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Separation of tetracycline and its potential impurities by liquid chromatography on XTerra RP‐18
Author(s) -
Capote Rául,
Diana José,
Roets Eugène,
Hoogmartens Jos
Publication year - 2002
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/1615-9314(20020501)25:7<399::aid-jssc399>3.0.co;2-4
Subject(s) - chromatography , silanol , chemistry , elution , impurity , acetonitrile , ethylenediaminetetraacetic acid , high performance liquid chromatography , volumetric flow rate , analytical chemistry (journal) , chelation , inorganic chemistry , biochemistry , organic chemistry , catalysis , physics , quantum mechanics
A linear gradient elution method is described for the separation of tetracycline, its fermentation impurities, and its degradation products. This method uses XTerra RP‐18, 5 μm (25 cm×4.6 mm ID), a silica‐based stationary phase with reduced silanol activity. Gradient elution with mobile phases containing acetonitrile, 0.3 M tetrabutylammonium hydrogen sulfate pH 7.5, 0.3 M ethylenediaminetetraacetic acid pH 7.5, water was used at a flow rate of 1.0 mL/min. The column temperature was set at 40°C. The UV detection was performed at 280 nm. A central composite experimental design was performed to optimize the separation. The method permits a good separation of TC and its impurities. Some of these were not reported before and have now been identified for the first time. Some unidentified impurities were also separated.