Capsule production
-20, LEAKAGE
The study persons provided a fresh faecal sample prior to study start. Sixty grams of the faecal sample was diluted with twice the amount of glycerol by weight, homogenized and filtered through a course sieve before freezing to -80°C in Falcon tubes. The procedure was finished within three hours after delivery of faeces.For capsulation, the frozen sample was thawed in room temperature for two hours, vortexed and then centrifuged at 300 g for 20 minutes. The supernatant was transferred to new Falcon tubes and was centrifuged at 4600 g for 40 minutes. The supernatant was then discarded, and the pellet was weighed and homogenized with the same amount of glycerol. Ten percent of olive oil and 1% of Tween 80 by weight were then added and the mixture homogenized. The mixture was added to white opaque, HPMC acid-resistant capsules nr 0, 0,6 mL (DRcaps™, Capsugel Corp.). The capsules were put into a second slightly larger capsule no 00 to avoid leakage of fecal material and to limit odor.Twenty capsules were packed in a plastic screw-top container and one drop of peppermint oil was placed on the inside of the lid. The containers were individually labeled for each study person and stored at -20°C until delivery to the subject. Reference samples were stored at -20°C.Placebo consisted of size 0 capsules filled with cocoa powder mixed in olive oil, placed inside size 00 capsules and packed and stored as study capsules.