Polyphosphates, determination

The most common way of phosphate determination is its separation with bicarbonate/carbonate or hydroxide eluent followed by suppressed conductivity detection. Yet this does not suffice for polyphosphates determination in food samples that have to be determined by means of gradient IC. When determining orthophosphate content, it should be noted that only free P04 ions, and not the total phosphate, are detected by IC. The reason is that some phosphate is bound by calcium especially in fruit juices. 

Cambella and Antia [385] determined phosphonates in seawater by fractionation of the total phosphorus. The seawater sample was divided into two aliquots. The first was analysed for total phosphorus by the nitrate oxidation method capable of breaking down phosphonates, phosphate esters, nucleotides, and polyphosphates. The second aliquot was added to a suspension of bacterial (Escherichia coli) alkaline phosphatase enzyme, incubated for 2h at 37 °C and subjected to hot acid hydrolysis for 1 h. The resultant hot acid-enzyme sample was assayed for molybdate reactive phosphate which was estimated as the sum of enzyme hydrolysable phosphate and acid hydrolysable... 

Calculation of Brine Components. In order to satisfy U.S.D.A. regulations, care must be taken during brine preparation so that finished product protein meets or exceeds 17 percent. The following formula can be used to determine brine composition (Isolated soy protein, salt, dextrose, polyphosphate, etc.) (6). 

A. Analysis of Wastewater and Natural Waters. The presence of certain anions in wastewater effluents can cause deterioration of natural water systems. Phosphorous and nitrogen can be present in several chemical forms in wastewaters. Phosphorous is usually present as phosphate, polyphosphate and organically-bound phosphorus. The nitrogen compounds of interest in wastewater characterization are ammonia, nitrite, nitrate and organic nitrogen. Analyses are often based on titrimetric, and colorimetric methods (3). These methods are time consuming and subject to a number of interferences. Ion Chromatography can be used to determine low ppm concentrations of these ions in less than thirty minutes with no sample preparation. 

Because many cells maintain ATP, ADP, and AMP concentrations at or near the mass action ratio of the adenylate kinase reaction, the cellular content of this enzyme is often quite high. A consequence of such abundance is that, even after extensive purification, many proteins and enzymes contain traces of adenylate kinase activity. The presence of this kinase can confound the quantitative analysis of processes that either require ADP or are carried out in the presence of both ATP and AMP. Furthermore, the equilibrium of any reaction producing ADP may be altered if adenylate kinase activity is present. To minimize the effect of adenylate kinase, one can utilize the bisubstrate geometrical analogues Ap4A and ApsA to occupy simultaneously both substrate binding pockets of this kinase . Typical inhibitory concentrations are 0.4 and 0.2 mM, respectively. Of course, as is the case for the use of any inhibitor, one must always determine whether Ap4A or ApsA has a direct effect on a particular reaction under examination. For example. Powers et al studied the effect of a series of o ,co-di-(adenosine 5 )-polyphosphates (e.g., ApnA, where n =... 

Even before the above facts and structural considerations were generally appreciated by chemists, some of the polyphosphates, such as Graham s salt and the triphosphate, for example, had acquired major technical importance 90, 91,127,129, 249). About thirty years ago the study of condensed phosphates was taken up from many sides in attempts to determine their structures and, from their structures, to understand their properties. Preparative methods, physico-chemical investigations and theoretical considerations were all brought into play in order to develop this branch of inorganic chemistry to a point where, today, the perspective is clear and we can regard it as well-investigated. 

Ions which do not have a rare gas configuration appear not to fit into this rule. They are much more firmly bound than the alkali and alkaline earth cations. A dependence on radius is not observed and for Hg(I) the binding is stronger with the diphosphate anion than with triphosphate 366). (See, also the binding of cations by highmolccular polyphosphates, Section IV,E,/,d.) For the analysis and determination of triphosphates, see Section VII,B. 

