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

被引:46
作者
Bystrowska, B [1 ]
Nowak, J [1 ]
Brandys, J [1 ]
机构
[1] Jagiellonian Univ, Collegium Medicum, Dept Toxicol, PL-30688 Krakow, Poland
关键词
5-aminosalicylic acid; 5-amino-2-hydroxybenzoic acid; mesalazine; N-acetyl-5-aminosalicylic acid; high-performance liquid chromatography; validation;
D O I
10.1016/S0731-7085(99)00292-7
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
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 (r(2) greater than or equal to 0,996) in the range 0.1 ng/ml to 8 mu g/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. (C) 2000 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:341 / 347
页数:7
相关论文
共 7 条
[1]   Validation of liquid chromatographic and gas chromatographic methods - Applications to pharmacokinetics [J].
Bressolle, F ;
BrometPetit, M ;
Audran, M .
JOURNAL OF CHROMATOGRAPHY B-BIOMEDICAL APPLICATIONS, 1996, 686 (01) :3-10
[2]   Validation of chromatographic methods in biomedical analysis - Viewpoint and discussion [J].
Causon, R .
JOURNAL OF CHROMATOGRAPHY B, 1997, 689 (01) :175-180
[3]  
DEKASKI MC, 1993, ALIMENT PHARM THERAP, V7, P567
[4]   SIMPLIFIED HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC METHOD FOR 5-AMINOSALICYLIC ACID IN PLASMA AND URINE [J].
FISCHER, C ;
MAIER, K ;
KLOTZ, U .
JOURNAL OF CHROMATOGRAPHY, 1981, 225 (02) :498-503
[5]  
KRINGLE RO, 1994, PHARC RES, V11
[6]  
SEGARS LW, 1992, CLIN PHARM, V11
[7]   A RAPID METHOD FOR THE SIMULTANEOUS DETERMINATION OF THE MAJOR METABOLITES OF SULPHASALAZINE IN PLASMA [J].
SHAW, PN ;
SIVNER, AL ;
AARONS, L ;
HOUSTON, JB .
JOURNAL OF CHROMATOGRAPHY, 1983, 274 (MAY) :393-397