A high-performance liquid chromatography method for hypoxanthine determination in vitreous humour: application to estimation of post mortem interval

Abstract

We have developed a new technique to determine the concentration of hypoxanthine [Hx] in a reverse phase column using a modified high-performance liquid chromatography (HPLC) method that is faster and more reliable than those previously described. In this paper we present a formula for estimating the post mortem interval (PMI) based on this HPLC method by applying the inverse prediction method. The regression line obtained by changing the variables gives PMI=0.183[Hx]+0.599 (PMI in hours, [Hx] in μmol/L, R²-0.531, P-0.05).

Keywords

HPLC hypoxanthine post mortem interval regression line vitreous humour

References

1 Bocaz-Beneventi G , Tagliari F , Bortolotti F , Manetto G , Havel J . Capillary zone electrophoresis and artificial neural networks for estimation of the post-mortem interval (PMI) using electrolytes measurements in human vitreous humour . Int J Legal Med 2002; 116: 5-11 .

2 Schafer AT . Colour measurements of pallor mortis . Int J Legal Med 2000; 113: 81–83 .

3 Muñoz JI , Suárez-Peñaranda JM , Otero XL , Rodrĺguez-Calvo MS , Costas E , Miguéns X , Concheiro L . Improved estimation of postmortem interval based on differential behaviour of vitreous potassium and hypoxanthine in death by hanging . Forensic Sci Int 2002; 125: 67–74 .

4 Madea B , Käferstein H , Hermann N , Sticht G . Hypoxanthine in vitreous humour and cerebrospinal fluid a marker of postmortem interval and prolonged (vital) hypoxia? Remarks also on hypoxanthine in SIDS . Forensic Sci Int 1994; 65: 19–31 .

5 Rognum TO , Hauge S , Oyasaeter S , Saugstad OD . A new biochemical method for estimation of postmortem time . Forensic Sci Int 1991; 51: 139–146 .

6 James RA , Hoadley PA , Sampson BG . Determination of postmortem interval by sampling vitreous humour . Am J Forensic Med Pathol 1997; 18: 158–162 .

7 Muñoz JI , Suárez-Peñ aranda JM , Otero XL , Rodrĺguez-Calvo MS , Costas E , Miguéns X , Concheiro L . A new perspective in the estimation of postmortem interval (PMI) based on vitreous [K] . J Forensic Sci Int 2001; 46: 209–214 .

8 Wung WE , Howell SB . Simultaneous liquid chromatography of 5-fluorouracil, uridine, hypoxanthine, xanthine, uric acid, allopurinol, and oxipurinol in plasma . Clin Chem 1980; 26: 1704–1708 .

9 Simmonds RJ , Harkness RA . High-performance liquid chromatographic methods for base and nucleoside analysis in extracellular fluids and in cells . J Chromatogr 1981; 226: 369–381 .

10.

10 Hansson L , Uotila U , Lindfors R , Laiho K . Potassium content of the vitreous body as an aid in determining the time of death . J Forensic Sci 1966; 11: 390–394 .