alexa Validation of a LC method for the determination of 5-aminosalicylic acid and its metabolite in plasma and urine.


Chemical Sciences Journal

Author(s): Bystrowska B, Nowak J, Brandys J

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Abstract The choice of a proper analytical method for the quantification of drugs and/or their metabolites in biological samples plays a significant role in the evaluation and interpretation of bioavailability, bioequivalence and pharmacokinetic data. The aim of this study was validation of a method for the identification and quantitative determination of 5-aminosalicylic acid (5-ASA) and its metabolite N-acetyl-5-aminosalicylic acid in human plasma and urine. According to previous studies on the disposition of 5-ASA (mesalazine) in a patient with inflammatory bowel diseases, we have developed a rapid, sensitive method for the determination of 5-ASA and its acetylated metabolite, N-acetyl-5-aminosalicylic acid (Ac-5-ASA). The advantage of this method is that it measures both compounds, and is more rapid, reproducible and credible than the previous studies [C. Fischer, K. Maier, U. Klotz, J. Chromatogr. Biomed. Appl. 225 (1981) 498-503; P.N. Shaw, A.L. Sivner, L. Aarons, J.B. Houston, J. Chromatogr. Biomed. Appl. 274 (1983) 393-397; B. Norlander, R. Gotthard, M. Strom, Aliment. Pharmacol. Ther. 3 (1989) 333-342; U. Klotz, G.L. Stracciari, Arzneim.-Forsch. Drug Res. II 43 (12) (1993) 1357-1359]. 5-ASA was quantitatively determined in human plasma and urine samples by liquid chromatography following prior derivatization to its acetylated metabolite (Ac-5-ASA). N-Acetyl-anthranilic acid was used as the internal standard. The detection was performed with a spectrofluorimetric detector, excitation at 311 nm, cut-off at 449 nm. The method was validated for the following parameters: linearity, recovery, sensitivity, precision, accuracy, selectivity and stability, limits of quantification and of detection. It showed good linearity (r2 > or = 0,996) in the range 0.1 ng/ml to 8 microg/ml using a Lichrospher 60 RP-select B column. The lower limit of detection was 20 ng/ml in plasma and urine. The within-run relative standard deviations (R.S.D.) were below 6.7\% at all concentration levels and the between-run R.S.D. were below 25.4\% at all concentration levels.
This article was published in J Pharm Biomed Anal and referenced in Chemical Sciences Journal

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