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

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 microg/mL range for perfusate; the lower limit of quantification (LLQ) is 0.05 microg/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 degrees 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.