6 g of glucose per hour and was continuously monitored Blutzuckermessger Ten. In each case, blood sugar was below 3.5 mmol / l concerning Gt One participant reported symptoms of gastrointestinal discomfort after consumption of controlled processing On. Tea drinking green tea TGX-221 extract consisted of 1.75 g of decaffeinated coffee in 300 ml of water gel St. This dose is equivalent to 4 5 cups of green tea. In the GT M group, 20% water was prepared by commercial skimmed milk, which corresponds to g to a protein content of 2.17 replaced. The other two treatments soy protein or casein was used in amounts equal to the green tea beverage Nk added. The exact compositions of green tea beverage drinks which are given in Table 2.
Sample preparation and analysis of fasting blood-tive Sen Blood samples were taken before and 30, 60, 90, 120, 150, 180, 210, 240 and BMS 777607 270 min after ingestion of tea. Blood was obtained through a cannula placed in the median cubital vein. The blood is in R Hrchen Collected with lithium heparin. The plasma was separated by centrifugation at 2.0009 g, 10 min at 4 C and aliquoted. Subsequently End, 1 ml of each plasma sample with 20 ll of an L Mixed solution of ascorbate EDTA and stored at 80 C until analysis. EGCG, EGC, ECG, EC, catechin, gallocatechin, and all other reagents and HPLC L Solvents were purchased from Carl Roth. The plasma concentrations of individual catechins were determined by HPLC as described by Lee et al. with some modifications. Briefly, total catechins after hydrolysis by incubating 0.5 ml plasma for 45 min with an enzyme mixture of glucuronidase and sulfatase D b determined.
Then, 1 ml of methylene chloride was added and the sample was shaken and centrifuged at 3.2209 g for 15 min at 4 C. The supernatant was in a Testr Transferred Hrchen and with new 1 ml of ethyl acetate. Thereafter, the sample was mixed and centrifuged again. A 800 ll volume of the organic phase was transferred into a fresh R Hrchen transferred and the ethyl acetate extraction was repeated twice. The combined whichever type Walls were coated with 10 ll of ascorbic Acid and 1% w Ssrigem dried by vacuum centrifugation. The dried sample was dissolved in 150 ll mobile phase by vortex mixing and sonication St. The sample was centrifuged, the supernatant and 30 ll injected into the HPLC. The analysis was performed on an LC-2000 Plus series HPLC system at a coulometric electrochemical detector 4 canals le.
The separation was maintained on reverse phase C18 Kromasil 100-S Column at 30 C, and performed by a Inertsil ODS-C18-S Column 2 Guard. Mobile phase A and B of the water, acetonitrile and trifluoroacetic acid, at a rate of 92: 8:0.1 respectively, and 65:35:0.1. The flowsheets rate was 0.9 ml / min and the eluate was monitored by electrochemical detection with m Adjusted settings at 0, 120, 240 and 360 mV. The st Strongest signals were used for quantification were 0 mV for EGC, 120 mV for GC, EC, and EGCG, and 240 mV for C and ECG. The detection limit was 10 nmol / L amounts to Gt The coefficient of variation was 2.4 2.6 6.5 4.7% for all the analyzed catechins. The individual plasma cells were re-catechins with external standards. The calibration curves were prepared by addition of C, GC, EGC, EGCG, ECG and EC plasma blank form finalpolyphenols st Produces amylopectin Ren Of