Journal of Biomedical and Translational Research

Research Institute of Veterinary Medicine Chungbuk National University (충북대학교 동물의학연구소)
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Development and validation of LC-MS/MS for bioanalysis of hydroxychloroquine in human whole blood

  • Park, Jung Youl (Department of Applied Chemistry, Daejeon University) ;
  • Song, Hyun Ho (Clinical & Bio Research Center, Korea Pharmaceutical Test & Research Institute (KPTR)) ;
  • Kwon, Young Ee (Clinical & Bio Research Center, Korea Pharmaceutical Test & Research Institute (KPTR)) ;
  • Kim, Seo Jin (Department of Marine Molecular Bioscience, College of Life Science, Gangneung-Wonju National University) ;
  • Jang, Sukil (Department of Marine Molecular Bioscience, College of Life Science, Gangneung-Wonju National University) ;
  • Joo, Seong Soo (Department of Marine Molecular Bioscience, College of Life Science, Gangneung-Wonju National University)
  • Received : 2018.11.22
  • Accepted : 2018.12.18
  • Published : 2018.12.31
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Abstract

This study aimed to analyze a high-performance liquid chromatography (HPLC) separation using a pentafluorophenyl column of parent drug hydroxychloroquine (HCQ) and its active metabolite, desethylhydroxchloroquine (DHCQ) applying to determine bioequivalence of two different formulations administered to patients. A rapid, simple, sensitive and specific liquid chromatography-tandem mass spectrometry (LC-MS/MS) method has been developed and validated for bioanalysis of HCQ and its metabolite DHCQ in human whole blood using deuterium derivative $hydroxychloroquine-D_4$ as an internal standard (IS). A triple-quadrupole mass spectrometer was operated using electrospray ionization in multiple reaction monitoring (MRM) mode. Sample preparation involves a two-step precipitation of protein techniques. The removed protein blood samples were chromatographed on a pentafluorophenyl (PFP) column ($50mm{\times}4.6mm$, $2.6{\mu}m$) with a mobile phase (ammonium formate solution containing dilute formic acid) in an isocratic mode at a flow rate of 0.45 mL/min. The standard curves were found to be linear in the range of 2 - 500 ng/mL for HCQ; 2 - 2,000 ng/mL for DHCQ in spite of lacking a highly sensitive MS spectrometry system. Results of intra- and inter-day precision and accuracy were within acceptable limits. A run time of 2.2 min for HCQ and 2.03 min for DHCQ in blood sample facilitated the analysis of more than 300 human whole blood samples per day. Taken together, we concluded that the assay developed herein represents a highly qualified technology for the quantification of HCQ in human whole blood for a parallel design bioequivalence study in a healthy male.

