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Examines the effect of hydrolysis time on amino acid analysis. Correction for the impact of hydrolysis duration on amino acid determination; Practicalities associated with correcting for hydrolysis interval; Application of correction factors.

The current recommendation, when calculating a protein digestibility-corrected amino acid score, is to determine the digestibility of a dietary protein across the entire digestive tract, using the rat as a model animal for humans. This fecal digestibility value is subsequently corrected for endogenous contributions of protein using a metabolic nitrogen value determined by feeding rats a protein-free diet. The limitations inherent with this method are well recognized, however, and determining the digestibility of a dietary protein to the end of the small intestine is the preferred alternative. Unlike the fecal digestibility assay, which has only one basic methodology, ileal digestibility values can be determined in a number of ways. We discuss the various methods available for determining ileal digestibility values and compare results obtained for dietary proteins using both fecal and ileal digestibility assays. The relative value of using individual amino acid digestibility values as opposed to nitrogen digestibility values is reviewed. In addition, we address issues surrounding measurement of endogenous nitrogen flows, and in particular, the relative merits of determining "true" versus "real" digestibility values. [ABSTRACT FROM AUTHOR]

Background The validation of a dual in vivo-in vitro digestibility assay ('dual digestibility assay') for separately predicting the upper-tract, hindgut and total tract digestibility of nutrients in humans, as estimated using organic matter digestibility ( OMD), is described. Human upper-tract OMD was predicted using an animal (rat) model with digesta from the terminal ileum collected from rats fed one of four complete human diets (wheat bran diet, pectin diet, mixed low-fibre diet, mixed high-fibre diet). Large intestinal OMD was predicted using an in vitro hindgut fermentation assay employing a human faecal inoculum and with the rat ileal digesta as the substrate. Results A comparison of total tract OMD of the four diets from a human balance study ( OMDhuman) with that predicted using the dual digestibility assay ( OMDdual) showed no significant differences ( P > 0.05). OMDhuman and OMDdual were highly correlated ( r = 0.953, P = 0.047). Conclusion The dual digestibility assay accurately predicts the uptake of dietary nutrients (as grams of organic matter) in humans over the total tract. The assay is able to separately quantify the digestibility of nutrients in the upper and lower digestive tracts. The validation of the dual digestibility assay needs to be extended to a wider range of human diets. © 2013 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

Abstract: The aim was to optimise inoculum concentration and incubation duration for a published in vitro hindgut digestibility assay using ileal digesta (sampled from the chicken or rat) pertaining to a mixed human diet as the substrate. The study also sought to investigate the digestibility of the inoculum itself and the importance of correcting for this in the in vitro hindgut digestion assay. For two assays, hindgut dry matter digestibility (DMD) generally increased with inoculum concentration. A sharp increase in DMD observed at high inoculum concentrations may have been related to problems with filtering the inoculum. An inoculum concentration of 160g/L was considered optimal based on close agreement of observed values with previously published in vivo hindgut dry matter digestibility for similar diets. One of the methods was chosen for optimisation of the duration of incubation. Ileal substrate organic matter digestibility (OMD) increased with increasing time of incubation for all diets. An incubation duration of 18h using a mean inoculum digestibility value for calculation purposes was considered optimal based on observed in vivo hindgut DMD values in humans, but there was little difference in estimated in vitro hindgut DMD between 18 and 24h incubation durations. Although considerably lower than the OM digestibility of the substrate (no less than 51% after 48h), the OM digestibility of the inoculum (13% after 48h) itself was of significance in calculating estimated digestibility. The optimised assay gave realistic hindgut OMD values ranging from 55% to 79% (Wheat Bran Diet and Pectin Diet, respectively) using an 18-h incubation duration. [Copyright &y& Elsevier]

Abstract: In vitro assays have been developed to predict dry matter digestibility (DMD) in the human colon, but there is little information on the effect of assay variables. The effect of altering pH, duration of incubation, presence of shaking during incubation and the concentration of faecal inoculum or digestive enzymes on DMD was investigated for three in vitro hindgut digestibility assays. Three mixed human diets varying in the type and ratio of soluble and insoluble dietary fibre were used as substrates. The pH, duration of incubation and the concentration of inoculum relative to substrate significantly (P >0.05) affected predicted DMD for the two in vitro methods employing faecal inocula. The method using synthetic enzymes showed little sensitivity to alteration of assay variables and gave highly variable results and for this reason was not pursued further. Shaking did not affect (P >0.05) digestibility for any method or diet. The different methods led to large differences in predicted hindgut DMD within each of the three diets. In vitro hindgut digestibility assay variables need to be optimised and the predicted DMD data validated against in vivo data. [ABSTRACT FROM AUTHOR]

The apparent digestibility of energy-yielding nutrients (carbohydrate, protein, and fat) was predicted in the human upper digestive tract and large bowel separately for 4 diverse diets containing either a single dietary fiber source [wheat bran and pectin (PE) diets] or mixed fiber sources [low-fiber (LF) and high-fiber (HF) diets). A human balance study was undertaken to determine fecal energy and nutrient excretion and a rat model was used to predict human ileal energy and nutrient excretion. Total tract energy digestibility ranged from 92 (HF diet) to 96% (PE diet and LF diet), while at the ileal level it ranged from 79 to 86% for the HF diet to the LF diet. The predicted upper-tract digestion of starch, sugars, and fat was high, with ileal digestibilities exceeding 90% for all diets. Nonstarch polysaccharides were poorly digested in the upper tract for all diets except in the PE diet. The daily quantity of protein excreted at the ileal level was between 2 (HF diet) and 5 (RE diet) times higher than that at the fecal level. The large differences between fecal and ileal nutrient loss highlight that fecal digestibility data alone provide incomplete information on nutrient loss. There is a need to be able to routinely determine the uptake of energy in the upper and lower digestive tracts separately. [ABSTRACT FROM AUTHOR]