Abstract

A simple, rapid, and highly sensitive liquid chromatography–mass spectrometry (LC–MS) method was successfully developed and validated for the simultaneous determination of Decitabine and Cedazuridine in plasma matrices. The method demonstrated excellent linearity across the selected calibration range, with correlation coefficients (R²) consistently exceeding 0.999 for both analytes, indicating strong analytical performance. Method validation was carried out in accordance with ICH Q2(R2) and US FDA bioanalytical method validation guidelines, ensuring compliance with internationally accepted standards. The results confirmed that the method meets the required criteria for precision, accuracy, recovery, and stability at all tested quality control levels. Furthermore, the method exhibited low limits of detection (LOD) and quantification (LOQ), highlighting its high sensitivity and suitability for detecting trace-level concentrations in biological samples. The optimized sample preparation procedure contributed to reproducible extraction efficiency while minimizing potential matrix interferences, thereby enhancing overall method reliability. In addition, the analytical conditions were carefully optimized to achieve rapid separation and accurate quantification within a short runtime, making the method both time-efficient and cost-effective. Overall, the validated LC–MS method proved to be robust, reproducible, and dependable for the simultaneous analysis of Decitabine and Cedazuridine. Its strong analytical performance and compliance with regulatory requirements make it a suitable choice for routine applications. The method can be effectively utilized in preclinical and clinical studies, including pharmacokinetic profiling, bioavailability assessment, and therapeutic drug monitoring, where accurate and sensitive quantification is essential.

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1. Garcia-Manero G, Abaza HE, Roboz JA, Stein AA, Klimek JM. Oral cedazuridine/decitabine for myelodysplastic syndromes and chronic myelomonocytic leukaemia: A phase 2 pharmacokinetic/pharmacodynamic randomized crossover study. Lancet Haematol. 2019;6:e237–46.
2. Kantarjian R, Montalban-Bravo E. Decitabine/cedazuridine in myelodysplastic syndromes and chronic myelomonocytic leukaemia: A phase 3 study. Blood Adv. 2021;5(9):2264–83.
3. Dhillon S. Decitabine/cedazuridine: First approval. Drugs. 2021;81(2):179–86.
4. Jabbour E, Garcia-Manero G, Fenaux JM. Pharmacokinetic and safety evaluation of oral cedazuridine/decitabine in myelodysplastic syndromes. Blood. 2020;136(6):674–84.
5. Song Y, Peterson L, Moynihan T. Quantitation of decitabine in human plasma by LC-MS/MS. J Pharm Biomed Anal. 2012;68:153–9.
6. Souryavong P, Hansen MS, Hinzman AL. LC-MS/MS quantification of decitabine and cedazuridine in biological samples. J Chromatogr B. 2023;1224:124007.
7. Liu Z, Marcucci G, Byrd JC. LC-MS/MS determination of decitabine in plasma for pharmacokinetic studies. J Chromatogr B. 2013;912:20–6.
8. Kaufmann B, Souverain S, Cherkaoui S, Christen P, Veuthey JL. Analysis of alkaloids by capillary electrophoresis and LC-MS. Chromatographia. 2002;56(3–4):137–42.
9. Lee MS, Kerns EH. LC/MS applications in drug development. Mass Spectrom Rev. 1999;18(3–4):187–279.
10. Prakash C, Shaffer CL, Nedderman A. Analytical strategies for characterizing drug metabolites. Mass Spectrom Rev. 2007;26(3):340–69.
11. Snyder A, Dolan K. Advances in liquid chromatography method development. J Chromatogr A. 2016;1472:30–43.
12. International Council for Harmonisation (ICH). ICH Q2(R2): Validation of analytical procedures—text and methodology. Geneva: ICH; 2023.
13. S. Food and Drug Administration (FDA). Bioanalytical method validation: Guidance for industry. Silver Spring (MD): FDA; 2018.
14. Bults P, Bischoff R, Bakker H, Gietema J, van de Merbel N. LC-MS/MS-based monitoring of in vivo protein biotransformation: Quantitative determination of trastuzumab and its deamidation products in human plasma. Anal Chem. 2016;88(3):1871–9.
15. Willeman T, Jourdil JF, Gautier-Veyret E, Bonaz B, Stanke-Labesque F. A multiplex liquid chromatography tandem mass spectrometry method for the quantification of seven therapeutic monoclonal antibodies: Application for adalimumab therapeutic drug monitoring in patients with Crohn’s disease. Anal Chim Acta. 2019;1067:63–75.
16. Jiang H, Zeng J, Titsch C, Voronin K, Akinsanya B, Luo L, et al. Fully validated LC-MS/MS assay for the simultaneous quantitation of coadministered therapeutic antibodies in cynomolgus monkey serum. Anal Chem. 2013;85(20):9859–67.
17. Albanell J, Montagut C, Jones ET, Pronk L, Mellado B, Beech J, et al. A phase I study of the safety and pharmacokinetics of the combination of pertuzumab (rhuMab 2C4) and capecitabine in patients with advanced solid tumors. Clin Cancer Res. 2008;14(9):2726–31.
18. Li H, Ortiz R, Tran L, Hall M, Spahr C, Walker K, et al. General LC-MS/MS method approach to quantify therapeutic monoclonal antibodies using a common whole antibody internal standard with application to preclinical studies. Anal Chem. 2012;84(3):1267–73.
19. Liu N, Tengstrand E, Boernsen O, Stephan B, Frank H. Validation of a multiplexed LC-MS/MS clinical assay to quantify insulin growth factor binding proteins in human serum and its application in a clinical study. Toxicol Appl Pharmacol. 2019;371:74–83.
20. Iwamoto N, Takanashi M, Hamada A, Shimada T. Validated LC/MS bioanalysis of rituximab CDR peptides using nano-surface and molecular-orientation limited (nSMOL) proteolysis. Biol Pharm Bull. 2016;39(7):1187–94.

