BIO-ANALYTICAL METHOD DEVELOPMENT AND VALIDATION FOR THE SIMULTANEOUS ESTIMATION OF DECITABINE AND CEDAZURIDINE IN HUMAN PLASMA USING LC-MS/MS
Keywords:Decitabine, Cedazuridine, LCMS analysis, Human plasma, Bio-analytical method
Objective: The present work aimed to develop a novel, reliable and accurate Liquid Chromatography-Mass Spectrometry/Mass spectrometry (LC-MS/MS) method for the simultaneous quantification of Decitabine and Cedazuridine a combined medication used for the treatment of chronic myelomonocytic leukemia in human plasma.
Methods: Talazoparib drug is used as an internal standard in the study. Both the analytes and internal standard were isolated from 100 ml plasma samples by liquid-liquid extraction and then chromatographed on Zorbax SB-CN (4.6 mm×75 mm, 3.5 µm) column with a mobile phase consisting of 0.1 % ammonium formate and methanol in the ratio of 65:45 (v/v) pumped at 0.5 ml/min. The method had a chromatographic total run time of 5 min.
Results: The developed method gave a symmetric peak at a retention time of 1.7 min for Decitabine, 2.2 min for Cedazuridine, 3.5 min for Talazoparib and satisfied all the peak properties as per USP guidelines. The mass spectral characterization of separated analytes in the LC method was performed using a mass detector operated at Multiple Reaction Monitoring mode with precursor-to-product ion transitions at m/z of 229 to m/z of 114 as MH+ion for Decitabine, m/z of 269 to m/z of 118 as MH+ion for Cedazuridine. A very sensitive limit of detection of 0.3 ng/ml was observed and showed a calibration curve linear over the concentration range of LLOQ (lower limit of quantification) to 500 ng/ml. The other validation parameters were found to have acceptable accuracy, precision, linearity, and selectivity. The mean extraction concentration was acceptable and very high for both the analytes in HQC (high-quality control concentration), MQC (medium quality control concentration) and LLOQ levels. The peak area response ratio of Decitabine and Cedazuridine with the internal standard in freeze-thaw, short term and long term stability studies was found to be acceptable confirms that the method is stable.
Conclusion: It can be concluded that the proposed method is specific, accurate, and precise and could be used for the simultaneous estimation of Decitabine and Cedazuridine in human plasma.
Christine BY, Peter AJ. Epigenetic therapy of cancer: past, present and future. Nat Rev Drug Discovery 2006;5:37-50.
Li LH, Olin EJ, Fraser TJ, Bhuyan BK. Phase specificity of 5-azacytidine against mammalian cells in tissue culture. Cancer Res 1970;30:2770-5.
Stanton LG, Paolo FC, Basem MW, Richard JC. Chapter 57-pharmacology and molecular mechanisms of antineoplastic agents for hematologic malignancies, hematology (Seventh Edition), Elsevier Inc; 2018. p. 849-912.
Michael RS, Olatoyosi O, Philip CA. An oral fixed-dose combination of decitabine and cedazuridine in myelodysplastic syndromes: a multicentre, open-label, dose-escalation, phase 1 study. Lancet Haematol 2019;6:194-203.
Guillermo GM, Elizabeth AG, David PS. Oral cedazuridine/decitabine for MDS and CMML: a phase 2 pharmacokinetic/pharmacodynamic randomized crossover study. Blood 2020;136:674-83.
Jesus DG, Samarpana C, Amit V, Aditi S. The evolution of epigenetic therapy in myelodysplastic syndromes and acute myeloid leukemia. Semin Hematol 2021;58:56-65.
Mohammed IB, Siva SR, Venu S, Sreenivasulu M, Vanitha PK. RP-HPLC-PDA method development, validation and stability studies of the novel antineoplastic drug combination-decitabine and cedazuridine. J Pharm Res Int 2020;32:10-6.
Glory H, Teja BB, Ashok KK, Ravindra RY. Stability indicating RP-HPLC method development and validation of decitabine drug in the formulation. Int J Pharmtech Res 2011;3:237-43.
Yub Raj N, Manish Srivastava, Nafees Ahmad, Kriti Soni, Kanchan Kohli. Stability indicating RP-HPLC method for the estimation of decitabine in bulk drug and lipid-based nanoparticles. Int J Pharm Sci 2014;5:294-302.
Sunilkumar Adupa, Satish K, Ravi J. Development and validation method for decitabine injection by RP-HPLC. Int J Pharm Sci Res 2014;5:3425-9.
Donthineni Kalyan, Swetha A, Arabinda Patnaik, Om Prakash Chary V. A RP-HPLC method development and validation for estimating decitabine with its stability studies. Int J Innov 2014;2:1495-506.
Yahdiana Harahap, Norma Andriyani, Harmita. Method development and validation of lercanidipine in human plasma by liquid chromatography-tandem mass spectrometry. Int J Appl Pharm 2018;10:87-91.
Prasad PBN, Satyanarayana K, Krishna Mohan G. Simultaneous determination of metformin, linagliptin in jentadueto and metformin, saxagliptin in kombiglyze by LC-MS method. Int J Pharm Pharm Sci 2018;10:110-1.
Validation of Analytical Procedures: Text and Methodology, Q R1, Step 4, ICH Tripartie Guidelines; 2005.
Validation of Compendial Methods, United States Pharmacopoeia 30, National Formulary 25, The United States Pharmacopoeial Convention, Inc., Rockville, Md, USA; 2007.
How to Cite
Copyright (c) 2021 Bikshal Babu Kasimala
This work is licensed under a Creative Commons Attribution 4.0 International License.