Pyrophosphate, determination

Pyrolusite, in oxidation of sulfurous acid to dithionate ion, 2 168 Pyrophosphates, determination of, in mixtures of phosphates, 3 93 Pyrophosphoric acid, 3 96, 97 Pyrosulfites, alkali metal, 2 162 Pyrosulfuryl chloride, 3 124, 126... 

Floridi A, Pupita M, Palmerini CA, Fini C, Alberti FA. Thiamine pyrophosphate determination in whole blood and erythrocytes by high performance liquid chromatography. Int J Vitam Nutr Res 1984 54 165-71. 

Function 3 affects performance as determined by the chemistry of alkylate, carboxyl, sulfonyl, phenolic, phosphate, pyrophosphate, and phosphite groups. 

In the normal process ( ), step (J) occurs very rapidly and step (/) is the rate-determining step, whereas in the inhibition process (B), step (3) occurs very slowly, generally over a matter of days, so that it is rate determining. Thus it has been demonstrated with AChE that insecticides, eg, tetraethyl pyrophosphate and mevinphos, engage in first-order reactions with the enzyme the inhibited enzyme is a relatively stable phosphorylated compound containing one mole of phosphoms per mole of enzyme and as a result of the reaction, an equimolar quantity of alcohoHc or acidic product HX is hberated. 

Simultaneous detenuination of Cu and Zn in the form of coloured PAR complexes is performed at pH 10 in the presence of pyrophosphate which binds the admixtures of Al, Fe and Mn into the inactive complexes. The measurements of the change in the optical density are made at 520 and 550 nm before and after the destmction of the complexes by EDTA, or at 530 nm before and after the destruction of the copper complexes by the thioglycolic acid and the destmction of the zinc complexes by EDTA. The detection limit for Cu is 2-5, for Zn - 3 p.g/diW. The application of these methodics at pH 8 enables one to determine simultaneously Cu and Zn at high excess of the latter. 

The simultaneous determination of Co and Ni is also made at pH 8 in the presence of pyrophosphate. The EDTA is added to the mixture of coloured complexes of these metals to bind the Cu and Zn admixtures into the inactive complexes. The optical density of the solution is measured at 530, 555 and 580 nm. The solution is heated to the boiling point to destmct the complex formed by Ni with PAR, and then is cooled. Again the measurements of optical density ai e performed at the same wavelengths. The Ni concentration is calculated from the variation in the optical density, and the Co concentration is calculated from the final values of optical density. The detection limits for these metals are 4 and 2 p.g/dm, respectively. 

Prepare also a saturated solution of the purest available sodium pyrophosphate (do not heat above 25 °C, otherwise appreciable hydrolysis may occur) 12 g of the hydrated solid Na4P2O7,10H2O will dissolve in 100-150 mL of water according to the purity of the compound. It is essential to employ freshly made sodium pyrophosphate solution in the determination. 

Kinetic data exist for all these oxidants and some are given in Table 12. The important features are (i) Ce(IV) perchlorate forms 1 1 complexes with ketones with spectroscopically determined formation constants in good agreement with kinetic values (ii) only Co(III) fails to give an appreciable primary kinetic isotope effect (Ir(IV) has yet to be examined in this respect) (/ ) the acidity dependence for Co(III) oxidation is characteristic of the oxidant and iv) in some cases [Co(III) Ce(IV) perchlorate , Mn(III) sulphate ] the rate of disappearance of ketone considerably exceeds the corresponding rate of enolisation however, with Mn(ril) pyrophosphate and Ir(IV) the rates of the two processes are identical and with Ce(IV) sulphate and V(V) the rate of enolisation of ketone exceeds its rate of oxidation. (The opposite has been stated for Ce(IV) sulphate , but this was based on an erroneous value for k(enolisation) for cyclohexanone The oxidation of acetophenone by Mn(III) acetate in acetic acid is a crucial step in the Mn(II)-catalysed autoxidation of this substrate. The rate of autoxidation equals that of enolisation, determined by isotopic exchange , under these conditions, and evidently Mn(III) attacks the enolic form. 

G.l.c. studies of tributylphosphine, dialkyl phosphites, and dialkyl alkylphosphonates are reported. Tributyl phosphate in nitric acid can be estimated by g.l.c. if a glass column is used. Tetraethyl pyrophosphate has been directly determined on a nanogram scale by g.l.c., whereas it was found most convenient to first convert the tetra-aryl pyrophosphates by methanolysis to diarylmethyl phosphates. Phosphorochloridates were converted by t-butyl alcohol into t-butyl chloride before analysis. G.l.c. studies of pesticides have been reported and the isomeric thiophosphates (138a) and (138b) have quite different retention times. ... 

Of the three organic phosphorus insecticides—hexaethyl tetraphosphate, tetraethyl pyrophosphate, and parathion—the first two have been shown to be mixtures (36) that contain tetraethyl pyrophosphate as the principal active ingredient. Several methods have been proposed for the determination of this compound in the commercial products (25, 35). All are based on the separation of the tetraethyl pyrophosphate from the related ethyl phosphates, followed by its hydrolysis to diethyl orthophosphoric acid and titration with standard alkali. Both hexaethyl tetraphosphate and tetraethyl pyrophosphate are soluble in water and are rapidly hydrolyzed to monoethyl and diethyl orthophosphoric acid. This rapid hydrolysis to nontoxic products greatly limits the duration of the in- secticidal effectiveness of tetraethyl pyrophosphate, but it also eliminates the danger of toxic residues on the crops treated. 

