A simple high-performance liquid chromatography (HPLC) assay for the simultaneous determination of guanase and aspartate aminotransferase (AST) activities in a single serum sample is described. The method is based on direct detection of enzymatically formed products xanthine and glutamate, respectively. The procedure is sensitive, precise (C.V. below 2% for guanase and 3% for AST), suitable for routine purposes and requires only 100 mu l of sample. Kinetic measurements have shown the guanase activity to have an apparent Michaelis constant of 24.5 mu M and the AST activity of 11.1 and 0.18 mM for aspartate and oxoglutarate, respectively, at 37 degrees C in Tris-HCl buffer (pH 7.5).
Simultaneous assay for aspartate aminotransferase and guanase in human serum by high-performance liquid chromatography / V., Carunchio; P., Castellano; Girelli, Anna Maria; A., Messina. - In: JOURNAL OF CHROMATOGRAPHY B. BIOMEDICAL SCIENCES AND APPLICATIONS. - ISSN 1387-2273. - STAMPA. - 689:2(1997), pp. 305-311. [10.1016/s0378-4347(96)00353-2]
Simultaneous assay for aspartate aminotransferase and guanase in human serum by high-performance liquid chromatography
GIRELLI, Anna Maria;
1997
Abstract
A simple high-performance liquid chromatography (HPLC) assay for the simultaneous determination of guanase and aspartate aminotransferase (AST) activities in a single serum sample is described. The method is based on direct detection of enzymatically formed products xanthine and glutamate, respectively. The procedure is sensitive, precise (C.V. below 2% for guanase and 3% for AST), suitable for routine purposes and requires only 100 mu l of sample. Kinetic measurements have shown the guanase activity to have an apparent Michaelis constant of 24.5 mu M and the AST activity of 11.1 and 0.18 mM for aspartate and oxoglutarate, respectively, at 37 degrees C in Tris-HCl buffer (pH 7.5).I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
