An LC-MS Assay with Isocratic Separation and On-Line Solid Phase Extraction to Improve the Routine Therapeutic Drug Monitoring of Busulfan in Plasma

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Summary

Background: Busulfan (Bu) requires therapeutic drug monitoring (TDM) in subjects undergoing a conditioning regimen for hematopoietic stem cell transplantation (HSCT). To speed up the procedure and increase reproducibility, we improved our routine LC-MS/MS assay using the on-line solid-phase extraction (SPE) of samples.

Methods: A protein precipitation (PP) step was performed before the on-line SPE of Bu from 200 μL of plasma spiked with octa-deuterated Bu (D8-Bu) as the internal standard. Bias was assessed with respect to our routine LC-MS/MS Bu assay with off-line extraction using the Passing-Bablok robust regression. Root cause of bias for individual samples was assessed by analyzing the regression residuals.

Results: The method was linear in the range 37.75-2,416 ng/mL (r2>0.999), with 19.74 ng/mL LLOQ and 10.5% CV at 20 ng/mL. Precision and accuracy were both within ±5%, and neither appreciable matrix nor carryover effects were observed. The Passing-Bablok regression analysis returned a 0.99 slope (95% Cl: 0.97 to 1.01) and -6.82 intercept (95% Cl: -15.23 to 3.53). Residuals analysis against the 2.5th-97.5th percentiles range showed four samples with significant bias individually.

Conclusions: The method presented can be successfully employed for the routine analysis of Bu in plasmatic samples, and can replace the LC-MS/MS method with off-line extraction without any statistically significant overall bias. In this regard, samples with individual significant bias were reasonably produced by preanalytical issues which had no relation with the conversion to the on-line SPE extraction.

1. Lucarelli G, Isgro A, Sodani P, Gaziev J. Hematopoietic stem cell transplantation in thalassemia and sickle cell anemia. Cold Spring Harb Perspect Med 2012; 2: a011825.

2. Russell JA, Kangarloo SB. Therapeutic drug monitoring of busulfan in transplantation. Curr Pharm Des 2008; 14: 1936-49.

3. Iwamoto T, Hiraku Y, Oikawa S, Mizutani H, Kojima M, Kawanishi S. DNA intrastrand cross-link at the 5’-GA-3’ sequence formed by busulfan and its role in the cytotoxic effect. Cancer Sci 2004; 95: 454-8.

4. Andersson BS, Madden T, Tran HT, Hu WW, Blume KG, Chow DS, et al. Acute safety and pharmacokinetics of intravenous busulfan when used with oral busulfan and cyclophosphamide as pretransplantation conditioning therapy: a phase I study. Biol Blood Marrow Transplant 2000; 6: 548-54.

5. Andersson BS, Thall PF, Madden T, Couriel D, Wang X, Tran HT, et al. Busulfan systemic exposure relative to regimen- related toxicity and acute graft-versus-host disease: defining a therapeutic window for i.v. BuCy2 in chronic myelogenous leukemia. Biol Blood Marrow Transplant 2002; 8: 477-85.

6. Juenke JM, Miller KA, McMillin GA, Johnson-Davis KL. An automated method for supporting busulfan therapeutic drug monitoring. Ther Drug Monit 2011; 33: 315-20.

7. Courtney JB, Harney R, Li Y, Lundell G, McMillin GA, Agarwal G, et al. Determination of busulfan in human plasma using an ELISA format. Ther Drug Monit 2009; 31: 489-94.

8. Kellogg MD, Law T, Sakamoto M, Rifai N. Tandem mass spectrometry method for the quantification of serum busulfan. Ther Drug Monit 2005; 27: 625-9.

9. Snyder ML, Ritchie JC. Quantification of busulfan in plasma using liquid chromatography electrospray tandem mass spectrometry (HPLC-ESI-MS/MS). Methods Mol Biol 2010; 603: 129-36.

10. Rauh M, Stachel D, Kuhlen M, Groschl M, Holter W, Rascher W. Quantification of busulfan in saliva and plasma in haematopoietic stem cell transplantation in children: validation of liquid chromatography tandem mass spectrometry method. Clin Pharmacokinet 2006; 45: 305-16.

11. Ansari M, Uppugunduri CR, Deglon J, Theoret Y, Versace F, Gumy-Pause F, et al. A simplified method for busulfan monitoring using dried blood spot in combination with liquid chromatography/tandem mass spectrometry. Rapid Commun Mass Spectrom 2012; 26: 1437-46.

