fatal pathologies do not exhibit clear postmortem macroscopic and/or microscopic features, such as in cases of ketoacidosis. Indeed, in these cases, the measurement of β-hydroxybutyrate (BHB) in femoral blood and/or vitreous humor is of particular importance. However, data on its in vitro stability remain scarce, especially in vitreous humor. In this context, the study reported here aims to assess the in vitro stability of BHB. BHB quantification was performed using a liquid chromatography coupled with tandem mass spectrometry method. To investigate BHB stability, two different postmortem matrices were considered: femoral blood and vitreous humor. These matrices were pooled, aliquoted and spiked with BHB at three different concentrations (50 mg/L, 100 mg/L, and 200 mg/L; n = 3). Initial BHB concentrations were established on day 1. Each sample was then divided into two aliquots for storage under two conditions: 20 °C and 4 °C. Analyses were performed on Day 3, 7, 14, and 28. The study revealed no significant degradation of BHB in femoral blood or vitreous humor over time (days 1-28), confirming the robustness and reliability of BHB measurement in these matrices as a postmortem biomarker of ketoacidosis under the tested temperature conditions (+4 °C or -20 °C). These results support a systematic integration of BHB measurement into the routine workflow of forensic toxicology laboratories.
死亡生物化学在确定死亡原因方面变得越来越重要，尤其是在可疑的致命病理学表现没有明显的死后宏观和/或微观特征时，例如酮症酸中毒的情况。实际上，在这些情况下，测量股血和/或玻璃体中的β-羟丁酸（BHB）尤为重要。然而，关于其体外稳定性的数据仍然很少，特别是在玻璃体中。在这种情况下，本研究旨在评估 BHB 的体外稳定性。使用液相色谱-串联质谱法进行 BHB 定量。为了研究 BHB 的稳定性，考虑了两种不同的死后基质：股血和玻璃体。将这些基质混合、等分并分别加入 BHB 三种不同浓度（50mg/L、100mg/L 和 200mg/L；n=3）。第一天建立初始 BHB 浓度。然后，将每个样品分为两份，在两种条件下储存：20°C 和 4°C。在第 3、7、14 和 28 天进行分析。研究结果表明，BHB 在股血或玻璃体中随时间（1-28 天）没有明显降解，证实了在测试温度条件下（+4°C 或-20°C），BHB 作为酮症酸中毒死后生物标志物在这些基质中的测量具有稳健性和可靠性。这些结果支持将 BHB 测量系统地纳入法医毒理学实验室的常规工作流程。
