A PERIODATE-RESORCINOL MICROASSAY FOR THE QUANTITATION OF TOTAL SIALIC ACID IN HUMAN SERUM

ABSTRACT

Abstract A periodate-resorcinol microassay for the quantitation of total sialic acid (TSA) has been developed. A 96-well microtiter plate format was used as a single reagent mixing well. The principle of this newly-developed assay is the periodate-resorcinol procedure. Conditions for the quantitation of sialic acid in serum, such as the concentration of periodate, resorcinol, incubation time and human serum volume, were optimized. It was found that the optimal concentration of periodic acid and resorcinol reagent were 1.3 mM and 0.6%, respectively. An incubation time of 60 minutes for the reaction of periodic acid and resorcinol with samples was found to give the highest absorbance. These new procedures were used for the quantitation of sialic acid in human serum. It was found that only 5 μL samples were needed to give a varied coefficient of the intraand inter-assay of 0.79% and 4.68%, respectively. Furthermore, this method demonstrated the recovery percentage of 94.25% by the addition of a known amount of pure sialic acid. This has been used to quantitate TSA in normal healthy and cancer serum and a significant difference between these two groups (p