Fluorometric Assay of Serum Acid or Alkaline Phosphatase, Either in Solution or on a Semisolid Surface

1975 ◽  
Vol 21 (12) ◽  
pp. 1791-1794 ◽  
Author(s):  
Bernd Rietz ◽  
George G Guilbault
Keyword(s):  
Alkaline Phosphatase ◽  
Enzyme Activity ◽  
Acid Phosphatase ◽  
Clinical Laboratory ◽  
Serum Alkaline Phosphatase ◽  
Buffer Solution ◽  
Citrate Buffer ◽  
Enzymatic Action ◽  
Stable Substrate ◽  
Serum Acid Phosphatase

Abstract We describe enzymatic fluorometric methods for determining activities of serum alkaline phosphatase and of serum acid phosphatase in solution and on silicone rubber pads. 4-Methylumbelliferone phosphate is used as substrate, in either tris(hydroxymethyl)aminomethane or citrate buffer. In solution, the reaction is measured at 37 °C in a 3-ml Pyrex cuvette. Measurements on the pads are also made at 37 °C, after establishing a stable substrate film by lyophilizing all reagents on the surface of the pads. Only 20 to 30 µl of substrate solution, 50 µl of buffer solution, and 1 to 10 µl of blood are necessary, making a total volume of 51 to 60 µl for each assay. The rate of appearance of the fluorescent 4-methylumbelliferone liberated from 4-methylumbelliferone phosphate by the enzymatic action is measured and equated to enzyme activity. Calibration plots of the change in fluorescence per minute vs. enzyme activity for measurements in solution and on pads show a good proportionality in the range of 30.8 to 633 U/liter for alkaline phosphatase and in the range of 0.265 to 5.3 King— Armstrong units for acid phosphatase, indicating the usefulness of these methods in the clinical laboratory.

Ammonium Thymolphthalein Monophosphate as a New Substrate for Alkaline and Acid Phosphatase Determinations in Serum

1973 ◽  
Vol 19 (10) ◽  
pp. 1135-1138 ◽  
Author(s):  
Leo G Morin
Keyword(s):  
Alkaline Phosphatase ◽  
Acid Phosphatase ◽  
Phosphatase Activity ◽  
Alkaline Phosphatase Activity ◽  
Sodium Salt ◽  
Prostatic Acid Phosphatase ◽  
Citrate Buffer ◽  
Coefficients Of Variation ◽  
Stable Substrate ◽  
Alkaline And Acid Phosphatase

Abstract A new substrate, ammonium thymolphthalein monophosphate, is proposed for use in alkaline and acid phosphatase determinations. The ammonium salt shares the advantages of the sodium salt, but is a more stable substrate and results in more stable working reagents. The substrate is buffered at pH 10.1 in diethanolamine for determining alkaline phosphatase activity in 5 min, and at pH 5.9 in citrate buffer for determining acid phosphatase activity in 30 min. The substrate is relatively specific for bone and liver alkaline phosphatase and prostatic acid phosphatase. Coefficients of variation were 2.5% and 6.6% for alkaline and acid phosphatase, respectively.

SIGNIFICANCE OF TRANSAMINASE ACTIVITY OF SERUM

PEDIATRICS ◽  
1959 ◽  
Vol 24 (3) ◽  
pp. 360-361
Author(s):  
SAMUEL P. BESSMAN
Keyword(s):  
Alkaline Phosphatase ◽  
Enzyme Activity ◽  
Acid Phosphatase ◽  
Transaminase Activity ◽  
Specific Approach ◽  
Normal Tissues ◽  
The Past ◽  
Glycolytic Activity ◽  
Enzyme Characteristic ◽  
Phosphatase Alkaline

THE MEASUREMENT of enzyme activity of serum as an indicator of disease has a long history in medicine. In the past, it has been the aim of the designers of these methods to make them as specific as possible for assay of an enzyme characteristic of a particular system or group of similar organs. Examples of these venerable tests are those for amylase, acid phosphatase, alkaline phosphatase and choline esterase in the serum. Warburg made the first departure from this specificity by demonstrating that the activity of triosephosphate dehydrogenase in the serum of animals with cancer was much greater than that of controls. This test was partially specific, for as Warburg had earlier shown, the glycolytic activity of tumors is much greater than that of normal tissues. The non-specific approach became extreme with the introduction of the measurement of the glutamic-oxalacetic transaminase reaction in the diagnosis of acute coronary disease.

