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Determination of Cycloserine in Human Plasma by High Performance Chromatography-Tandem Mass Spectrometry


Affiliations
1 School of Pharmacy and Technology Management, Narsee Monjee Institute of Management and Higher Studies University, Vile Parle (West), Mumbai-400 056, India
2 Macleods Pharmaceuticals Ltd., G-2, Mahakali Caves Road, Shanti Nagar, Andheri West, Mumbai-400093, India
     

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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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  • Determination of Cycloserine in Human Plasma by High Performance Chromatography-Tandem Mass Spectrometry

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Authors

Halde Supriya
School of Pharmacy and Technology Management, Narsee Monjee Institute of Management and Higher Studies University, Vile Parle (West), Mumbai-400 056, India
Mungantiwar Ashish
Macleods Pharmaceuticals Ltd., G-2, Mahakali Caves Road, Shanti Nagar, Andheri West, Mumbai-400093, India
Chintamaneni Meena
School of Pharmacy and Technology Management, Narsee Monjee Institute of Management and Higher Studies University, Vile Parle (West), Mumbai-400 056, India

Abstract


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.