Abstract
A simple and highly sensitive fluorometric high-performance liquid chromatographic method was developed for the determination of busulfan in human serum. After busulfan and 1,6-bis(methanesulfonyloxy)hexane as an internal standard were extracted from serum with ethyl acetate, they were derivatized with 2-naphthalenethiol in an alkaline medium. The derivatives were separated by reversed-phase chromatography on a YMC-Pack C4 column with a mixture of methanol–0.1 M sodium acetate buffer (pH 7.0) (8:2, v/v) as a mobile phase, and were then detected spectrofluorometrically at 370 nm with excitation at 255 nm. Extraction and derivatization efficiencies were 73.9–75.1% and greater than 91.1%, respectively. The detection limit for busulfan added to serum was 2 ng (8 pmol) ml–1 serum (330 fmol on column) at a signal-to-noise ratio of three with a linear relationship over the 10 ng–3.0 µg ml–1 (0.04–12 µM) concentration range. The accuracy and precision of this method were within 7.0% and 9.3% even at low concentration (10 ng ml–1). The within- and between-day variations were lower than 9.3% and 10.9%, respectively.
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G. P. Canellos, Clin. Haematol., 1977, 6, 113.
H. D. Preisler and D. J. Higby, N. Y. J. Med., 1982, 82, 703.
J. R. Hobbs, K. Hugh-Jones, P. J. Shaw, C. J. C. Dowme, and S. Williamson, Bone Marrow Transplant., 1986, 1, 201.
P. J. Tutschka, E. A. Copelan, and J. P. Klein, Blood, 1987, 70, 1382.
G. Vassal, A.Gouyette, O. Hartmann, J. L. Pico, and J. Lemerle, Cancer Chemother. Pharmacol., 1989, 24, 386.
W. Emminger, W. Emminger-Schmidmeier, C. Peters, and H. Gadner, Blood, 1991, 77, 1621.
T. J. Nevill, M. J. Barnett, H. G. Klingemann, D. E. Reece, J. D. Shepherd, and G. L. Phillips, J. Clin. Oncol., 1991, 9, 1224.
M. Hassan and H. Ehrsson, J. Chromatogr., 1983, 277, 374.
T. L. Chen, L. B. Grochow, L. A. Hurowitz, and R. B. Brundrett, J. Chromatogr., 1988, 425, 303.
L. Embree, R. B. Burns, J. R. Heggie, G. L. Phillips, D. E. Reece, J. J. Spinelli, D. O. Hartley, N. J. Hudon, and J. H. Goldie, Cancer Chemother. Pharmacol., 1993, 32, 137.
R. B. Burns, J. R. Heggie, and L. Embree, J. Pharm. Biomed. Anal., 1995, 13, 1073.
G. Vassal, M. Re, and A. Gouyette, J. Chromatogr., 1988, 428, 357.
M. H. Quernin, B. Poonkuzhali, C. Montes, R. Krishnamoorthy, D. Dennison, A. Srivastava, E. Vilmer, M. Chandy, and E. Jacqz-Aigrain, J. Chromatogr. B, 1998, 709, 47.
W. D. Henner, E. A. Furlong, M. D. Flaherty, and T. C. Shea, J. Chromatogr., 1987, 416, 426.
J. J. MacKichan and T. P. Bechtel, J. Chromatogr., 1990, 532, 424.
J. Blanz, C. Rosenfeld, B. Proksch, and G. Ehninger, J. Chromatogr., 1990, 532, 429.
K. Funakoshi, K. Yamashita, W. Chao, M. Yamaguchi, and T. Yashiki, J. Chromatogr. B, 1994, 660, 200.
D. S. L. Chow, H. P. Bhagwatwar, S. Phadungpojna, and B. S. Andersson, J. Chromatogr. B, 1997, 704, 277.
J. R. Heggie, M. Wu, R. B. Burns, C. S. Ng, H. C. Fung, G. Knight, M. J. Barnett, J. J. Spinelli, and L. Embree, J. Chromatogr. B, 1997, 692, 437.
A. G. Kazemifard and D. J. Morgan, J. Chromatogr., 1990, 528, 274.
S. Pichini, I. Altieri, A. Bacosi, S. DiCarlo, P. Zuccaro, P. Iannetti, and R. Pacifici, J. Chromatogr., 1992, 581, 143.
P. Ljungman, M. Hassan, A. N. Bekassy, O. Ringden, and G. Oberg, Bone Marrow Transplant., 1995, 15, 869.
G. Vassal, S. Koscienlny, D. Challine, D. Valteau-Couanet, I. Boland, A. Deroussent, J. Lemerle, A. Gouyette, and O. Hartman, Cancer Chemother. Pharmacol., 1996, 37, 247.
A. G. Kazemifard and D. J. Morgan, J. Chromatogr., 1990, 528, 274.
M. H. Quernin, B. Poonkuzhali, Y. Medard, D. Dennison, A. Srivastava, R. Krishnamoorthy, M. Chandy, and E. Jacqz-Aigrain, J. Chromatogr. B, 1999, 721, 147.
J. E. Peris, J. A. Latorre, V. Castel, A. Verdeguer, S. Esteve, and F. Torres-Molina, J. Chromatogr. B, 1999, 730, 33.
M. Hassan and H. Ehrsson, Drug Metab. Dispos., 1987, 15, 399.
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Hara, S., Tsuchie, M., Tsujioka, R. et al. High-Performance Liquid Chromatographic Quantification of Busulfan in Human Serum after Fluorescence Derivatization by 2-Naphthalenethiol. ANAL. SCI. 16, 287–291 (2000). https://doi.org/10.2116/analsci.16.287
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DOI: https://doi.org/10.2116/analsci.16.287