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Sample preparation for the measurement of non-protein-bound platinum was evaluated by precipitation of plasma proteins with cold ethanol. The method was compared with the routinely used plasma ultrafiltration and with trichloroacetic acid (TCA) protein precipitation. After incubation of human plasma samples with cisplatin or carboplatin, unbound platinum concentrations were determined applying Amicon Diaflo ultrafiltration membranes and Millipore ultrafree-MC filters. For protein precipitation, 1 ml of cold (-20 degrees C) pure ethanol was added to 0.5 ml of human plasma and the supernatant was collected after 2 h, or 0.5 ml of cold 20% TCA was added to 0.5 ml of plasma. Platinum was analyzed by atomic absorption spectrophotometry (AAS). There was no significant difference between the ethanol and ultrafiltration methods in the unbound platinum concentration. The protein content in the supernatant (1.00 +/- 0.20%) was slightly higher than that in the Amicon (0.58 +/- 0.05%) and Millipore (0.55 +/- 0.04%) ultrafiltrates. On average, the TCA and ethanol method seemed to be equally appropriate. The ethanol precipitation method is concluded to be simple, convenient, and reproducible and has negligible costs.

cancer chemotherapy and pharmacology

Comparison of ethanol plasma-protein precipitation with plasma ultrafiltration and trichloroacetic acid protein precipitation for the measurement of unbound platinum concentrations