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. 1971 Sep;24(6):493–500. doi: 10.1136/jcp.24.6.493

Colorimetric determination of serum acid phosphatase activity using adenosine 3′-monophosphate as substrate

Graham Ellis 1, Alan Belfield 1, David M Goldberg 1
PMCID: PMC477079  PMID: 4106191

Abstract

The hydrolysis of adenosine 3′-monophosphate by serum acid phosphatase has been coupled to the liberation of ammonia from the adenosine generated through the action of exogenous adenosine deaminase. The ammonia is measured at the end of the incubation by a modification of the phenol-hypochlorite reaction of Berthelot. Optimum conditions for the enzyme reaction have been defined. Inhibition of the Berthelot reaction by the serum used in the assay is small, and may be compensated by a correction factor. Although the value for the control is high in relation to the test over the normal range, this is largely outweighed by the good sensitivity and precision of the method.

The substrate is not significantly hydrolysed by erythrocyte acid phosphatase within the limits encountered in haemolysed sera. Experience of the method in routine hospital diagnosis compared favorably with that of a standard method employing disodium phenyl phosphate as substrate. It is suggested that activities greater than 3·1 IU/l should be further investigated and those greater than 3·7 IU/l should be regarded as definitely raised. The stability of human serum AcPase when promptly separated and held at 4°C or − 20°C was confirmed. At room temperature, acidification to pH 6·0 greatly improved stability.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. ALTSCHULE M. D., PARKHURST B. H., ZAGER G. R. Measurement of acid phosphatase activity of urine. Am J Clin Pathol. 1951 May;21(5):480–483. doi: 10.1093/ajcp/21.5_ts.480. [DOI] [PubMed] [Google Scholar]
  2. Abul-Fadl M. A., King E. J. Properties of the acid phosphatases of erythrocytes and of the human prostate gland. Biochem J. 1949;45(1):51–60. [PMC free article] [PubMed] [Google Scholar]
  3. Amador E., Price J. W., Marshall G. Serum acid alpha-naphthyl phosphatase activity. Am J Clin Pathol. 1969 Feb;51(2):202–206. doi: 10.1093/ajcp/51.2.202. [DOI] [PubMed] [Google Scholar]
  4. BABSON A. L., READ P. A. A new assay for prostatic acid phosphatase in serum. Am J Clin Pathol. 1959 Jul;32(1):88–91. doi: 10.1093/ajcp/32.1_ts.88. [DOI] [PubMed] [Google Scholar]
  5. Babson A. L., Phillips G. E. An improved acid phosphatase procedure. Clin Chim Acta. 1966 Feb;13(2):264–265. doi: 10.1016/0009-8981(66)90303-2. [DOI] [PubMed] [Google Scholar]
  6. Belfield A., Ellis G., Goldberg D. M. A specific colorimetric 5'-nucleotidase assay utilizing the Berthelot reaction. Clin Chem. 1970 May;16(5):396–401. [PubMed] [Google Scholar]
  7. Belfield A., Goldberg D. M. Activation of serum 5' nucleotidase by magnesium ions and its diagnostic applications. J Clin Pathol. 1969 Mar;22(2):144–151. doi: 10.1136/jcp.22.2.144. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Belfield A., Goldberg D. M. Human serum glucose-6-phosphatase activity: confirmation of its presence and lack of diagnostic value. Clin Chim Acta. 1971 Jan;31(1):81–85. doi: 10.1016/0009-8981(71)90364-0. [DOI] [PubMed] [Google Scholar]
  9. Belfield A., Goldberg D. M. Hydrolysis of adenosine monophosphates by acid phosphatases as measured by a continuous spectrophotometric assay. Biochem Med. 1970 Sep;4(2):135–148. doi: 10.1016/0006-2944(70)90090-6. [DOI] [PubMed] [Google Scholar]
  10. Brightwell R., Tappel A. L. Lysosomal acid pyrophosphatase and acid phosphatase. Arch Biochem Biophys. 1968 Mar 20;124(1):333–343. doi: 10.1016/0003-9861(68)90335-4. [DOI] [PubMed] [Google Scholar]
  11. CAMPBELL D. M., MOSS D. W. Spectrofluorimetric determination of acid phosphatase activity. Clin Chim Acta. 1961 May;6:307–315. doi: 10.1016/0009-8981(61)90055-9. [DOI] [PubMed] [Google Scholar]
  12. CHANEY A. L., MARBACH E. P. Modified reagents for determination of urea and ammonia. Clin Chem. 1962 Apr;8:130–132. [PubMed] [Google Scholar]