Acid phosphatase, alkaline phosphatase, and nitrate reductase activity of selected ectomycorrhizal fungi

1989 ◽  
Vol 67 (3) ◽  
pp. 750-753 ◽  
Author(s):  
Iwan Ho
Keyword(s):  
Alkaline Phosphatase ◽  
Enzyme Activity ◽  
Acid Phosphatase ◽  
Nitrate Reductase ◽  
Phosphatase Activity ◽  
Ectomycorrhizal Fungi ◽  
Acid Phosphatase Activity ◽  
Nitrate Reductase Activity ◽  
Reductase Activity ◽  
Phosphatase Alkaline

Seventeen isolates, encompassing five genera and eight species of ectomycorrhizal fungi, were compared for acid phosphatase, alkaline phosphatase, and nitrate reductase activity. Isolates within species differed in enzyme activity and isozyme patterns by host specificity and site (as exemplified by the genus Suillus). Host and site may have affected phosphatase enzyme activity. Generally, the Douglas-fir associates, which dominate in mesic sites, have higher acid phosphatase activity than pine associates, which mostly occupy xeric sites; however, pine associates from mesic sites also have higher acid phosphatase activity (e.g., S. tomentosus). In four isolates of Amanita muscaria, the effect of site was also apparent. Two of them, which have significantly higher acid phosphatase activity than the others, were isolated from mesic sites. The isozyme pattern of the genus Suillus appeared to be separated by host groups. Other isolates with only one species also differed more or less by host groups. They shared at least one band within host groups, except for the two isolates of Paxillus involutus from different hosts. The P. involutus S-403 isolated from an orchard showed much higher nitrate reductase activity than all other isolates. No apparent differences in nitrate reductase activity were found between the other isolates.

Serum alkaline phosphatase: normal values by sex and age.

1977 ◽  
Vol 23 (9) ◽  
pp. 1769-1770 ◽  
Author(s):  
J R Eastman ◽  
D Bixler
Keyword(s):  
Alkaline Phosphatase ◽  
Enzyme Activity ◽  
Statistical Analysis ◽  
Normal Values ◽  
Serum Alkaline Phosphatase ◽  
Blood Type ◽  
Sample Population ◽  
Normal Range ◽  
Larger Sample ◽  
Alkaline Phophatase

Abstract We report here the normal range of serum alkaline phophatase activity as measured by the method proposed by Hausamen et al. [Clin. Chim. Acta 15, 241 (1967)] with a much larger sample size than used in previous investigations. In the statistical analysis the sample population is subdivided by sex and age, two variables which are known to influence enzyme activity. No statistically significant influence of blood type on enzyme activity was observed. The normal range of enzyme activity is reported in percentiles.

PHOSPHATASE OF RABBIT POLYMORPHONUCLEAR LEUCOCYTES

1949 ◽  
Vol 27e (5) ◽  
pp. 290-307 ◽  
Author(s):  
D. M. Cram ◽  
R. J. Rossiter
Keyword(s):  
Alkaline Phosphatase ◽  
Enzyme Activity ◽  
Acid Phosphatase ◽  
Bile Salts ◽  
Polymorphonuclear Leucocytes ◽  
Alkyl Sulphate ◽  
Low Concentrations ◽  
Activity Curve ◽  
Surface Active ◽  
Active Substances

