Employment of bioluminescence for the quantification of adenosine phosphates in the human cornea.
BACKGROUND
Quantification of adenosine phosphates in human corneal extracts has been performed using spectrophotometry. We employed the bioluminescence technique to obtain a more sensitive assay for adenosine phosphates and to reduce the volume of the test sample.


METHODS
The bioluminescence assay for ATP, already known from sterility control, was modified and expanded. Standard curves were established using a standard solution with equimolar concentrations of ATP, ADP and AMP. To monitor the method, adenosine phosphates were measured in 35 human corneal extracts using both spectrophometry and bioluminescence.


RESULTS
Linear standard curves ranging from 1 to 45 pmol were established. The two methods yielded comparable results despite the use of a basic dilution of 1:100 for the new technique.


CONCLUSIONS
Bioluminescence provides a highly sensitive quantification of adenosine phosphates in the human cornea and facilitates an extremely detailed evaluation of the metabolic status of the cornea.
Salla
S
S
Department of Ophthalmology, RWTH Aachen, Germany.
Redbrake
C
C
Frantz
A
A
D000227:Adenine Nucleotides / Q000032:analysis*; D003315:Cornea / Q000737:chemistry*; D049454:Firefly Luciferin; D006801:Humans; D008156:Luciferases; D008163:Luminescent Measurements; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013053:Spectrophotometry / Q000379:methods
D000227:Adenine Nucleotides; D049454:Firefly Luciferin; D008156:Luciferases