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毛细管电泳法测定人玻璃体房水中的乳酸与死亡时间推断。

Lactate determination in human vitreous humour by capillary electrophoresis and time of death investigation.

机构信息

Department of Diagnostics and Public Health, Unit of Forensic Medicine, University of Verona, Verona, Italy.

Institute of Translational Medicine and Biotechnology, Sechenov First Moscow State Medical University, Moscow, Russia.

出版信息

Electrophoresis. 2020 Jun;41(12):1039-1044. doi: 10.1002/elps.201900462. Epub 2020 May 12.

Abstract

Forensic inquests, particularly, in assessing time since death currently recognize the importance of the analysis of vitreous humour (VH) biomarkers. Present research, studies, and validates the determination of lactate (La) in VH by CZE with indirect UV detection. The BGE (pH 8.9) consisted of Tris buffer (37 mM) containing 4-methoxybenzoic acid (4 mM) and alkyl-trimethyl-ammonium bromide (1.2 mM). Each VH specimen was diluted with a butyric acid solution (internal standard 0.057 mM) and La and butyrate were separated within 3-5 min (30 kV). The La LOQ and LOD were 4 and 2 mM, respectively. The calibration curve linearity ranged from 4 to 80 mM; intra- and interruns precisions were less than 10% for standard as well as for VH specimen, respectively. To investigate postmortem interval (PMI) and VH lactate level correlation, human VH specimens were collected during autopsy (n = 40) and stored at -20°C until assay. La levels ranged from 16 to 42 mM; PMI values ranged from 10 to 141 h. La (mM) and PMI (h) correlation was statistically significant (r = 0.527; p < 0.05). In conclusion, the present CZE analysis is efficacious to determine VH La as a biomarker for PMI investigation.

摘要

法医尸检,特别是在评估死亡时间时,目前已经认识到分析玻璃体液 (VH) 生物标志物的重要性。目前的研究、研究和验证了通过 CZE 间接紫外检测法测定 VH 中乳酸 (La) 的方法。BGE(pH 8.9)由 Tris 缓冲液(37 mM)组成,其中含有 4-甲氧基苯甲酸(4 mM)和烷基三甲基溴化铵(1.2 mM)。每个 VH 标本均用丁酸溶液(内标 0.057 mM)稀释,La 和丁酸在 3-5 分钟内(30 kV)分离。La 的 LOQ 和 LOD 分别为 4 和 2 mM。校准曲线的线性范围为 4 至 80 mM;标准和 VH 标本的日内和日间精密度均小于 10%。为了研究死后间隔时间 (PMI) 和 VH 乳酸水平的相关性,在尸检期间收集了人类 VH 标本(n = 40),并储存在 -20°C 直至检测。La 水平范围为 16 至 42 mM;PMI 值范围为 10 至 141 小时。La(mM)和 PMI(h)之间的相关性具有统计学意义(r = 0.527;p < 0.05)。总之,本研究中的 CZE 分析能够有效地确定 VH 中的 La 作为 PMI 研究的生物标志物。

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