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Research Journal of Pharmacy and Technology
Year : 2020, Volume : 13, Issue : 7
First page : ( 3274) Last page : ( 3278)
Print ISSN : 0974-3618. Online ISSN : 0974-360X.
Article DOI : 10.5958/0974-360X.2020.00580.6

Sensitive method for the Determination of Colchicine in human plasma by gradient UPLC-ESI-MS/MS: separation of endogenous Interference

Puram Sambasiva Rao, Nithya G*

Department of Chemistry, School of Basic Sciences, Vels Institute of Science, Technology and Advanced Studies, Pallavaram, Chennai, India-600117

*Corresponding Author E-mail: nithyaranju@gmail.com

Online published on 10 August, 2020.

Abstract

The objective of the current research work was to develop a sensitive, specific, accurate and precise method for the determination of Colchicine in human plasma using Liquid Chromatography Electron Spray Ionization-Tandem Mass Spectrometry. Extraction was done using solid phase extraction technique. Ammonium Acetate 10mM with Acetonitrile was used as mobile phase with gradient programme of 6 minutes on Innoval, C18, 125mm x 4.6mm, 5μ column. Detection was performed on ESI positive mode using MRM transitions 400.400/358.300 for colchicine and 406.400/362.000 for colchicine D6 (ISTD) on API 4000 coupled with Shimazdu Nexara UPLC. Developed method was found to be selective, specific and the Calibration Curve was found to be linear over the range 0.075 to 10.091ng/mL. Endogenous interferences were separated from the peak of interest. Method validation was conducted as per EMEA guideline and the developed method can be applied to Pharmacokinetic/Bioequivalence studies.

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Keywords

LC-MS/MS method, Human plasma, gradient elution.

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