Methodological problems of direct bioluminescent ADP assay in platelets and erythrocytes.
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 linea 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 mumol/10(11) PLT for ATP determination and 1.9-3.7 mumol/10(11) PLT for ADP determination in platelets, and 3.2-3.8 mumol/g Hgb for ATP determination and 0.56-0.73 mumol/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.
Girotti
S
S
Istituto di Scienze Chimiche, Università di Bologna, Italy.
Ferri
E
E
Cascione
M L
ML
Orlandini
A
A
Farina
L
L
Nucci
S
S
Di Graci
F
F
Budini
R
R
D000244:Adenosine Diphosphate / Q000032:analysis*; D000255:Adenosine Triphosphate / Q000032:analysis; D001792:Blood Platelets / Q000737:chemistry*; D001793:Blood Preservation; D004912:Erythrocytes / Q000737:chemistry*; D005615:Freezing; D006801:Humans; D008163:Luminescent Measurements; D008722:Methods
D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate