Abstract
It is desirable to develop a fast method for quantification of melphalan due to its instability. Here we report a method for quantification of melphalan (MPL) in human plasma using a UPLC–PDA system. Briefly, 50 µL plasma sample was mixed with 25 µL internal standard (2500 ng/mL acetylmelphalan in methanol) and 25 µL 20% trichloroacetic acid, and centrifuged at 21,000 g (15,000 rpm) at 4 °C for 3 min. The supernatant (5 µL) was injected onto an Acquity™ BEH C18 LC column (2.1 × 50 mm, 1.7 µm) and eluted with 25 mM NH4AC (pH 4.7)—acetonitrile in a gradient mode at a flow rate of 0.6 mL/min. The column kept at 40 ± 5 °C and the autosampler kept at 4 ± 5 °C. The detector set at 261 nm, and sampling rate was 40points/sec. The retention times were typically 2.11 min for melphalan and 2.38 min for the internal standard. Total run time is 4 min per sample. Calibration range was 100–40,000 ng/mL. The lower limit of quantification was 100 ng/mL. The method was validated based on the FDA guidelines, and applied to a clinical pharmacokinetic study in pediatric patients.
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Acknowledgements
This work was supported by the National Center for Research Resources and the National Center for Advancing Translational Sciences, National Institutes of Health, through UCSF-CTSI Grant Number UL1 RR024131 (a shared instrument award to Dr. Huang) and UCSF-CTSI Grant number KL2 TR000143 (Dr. Long-Boyle). Its contents are solely the responsibility of the authors and do not necessarily represent the official views of the NIH.
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Huang, L., Cheah, V., Chan, D. et al. Determination of melphalan in human plasma by UPLC–UV method. Cancer Chemother Pharmacol 83, 905–910 (2019). https://doi.org/10.1007/s00280-019-03786-6
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DOI: https://doi.org/10.1007/s00280-019-03786-6