Colipase

The peptide AVFQHNCQE was synthesized by CASLO Aps. (Kongens Lyngby, Denmark) and its purity was 98.94%. Captopril was provided by Santa Cruz Biotechnology (Dallas, TX, United States). Pepsin, bile salts mixture, trypsin, α-chymotrypsin, porcine pancreatic lipase, colipase, trifluoroacetic acid (TFA), formic acid (FA) LC-MS grade, and bovine serum albumin (BSA) were obtained from Sigma-Aldrich (Madrid, Spain). Acetonitrile and methanol LC-MS grade were purchased from Merck (Darmstadt, Germany). The synthetic isotopically labeled peptide IFV*TGQDYNDK (V* = Val-13C5,15N) was obtained from Biomatik (Ontario, Canada) and used as internal standard (IS) for MS experiments. Chicken foot hydrolysate Hpp11 was obtained by our group following the procedure described by Bravo et al. [22 ,24 (link)]. All other chemical solvents used were of analytical grade.

Sepharose 2B, methyl orange (MO), dextran blue (DB), sodium chloride, sodium azide, Tris, methanol, acetone, colipase from porcine pancreas ≥ 95% protein (Prod No: C3028), lipase from porcine pancreas type II 100–500 units/mg protein using olive oil as a substrate (Product No: L3126) and orlistat (tetrahydrolipstatin) were purchased from Sigma–Aldrich (Poole, UK). Aluminium oxide was purchased from Fisher Scientific (Loughborough, UK), and olive oil was purchased from Co-operative Foods (Manchester, UK). Deoxycholic acid sodium salt and taurodeoxycholic acid sodium salt were purchased from Fluka (Buchs, Switzerland). Alginates LFR 5/60 which is a low viscosity and low molecular weight (40,000) sodium alginate rich in guluronate F_g 0.64, SF200 which is a high viscosity and high molecular weight (380,000) sodium alginate rich in guluronate F_g 0.69 and Manucol^® DM which is a high viscosity sodium alginate with a molecular weight ranging from 250,000 to 320,000 were a gift from FMC BioPolymer AS, Drammen, Norway and were stored at 4 °C in tightly-sealed containers and all alginate weights were corrected for water content. The control bread (CB) and alginate bread (AB) were produced by Greggs Plc and ingredients are presented in Table 1. The AB was 4% alginate MANUCOL^® DM (w/w) wet dough as this made the most palatable bread.

All alginate samples were kindly provided by Technostics Limited (Hull, UK) (Table 1). The bile acids (deoxycholate sodium salt and taurodeoxycholate sodium salt) were both purchased from Fluka (Buchs, Switzerland). The lipase, colipase and orlistat (tetrahydrolipstatin), tris(hydroxymethyl)-methylamine, 1,2 Di-o-lauryl-rac-glycero-3-(glutaric acid 6-methyl resorufin ester) (DGGR), sodium acetate, calcium chloride and acetone were all purchased from Sigma–Aldrich (Poole, UK). The olive oil was purchased from a local supermarket (Cooperative Foods, UK) and the aluminium oxide was purchased from Fisher Scientific (Loughborough, UK).