Thomas Erk

• A positive affection of human health by nutrition is of high interest, especially for bioactive compounds which are consumed daily in high amounts. This is the case for chlorogenic acids (CGA) ingested by coffee. This molecule class is associated with several possible beneficial health effects observed in vitro that strongly depend on their bioavailability. So far factors influencing bioavailability of CGA such as dose, molecule structure and site of absorption haven´t been investigated sufficiently. Therefore we performed an in vivo dose-response study with ileostomists, who consumed three different nutritional doses of CGA ingested as instant coffee (4,525 (HIGH); 2,219 (MEDIUM); 1,053 (LOW) μmol CGA). CGA concentrations were determined in ileal fluid, urine and plasma. Furthermore, we conducted an ex vivo study with pig jejunal mucosa using the Ussing chamber model to confirm the in vivo observations. Individual transfer rates of CGA from coffee were investigated, namely: caffeoylquinic acid (CQA), feruloylquinic acid (FQA), caffeic acid (CA), dicaffeoylquinic acid (diCQA) and QA at physiological concentrations (0.2–3.5 mM). Samples were analyzed by HPLC-DAD, -ESI-MS and -ESI-MS/MS. About ⅔ of the ingested CGA by coffee consumption were available in the colon dose independent. Nevertheless, the results showed that the consumption of higher CGA doses leads to a faster ileal excretion. This corresponds to a plasma AUC0-8h for CGA and metabolites of 4,412 ± 751 nM*h0-8-1 (HIGH), 2,394 ± 637 nM*h0-8-1 (MEDIUM) and 1,782 ± 731 nM*h0-8-1 (LOW) respectively, and a renal excretion of 8.0 ± 4.9% (HIGH), 12.1 ± 6.7% (MEDIUM) and 14.6 ± 6.8% (LOW). Moreover interindividual differences in gastrointestinal transit times were related to differences in total CGA absorption. Thus the variety of patient´s physiology is a decisive bioavailability factor for CGA uptake. This is corroborated ex vivo by a direct proportional relationship of incubation time with absorbed CGA amount. The consumption of high CGA doses influences the metabolism pattern as an increasing glucuronidation was observed with consumption of increasing CGA doses. However, the different CGA doses have only minor effects on the overall bioavailability which was confirmed ex vivo by a non-saturable passive diffusion of 5-CQA. Furthermore, we identified in the Ussing chamber an active efflux secretion for 5-CQA that decreases its bioavailability and the physicochemical properties of the CGA subgroups as an important bioavailability factor. Transferred amount in increasing order: diCQA, trace amounts; CQA ≈ 1%; CA ≈ 1.5%; FQA ≈ 2%; and QA ≈ 4%. Altogether, the consumption of increasing CGA doses by coffee had a minor effect on oral bioavailability in ileostomists, such as a slightly increased glucuronidation. Thus, the consumption of high amounts of CGA from coffee in the daily diet is not limiting the CGA concentrations at the site of possible health effects in the human body. However, according to the patient´s physiology the interindividual gastrointestinal transit time which is possibly influenced by dose is influencing CGA bioavailability. Moreover, ex vivo CGA absorption is governed by diffusion as an absorption mechanism corroborating an unsaturable uptake in vivo and by the individual physicochemical properties of CGA.