Keywords

References

  1. Bell CL. Hyroxychloroquine sulfate in rheumatoid arthritis: long-term response rate and predictive parameter. Am J Med 1983;75:46-51.
  2. Steere AC, Angelis SM. Therapy for Lyme arthritis: strategies for the treatment of antibiotic-refractory arthritis. Arthritis Rheum 2006;54:3079-3086. https://doi.org/10.1002/art.22131
  3. Nayak V, Esdaile JM. The efficacy of antimalarials in systemic lupus erythematosus. Lupus 1996;5:S23-S27. https://doi.org/10.1177/0961203396005001061
  4. Kapinska-Mrowiecka M, Czubak-Macugowska M, Michcik A, Chomik P, Wlodarkiewicz A. Chronic ulcerative stomatitis (CUS). Lichen planus disseminatus, unguium pedis utriusque and pseudopelade. Am J Case Rep 2010;11:252-257.
  5. McChesney EW. Animal toxicity and pharmacokinetics of hydroxychloroquine sulphate. Am J Med 1983;75:11-18. https://doi.org/10.1016/0002-9343(83)91265-2
  6. McLachlan AJ, Cutler DJ, Tett SE. Plasma protein binding of the enantiomers of hydroxychloroquine and metabolites. Eur J Clin Pharmacol 1993;44:481-484. https://doi.org/10.1007/BF00315548
  7. McLachlan AJ, Tett SE, Cutler DJ, Day RO. Disposition and absorption of hydroxychloroquine enantiomers following a single dose of the racemate. Chirality 1994;6:360-364. https://doi.org/10.1002/chir.530060421
  8. Ducharme J, Fieger H, Ducharme MP, Khalil SK, Wainer IW. Enantioselective disposition of hydroxychloroquine after a single oral dose of the racemate to healthy subjects. Br J Clin Pharmacol 1995;40:127-133. https://doi.org/10.1111/j.1365-2125.1995.tb05768.x
  9. Munster T, Gibbs JP, Shen D, Baethge BA, Botstein GR, Caldwell J, Dietz F, Ettlinger R, Golden HE, Lindsley H, McLaughlin GE, Moreland LW, Roberts WN, Rooney TW, Rothschild B, Sack M, Sebba AI, Weisman M, Welch KE, Yocum D, Furst DE. Hydroxychloroquine concentration-response relationships in patients with rheumatoid arthritis. Arthritis Rheum 2002;46:1460-1469. https://doi.org/10.1002/art.10307
  10. Carmichael SJ, Charles B, Tett SE. Population pharmacokinetics of hydroxychloroquine in patients with rheumatoid arthritis. Ther Drug Monit 2003;25:671-681. https://doi.org/10.1097/00007691-200312000-00005
  11. Miller DR, Fiechtner JJ, Carpenter JR, Brown RR, Stroshane RM, Stecher VJ. Plasma hydroxychloroquine concentrations and efficacy in rheumatoid arthritis. Arthritis Rheum 1987;30:567-571. https://doi.org/10.1002/art.1780300512
  12. Costedoat-Chalumeau N, Amoura Z, Hulot JS, Aymard G, Leroux G, Marra D, Lechat P, Piette JC. Very low blood hydroxychloroquine concentration as an objective marker of poor adherence to treatment of systemic lupus erythematosus. Ann Rheum Dis 2007;66:821-824. https://doi.org/10.1136/ard.2006.067835
  13. Costedoat-Chalumeau N, Amoura Z, Hulot JS, Hammoud HA, Aymard G, Cacoub P, Frances C, Wechsler B, Huong du LT, Ghillani P, Musset L, Lechat P, Piette JC. Low blood concentration of hydroxychloroquine is a marker for and predictor of disease exacerbations in patients with systemic lupus erythematosus. Arthritis Rheum 2006,54:3284-3290. https://doi.org/10.1002/art.22156
  14. Lim HS, Im JS, Cho JY, Bae KS, Klein TA, Yeom JS, Kim TS, Choi JS, Jang IJ, Park JW. Pharmacokinetics of hydroxychloroquine and its clinical implications in chemoprophylaxis against malaria caused by Plasmodium vivax. Antimicrob Agents Chemother 2009;53:1468-1475. https://doi.org/10.1128/AAC.00339-08
  15. Tett SE, Cutler DJ, Day RO, Brown KF. Bioavailability of hydroxychloroquine tablets in healthy volunteers. Br J Clin Pharmacol 1989;27:771-779. https://doi.org/10.1111/j.1365-2125.1989.tb03439.x
  16. Brown RR, Stroshane RM, Benziger DP. High-performance liquid chromatographic assay for hydroxychloroquine and three of its major metabolites, desethylhydroxychloroquine, desethylchloroquine and bidesethylchloroquine, in human plasma. J Chromatogr 1986;377:454-459. https://doi.org/10.1016/S0378-4347(00)80809-9
  17. Tett SE, Cutler DJ, Brown KF. High performance liquid chromatographic assay for hydroxychloroquine and metabolites in blood and plasma, using a stationary phase of poly (styrenedivinylbenzene) and a mobile phase at pH 11, with fluorimetric detection. J Chromatogr 1985;344:241-248. https://doi.org/10.1016/S0378-4347(00)82024-1
  18. Morris RG. Estimation of plasma hydroxychloroquine by high-performance liquid chromatography with ultraviolet detection. J Chromatogr 1985;338:422-427. https://doi.org/10.1016/0378-4347(85)80115-8
  19. Volin P. Simple and specific reversed-phase liquid chromatographic method with diode-array detection for simultaneous determination of serum hydroxychloroquine, chloroquine and some corticosteroids. J Chromatogr 1995;666:347-353. https://doi.org/10.1016/0378-4347(94)00584-R
  20. Midha KK, Hubbard JW, Rawson MJ, McKay G, Schwede R. The roles of stereochemistry and partial areas in a parallel design study to assess the bioequivalence of two formulations of hydroxychloroquine: a drug with a very long half-life. Eur J Pharm Sci 1996;4:283-292. https://doi.org/10.1016/0928-0987(96)00178-9
  21. Wang LZ, Ong RY, Chin TM, Thuya WL, Wan SC, Wong AL, Chan SY, Ho PC, Goh BC. Method development and validation for rapid quantification of hydroxychloroquine in human blood using liquid chromatography-tandem mass spectrometry. J Pharm Biomed Anal 2012;61:86-92. https://doi.org/10.1016/j.jpba.2011.11.034
  22. Lacy CF, Armstrong LL, Goldman MP, Lance LL. Drug information handbook with international trade names index. 19th ed. Hudson: Lexi-Comp Inc; 2010, p. 818-820.
  23. Tett SE, Cutler DJ, Day RO, Brown KF. A dose-ranging study of the pharmacokinetics of hydroxychloroquine following intravenous administration to healthy volunteers. Br J Clin Pharmacol 1988;26:303-313. https://doi.org/10.1111/j.1365-2125.1988.tb05281.x
  24. Tett S, Cutler D, Day R. Antimalarials in rheumatic diseases. Baillieres Clin Rheumatol 1990;4:467-489. https://doi.org/10.1016/S0950-3579(05)80004-4

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