Development of a liquid chromatographic method for the determination of related substances and assay of d-cycloserine.
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.
Pendela
Murali
M
Laboratorium voor Farmaceutische Analyse, Faculteit Farmaceutische Wetenschappen, Katholieke Universiteit Leuven, O & N 2, PB 923, Herestraat 49, B-3000 Leuven, Belgium.
Dragovic
Sanja
S
Bockx
Lien
L
Hoogmartens
Jos
J
Van Schepdael
Ann
A
Adams
Erwin
E
D000097:Acetonitriles / Q000737:chemistry; D000904:Antibiotics, Antitubercular / Q000032:analysis* / Q000737:chemistry; D001681:Biological Assay; D002021:Buffers; D002210:Caprylates / Q000737:chemistry; D002214:Capsules / Q000032:analysis / Q000737:chemistry; D002853:Chromatography, Liquid / Q000295:instrumentation / Q000379:methods*; D003523:Cycloserine / Q000032:analysis* / Q000737:chemistry; D019281:Dimerization; D004355:Drug Stability; D006863:Hydrogen-Ion Concentration; D015394:Molecular Structure; D010710:Phosphates / Q000737:chemistry; D017680:Potassium Compounds / Q000737:chemistry; D012044:Regression Analysis; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012694:Serine / Q000032:analysis* / Q000737:chemistry; D012997:Solvents / Q000737:chemistry; D013056:Spectrophotometry, Ultraviolet; D013678:Technology, Pharmaceutical / Q000379:methods*; D013696:Temperature; D013997:Time Factors; D014867:Water / Q000737:chemistry
D000097:Acetonitriles; D000904:Antibiotics, Antitubercular; D002021:Buffers; D002210:Caprylates; D002214:Capsules; D010710:Phosphates; D017680:Potassium Compounds; D012997:Solvents; D014867:Water; D012694:Serine; D003523:Cycloserine; C013216:potassium phosphate; C031492:octanoic acid; C032159:acetonitrile