Lipase activity measured in serum by a continuous-monitoring pH-Stat technique--an update.
Using recent knowledge regarding the roles of colipase, bile acids, Ca2+, and emulsifiers, we optimized a previously published pH-Stat method for lipase (EC 3.1.1.3) activity measurements. The recommended assay conditions are: olive oil/triolein, 100 mL/L; sodium glycocholate, 35 mmol/L; Ca2+, 8.5 mmol/L; and colipase, 6.0 mg/L. The sample volume is 0.10 mL, the reaction pH 9.0, the temperature 30 degrees C, and the concentration of titrant 15 mmol/L. Hydroxypropyl methylcellulose, 20 g/L, replaces acacia as emulsifier to avoid inhibition by excess Ca2+. The standard curve is linear to greater than 4566 U/L. The reference interval with olive oil as substrate is 30-235 U/L. Lipase activities with triolein substrate are 9.9% greater than with olive oil. Interference by pancreatic carboxylesterase (EC 3.1.1.1) activity is inhibited by incubating the sample with diisopropylfluorophosphate. Results correlate well with those by the optimized SingleVial method of Boehringer Mannheim Diagnostics (r = 0.997) and the immunochemical assay of Beckman Instruments, Inc. (r = 0.995). Correlation with the aca method (E.I. DuPont de Nemours & Company) is less satisfactory (r = 0.892), probably owing to lack of colipase in the latter method.
Department of Pathology, University of Kentucky Medical Center, Lexington.
N W
NW
Tietz
J R
JR
Astles
D F
DF
Shuey
D000328:Adult; D001322:Autoanalysis; D002118:Calcium / Q000494:pharmacology; D043182:Carboxylesterase; D002265:Carboxylic Ester Hydrolases / Q000037:antagonists & inhibitors; D003089:Colipases; D006000:Glycocholic Acid; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008049:Lipase / Q000097:blood*; D000069463:Olive Oil; D010938:Plant Oils; D012965:Sodium Chloride / Q000494:pharmacology; D014304:Triolein
D003089:Colipases; D000069463:Olive Oil; D010938:Plant Oils; D014304:Triolein; D012965:Sodium Chloride; D002265:Carboxylic Ester Hydrolases; D043182:Carboxylesterase; D008049:Lipase; D006000:Glycocholic Acid; D002118:Calcium