High-performance liquid chromatographic separation of a complex mixture of diuretics using a micellar mobile phase of sodium dodecyl sulphate: Application to human urine samples

• Published in: Journal of chromatography. B, Biomedical applications Vol. 748; no. 2; pp. 415 - 424
• Main Authors: Rosado-Maria, A, Gasco-Lopez, A.I, Santos-Montes, A, Izquierdo-Hornillos, R
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 10.10.2000; Elsevier Science
• Subjects: Analysis; Biological and medical sciences; Calibration; Chromatography, High Pressure Liquid - methods; Diuretics; Diuretics - urine; General pharmacology; Medical sciences; Micelles; Pharmacology. Drug treatments; Reproducibility of Results; Sensitivity and Specificity; Sodium Dodecyl Sulfate - chemistry; Spectrophotometry, Ultraviolet; Temperature; Diuretics; Human; Urine; Validation; Biological fluid; Multicomponent mixture; Oral administration; HPLC chromatography; Normal; Optimization; Lauryl sulfate; Diuretic; Separation method; Ultraviolet detector; Dosage form; Tablet; Pharmacokinetics; Micellar solution; Quantitative analysis
• ISSN: 0378-4347; 1387-2273
• DOI: 10.1016/S0378-4347(00)00367-4

A systematic optimization of the HPLC separation of a complex mixture containing 19 diuretics by micellar liquid chromatography using sodium dodecyl sulphate (SDS), a Hypersil (150 mm×3.0 mm I.D., 5 μm) C 18 column, a flow-rate of 0.5 ml min −1 and UV absorbance detection has been carried out. Several mobile phases consisting of SDS and organic modifiers such as acetonitrile, tetrahydrofuran, propanol, butanol or pentanol, and the pH adjusted to 3.2, were tested. The effect of the organic modifier and SDS concentration on the retention behavior and separation of the diuretics was investigated. A mobile phase containing 40 m M SDS and 4% tetrahydrofuran was finally selected. Under these conditions, 14 out of 19 diuretics were separated in about 31 min. A bivariant optimization method for the mobile phase SDS–tetrahydrofuran corroborated the above results. The effect of temperature on the retention was also studied, and 50°C was selected. The optimized method was applied to human urine samples of subjects administered Diurex ® (tablets containing 20 mg of the active ingredient xipamide) without sample preparation.