TY - JOUR
T1 - 1,2-Dioleoylglycerol method for pancreatic lipase catalytic activity in serum
AU - Iizuka, Yoshiaki
AU - Ueda, Shigeru
AU - Hanada, Toshiro
AU - Tani, Wataru
AU - Adachi, Hiroshi
AU - Haga, Riichi
AU - Yamaguchi, Mari
AU - Kurotani, Wataru
AU - Sekiguchi, Mitsuo
AU - Kang, Dongchon
AU - Sakasegawa, Shin Ichi
PY - 2012/3/1
Y1 - 2012/3/1
N2 - 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.
AB - 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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U2 - 10.1515/cclm.2011.811
DO - 10.1515/cclm.2011.811
M3 - Article
C2 - 22126376
AN - SCOPUS:84860475370
SN - 1434-6621
VL - 50
SP - 475
EP - 481
JO - Clinical Chemistry and Laboratory Medicine
JF - Clinical Chemistry and Laboratory Medicine
IS - 3
ER -