Rabbit polymorphonuclear leucocytes contain an active phosphatase that readily hydrolyzes disodium phenyl phosphate. The pH activity curve of the enzyme was found to have two maxima, one in the region of pH 10 and the other in the region of pH 5. The alkaline phosphatase was much more active than the acid phosphatase. The concentration of alkaline phosphatase in rabbit white cells was approximately one thousand times that of the enzyme in the serum. Under the conditions of study, the alkaline phosphatase activity was proportional to the concentration of the enzyme. The effect of substrate concentration on the enzyme activity was studied and the Michaelis constant (Ks) determined. An excess of substrate inhibited the enzyme. The course of the reaction was linear with time for the first 60 min.; after 90 min. the activity fell off faster than would be expected if the reaction were of the first order.Magnesium and glycine, in low concentrations, caused an increase in the enzyme activity, whereas zinc, cyanide, borate, phosphate, bile salts, and glycine, in higher concentrations, were inhibitory. Fluoride had no demonstrable effect. Surface-active substances, such as saponin, bile salts, or alkyl sulphate, liberated the enzyme from the cells. Similar results were obtained when α-glycerophosphate or β-glycerophosphate was used as the substrate.The alkaline phosphatase can be considered to belong to Class AI of Folley and Kay (22) and the acid phosphatase to Class AII. The alkaline phosphatase can also be considered to be a Phosphatase II of Cloetens (9).

Coulometric Determination of Activity of Acid or Alkaline Phosphatases in Serum

1972 ◽  
Vol 18 (6) ◽  
pp. 503-508 ◽  
Author(s):  
Marvin A Brooks ◽  
William C Purdy
Keyword(s):  
Alkaline Phosphatase ◽  
Phosphatase Activity ◽  
Alkaline Phosphatase Activity ◽  
Serum Alkaline Phosphatase ◽  
Detection System ◽  
Constant Current ◽  
Coulometric Determination ◽  
Citrate Buffer ◽  
Serum Alkaline Phosphatase Activity ◽  
Clin Pathol

Abstract A constant-current coulometric technique is described for measuring acid or alkaline phosphatase activity in serum. Electrogenerated bromine is used as the titrant of phenol enzymatically released from phenyl phosphate. A biamperometric end-point detection system is used, with an applied potential of 135 mV. Titrations are performed at pH 1 with use of a generating electrolyte of 0.5 molar KBr in 0.05 molar H2SO4. A procedure is described for determining serum alkaline phosphatase activity by use of 2-amino-2-methyl-1-propanol (0.25 mol/liter, pH 10.25) with Mg2+ activator (1 mmol/liter), and for determining serum acid phosphatase activity in citrate buffer (0.1 mol/liter, pH 4.9). Results compare favorably with those of the method of Kind and King [J. Clin. Pathol. 7, 322 (1954)].

Relationship of plasma tartrate resistant acid phosphatase to the bone isoenzyme of serum alkaline phosphatase in hyperparathyroidism

1983 ◽  
Vol 133 (2) ◽  
pp. 189-200 ◽  
Author(s):  
J.J. Štěpán ◽  
E. Šilinková-Málková ◽  
T. Havránek ◽  
J. Formánková ◽  
M. Zichová ◽  
...  
Keyword(s):  
Alkaline Phosphatase ◽  
Acid Phosphatase ◽  
Serum Alkaline Phosphatase ◽  
Tartrate Resistant Acid Phosphatase ◽  
Relationship Of

Biochemical and histochemical studies of alkaline and acid phosphatases in a digenetic trematode,Pegosomum egretti

1986 ◽  
Vol 60 (4) ◽  
pp. 293-298 ◽  
Author(s):  
Indra Rajvanshi ◽  
K. L. Mali
Keyword(s):  
Alkaline Phosphatase ◽  
Enzyme Activity ◽  
Acid Phosphatase ◽  
Raw Materials ◽  
Digenetic Trematode ◽  
Acid Phosphatases ◽  
Alkaline Phosphatases ◽  
Optimum Ph ◽  
Standard Techniques ◽  
Alkaline And Acid Phosphatases

ABSTRACTThe biochemistry and histochemistry ofPegosomum egrettihave been studied using standard techniques. Phosphatases were analysed colorimetrically; the optimum pH for acid phosphatase activity was 5·0 and for alkaline phosphatase was 10·0. The results were compared with those of other trematodes. Histochemical localization of acid and alkaline phosphatases revealed differences in enzyme activity in various tissues. These differences in the site and pattern of distribution of the two enzymes have been discussed in relation to transport of raw materials and the metabolism of the cell concerned.

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