Influence of assay conditions on the in vitro hindgut digestibility of dry matter

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.