Abstract

A simple, rapid, and highly sensitive liquid chromatography–mass spectrometry (LC–MS) method was successfully developed and validated for the simultaneous determination of Decitabine and Cedazuridine in plasma matrices. The method demonstrated excellent linearity across the selected calibration range, with correlation coefficients (R²) consistently exceeding 0.999 for both analytes, indicating strong analytical performance. Method validation was carried out in accordance with ICH Q2(R2) and US FDA bioanalytical method validation guidelines, ensuring compliance with internationally accepted standards. The results confirmed that the method meets the required criteria for precision, accuracy, recovery, and stability at all tested quality control levels. Furthermore, the method exhibited low limits of detection (LOD) and quantification (LOQ), highlighting its high sensitivity and suitability for detecting trace-level concentrations in biological samples. The optimized sample preparation procedure contributed to reproducible extraction efficiency while minimizing potential matrix interferences, thereby enhancing overall method reliability. In addition, the analytical conditions were carefully optimized to achieve rapid separation and accurate quantification within a short runtime, making the method both time-efficient and cost-effective. Overall, the validated LC–MS method proved to be robust, reproducible, and dependable for the simultaneous analysis of Decitabine and Cedazuridine. Its strong analytical performance and compliance with regulatory requirements make it a suitable choice for routine applications. The method can be effectively utilized in preclinical and clinical studies, including pharmacokinetic profiling, bioavailability assessment, and therapeutic drug monitoring, where accurate and sensitive quantification is essential.

🔒 This is a subscription article

Full text is available to subscribers and institutional members. Please choose an option below to access it.

