It was hypothesized that ester-linked gallic acid glycosides could be absorbed and metabolized in vivo due to the instability of an ester-linked glycosides at neutral pH. To evaluate in vivo absorption of Mangifera Indica, L. var. Keitt polyphenolics, it was first necessary to chemically characterize the compounds present using HPLC-MSn analysis. Mango pulp and extracts were also incubated with a pectinase, cellulase, pectinase with ß-glucosidase activity, and a pure ß-glucosidase to learn the extent of hydrolysis with potential application to enhancing bioavailability as a result of incubation to increase mango juice yield. After which the same extracts were assessed in differentiated Caco-2 cells to discern stability at physiological pH and to characterize metabolite formation in vitro. Finally, human urinary metabolites were characterized after 10 day consumption of 400 g in 11 individuals. Mass spectroscopic characterization and HPLC quantification of mango pulp revealed for the first time two monogalloyl glucosides (MGGs) with distinct differences in their glycoside linkages, with the ester form dominating, as well as the presence of five other phenolic acid glycosides; hydroxybenzoic acid glucoside, courmaric glucoside, ferulic acid glucoside, and sinapic acid mono and di-glucosides. Six oxygenated carotenoid derivatives were identified for the first time in a phytochemical extract, namely, a phytohormone, abscisic acid and its glycoside, two catabolism products of abscisic acid, dihydrophasic acids, and two hydroxy-dimethyl decadiene-dioic acid glucopiranosylesters. Gallotannins ranging from tetra-galloyl glucosides to nona-galloyl glucosides were also identified in the pulp, but not quantified. Clearzyme 200XL and Rapidase AR2000 were the most effective at increasing juice yield of mango pulp due to their pectinase action. Cz reduced the amount of ester-linked MGG by 70% after 4 hours of incubation while Rap hydrolyzed the ether linked MGG. The instability of ester-linked galloyl-glycosides at pH 7.4 was characterized by HPLC-MS and after only four hours of incubation a shift from HWM tannins (>8GG) to LMW (<8GG) occurred and resulted in 25 mg/L free gallic acid. Caco-2 cells metabolized gallic acid, MGG from a mango extract, and a gallotannin extract into O-methyl gallic acid indicating catechol-O-methyl transferase (COMT) as a major metabolizing enzyme for gallic acid. Urinary metabolites were identified by HPLC-MSn in dependant scans. O-methylgallic acid-O-sulfate was identified as the major metabolite 0-6 hours post consumption, followed by O-methylgallic acid at a lower concentration. The presence of gallic acid metabolites in the urine indicates absorption of ester-linked glycosides. Colonic metabolites were detected beginning 3-6 hours after consumption of mango, and were identified as pyrogalloyl derivatives. They are hypothesized to be the products of microbial breakdown of gallotannins. Daily consumption of mango for 10 days increased the concentration of O-methylgallic acid-O-sulfate, but was not significant.

For many years, substitutes for butterfat have been the subject of much controversy. Their acceptance as being nutritionally equal to butterfat has often been question. The importance of this problem lies primarily in its economic and nutritional significance. A recent study of pulic attitudes toward dairy products conducted by Alfred Politz Research, Inc. for the American Dairy Association, shows that 48.4 per cent of the poulation believed that butter is a more nutritions food than oleomargarine. Only 6 per cent thought that oleomargarine is more nutritious while 27.2 per cent believed they are about the same. The remainder, 18.4 per cent, stated that they did not know. Margarine tax laws were in force for some time while vegetable oils were being tested as a human food. In addition to chemical examination, many investigations were conducted with rats, calves, and humans. Although there were still conflicting results, the tax laws were repealed and substitutes wen on the market to compete with butter. Many investigators still question the decision that substitutes are qual to butter, nutritionwise. Many investigations (4, 6, 21, 34) have shown that butter contains one or more growth factors which have not, as yet been identified. Chemical methods have failed to identify any of these factors, and so other methods have been considered. It is a well known fact that interrelationships exist between fat metabolism and the metabolism of other substances. It has been shown (11) that dietary fat affects the metabolism of other nutrients such as calcium and phosphorus. Although not clearly understood, it has been demonstrated (25, 37) that the kidney plays an important role in fat transport and metabolism. The value of urinalysis as a method of indicating the metabolism as a whole is well known, as well as is the high sensitivity of metabolic systems to minute quantities of some substances. The possibility of detecting growth factors or other substances through a study of excreted metabolites seems worthy of consideration in this study. Recent work with paper chromatography at the University of Texas (48) has indicated that each individual has a specific urinary metabolic pattern. Similarities have been found in the patterns of identical twins and schizophrenics. It was also found that the diet changed this pattern to a certain extent, although this was not studied extensively. The problem reported here is a study of urine obtained from highly inbred rats fed animal and vegetable fats analyzed by use of paper chromatography.