Hypoxanthine-guanine phosphoribosyl transferase: assay using high performance liquid chromatography.
A method is described for the estimation of hypoxanthine-guanine phosphoribosyltransferase using high performance liquid chromatography. The inosine monophosphate (IMP) generated from hypoxanthine is determined, after separation on a C18 reversed phase silica column with a buffer-methanol gradient, by the absorbance at 254 nm. Simultaneous reciprocal measurement of hypoxanthine consumption is made. The assay is suitable for screening red cell lysates for hypoxanthine-guanine phosphoribosyltransferase deficiency; the results being expressed as nmol inosine monophosphate . h-1. mg-1 haemoglobin. The normal range found was 94 +/- 15 nmol IMP . h-1 . mg-1 haemoglobin and hypoxanthine-guanine phosphoribosyltransferase activities down to 1% of normal can be assayed accurately.
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004912:Erythrocytes / Q000201:enzymology; D006454:Hemoglobins / Q000032:analysis; D006801:Humans; D007041:Hypoxanthine Phosphoribosyltransferase / Q000097:blood*; D007291:Inosine Monophosphate / Q000032:analysis
D006454:Hemoglobins; D007291:Inosine Monophosphate; D007041:Hypoxanthine Phosphoribosyltransferase