  13. Gutman A. B., Gutman E. B. AN " ACID " PHOSPHATASE OCCURRING IN THE SERUM OF PATIENTS WITH METASTASIZING CARCINOMA OF THE PROSTATE GLAND. J Clin Invest. 1938 Jul;17(4):473–478. doi: 10.1172/JCI100974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hoy T. G., Goldberg D. M. An ALGOL program for the computer-assisted calculation of enzyme kinetic data. Int J Biomed Comput. 1971 Jan;2(1):71–77. doi: 10.1016/0020-7101(71)90020-1. [DOI] [PubMed] [Google Scholar]
  15. JACOBSSON K. The determination of tartrate-inhibited phosphatase in serum. Scand J Clin Lab Invest. 1960;12(4):367–380. doi: 10.3109/00365516009065399. [DOI] [PubMed] [Google Scholar]
  16. JEGATHEESAN K. A., JOPLIN G. F. Correlation of serum glycolytic enzymes and acid phosphatases with sites of metastases in mammary carcinomatosis. Br Med J. 1962 Mar 24;1(5281):831–834. doi: 10.1136/bmj.1.5281.831. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. JOPLIN G. F., JEGATHEESAN K. A. Serum glycolytic enzymes and acid phosphatases in mammary carcinomatosis. Br Med J. 1962 Mar 24;1(5281):827–831. doi: 10.1136/bmj.1.5281.827. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. KILSHEIMER G. S., AXELROD B. Inhibition of prostatic acid phosphatase by alpha-hydroxycarboxylic acids. J Biol Chem. 1957 Aug;227(2):879–890. [PubMed] [Google Scholar]
  19. KING E. J., JEGATHEESAN K. A. A method for the determination of tartratelabile, prostatic acid phosphatase in serum. J Clin Pathol. 1959 Jan;12(1):85–89. doi: 10.1136/jcp.12.1.85. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Marshall G., Amador E. Diagnostic usefulness of serum acid beta-glycerophosphatase activities in prostatic disease. Am J Clin Pathol. 1969 May;51(5):551–554. doi: 10.1093/ajcp/51.5.551. [DOI] [PubMed] [Google Scholar]
  21. Mobley T. L., Frank I. N. Influence of tumor grade on survival and on serum acid phosphatase levels in metastatic carcinoma of the prostate. J Urol. 1968 Mar;99(3):321–323. doi: 10.1016/S0022-5347(17)62701-0. [DOI] [PubMed] [Google Scholar]
  22. Murphy G. P., Reynoso G., Kenny G. M., Gaeta J. F. Comparison of total and prostatic fraction serum acid phosphatase levels in patients with differentiated and undifferentiated prostatic carcinoma. Cancer. 1969 Jun;23(6):1309–1314. doi: 10.1002/1097-0142(196906)23:6<1309::aid-cncr2820230611>3.0.co;2-m. [DOI] [PubMed] [Google Scholar]
  23. Nelson B. D. Rat liver acid phosphatase: differences in lysosomal and cytoplasmic forms. Proc Soc Exp Biol Med. 1966 Apr;121(4):998–1001. doi: 10.3181/00379727-121-30947. [DOI] [PubMed] [Google Scholar]
  24. Ockerman P. A., Kohlin P. Acid hydrolases in plasma in Gaucher's disease. Clin Chem. 1969 Jan;15(1):61–64. [PubMed] [Google Scholar]
  25. Prout G. R., Jr, Brewer W. R. Response of men with advanced prostatic carcinoma to exogenous administration of testosterone. Cancer. 1967 Nov;20(11):1871–1878. doi: 10.1002/1097-0142(196711)20:11<1871::aid-cncr2820201112>3.0.co;2-d. [DOI] [PubMed] [Google Scholar]
  26. Prout G. R., Jr Chemical tests in the diagnosis of prostatic carcinoma. JAMA. 1969 Sep 15;209(11):1699–1700. [PubMed] [Google Scholar]
  27. REYNOLDS O. B., REYNOLDS M. D., WALKER B. S. Yeast adenylic acid as substrate for serum acid phosphatase. Clin Chem. 1956 Apr;2(2):117–124. [PubMed] [Google Scholar]
  28. Rozenszajn L., Epstein Y., Shoham D., Arber I. The acid phosphatase isoenzymes in normal and pathological sera and in tissue homogenates. J Lab Clin Med. 1968 Nov;72(5):786–793. [PubMed] [Google Scholar]
  29. TSUBOI K. K., HUDSON P. B. Acid phosphatase. I. Human red cell phosphomonoesterase; general properties. Arch Biochem Biophys. 1953 Apr;43(2):339–357. doi: 10.1016/0003-9861(53)90129-5. [DOI] [PubMed] [Google Scholar]
  30. TSUBOI K. K., HUDSON P. B. Acid phosphatase. III. Specific kinetic properties of highly purified human prostatic phosphomonoesterase. Arch Biochem Biophys. 1955 Mar;55(1):191–205. doi: 10.1016/0003-9861(55)90557-9. [DOI] [PubMed] [Google Scholar]
  31. WOODARD H. Q. The clinical significance of serum acid phosphatase. Am J Med. 1959 Dec;27:902–910. doi: 10.1016/0002-9343(59)90174-3. [DOI] [PubMed] [Google Scholar]

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