Methodological problems of direct bioluminescent ADP assay in platelets and erythrocytes.

Abstract We have developed a method for ADP bioluminescent measurement in platelets and erythrocytes which complements our previous method for ATP assay. When the different parameters of the system under investigation are taken into account, a linear range between 10 −9 and 10 −7 g/ml can be obtained without incubation or troublesome extraction. This makes the method easy and useful for identifying any disease-induced alterations in ATP and/or ADP levels in these blood cells. The data obtained correlate well with those of a bioluminescent method requiring extraction with ethanol/EDTA and incubation, giving the reference intervals of 3.5 – 5.5 μ mol 10 11 PLT for ATP determination and 1.9 – 3.7 μ mol 10 11 PLT for ADP determination in platelets, and 3.2–3.8 μmol/g Hgb for ATP determination and 0.56 – 0.73 μmol/g Hgb for ADP in erythrocytes. This assay was applied to quality control on blood bags in transfusion centers and proved to be a rapid and reliable method for testing the viability of stored blood cells.