A UPLC-MS/MS METHOD DEVELOPMENT AND VALIDATION FOR THE ESTIMATION OF POMALIDOMIDE FROM HUMAN PLASMA
DOI:
https://doi.org/10.22159/ijap.2017v9i1.15653Keywords:
Pomalidomide, LC-MSMS, Human plasma, Liquid-liquid extractionAbstract
Objective: The present work aimed to develop a simple, rapid, specific and precise liquid chromatography-tandem mass spectrophotometric (LC–MS/MS) validated method for quantification of pomalidomide and internal standard (ISTD) Fluconazole in human plasma.
Methods: 50 µl of 0.1% formic acid was added to plasma samples prior to liquid-liquid extraction (LLE) using 2.5 ml of ethyl acetate. Chromatographic separation was achieved on Xterra, RP18, 5 µ (50 x 4.6 mm) column using a mixture of 0.1% (v/v) formic acid in water to methanol at a ratio of 12:88, v/v as the mobile phase. The flow rate was 0.50 ml/min. The LC eluent was split, and approximately 0.1 ml/min was introduced into Tandem mass spectrometer using turbo Ion Spray interface at 325 °C. Quantitation was performed by transitions of m/z 260.1 precursor ion to the m/z 148.8 for pomalidomide and m/z 307.1/238.0 for fluconazole.
Results: The concentrations of nine working standards showed linearity between 9.998 to 1009.650 ng/ml (r2 ≥ 0.9968). Chromatographic separation was achieved within 2 min. The average extraction recoveries of three quality control concentrations were 53.86% for pomalidomide and were within the acceptance limits. The coefficient of variation was ≤15% for intra-and inter-batch assays. The %CV of ruggedness ranges 1.32 to 4.03. The % stability of short term and long term stock solution stability studies was found to be 99.01% and 98.49% respectively.
Conclusion: The results obtained for specificity, linearity, accuracy, precision, ruggedness and stability studies were within limits. Thus the validated economical method was applied for pharmacokinetic studies of pomalidomide.
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References
Lacy MQ, McCurdy AR. Pomalidomide. Blood 2013;122:2305-9.
https://pubchem.ncbi.nlm.nih.gov/compound/134780. [Last accessed on 12 Jun 2016]
Amato D, Robert J, Loughnan, Michael S, Flynn, Evelyn, et al. Thalidomide is an inhibitor of angiogenesis. Proc Natl Acad Sci USA 1994;91:4082-5.
Amato D, Lentzsch R, Anderson S, Rogers KCMS. Mechanism of action of thalidomide and 3-aminothalidomide in multiple myeloma. Semin Oncol 2001;28:597-601.
Teo SK, Chen Y, Muller GW, Chen RS, Thomas SD, Stirling DI, et al. Chiral inversion of the second generation IMiD CC-4047 (ACTIMID) in human plasma and phosphate-buffered saline. Chirality 2003;15:348-51.
Meiler SE, Wade M, Kutlar F, Yerigenahally SD, Xue Y, Moutouh-de Parseval LA, et al. Pomalidomide augments fetal hemoglobin production without the myelosuppressive effects of hydroxyurea in transgenic sickle cell mice. Blood 2011;118:1109-12.
Hoffmann M, Kasserra C, Reyes J, Schafer P, Kosek J, Capone L, et al. Absorption, metabolism and excretion of [14C] pomalidomide in humans following oral administration. Cancer Chemother Pharmacol 2013;71:489-501.
Iqbal M, Ezzeldin E, Al-Rashood KA, Shakeel F. A validated UPLC-MS/MS assay using negative ionisation mode for high-throughput determination of pomalidomide in rat plasma. J Chromatogr B: Anal Technol Biomed Life Sci 2015;983:76-82.
Analytical Procedures and Methods Validation for Drugs and Biologics Guidance for Industry. U. S. Department of Health and Human Services, Food and Drug Administration Center for Drug Evaluation and Research (CDER), Center for Biologics Evaluation and Research (CBER); 2015.
Sindhusri M, Swetha T, Ramadevi A, Ashok Kumar A. A novel rapid rp-hplc method development and validation for the quantitative estimation of balofloxacin in tablets. Int J Pharm Pharm Sci 2014;7:319-22.
Raveendra Babu G, Lakshmana Rao A, Venkateswara Rao J. A rapid RP-HPLC method development and validation for the quantitative estimation ribavirin in tablets. Int J Pharm Pharm Sci 2014;7:60-3.
Srinidhi M, Mushabbar Basha MD, V Raj Kumar, Rajendra Kumar J. Stability indicating RP-HPLC method development and validation for the estimation of sumatriptan in bulk and pharmaceutical dosage form. J Appl Pharm Sci 2016;6:20-5.
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