The purpose of this work was to determine the toxicity to mosquito larvae of insecticide spray residues. That certain insecticides are translocated in plants (4, 5) adds impetus to this study. Fresh orchard fruit sprayed or dusted with preparations containing parathion (0,0-diethyl O-p-nitrophenyl thiophosphate), tetraethyl pyrophosphate (TEPP, HEPP), DDD [2,2-bis(p-chlorophenyl)-l,l-dichloroethane], DDT [2,2-bis(p-chlorophenyl)-l,l,l-trichloroethane], chlorinated camphene, and basic lead arsenate were shipped from California to Yonkers, N. Y., by air express for bioassay. 

The purified tetraethyl pyrophosphate is a colorless, odorless, water-soluble, hygroscopic liquid (24, 4 )- It possesses a very high acute toxicity (28), exceeding that of parathion, and is rapidly absorbed through the skin. There is no spray-residue problem, however, for tetraethyl pyrophosphate hydrolyzes even in the absence of alkali to nontoxic diethyl phosphoric acid. Hall and Jacobson (24) and Toy (47) have measured its rate of hydrolysis, which is a first-order reaction. Its half-life at 25° C. is 6.8 hours and at 38° C. is 3.3 hours. Coates (10) determined the over-all velocity constant at 25° C. k = 160 [OH-] + 1.6 X 10 3 min.-1 Toy (47) has described an elegant method for preparing this ester as well as other tetraalkyl pyrophosphates, based upon the controlled hydrolysis of 2 moles of dialkyl chlorophosphate ... 

As a solvent for this reaction, anhydrous methyl ethyl ketone was found satisfactory. Coates (10) determined the rate of hydrolysis of the monothio analog as approximately one fifth that of tetraethyl pyrophosphate under similar conditions. The dithio analog has been prepared (22) in 90% yield from diethyl chlorothiophosphate, water, and pyridine in a modification of the reaction Toy (47) used to make tetraethyl pyrophosphate ... 

Marcus, S.L., and Balbinder, E. (1972) Use of affinity matrices in determining steric requirements for substrate binding Binding of anthranilate 5-phosphoribosyl-pyrophosphate phosphoribosyltransferase from Salmonella typhimurium to Sepharose-anthranilate derivatives. Anal. Biochem. 48, 448-459. 

A flow assay was reported for determination of inorganic pyrophosphate a pyrophosphatase was coimmobilized with luciferase on Sepharose beads with continuous flow of saturating concentrations of substrates. The instrument allowed automation with a throughput of approximately one sample every 4 min. 

Less than 1 pmol pyrophosphate was determined in a volume of 20 pL with a coefficient of variation approximately equal to 4% [230],... 

Phosphorus is a common component of additives and appears most commonly as a zinc dialkyl dithiophosphate or triaryl phosphate ester, but other forms also occur. Two wet chemical methods are available, one of which (ASTM D1091) describes an oxidation procedure that converts phosphorus to aqueous orthophosphate anion. This is then determined by mass as magnesium pyrophosphate or photochemically as molybdivanadophosphoric acid. In an alternative test (ASTM D4047), samples are oxidized to phosphate with zinc oxide, dissolved in acid, precipitated as quinoline phosphomolybdate, treated with excess standard alkali, and back-titrated with standard acid. Both of these methods are used primarily for referee samples. Phosphorus is most commonly determined using x-ray fluorescence (ASTM D4927) or ICP (ASTM D4951). 

One of the important consequences of studying catalysis by mutant enzymes in comparison with wild-type enzymes is the possibility of identifying residues involved in catalysis that are not apparent from crystal structure determinations. This has been usefully applied (Fersht et al., 1988) to the tyrosine activation step in tyrosine tRNA synthetase (47) and (49). The residues Lys-82, Arg-86, Lys-230 and Lys-233 were replaced by alanine. Each mutation was studied in turn, and comparison with the wild-type enzyme revealed that each mutant was substantially less effective in catalysing formation of tyrosyl adenylate. Kinetic studies showed that these residues interact with the transition state for formation of tyrosyl adenylate and pyrophosphate from tyrosine and ATP and have relatively minor effects on the binding of tyrosine and tyrosyl adenylate. However, the crystal structures of the tyrosine-enzyme complex (Brick and Blow, 1987) and tyrosyl adenylate complex (Rubin and Blow, 1981) show that the residues Lys-82 and Arg-86 are on one side of the substrate-binding site and Lys-230 and Lys-233 are on the opposite side. It would be concluded from the crystal structures that not all four residues could be simultaneously involved in the catalytic process. Movement of one pair of residues close to the substrate moves the other pair of residues away. It is therefore concluded from the kinetic effects observed for the mutants that, in the wild-type enzyme, formation of the transition state for the reaction involves a conformational change to a structure which differs from the enzyme structure in the complex with tyrosine or tyrosine adenylate. The induced fit to the transition-state structure must allow interaction with all four residues simultaneously. 

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