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Determination of Cycloserine in Human Plasma by High Performance Chromatography-Tandem Mass Spectrometry
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A selective, sensitive, rugged and high throughput high performance liquid chromatography tandem mass spectrometric method was developed for the estimation of Cycloserine in human plasma using Acyclovir as an internal standard (IS). Cycloserine and IS were extracted from human plasma by solid phase extraction using Water Oasis MCX 1cc/30mg cartridges. The samples were chromatographed on Inertsil ODS-3V, 4.6 × 250 mm, 5μ column using a mobile phase consisting of Milli-Q water: Acetonitrile: formic acid (30:70:0.3 v/v). The chromatographic separation is achieved in 2.6 minutes. The protonated precursor product ion transition for cycloserine and internal standard were monitored in multiple reaction monitoring (MRM) and positive ion mode. The method was validated over a concentration range of 0.200 μg/ml to 16.000 μg/ml. The method was validated for its sensitivity, selectivity, accuracy and precision, matrix effect recovery, various stabilities and dilution integrity.
Keywords
LC-MS/MS, Cycloserine, Tuberculosis, Solid Phase extraction and Human Plasma.
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