12. Adaway JE, Keevil BG. Therapeutic drug monitoring and LC-MS/MS. J Chromatogr B Analyt Technol Biomed Life Sci 2012; 883-884: 33-49.

13. Chen L, Wang H, Zeng Q, Xu Y, Sun L, Xu H, et al. Online coupling of solid-phase extraction to liquid chromatography - a review. J Chromatogr Sci 2009; 47: 614-23.

14. Gaziev J, Nguyen L, Puozzo C, Mozzi AF, Casella M, Per - rone Donnorso M, et al. Novel pharmacokinetic behavior of intravenous busulfan in children with thalassemia undergoing hematopoietic stem cell transplantation: a prospective evaluation of pharmacokinetic and pharmacodynamic profile with therapeutic drug monitoring. Blood 2010; 115: 4597-604.

15. Annesley TM. Ion suppression in mass spectrometry. Clin Chem 2003; 49: 1041-4.

16. Institute CaLS. Evaluation of linearity of quantitative measu rement procedures: a statistical approach; approved guide line (EP6-A) Wayne, PA: CLSI 2003.

17. Use ICoHoTRfRoPfH. Validation of analytical procedures: text and methodology Q2(R1); 2005.

18. Institute CaLS. Preliminary evaluation of quantitative clinical laboratory measurement procedures; approved guide line (EP10-A3). Wayne, PA CLSI; 2006.

19. Department of Health and Human Services USFaDA, Center for Drug Evaluation and Research. Guidance for the industry: bioanalytical method validation. Rockville, MD2001.

20. Matuszewski BK, Constanzer ML, Chavez-Eng CM. Strate gies for the assessment of matrix effect in quantitative bioanalytical methods based on HPLC-MS/MS. Anal Chem 2003; 75: 3019-30.

21. Gaziev J, Isgro A, Mozzi AF, Petain A, Nguyen L, Ialongo C, et al. New insights into the pharmacokinetics of intravenous busulfan in children with sickle cell anemia undergoing bone marrow transplantation. Pediatr Blood Cancer 2015; 62: 680-6.

22. Theodorsson E. Quality assurance in clinical chemistry: a touch of statistics and a lot of common sense. J Med Biochem 2016; 35: 103-112.

23. Moon SY, Lim MK, Hong S, Jeon Y, Han M, Song SH, et al. Quantification of human plasma-busulfan concentration by liquid chromatography-tandem mass spectrometry. Ann Lab Med 2014; 34: 7-14.

24. Spasojevic I, da Costa LR, Horwitz ME, Long GD, Sulli - van KM, Chute JP, et al. Mini test dose of intravenous busulfan (busulfex (R)) in allogeneic non-myeloablative stem cell transplantation, followed by liquid chromatography tandem-mass spectrometry. Cancer Invest 2012; 30: 679-82.

25. French D, Sujishi KK, Long-Boyle JR, Ritchie JC. Development and validation of a liquid chromatographytandem mass spectrometry assay to quantify plasma busulfan. Ther Drug Monit 2014; 36: 169-74.

26. Danso D, Jannetto PJ, Enger R, Langman LJ. High- Throughput Validated Method for the Quantitation of Busulfan in Plasma Using Ultrafast SPE-MS/MS. Ther Drug Monit 2015; 37: 319-24.

27. Bunch DR, Heideloff C, Ritchie JC, Wang S. A fast and simple assay for busulfan in serum or plasma by liquid chromatography-tandem mass spectrometry using turbulent flow online extraction technology. J Chromatogr B Analyt Technol Biomed Life Sci 2010; 878: 3255-8.

28. Eksborg S. Evaluation of method-comparison data. Clin Chem 1981; 27: 1311-2.

29. Bland JM, Altman DG. Statistical methods for assessing agreement between two methods of clinical measurement. Lancet 1986; 1: 307-10.

30. Euser AM, Dekker FW, le Cessie S. A practical approach to Bland-Altman plots and variation coefficients for log transformed variables. J Clin Epidemiol 2008; 61: 978-82.

31. Balasubramanian P, Srivastava A, Chandy M. Stability of busulfan in frozen plasma and whole blood samples. Clin Chem 2001; 47: 766-8.

32. Murdter TE, Coller J, Claviez A, Schonberger F, Hofmann U, Dreger P, et al. Sensitive and rapid quantification of busulfan in small plasma volumes by liquid chromatography- electrospray mass spectrometry. Clin Chem 2001; 47: 1437-42.

Journal of Medical Biochemistry

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