[Simultaneous determination of salicylic acid and its hydroxylated metabolites in poultry serum by HPLC]

ABSTRACT

To introduce an HPLC method for indirect measuring of hydroxyl radicals in poultry serum. Chromatographic study. Poultry serum. By acidification of serum with 1.0 M HCl, salicylic acid [SA] and its hydroxylated metabolites were extracted into diethylether, dried and redissolved in mobile phase buffer and HCl. It was filtered and 20 micro L of the filtrate injected into the HPLC system. The chromatographic condition was as follows acetate-citrate buffer [94%] methanol [6%] as mobile phase run at 1.2 ml/min through a C18 column using UV and EC detectors concurrently. Descriptive statistics good. Separations among these compounds obtained without any interference with poultry serum elements. Calibration curves for SA and Dihydroxy benzoic acids [DHBAs] were linear at 0.28-88 micro g/ml [R[2] =0.996] and 0.3 -12.3 micro g/ml [R[2] = 0.998], respectively. Detection limit for SA and DHBAs were 0.56 micro g/ml and 0.3 micro g/ml, respectively. Precision of the method [CV%] as intra- and inter-day variations were defined for SA [2.5 -6.5%, 4.4-6.8%], 2,3-DHBA [4.2-8.5%, 5.8-9.0%] and 2, 5-DHBA [4.1-6.8%, 5.9- 7.4%]. Recovery of the method for SA and DHBA were 87 +/- 5% and 92 +/- 4%, respectively. This method is useful for simultaneous determination of SA and its hydroxylated metabolites in poultry serum as an indirect measurement of hydroxyl radicals