Enzymic hydrolysis, analysis

In examining the structure of a polysaccharide, it is convenient to consider the methods involved under the three main headings (a) quantitative analysis, (b) methylation, and (c) periodate oxidation. These techniques may be supplemented by partial or enzymic hydrolysis as the circumstances indicate. Each of these aspects of polysaccharide chemistry may be aided by the application of gas-liquid chromatography, either qualitative or quantitative, or both. Thus, separations impossible by other techniques may often be achieved, and analytical data obtained in a fraction of the time demanded by other methods. 

DS Forsyth, JR Lyengar. Enzymic hydrolysis of biological and environmental samples as pretreatment for analysis. J Assoc Off Anal Chem 72 997-1001, 1989. 

Enzyme hydrolysis, with papain, diastase, clarase, takadiastase, intestinal phosphatase, or combinations thereof is most commonly used to release pantothenate from food proteins (186). A cold perchloric acid extraction was used to release pantothenic acid from tissue samples (187). Food spoilage prior to analysis may lead to inflated pantothenic acid levels (19). 

Fractionation Data and Distribution Analysis of HEC After One Hour of Cellulase Attack. The results of the gel chromatographic separation of HEC after one hour of enzymic hydrolysis are given in Table II. These fractionation data did not correspond to any of the distribution functions mentioned by Peebles (41) and by Tung (42). In the middle of the distribution it corresponded to the Lansing-Kraemer distribution functions, but deviations occurred at the low- and high-molecular-weight ends. 

Fractionation Data and Distribution Analysis of HEC After One Day of Cellulase Attack. In Table III, the fractionation data of HEC are given after one day of enzymic hydrolysis. As after one hour of enzymic hydrolysis, no theoretical distribution function accorded well with the fractionation data, but we evaluated the parameters by numerical analysis, using the Gauss-Laguerre method (43,44). This method has one advantage over other numerical methods, e.g., (45)—all the calculations involved can be done manually without the need of high-speed computers. 

Macromolecule therapeutic agents tend to be more reactive than small-molecule drags under various circumstances, from sample collection to analysis. Biological and chemical transformation may occur as a result of enzyme hydrolysis, heat or... 

Figure 13. Chromatogram of silylated ether extract (pH 5.5) of human urine prior to enzyme hydrolysis. (Chromatographic conditions same as Figure 4). Shaded area denotes retention time window during which the Olfax searched for the 9-THC-ll-oic acid/TMS contracted spectrum. Inset Auto-Assay analysis for THC panel of 8 compounds in which 240 ng (2.40 X 10 1 fig) of A9-THC-ll-oic acid was found with a Confidence Index of 62.

Mottram, H.R. (1999) The Application of HPLC-APCI MS to the Regiospecific Analysis of Triacylglycerols in Edible Oils and Fats. PhD thesis, Department of Chemistry, University of Bristol, UK. Movia, E. and Remoli, S. (1977) Application of enzymic hydrolysis to determine the genuineness of butter., Bollettino dei Chimici dei Laboratori Provinciali, 3, 187-192. 

Extraction of Active Compounds from Food. Vitamins are the group of compounds more usually extracted from foods using SFE (83). A method for the analysis of the natural contents of vitamins A and E in milk powder based on SFE, a miniaturized alkaline saponification procedure, and HPLC was proposed by Turner and Mathiasson (84). Modifications of the sample matrix, the combination of static and dynamic extraction modes, and the effect of changes in extraction parameters such as temperature, flow rate, time, collection solvent, and collection temperature were optimized, obtaining recoveries of 99% and 96% for vitamins A and E, respectively. Another method for the determination of vitamins A and E based on the coupling of SFE-enzymic hydrolysis-HPLC has also been proposed providing recoveries between 79% and 152% (85). 

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