Effects of temperature on measurement of alkaline phosphatase activity.

We examined the effects of temperature on the activity and steady-state kinetic properties of alkaline phosphatase (EC 3.1.3.1). Purified isoenzymes from human liver, intestine, and placenta were used, as was human serum, and the enzyme from porcine kidney. Phosphatase activity was estimated by two different assay techniques. For all isoenzymes, apparent Michaelis constants for the substrate 4-nitrophenyl phosphate decreased with increased temperature; Km at 37 degrees C was typically half that determined at 25 degrees C. All enzymes of human origin exhibited similar linear Arrhenius relationships over the range examined, 20-37 degrees C (Ea of 30-36 kJ X mol-1). The porcine kidney enzyme obeyed an Arrhenius relationship that was slightly, but significantly, different from the isoenzymes of human origin. Temperature relationships based upon Arrhenius behavior and individual activity measurements are presented. For human alkaline phosphatases, they differed by no more than 10%.

Proficiency testing: purification of lactate dehydrogenase 1 and results of its use as a reference material in the New York State program.

Highly purified human lactate dehydrogenase 1 has been used in an interlaboratory evaluation and improvement progran in clinical chemistry in New York State since 1971. Although there are difficulties in determining and assigning the most nearly accurate values for test samples in the absence of a reference method and a reference material, we have minimized these difficulties by using human lactate dehydrogenase preparations purified as we describe here and suspended in the same matrix, and by utilizing "reference laboratories" that routinely are doing multiple assays to determine the most nearly accurate value. The lactate dehydrogenase used in the program is stable for longer than 1.5 years. Conversion factors were used to convert all results to U/liter at 30 degrees C. Review of the data for 1972-75 shows a marked improvement in the accuracy of virtually all methods used to determine this enzyme.