Estimating time since death (TSD) with fair accuracy from postmortem changes still remains an important but difficult task to be performed by every autopsy surgeon in medicolegal scenario. The aim of the present study was to estimate TSD from electrolyte analysis of postmortem vitreous humour collected from samples under semi-arid climate. Vitreous humour was collected from 210 dead bodies brought to University College of Medical Sciences and Guru Teg Bahadur Hospital, Delhi, during the period of November 2010 to April 2012. The vitreous samples were analysed for sodium, potassium, calcium, chloride and glucose. The results showed a significant positive relationship between TSD and potassium (r = 0.841, p = 0.000) and a weak negative relationship between TSD and sodium (r = –0.137, p = 0.048) and glucose (r = –0.241, p = 0.000), whereas no significant relationship could be established between TSD and calcium (r = 0.055, p = 0.429) and chloride (r = 0.075, p = 0.11).