Gas chromatographic method for determining very long chain fatty acids that compose D003 in 1 to 10 mg/ml suspensions used in pharmacological and toxicological studies

Abstract

D003 is a mixture of fatty acids (C240 to C36.0), that shows antiplatelet, antithrombotic and cholesterol-lowering effects in experimental models. A gas chromatographic method using a DB-5 Widebore column and 1-nonadecanoic acid as internal standard was developed and validated in order to determine D003 in the aqueous suspensions, with concentrations from I to 10 mg/mL, used in pharmacological and toxicological studies . Fatty acids were analyzed as methyl esters derivatives, prepared using 5 aqueous HCI-methanol . This method was specific for D003 determination in these suspensions . The determination of total content of fatty acids showed a CV of 0.39%. The values of the correlation coefficients (r) were higher than that of the acceptation limit (0 .99) for the individual fatty acids and for the total content of them, indicating that correlations weree positives for a probability higher than 99.9%. In the repeatability study the CV for total content of D003 (1.55, 1.68 and 0.62%, respectively) were lower, indicating that the method has repeatability. The inter-mean precision for the total content of fatty acids (CV 1 .47%), demonstrated that the method is reproducible . The average recovery obtained for the total content of acids was greater than 99% for all cases. Also, for all the fatty acids 4,o < tab for a 95% of releability, there were no significant differences between the average recovery and 100%, then, it can be considered that this method is exact. The method was suitable for quality control and stability studies of these suspensions.