Analysis of Levonorgestrel in Human Plasma by UPLC-MS/MS: Application to a Bioequivalence Study in Healthy Volunteers

Research Article

Analysis of Levonorgestrel in Human Plasma by UPLC-MS/MS: Application to a Bioequivalence Study in Healthy Volunteers

Sneha G. Nair, Daxesh P. Patel, Frank J. Gonzalez, Puran Singhal, Darshan V. Chaudhary

Corresponding Author : Darshan V. Chaudhary

Department of Chemistry, School of Sciences, Gujarat University, Navrangpura, Ahmedabad 380009, Gujarat, India

Email ID : darshan.msc98@gmail.com

Received : 2017-12-25 Accepted : 2018-01-30 Published : 2018-01-30

Abstract : The metabolic and the pharmacokinetics studies in the pharmaceutical industry at different development stages of drugs have gained a tremendous momentum by using a liquid chromatography interfaced with a tandem mass spectrometry. However, due to the advancement in the drugs analytical techniques like ultra-performance liquid chromatography linked with tandem mass spectrometry (UPLC-MS/MS) have emerged immensely in comparison to the conventional techniques due to advantages in speed, sensitivity, selectivity and resolution with a small particle size. Levonorgestrel (LNG) was determined in human plasma by UPLC-MS/MS while the determination of the internal standard LNG-d6 was done using the SPE mode of extraction using a Phenomenex Strata-X (30 mg, 1cc) cartridge. Waters Acquity UPLC BEH C18 column with the internal diameter of 1.7 µm and mobile phase composition as acetonitrile-2 mM ammonium formate, pH 3.0 (90:10, v/v) was used for the analysis which was then detected with the tandem mass spectrometer equipped with electrospray ionization (ESI) in positive ion mode with the multiple reaction monitoring (MRM). Calibration curve was linear over the range 0.025-25.0 ng/mL with the correlation coefficient value as ≥ 0.99. Precision and accuracy value of intra and inter-batch fall within the range of 0.96-2.84 % and 98.9-101.0 % respectively. Various parameters like sensitivity, selectivity, stability, ruggedness, carryover, dilution integrity, ion suppression/ enhancement was further validated to perk the developed method. The overall recoveries for LNG were within 93.2 to 95.0 %. Stability assessment was done for varied conditions like bench top, freeze and thaw, wet extract, dry extract and long-term stability. The propound method offers greater sensitivity, lower sample consumption, rapid and can be relevantly used for the pharmacokinetic study of 34 human volunteers with the reproducibility value as 122.

Keywords : Levonorgestrel, sensitive, high throughput, UPLC-MS/MS, incurred sample reanalysis.

Citation : Darshan V. Chaudhary et al. (2018). Analysis of Levonorgestrel in Human Plasma by UPLC-MS/MS: Application to a Bioequivalence Study in Healthy Volunteers. J. of Advancement in Medical and Life Sciences. V6I2. DOI : 10.5281/zenodo.1162427

Copyright : © 2018 Darshan V. Chaudhary. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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