Abstract
A methodology for the determination of lipase, based on the coupled processes of energy transfer and enhancement of the chemiluminescence of the luminol-H2O2-horseradish peroxidase (HRP) system has been developed. Fluorescein diacetate (FDA) was hydrolyzed to fluorescein by the action of the enzyme lipase, and this compound acted as an enhancer of the chemiluminescent process and acceptor of the chemiluminescent emission from the luminol-H2O2-HRP system. By measuring the transferred emission to fluorescein at 525 nm, lipase (range 0.2–1.5 U/mL, RSD 2.3%) was determined. This methodology permited the determination of every compound of the system, thus, H2O2 (range 0.5–2 mM, RSD 6.9%) and HRP (range 5.5–49.5 U/mL, RSD 3.6%) could also be determined. Lipase was determined in rabbit serum with 96.7 ± 3.3% and 102.9 ± 5.4% recoveries for two different lipase concentrations. Besides, H2O2 was determined in the disinfectant solution for contact lenses.
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Received: 1 March 1999 / Revised: 6 May 1999 / Accepted: 12 May 1999
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Navas Díaz, A., Sanchez, F., Torijas, M. et al. Chemiluminescent lipase determination based on the enhanced luminol/H2O2/horseradish peroxidase/fluorescein diacetate energy transfer system. Fresenius J Anal Chem 365, 537–540 (1999). https://doi.org/10.1007/s002160051518
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DOI: https://doi.org/10.1007/s002160051518