Hydrolysis

Sample Pseudo Dating Site

Due to volatilisation of the test material during test system preparation, the concentration of the test solutions within each pH gradually decreased, as they were dispensed into their individual test vessels.

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The actual concentration of the test solutions used for the determination of mass balance was taken from the average of the concentration of the time 0 samples for each pH. Randomisation of the test systems, specifically the averaged time 0 samples, provided a test solution concentration that was representative of all of the samples within each pH. For test 1 pH 4, 7, and 9, the test solution concentrations were For test 2 pH 4, 7, and 9, the concentrations were Next, the material balance for pH 7 samples ranged from Finally, the material balance for pH 9 samples ranged from Overall, the material balance for individual replicates in test 1 ranged from Overall, the material balance for individual replicates in test 2 ranged from Hydrolysis product methanol correspondingly increased from average values of 2.

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Other unknown metabolites ranged from 0. Next, in the pH 7 test systems, the test material decreased from average values of One other compound gradually increased from an average of 0.

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Additional experiments were performed to show that over time at 50 °C, this artefact will slowly degrade to methanol. The percent of applied radioactivity as the test material was further adjusted to subtract out the effects of its reaction with the TRIS buffer, to determine a more accurate hydrolysis half-life.

All other unknown metabolites ranged from 0. Finally, in the pH 9 test systems the test material decreased, with average values of Hydrolysis product methanol correspondingly increased from average values of 0.

This compound resulted from the direct reaction of the test material with the TRIS buffer, and was not a hydrolysis product. Additional experiments were performed to show that over time this artefact will slowly degrade to methanol. Hydrolysis product methanol correspondingly increased from average values of 1.

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The experimental half-lives of the test material at pH 4, 7, and 9 were directly calculated to be 3. The rate of hydrolysis for the pH 7-test systems was further adjusted to subtract out the reaction of test material with TRIS. Days 1 to 5 were used for this analysis because they follow the same general trend.

Results from days 6 and 7 begin to plateau off and decline, and were therefore not used for the analysis. Next, the rate constant from this data was multiplied by the sampling day and added to the value for the y-intercept 1. These theoretical amounts increase from 3.

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The first-order rate constants for the theoretical amount of non-hydrolysed test material remaining was determined by plotting the natural logarithm In of the average percent of applied radioactivity versus time. The linear regression equation slopes of the best fit line through the data, determined by least squares analysis, was The adjusted experimental half-life of the test material at pH 7 was calculated to be 3. The linear regression equation slopes of the best fit line through each set of data, determined by least squares analysis, was The experimental half-lives of the test material at pH 4, 7, and 9 were calculated to be The rate of hydrolysis for the pH 7 test systems was further adjusted to subtract out the reaction of the test material with TRIS.

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The first-order rate constant for the formation of the TRIS artefact was determined from day 7 to day 30 Sample Pseudo Dating Site plotting the In of the average percent of applied radioactivity of the artefact versus time.

Results from time 0 and day 3 were both 0, and were therefore not used for the analysis.

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Next, the rate constant from the data was multiplied by the sampling day and added to the value for they-intercept These theoretical amounts increase from 0. Since both time 0 and day 3 had no TRIS artefact present, their values were not adjusted.

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The first-order rate constants for the theoretical amount of non-hydrolysed test material remaining was determined by plotting the natural logarithm ln of the average percent of applied radioactivity versus time. The adjusted experimental half-life of the test material at pH 7 was calculated to be The extrapolated rate constants Kh for pH 4, 7, and 9 at 20 °C are 0. The calculated half-lives for pH 4, 7, and 9 at 20 °C are The adjusted rate constant for pH 7 was determined to be 0.

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The fragment Sample Pseudo Dating Site at was due to the loss of CH3. The results of this study show that by co-injecting the standard and sample together that there is an increase in the percentage peak area of methanol, thus confirming that the two peaks are from the same compound.

Validity criteria fulfilled: yes Conclusions: Under the conditions of this study, the Half-lives of the test material in sterile aqueous buffer solutions are: A hydrolysis study was conducted with [14C] labelled test material at a concentrations ranging from For the 50 °C test, duplicate test systems for pH 4, 7, and 9 were measured at time 0 and days 1, 2, 3, 4, 5, 6, and 7.

The material balance ranged from The experimental half-lives for pH 4, 7, and 9 were calculated to be 3. Methanol was the major degradate, reaching an average of A minor artefact from the reaction of the test material with the pH 7 TRIS buffer resulted in the slightly lower half-life.

See table 'Overall remarks' below for full discussion Other kinetic parameters: None. Details on results: The test material was demonstrated to undergo hydrolysis, evident by the formation of orthophosphate, and also by assessment of individual components. However, it was not possible to obtain a definitive result for hydrolytic stability since the test material is a complex mixture of phosphates and the concentration of any individual component monitored may be influenced by the hydrolysis of larger complex phosphates.

The half-life value was adjusted to add back in the theoretical amount of test material that reacted with TRIS to get a better idea of the direct hydrolysis half-life. The adjusted half-life value for pH 7 was calculated to be 3. Tests were also performed using acetate and phosphate as a buffer at pH 7 in place of TRIS, but results showed that acetate could not buffer the system well enough at pH 7 and phosphate also produced artefacts.

TRIS was determined to be the best choice for the pH 7 buffer since it buffers well at that pH and produced only one minor artefact. For the 25 °C test, duplicate test systems for pH 4, 7, and 9 were measured at time 0 and days 3, 7, 14, 21, 28, and The overall material balance for test 2 ranged from The experimental half-lives of the test material at pH 4, 7, and 9, were calculated to be The adjusted half-life for the pH 7 buffer was Sample Pseudo Dating Site The mass spectral and HPLC co-chromatographic data confirmed that the major metabolite was methanol.

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Under the conditions of this study, the half-lives of the test material in sterile aqueous buffer solutions are: Description of key information Under the conditions of this study, the half-lives of the test material in sterile aqueous buffer solutions are: Key value for chemical safety assessment Half-life for hydrolysis:.