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Employment of bioluminescence for the quantification of adenosine phosphates in the human cornea

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Abstract

• 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.

• Conclusion: 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.

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Authors and Affiliations

  1. Department of Ophthalmology, RWTH Aachen, Pauwelsstrasse 30, D-52057, Aachen, Germany

    Sabine Salla, Claudia Redbrake & Andrea Frantz

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  1. Sabine Salla
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  2. Claudia Redbrake
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  3. Andrea Frantz
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Dedicated to Prof. Martin Reim on the occasion of his 65th birthday

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Salla, S., Redbrake, C. & Frantz, A. Employment of bioluminescence for the quantification of adenosine phosphates in the human cornea. Graefe's Arch Clin Exp Ophthalmol 234, 521–526 (1996). https://doi.org/10.1007/BF00184862

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  • DOI: https://doi.org/10.1007/BF00184862

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