Bodansky method

In practice it is often more convenient to measure the release of a phenol from an aryl phosphomonoester. Standard serum phosphatase methods employ phenyl phosphate (188), p-nitrophenyl phosphate (189), phenolphthalein monophosphate (140), or thymolphthalein monophosphate (141) where the phenol released can be determined spectrophoto-metrically [only the Bodansky method (13) uses a Pi determination]. A number of fluorogenic substrates have been used for phosphatase studies, e.g., jS-naphthyl phosphate (30, 148), 4-methylumbelliferyl phosphate (143), and 3-O-methylfluorescein phosphate (144) The main advantage here is the much greater sensitivity of fluorescence as compared with spectrophotometric assays as little as 1 pmole of 4-methyl-umbelliferone can be detected in continuous assay. 

The serum acid phosphatase activity was determined by the Bodansky method (B18, J2) with 3-glycerophosphate as substrate. The mean value for the series of 12 patients was 0.983 0.122 Bodansky unit, consider-... 

The Bodansky method uses -glycerophosphate as the substrate and measures the amount of free phosphate liberated. 

M. Bodansky, Peptide Synthesis, 2nd ed., John Wiley Sons, Inc., New York, 1976 J. Meinhofer in Ref. 1, Chapt. 9, p. 297 G. R. Pettit, Synthetic Peptides, Vols. 1—4, Van Nostrand Reinhold, New York, 1980, Vols. 5, 6, Elsevier New York, 1982 E. Shroeder and K. Luebke, The Peptide, Vol. 1, Methods of Peptide Synthesis, Academic Press, New York, 1965 N. Izumiya and co-workers. Fundamentals and Experiments of Peptide Synthesis (in Japanese), Mamzen, Tokyo, Japan, 1987 R. B. Merriheld,/ Mm. Chem. Soc. 85, 2149 (1963) G. Barany and R. B. Merriheld in E. Gross andj. Meinenhofer, eds.. The Peptides Mnalysis, Synthesis, Biology, Vol. 2, Academic Press, New York, 1980, pp. 1—284 G. R. Marshall, Peptides Chemistry and Biology, Escom, Leiden, The Netherlands, 1988. 

Enzyme Reference Serums. Several companies sell lyophilized or stabilized reference serums for the calibration of instruments and for quality control. The label values given for the enzymatic activity of these serums should never be taken at face value, as at times they may be quite erroneous (19,33). Also, these values should only be used for the assay with which they were standardized, as interconversion of activity from one method to another for the same enzyme may often lead to marked errors. For instance, it is not recommended that alkaline phosphatase expressed in Bodansky units be multiplied by a factor to convert it to the units of the Ring-Armstrong method, or any other method for that matter. 

Schwartz, Morton K, and Bodansky, Oscar, Automated Methods for... 

This procedure was based on the use of, 0-glycerophosphate as substrate as in A. Bodansky s method for alkaline phosphatase (B17). In this method the mixture, to which the serum was added to start the reaction, had concentrations of 0.016 Af sodium /3-glycerophosphate and 0.021 M sodium diethylbarbiturate. The addition of 1.0 ml serum to 10 ml of this mixture or, as in a later version, of 0.5 ml to 4.5 ml of the mixture led to concentrations in the final reaction mixture of 0.0144 M sodium, 3-glycerophosphate and 0.019 barbiturate buffer. 

The question may arise as to which is the preferred method. In the author s experience, and this will be documented more completely later, the use of the substrate, sodium (S-glycerophosphate, as in the Bodansky procedure (B18, 32), is more specific for elevations of serum acid phosphatase activity due to prostatic carcinoma. However, the use of other substrates, such as sodium phenyl phosphate in the Gutman method (GIO, G14), may elicit alterations of activity in the serum that reflect diseases in other tissues. 

The next most common method for determination of serum acid phosphatase activity was based on the use of sodium )3-glycerophosphate as substrate. This method and its modifications have been described in Section 2.3. Values obtained by these methods in normal males are shown in Table 7 a liberal summary of these values indicates a mean value of about 0.4 0.2 Bodansky units. 

Values fob Sebum Acid Phosphatase in Nobmal Males by the Method OF A. Bodansky (J2) OB Slight Modifications Thebeof... 

The possibilities that cryptic prostatic carcinoma may coexist with other diseases or that a high serum alkaline phosphatase activity possesses some residual activity at pH 5.0 tend to be negated by an analysis of these five patients. One female with carcinoma of the breast and osteolytic metastases of the femur, pelvis, and spine had an elevated serum acid phosphatase activity of 4.2 K.A. units, and a second female with an unknown primary but with osteolytic lesions of the ribs and scapula had an activity of 4.1 K.A. units. The serum alkaline phosphatase activities were 17.8 Bodansky units in the first case and 5.2 Bodansky units in the second case, both above 4.2 Bodansky units, the upper limit of normal values by this method. 

Using the Bodansky (B18, 52) procedure with 8-glycerophosphate as substrate, Woodard (W8) was unable to obtain such elevations. She determined the serum acid phosphatase activities in 83 females and 342 males, or a total of 425 patients with miscellaneous diseases. Of these, 61 had various types of infectious or metabolic disorders, including 11 cases of inflammatory disease of bone and 12 cases of hepatic cirrhosis. The remainder had some type of neoplastic disease and about one-third had metastases to bone from cancer of various primary sites. There were 15 cases of osteogenic sarcoma and 32 cases of osteitis deformans. All these cases, whether their serum alkaline phosphatase activities were elevated or not, had serum acid phosphatase values that were essentially within the normal range, 0.06-0.89 Bodansky unit for females and 0.11-0.88 unit for males. In contrast to the Gutman method (GIO, G14), there-... 

Schwartz, M. K., and Bodansky, O., Utilisation of automation for studies of enz3Tne kinetics. Methods Biochem. Anal. 16, 183-218 (1968). 

Since a number of methods are in widespread use, it is worthwhile to review comparative studies of alkaline phosphatase activity measured by these different methods. Schwartz et al. (S24) established that a linear relation exists between the results of the Bodansky and King-Armstrong methods. Laurent and Norberg (L5) also demonstrated a linear relation between values on serum performed with a modified Bessey-Lowry compared to a modified King-Armstrong method. However, Gelb et al. 

Fig. 5. Relation between serum alkaline phosphatase determined in the Auto-Analyzer and by the manual method. AutoAnalyzer units are King-Armstrong, and manual units are Bodansky-Shinowara (F14).

Bodansky (B32a) has dealt in detail with considerations of pH in the Shinowara (S30) and Bodansky (B23) methods. 

B23. Bodansky, A., Phosphatase studies. I. Determination of inorganic phosphate. Beer s Law and interfering substances in the Kuttner-Lichtenstein Method. J. Biol. Chem. 99, 197-206 (1932). 

Tietz, N. W., Woodrow, D., and Woodrow, B., A comparative study of the Bodansky and the Bessey, Lowry and Brook methods for alkaline phosphatase in serum. Clin. Chim. Acta 15, 365-367 (1967),... 

Further reading Grant, G.H. and Butt, W.R. (1970). Immunochemical methods in clinical chemistry. In Bodansky, O. and Stewart, C.P. (eds.) Advances in Clinical Chemistry Vol. 13, p. 383. (New York Academic Press)... 

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