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KMID : 0369919970120000013
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Journal of Pharmaceutical Sciences (C.B.N.)
1997 Volume.12 No. 0 p.13 ~ p.18
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Improved Simple and Micro High Performance Liquid Chromatographic Assay of Furosemide and Its Metabolite in Biological Fluids
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Lee, Myung Gull
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Abstract
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High performance liquid chromatographic (HPLC) methos for the determination of furosemide, a diuretic, in plasma and urine have been developed (1~+4) earlier. However, extraction method (1~3) was employed and 1 ml (1, 2) or 2 ml (3) of the sample was needed. Furthermore, 4-chloro-5-sulfamoylanthranilic acid (CSA), a metabolite was not completely resolved from furosemide (3, 4) in their HPLC assay and the detection limit was greater than 0. 1 ㎍/ml (1~4). Although some additional methods (5~11) have been appeared since thcse publications to improve deteption limit aud to decrease sample volumes, there were potential limitations as discussed by Rapaka et al. (8). The method reported by Smith et al. (7) using phenobarbital as an internal standard required two detectors (a fluorescence detector for furosemide and a UV detector for the internal standard), as well as two different mobile phases for plasma and urine samples. Moreover, the method (7) needed the evaporation under nitrogen of the supernatant fluids after deproteinization. The improved method of Rapaka et al. (8) required the use of a pre-column, 0.2 ml of plasma, and injection of up to 0. 3 ml of the final solution onto the column in order to achieve an assay limit of 0.02 ㎍/ml. Its application to the analyses of urine samples, often required in bioavailability and pharmacokinetic studies, was not demonstrated. The drawbacks of the method of Nation a cxl. (5) were lack of internal standard and of application to urine sample. In other publications, similar problems could be found; extraction method (9~11) was used and 0. 2 ml (6, 7, 10) or 0.5 ml (9, 11) of the sample was also required. Snedden et cxl. (12) reported a sensitive HPLC assay method for furosemide using different mobile phases and internal standards in plasma and urine. However, the detection limit was 0. 2 and 5 ㎍/ml for plasma and urine, respectively, and an extraction method was employed.
Smith et cxl. (7) analyzed CSA in plasma and urine using two detectors (UV and fluorescence detectors), different mobile phases and internal standards for plasma and urine.
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