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A Rapid and Sensitive UPLC–ESI-MS/MS Method for Determination of Levonorgestrel by Chemical Derivatization in Human Plasma and its Application to Pharmacokinetic Study | Abstract
ISSN: 2155-9872

Journal of Analytical & Bioanalytical Techniques
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Research Article

A Rapid and Sensitive UPLC–ESI-MS/MS Method for Determination of Levonorgestrel by Chemical Derivatization in Human Plasma and its Application to Pharmacokinetic Study

Praveen Kumar V1,2*, Ashish Saxena1,3, Amol Pawar1, Sundara Moorthi Nainar M1, Ravikiran V1, Ravisekhar Kashibhatta1, Pratima Ashawat2and Ashawat MS4

1Bioanalytical Research Department, Lupin Bio-Research Center, Maharastra State, India

2Department of Science, Pacific University, Rajasthan State, India

3Department of Pharmacy, Pacific University, Rajasthan State, India

4Laureate Institute of Pharmacy, Himachal Pradesh, India

*Corresponding Author:
Praveen Kumar
Bioanalytical Research Department
Lupin Bio-Research Center
Pashan, Pune–411021,
Maharastra State, India
Tel: +91-020-66219242
Fax: +91-020-66219224
E-mail: [email protected]

Received date: June 06, 2014; Accepted date: June 23, 2014; Published date: June 25, 2014

Citation: Kumar VP, Saxena A, Pawar A, Nainar MSM, Ravikiran V, et al.(2014) A Rapid of Levonorgestrel by Chemical Derivatization in Human Plasma and its Application to Pharmacokinetic Study. J Anal Bioanal Techniques S6:003. doi: 10.4172/2155-9872.S6-003

Copyright: © 2014 Kumar VP, et al. 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.


A rapid and sensitive liquid chromatography–tandem mass spectrometry (LC–MS/MS) method in human plasma was developed for determination of levonorgestrel using ultra performance liquid chromatography aligned with tandem mass spectrometric detection. The analytes were extracted using solid–phase extraction technique followed by rapid derivatization. Levonorgestrel and levonorgestrel d6 were resolved on Kromasil C18 (50 × 4.6 mm) using gradient flow acetonitrile and 0.1% formic acid solution with a total run time of 5 minutes. The detection was achieved using Waters XEVO–TQ–S mass spectrometry system with a mass transition ion-pair of m/z 328.2 → 90.9 for levonorgestrel and m/z 334.1 → 91.0 for levonorgestrel D6. The method has been validated for a linear range of 100–30000 pg/ml with a correlation coefficient ≥ 0.99. The precision (%RSD) was less than 6.50% and accuracy (%RE) was within ± 5%. The overall recoveries for levonorgestrel and levonorgestrel D6 were 93.69% and 93.88% respectively. The validated method was applied for the pharmacokinetic study of levonorgestrel following a single oral dose of levonorgestrel/ethinyl estradiol (0.15 mg/0.03 mg) tablets in 36 healthy female volunteers.


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