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A sensitive assay for the quantification of glucose and lactate in the human cornea using a modified bioluminescence
A Frantz1, S Salla, C Redbrake
1Department of Ophthalmology, RWTH Aachen, Germany.
Background:
Quantification of glucose and lactate concentrations in human corneal extracts has been performed using spectrophotometry. We employed a bioluminescence technique to obtain a more sensitive assay for glucose and lactate and to reduce the volume of the test sample.
Materials And Methods:
The NAD(P)H bioluminescence assay (Boehringer Mannheim, Germany) was modified for glucose and lactate. Standard curves were established using a standard solution with 0.004 mM and 0.01 mM concentrations of glucose and lactate, respectively.
Results:
Linear standard curves ranging from 0 to 200 pmol for glucose and from 0 to 250 pmol for lactate were established. The sample volume was reduced from 100 microliters to 25 microliters compared with spectrophotometry.
Discussion:
The modified bioluminescence technique provides a highly sensitive quantification of glucose and lactate in the human cornea and thus reveals more details of the overall metabolic status of the tissue.
