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

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 (37mM) containing 4-methoxybenzoic acid (4mM) and alkyl-trimethyl-ammonium bromide (1.2mM). Each VH specimen was diluted with a butyric acid solution (internal standard 0.057mM) and La and butyrate were separated within 3-5min (30kV). The La LOQ and LOD were 4 and 2mM, respectively. The calibration curve linearity ranged from 4 to 80mM; 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 42mM; PMI values ranged from 10 to 141 h. La (mM) and PMI (h) correlation was statistically significant (r2 = 0.527; p < 0.05). In conclusion, the present CZE analysis is efficacious to determine VH La as a biomarker for PMI investigation.