1. Garcia-Manero G, Abaza HE, Roboz JA, Stein AA, Klimek JM. Oral cedazuridine/decitabine for myelodysplastic syndromes and chronic myelomonocytic leukaemia: A phase 2 pharmacokinetic/pharmacodynamic randomized crossover study. Lancet Haematol. 2019;6:e237–46.
2. Kantarjian R, Montalban-Bravo E. Decitabine/cedazuridine in myelodysplastic syndromes and chronic myelomonocytic leukaemia: A phase 3 study. Blood Adv. 2021;5(9):2264–83.
3. Dhillon S. Decitabine/cedazuridine: First approval. Drugs. 2021;81(2):179–86.
4. Jabbour E, Garcia-Manero G, Fenaux JM. Pharmacokinetic and safety evaluation of oral cedazuridine/decitabine in myelodysplastic syndromes. Blood. 2020;136(6):674–84.
5. Song Y, Peterson L, Moynihan T. Quantitation of decitabine in human plasma by LC-MS/MS. J Pharm Biomed Anal. 2012;68:153–9.
6. Souryavong P, Hansen MS, Hinzman AL. LC-MS/MS quantification of decitabine and cedazuridine in biological samples. J Chromatogr B. 2023;1224:124007.
7. Liu Z, Marcucci G, Byrd JC. LC-MS/MS determination of decitabine in plasma for pharmacokinetic studies. J Chromatogr B. 2013;912:20–6.
8. Kaufmann B, Souverain S, Cherkaoui S, Christen P, Veuthey JL. Analysis of alkaloids by capillary electrophoresis and LC-MS. Chromatographia. 2002;56(3–4):137–42.
9. Lee MS, Kerns EH. LC/MS applications in drug development. Mass Spectrom Rev. 1999;18(3–4):187–279.
10. Prakash C, Shaffer CL, Nedderman A. Analytical strategies for characterizing drug metabolites. Mass Spectrom Rev. 2007;26(3):340–69.
11. Snyder A, Dolan K. Advances in liquid chromatography method development. J Chromatogr A. 2016;1472:30–43.
12. International Council for Harmonisation (ICH). ICH Q2(R2): Validation of analytical procedures—text and methodology. Geneva: ICH; 2023.
13. S. Food and Drug Administration (FDA). Bioanalytical method validation: Guidance for industry. Silver Spring (MD): FDA; 2018.
14. Bults P, Bischoff R, Bakker H, Gietema J, van de Merbel N. LC-MS/MS-based monitoring of in vivo protein biotransformation: Quantitative determination of trastuzumab and its deamidation products in human plasma. Anal Chem. 2016;88(3):1871–9.
15. Willeman T, Jourdil JF, Gautier-Veyret E, Bonaz B, Stanke-Labesque F. A multiplex liquid chromatography tandem mass spectrometry method for the quantification of seven therapeutic monoclonal antibodies: Application for adalimumab therapeutic drug monitoring in patients with Crohn’s disease. Anal Chim Acta. 2019;1067:63–75.
16. Jiang H, Zeng J, Titsch C, Voronin K, Akinsanya B, Luo L, et al. Fully validated LC-MS/MS assay for the simultaneous quantitation of coadministered therapeutic antibodies in cynomolgus monkey serum. Anal Chem. 2013;85(20):9859–67.
17. Albanell J, Montagut C, Jones ET, Pronk L, Mellado B, Beech J, et al. A phase I study of the safety and pharmacokinetics of the combination of pertuzumab (rhuMab 2C4) and capecitabine in patients with advanced solid tumors. Clin Cancer Res. 2008;14(9):2726–31.
18. Li H, Ortiz R, Tran L, Hall M, Spahr C, Walker K, et al. General LC-MS/MS method approach to quantify therapeutic monoclonal antibodies using a common whole antibody internal standard with application to preclinical studies. Anal Chem. 2012;84(3):1267–73.
19. Liu N, Tengstrand E, Boernsen O, Stephan B, Frank H. Validation of a multiplexed LC-MS/MS clinical assay to quantify insulin growth factor binding proteins in human serum and its application in a clinical study. Toxicol Appl Pharmacol. 2019;371:74–83.
20. Iwamoto N, Takanashi M, Hamada A, Shimada T. Validated LC/MS bioanalysis of rituximab CDR peptides using nano-surface and molecular-orientation limited (nSMOL) proteolysis. Biol Pharm Bull. 2016;39(7):1187–94.