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 3.8026.4 (CV) for samples containing about 20290 U/L LIP activity; when assayed using an automated analyzer, the range was 1.864.86 (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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