• Supplier: Nutrition Innovation • Amount in TIM-2: 7 g and 3.5 g Test product #2 Negative Control •Name: Cellulose •Description: Cellulose, microcrystalline, extra pure, CAS: 9004-34-6 •Batch number: A0381663 •Form: powder, particle size 90 pm • Storage conditions: ambient temperature (15-25°C) •Expiry date: 30 November 2022 • Supplier: ACROS organics • Amount in TIM-2: 7 g SIEM and dialysis liquid SIEM (standardized ileal effluent medium) simulates material passing the ileocecal valve in humans, or in other words material reaching the colon. The SIEM in the TIM-2 experiments contained the major non-digestible carbohydrates (pectin, xylan, arabinogalactan, amylopectin, starch) found in a normal western diet as well as protein (bactopepton, casein), some ox-bile, Tween 80 as well as vitamins and minerals. SIEM does not require pre-digestion and was added to the system at a speed of 2.5 ml/hour. The speed of the dialysis liquid was 1.5 ml/min. Test system: TIM-2 The TNO intestinal model TIM-2 (Figure 9) is a dynamic in vitro model of the proximal large intestine. The conditions in the proximal large intestine are mostly determined by the composition of the microbiota. The TIM-2 system (Figure 9) was inoculated with a dense and highly metabolic active colon microbiota of human origin. In the system the following standardized conditions are simulated: body temperature; pH in the lumen of the proximal colon (pH 5.8); anaerobiosis; delivery of a predigested substrate from the ‘ileum’ (SIEM); mixing and transport of the intestinal contents; absorption of water and absorption of metabolic products (via dialysis). The content of the system was kept under strict anaerobic conditions (flushing with nitrogen). Fermentation products, metabolites and other low molecular weight