Simple high-performance liquid chromatographic assay for melphalan in perfusate, rat liver and tumour tissue

• Published in: Biomedical chromatography 2003-10, Vol.17 (7), p.458-464
• Main Authors: Sparidans, Rolf W., Silvertand, Linda, Dost, Frits, Rothbarth, Joost, Mulder, Gerard J., Schellens, Jan H. M., Beijnen, Jos H.
• Format: Article
• Language: English
• Subjects: Animals; Antineoplastic Agents, Alkylating - analysis; biological matrix; Chromatography, High Pressure Liquid - methods; isolated liver perfusion; Liver - chemistry; Liver Neoplasms, Experimental - chemistry; Male; Melphalan - analysis; nitrogen mustard; Perfusion; Rats; Reproducibility of Results; Sensitivity and Specificity; Spectrophotometry, Ultraviolet; stability; ultraviolet detection
• ISSN: 0269-3879; 1099-0801
• DOI: 10.1002/bmc.273

A simple, sensitive and selective reversed‐phase liquid chromatographic assay has been developed and validated for the anti‐cancer agent melphalan in perfusate, liver and tumour tissue originating from isolated rat liver perfusion studies. Melphalan was extracted from the matrix using ice‐cold methanol. The drug and the internal standard, propylparaben, were detected using ultraviolet absorbance at 262 nm. The assay has been validated in the 0.05–25 µg/mL range for perfusate; the lower limit of quantification (LLQ) is 0.05 µg/mL in perfusate and 0.25 ng/mg in liver and tumour tissues. Accuracies ranged from 89 to 110% and the inter‐assay precisions were all below 15% (20% at the LLQ). Melphalan in a biological matrix has to be processed between 0 and 4°C and is stable under all relevant processing and storage conditions tested. The assay has been exhaustively used in isolated liver perfusion studies with the drug demonstrating its applicability. Copyright © 2003 John Wiley & Sons, Ltd.