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J Pharm Biomed Anal. 2008 Aug 5;47(4-5):807-11. doi: 10.1016/j.jpba.2008.03.012. Epub 2008 Mar 22.

Development of a liquid chromatographic method for the determination of related substances and assay of d-cycloserine.

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  • 1Laboratorium voor Farmaceutische Analyse, Faculteit Farmaceutische Wetenschappen, Katholieke Universiteit Leuven, O & N 2, PB 923, Herestraat 49, B-3000 Leuven, Belgium.


d-cycloserine or d-4-amino-3-isoxazolidinone is an antibiotic produced by Streptomyces garyphalus and Streptomyces orchidaceus. d-Cycloserine is used in the second line treatment of tuberculosis and is often used in developing countries. Therefore, expensive high-tech techniques are not recommended for analysis. Here, a liquid chromatography method with ultraviolet detection (LC-UV) is described using a base deactivated column (Hypersil BDS column; 25 cm x 4.6 mm I.D.) kept at 45 degrees C. The gradient method uses mobile phases containing acetonitrile (ACN), 20mM sodium octane sulphonate (SOS), 0.2M potassium dihydrogen phosphate buffer pH 2.8, water: A: (4:70:10:16v/v/v/v) and B: (17:70:10:3v/v/v/v). The method proved to be robust, linear, repeatable, sensitive, selective and easy to perform. For the related substances test 50 microl of a 0.5 mg/ml d-cycloserine solution is injected. For assay, a concentration of 0.1 mg/ml is proposed to avoid overloading of the detector.

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