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Postmortem vitreous Postmortem vitreous

chemistry chemistry –– An evaluation of An evaluation of

sodium, potassium and sodium, potassium and

chloride levels in estimation chloride levels in estimation

of time since death ( during of time since death ( during of time since death ( during of time since death ( during

the first 36hours after death the first 36hours after death

))

Principal investigator: Dr Chandrakanth H.V.Principal investigator: Dr Chandrakanth H.V.

Associate professorAssociate professor

CoCo--investigator: Dr B.M. Balaraj investigator: Dr B.M. Balaraj

Professor & HeadProfessor & HeadProfessor & HeadProfessor & Head

Dr H.S.Virupaksha Dr H.S.Virupaksha

Professor & HeadProfessor & Head

Dr T. N. Chandrasekhar Dr T. N. Chandrasekhar

Professor & Head Professor & Head

IntroductionIntroductionEstimation of time since death is a Estimation of time since death is a

paramount medicoparamount medico--legal issue in autopsylegal issue in autopsy

An appropriate range of postmortem An appropriate range of postmortem

interval can be deduced by intelligent interval can be deduced by intelligent

interpretation of various changes that interpretation of various changes that interpretation of various changes that interpretation of various changes that

occurs after death.occurs after death.

FluidsFluids-- Blood, CSF & Vitreous humor Blood, CSF & Vitreous humor

show biochemical changes after death.show biochemical changes after death.

Vitreous is gained from the vitreous body and is Vitreous is gained from the vitreous body and is

accompanied by decelerated diffusion and accompanied by decelerated diffusion and

hardly contaminated even in late postmortem hardly contaminated even in late postmortem

intervals.intervals.

This study was intended to study correlation This study was intended to study correlation

between postmortem interval and vitreous between postmortem interval and vitreous

humor chemistry for sodium, potassium and humor chemistry for sodium, potassium and humor chemistry for sodium, potassium and humor chemistry for sodium, potassium and

chlorides.chlorides.

Aimed to find male tp female differences and Aimed to find male tp female differences and

differences between right and left eyes in differences between right and left eyes in

vitreous chemistryvitreous chemistry

VOLUNTEER’S CONSENT FORM

Materials & methodsMaterials & methods

�� Study was carried out in mortuary, Department Study was carried out in mortuary, Department

of Forensic medicine in association with Dept of Forensic medicine in association with Dept

of Biochemistry along with MMC&RI of Biochemistry along with MMC&RI

Forensic Medicine deptForensic Medicine dept

�� Vitreous was collected from 100 deceased Vitreous was collected from 100 deceased �� Vitreous was collected from 100 deceased Vitreous was collected from 100 deceased

subjected for autopsy.subjected for autopsy.

�� All the cases where exact time of death was All the cases where exact time of death was

known and where the time since death ranged known and where the time since death ranged

between 0 and 36 hours were included in the between 0 and 36 hours were included in the

study study

Inclusion criteriaInclusion criteria

Crystal clear fluid will be used for analysisCrystal clear fluid will be used for analysis

Exclusion criteriaExclusion criteria

�� Cases with known or suspected ocular disease, Cases with known or suspected ocular disease,

localized trauma, head injury involving orbit, localized trauma, head injury involving orbit,

burns.burns.

�� Vitreous contaminated with blood and tissue.Vitreous contaminated with blood and tissue.�� Vitreous contaminated with blood and tissue.Vitreous contaminated with blood and tissue.

�� 18 gauge needle will be inserted obliquely 18 gauge needle will be inserted obliquely

through sclera, 5mm lateral to limbus, this through sclera, 5mm lateral to limbus, this

traverse the pars plana and enter vitreous body traverse the pars plana and enter vitreous body

and gently 2ml of vitreous will be drawn into and gently 2ml of vitreous will be drawn into and gently 2ml of vitreous will be drawn into and gently 2ml of vitreous will be drawn into

10ml sterile syringe and later transferred to 10ml sterile syringe and later transferred to

borosil 5ml test tube, the same is repeated on borosil 5ml test tube, the same is repeated on

the other eyethe other eye

�� Collected Vitreous humor was centrifuged at Collected Vitreous humor was centrifuged at

3000 g and supernatant 3000 g and supernatant -- transferred to another transferred to another

test tube and sent to Laboratory for test tube and sent to Laboratory for

estimation. Care was ensured to prevent any estimation. Care was ensured to prevent any estimation. Care was ensured to prevent any estimation. Care was ensured to prevent any

contamination with blood/RBC’scontamination with blood/RBC’s

�� In the Laboratory, Potassium will be estimated In the Laboratory, Potassium will be estimated by Ion Selective Electrode Methods on AVL by Ion Selective Electrode Methods on AVL Roche Electrolyte Analyzer. Roche Electrolyte Analyzer.

�� Potassium electrodes were fitted with liquid Potassium electrodes were fitted with liquid �� Potassium electrodes were fitted with liquid Potassium electrodes were fitted with liquid ion exchange membranes that incorporate ion exchange membranes that incorporate valinomycin.valinomycin.

�� The analyzer can measure Potassium levels in The analyzer can measure Potassium levels in

the range of 1.8 to 12 mmol/litre. the range of 1.8 to 12 mmol/litre.

�� Daily Quality checks will be performed using Daily Quality checks will be performed using

level 2 and level 3 quality control sera from level 2 and level 3 quality control sera from level 2 and level 3 quality control sera from level 2 and level 3 quality control sera from

BioBio--Rad Laboratories.Rad Laboratories.

Statistical methodStatistical method

�� Data obtained was analyzed statistically using Data obtained was analyzed statistically using

SPSS version 11.0 SPSS version 11.0

�� The present research did not find a significant The present research did not find a significant

correlation between vitreous chemistry and correlation between vitreous chemistry and

postmortem interval. The differences in postmortem interval. The differences in

vitreous sodium, potassium, chloride levels vitreous sodium, potassium, chloride levels vitreous sodium, potassium, chloride levels vitreous sodium, potassium, chloride levels

and the sodium potassium ratio among males and the sodium potassium ratio among males

and females and between right and left eyes and females and between right and left eyes

was not found to be statistically significant was not found to be statistically significant

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