1,2-Dioleoylglycerol method for pancreatic lipase catalytic activity in serum
Background: There is a need for a pancreatic lipase (LIP) reference assay to provide an accurate base to which routine methods can be traceable. Methods: This study developed a novel LIP assay method in which 1,2-dioleoylglycerol (DODG) is the substrate and LIP activity is measured in a coupled enzy...
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Veröffentlicht in: | Clinical chemistry and laboratory medicine 2012-03, Vol.50 (3), p.475-481 |
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Sprache: | eng |
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Zusammenfassung: | Background: There is a need for a pancreatic lipase (LIP) reference assay to provide an accurate base to which routine methods can be traceable. Methods: This study developed a novel LIP assay method in which 1,2-dioleoylglycerol (DODG) is the substrate and LIP activity is measured in a coupled enzymatic reaction from the increase in absorbance at 340 nm with production of NADPH. Results: With this method, LIP activity was linear up to 440 U/L (8-times expected upper limit of physiological concentration). When assayed manually, the between-laboratory variation for six samples surveyed at five laboratories was 3.80–26.4% (CV) for samples containing about 20–290 U/L LIP activity; when assayed using an automated analyzer, the range was 1.86–4.86% (four laboratories). Interference by >5 mmol/L glycerol and low specificity with post-heparin samples were noted, but in practice these are avoidable. Precision analyzed by automated assay of 49 samples twice in random order produced a covariance of 2.27 U/L, which is comparable to routine methods, and good correlations were obtained with five routine methods. Conclusions: Although further studies are required, the DODG method may be likely applicable as one candidate reference method. |
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ISSN: | 1434-6621 1437-4331 |
DOI: | 10.1515/cclm.2011.811 |