Perhaps the formation of trimetaphosphate and small quantities of tetrametaphosphate from dissolved polyphosphates, which can amount to 70% in presence of Mg++ ions, is an indication of the form in which the anion chains are present in the solution. In any case this formation of trimetaphosphates is not, contrary to what was proposed initially (293, 326), an argument for the assumption that trimetaphosphate rings constitute a structural unit in the polyphosphates. Thus in solutions of Maddrell s salt, the anions of which are known to be linear chains from crystal structure determination (78) (see Section IV,E,2), up to 50% yields of trimeta-... 

Since all condensed phosphates are ultimately degraded to monophosphate in hot solution, especially at low pH, the total phosphorus(V) content of a substance may readily be determined after hydrolysis either gravimetrically or titrimetrically (109). However, as soon as it is a question of estimating the content of separate components in mixtures of condensed phosphates insuperable difficulties are encountered if methods depending on precipitation, titration, or a combination of the two are used. Even a quantitative precipitation of monophosphate is impossible if polyphosphates with chain length of n = 3 or more arc present in the solution. The precipitating cation and the compound to be precipitated by it are partly kept in solution by the polyphosphate part of the polyphosphate is also carried down by the precipitate. Both of these effects depend in their extent in different ways on the nature and quantity of the substances present and the analysis gives a correct quantitative result only in isolated instances... 

Provided only polyphosphates are present it is possible to determine titrimetrically (1) the end-group content of mixtures (2) the content of groups within the chain, after hydrolysis and by estimating the total phosphate content (3) the monophosphate content after precipitation with silver nitrate (347). These methods are likewise of no use in presence of meta- and cross-linked phosphates. 

Highly condensed polyphosphates, which cannot be separated by paper chromatography may be separated into groups by paper electrophoresis 255). Paper chromatography allows the complete separation and quantitative determination of all condensed phosphates with n up to 10, but its use is limited to 7-quantities. 

Addition of sodium polyphosphate appreciably altered the rate constants for reactions (19)—(21) and stabilized the small non-metallic silver clusters [512, 513]. Advantages of the steady-state and pulse-radiolytic approaches to silver-cluster formation are manifold. Firstly, experimental conditions can be precisely adjusted such that the reactive species is exclusively e or, alternatively, that it is a known alcohol radical. Secondly, the concentration of the reducing species (the number of reducing equivalents generated) is readily calculable. Thirdly, in time-resolved experiments, rate constants for the individual reaction steps can be determined by monitoring absorption and/or conductivity changes. These latter determinations permitted the assessment of agglomeration numbers [512,513]. 

Saiardi, A. Caffrey, J.J. Snyder, S.H. Shears, S.B. Inositol polyphosphate multikinase (ArgRIII) determines nuclear mRNA export in Saccharomyces cerevisiae. [Erratum to document cited in CA132 290859]. FEBS Lett., 469, 213 (2000)... 

Robinson, N.A. Clark, J.E. Wood, H.G. Polyphosphate kinase from Pro-pionibacterium shermanii. Demonstration that polyphosphates are primers and determination of the size of the synthesized polyphosphate. J. Biol. Chem., 262, 5216-5222 (1987)... 

Tzeng, C.M. Kornberg, A. The multiple activities of polyphosphate kinase of Escherichia coli and their subunit structure determined by radiation target analysis. J. Biol. Chem., 275, 3977-3983 (2000)... 

The whole question of the specificity was reopened with the discovery that E. coli phosphatase, contrary to an earlier statement (114), hydrolyzed a variety of polyphosphates including metaphosphate of average chain length 8 (97). It was subsequently reported that partially purified phosphatases from several mammalian tissues had appreciable PPi-ase activity at pH 8.5 (115). This was confirmed (116) and extended to include ATPase and fluorophosphatase activities (117). Proof that the same enzyme is responsible for the monoesterase and PPi-ase activities was afforded by heat inactivation studies, cross inhibition experiments, and inhibition of PPi-ase activity by L-phenylalanine, a specific inhibitor of intestinal phosphatase. It was also found that calf intestinal phosphatase couid be phosphorylated by 32P-PP and the number of sites so labeled agreed with the number of active sites determined with a monoester substrate using a stopped-flow technique (118). It would seem that the main reason for the confusion with regard to the PPi-ase activity results from the inclusion of Mg2+ in the assay. This stimulates the monoesterase activity but almost completely inhibits PPi-ase activity (117). 

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