DEVELOPMENT AND VALIDATION OF A LIQUID CHROMATOGRAPHY-ELECTRO-SPRAY IONIZATION-MASS SPECTROMETRY METHOD FOR THE QUANTITATION OF ERGOCALCIFEROL IN HUMAN PLASMAAbstract
A LC-MS method was developed and validated for determination of ergocalciferol in human plasma. n-hexane was used as extraction solvent while kinetex C18 50*2.1 mm, 1.7µm was used as analytical column for chromatographic separation. Mobile phase was acetonitrile: 0.1% formic acid in methanol (90:10). Turbo ion spray with positive polarity was used as an interface with API QTRAP 5500 LC/MS/MS system. The calibration range was validated for 2.03 ng/mL to 150.69 ng/mL. Limit of quantification was 2.03 ng/mL. The recovery of Analyte was 97.04% to 99.27%. Labeled erythromycin was used as an internal standard for this method. Inter-batch and intra-batch accuracy and precision were calculated to check method’s performance. Stability was established for all anticipated working conditions for analyte as well as internal standard. Apart from stability experiments, other experiments were also performed to simulate the conditions of subject sample analysis. This method was applied successfully on bioequivalence study to evaluate pharmacokinetic parameters.
A. Tiwari *, B. Dawkhar, M. Bagul and S. Goswami
Department of Doctoral Studies, R. K. University, Rajkot, Gujarat, India.
12 December, 2017
10 March, 2018
18 March, 2018
01 September, 2018