Porcine bile extract b8631

All solvents were HPLC grade and purchased from Fisher Scientific (Loughborough, UK). Standards (β-carotene and α-tocopherol) were HPLC grade and purchased from Sigma Aldrich (Merck, Darmstadt, Germany), except lutein which was purchased from PanReac AppliChem (Darmstadt, Germany). Enzymes, including α-amylase from human saliva type XIII-A, 300-1500 U mg -1 protein (A1031), pepsin from porcine gastric mucosa (P6887), pancreatin from porcine pancreas (8xUSP; P7545), and porcine bile extract (B8631), were purchased from Sigma Aldrich (Merck, Darmstadt, Germany). Other chemicals used were of analytical grade.

Fresh raw bovine milk was obtained from a local dairy farm, stored overnight at 4°C, and then skimmed or standardized to 3.25% (wt/wt) fat content. One-liter portions underwent homogenization at 60°C (Universal Pilot Plant, Waukesha Cherry-Burrell, Philadelphia, PA) and (or) HTST pasteurization at 72°C for 15 s, and UHT processing at 135°C for 2 s in an Armfield model FT74P/T HTST/UHT (Armfield Inc., Denison, IA) plate-and-frame continuous pasteurizer (Tomasula and Kozempel, 2004) and were then stored at 4°C. Fresh sample was used within 1 d and processed samples were used in <3 d. The abbreviations used throughout the text to define the milk samples are as follows: R = raw unprocessed milk, RS = raw skim milk, RW = raw whole milk, H = homogenized milk, P = HTSTpasteurized milk, SP = HTST-pasteurized skim milk, HP = homogenized and HTST-pasteurized milk, SU = UHT-processed skim milk, and HU = homogenized and UHT-processed milk.
Fat, lactose, and protein were determined by a MilkoScan Minor (Foss, Eden Prairie, MN). Pepsin from porcine gastric mucosa (P7000; 250 U/mg of solid), porcine bile extract (B8631), and porcine pancreatin (P1750; 4 × USP) were purchased from Sigma-Aldrich Corp. (St. Louis, MO). All other chemicals were of analytical grade and were purchased from Sigma-Aldrich unless specified otherwise.

Sodium caseinate (Nacas) was obtained from Fonterra Co-operative Group Ltd, Auckland, New Zealand. Porcine bile extract B8631 and porcine pancreatin (P1750, 4 × USP) were purchased from Sigma-Aldrich Chemical Company (St. Louis, MO, USA).
Porcine BE used in this study had a total bile salt content of 49 wt%, of which the majority of the bile acid species were glycodeoxycholic acid (10 15 wt%) followed by taurodeoxycholic acid (3 9 wt%) and deoxycholic acid (0.5 7 wt%) (Zangenberg, Müllertz, Kristensen, & Hovgaard, 2001) . The key phospholipid was phosphatidyl choline (6 wt%) and the content of Ca 2+ was less than 0.06% (w%). Based on the phospholipid/ bile acid ratio, it can be suggested that the phospholipid was present as mixed micelles in conjunction with bile salt (Wickham, et al., 1998) The pH was adjusted to 7.0 using 1 M NaOH or 1 M HCl. Initially, pre-emulsions were prepared by blending 20.0 wt% soy oil with 80.0 wt% aqueous Nacas solution or BE solution using a conventional high speed mixer (Silverson L4RT, OFI Testing Equipment, Inc., Houston, TX, USA) at 6500 rev/min for 3 min. These coarse emulsions were then passed twice through a mini two-stage valve homogenizer (12.5H, Rannie, Copenhagen, Denmark) operating at 250 bar and 50 bar in the first and second stages respectively. The Nacas and BE emulsions were prepared at least in duplicate.

Whey protein isolate (WPI) powder containing 96.3 wt% protein was kindly gifted by Fonterra Limited (Auckland, New Zealand). Cellulose nanocrystal powder (CNC, sulphated) was purchased from CelluForce™, Canada. Sunflower oil was purchased from a local supermarket (Morrisons, UK). Porcine bile extract B8631, porcine pancreatin (P7545, 8 × USP) and sodium azide were purchased from Sigma-Aldrich Company Ltd, Dorset, UK. All other chemicals used were of analytical grade and were obtained from Sigma-Aldrich Chemical Company unless otherwise specified.
Milli-Q water having an ionic purity of 18.2 M cm at 25 °C (water purified by treatment with a Milli-Q apparatus, Millipore Corp., USA) was used for all the experiments.

Raw goat milk samples from Murciano-Granadina breed were collected from a farm in the region of Granada (Spain). They were skimmed by centrifugation and concentrated by ultrafiltration through a 50 kDa membrane (Vivaflow 2000, Sartorius Stedin Biotech, Madrid, Spain). Fermentation was conducted with (i) the classical starter bacteria Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus salivarius subsp. thermophile (St) and (ii) St + the probiotic strain Lactobacillus plantarum C4 [8] (link) (St+ LP). Enzymes and bile salts were purchased from Sigma Chemical Co (St Louis, MO, USA), porcine pepsin (P-7000), porcine pancreatin (P-1500) and porcine bile extract (B-8631). All other reagents such as HCl, ammonia, NaHCO3, formic acid, acetonitrile, were purchased from Sigma Chemical.

Pure standard analytes, including gallic acid, catechin, chlorogenic acid, syringic acid, epicatechin, ferulic acid, rutin, quercetin, hydrochloric acid (HCl) and formic acid were obtained from Sigma-Aldrich (Zwijndrecht, The Netherlands). Kaempferol 3-glucoside and delphinidin 3-glucoside were provided by Phytolab (PhytoLab GmbH & Co, Vestenbergsgreuth, Germany). 4-hydroxy-benzoic acid and ethyl acetate were purchased from Merck (Merck GmbH, Darmstadt, Germany). Cyanidin 3,5-diglucoside, cyanidin 3-glucoside, pelargonidin 3,5-diglucode and cyanidin chloride were obtained from Extrasynthese (Genay, France). Porcine pepsin (P6887, 3.200-4.500 U/mg protein), pancreatin (P1750, 4X USP) and porcine bile extract (B8631) were purchased from Sigma-Aldrich Ltd. (St. Louis, MO, USA). HPLC grade methanol, acetonitrile, diethyl ether was acquired from Actu-All chemical (Oss, The Netherlands). Deionised water (< 8 M cm resistivity) was obtained from Milli-Q PureLab Ultra (Veolia Water Technologies, Ede, The Netherlands).