Electrochemical determination of hemoglobin, hematocrit, and hemolysis.

Novel electrochemical methods have been developed for determination of total hemoglobin, hematocrit, and detection of hemolysis in whole blood. Hemoglobin is measured through its peroxidase activity, a fluoride ion-selective electrode being used to monitor the rate of fluoride ion production from the oxidation of an organofluorine compound. Results agree well with those obtained with the cyanmethemoglobin method (r = 0.970). Hematocrit is determined from the ratio of the sodium ion concentrations measured with an ion-selective electrode before and after lysis of the erythrocytes. Results by this and the microhematocrit method correlated well (r = 0.987). Hemolysis in a whole-blood sample is detected by using an oxygen electrode to measure the oxygen released when hemoglobin in plasma is oxidized.

Platelet metabolism during storage of platelet concentrates at 22 degrees C.

The development of methods for storing platelet concentrates (PCs) at 22 degrees C for transfusion has been predominantly empiric, with minimal knowledge of metabolic events occurring during storage. It is known that a decrease in pH due to accelerated production of lactic acid in hypoxic conditions is a major cause for loss of platelet viability. In the current studies, we have measured metabolic parameters such as O2 and glucose consumption rates and CO2 and lactic acid production rates. We have also determined the O2 and CO2 transport capacities of various containers and the buffering capacity of plasma. The O2 consumption rate was 1.10 +/- 0.16 (SD) nmol/min/10(9) platelets. In well-oxygenated systems, lactic acid formation was 1.74 +/- 0.12 nmol/min/mL PC for PCs with a platelet count of 1 to 2 X 10(9)/mL; and 0.52 mol of glucose was consumed per 1 mol lactic acid produced. In a completely oxygen-free system, lactic acid production increased 5-8-fold. These calculations suggest that 85% of energy generation is derived through oxidative metabolism and that glucose may not be the primary substrate for this metabolism. Bicarbonate concentration, initially 22.1 +/- 1.6 mEq/L, decreased 1.41 +/- 0.18 nEq/min/mL PC for PCs with counts 1 to 2 X 10(9) platelets/mL. The loss of bicarbonate was caused by displacement by lactic acid and as a consequence of spontaneous CO2 efflux from the container. CO2 production, 2.3 +/- 0.4 nmol/min/10(9) platelets, was derived from oxygen consumption and the CO2 liberated from bicarbonate as it was consumed. A rapid fall in pH to levels below 7.0 (22 degrees C) took place when the bicarbonate concentration fell below 5 mEq/L as lactate concentrations reached 20 to 25 mmol/L. A further increase in lactate concentration from 25 mmol/L to 40 mmol/L correlated with a further fall in pH to 5.8. Thus, the ultimate storage life of a PC is determined by continuous lactate production and the fixed buffering capacity of plasma and by the glucose concentration of the PC. With knowledge of these parameters, methods for predicting pH as a function of time, platelet count, and O2 and CO2 transport capability of the container have been developed as guidelines for future work.