Thin Layer Coulometry Based on Ion-Exchanger Membranes for Heparin Detection in Undiluted Human Blood
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文摘
We explore here for the first time a potentially calibration-free methodology for the detection of protamine (and, by titration, heparin) in undiluted human blood in the therapeutic concentration range from 20 to 120 mg L鈥?. The use of a thin layer sample (5.8 渭L) confined between a tubular protamine selective membrane (inner diameter, 600 渭m) and a Ag/AgCl wire (diameter 400 渭m) achieves an exhaustive depletion from the sample. Coulometry detection was chosen for the interrogation of the thin layer, employing a double pulse technique with 120 s for each pulse. Protamine calibration curves were recorded at physiological concentrations and in undiluted human blood. A linear relationship was obtained in both cases, but a diminished sensitivity was observed in contact with blood, which is explained with a partial passivation of the inner Ag/AgCl element. Heparin鈥損rotamine titrations were performed in undiluted human blood samples, mimicking the final application with patients undergoing critical care. The observed values correlate satisfactorily with those of an alternative technique, so-called flash-chronopotentiometry on planar membranes.
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