Mgcl2 h2o 6

All 36 investigated drugs and the two standard solutions (thiorphan and S-acetylthiorphan) were commercially purchased from Sigma-Aldrich (St. Louis, MO, United States), and the detailed information about these compounds is listed in Supplementary Table S1. Acetonitrile, methanol, dimethyl sulfoxide, formic acid, and ammonium acetate were HPLC grade, and along with KCl, KH₂PO₄, NaHCO₃, NaCl, MgCl₂(H2O)₆, (NH₄)₂CO₃, CaCl₂(H₂O)₂, L-cysteine, ethyl acetate, HCl, and NaOH were bought from Merck (Darmstadt, Germany). Bryant and Burkey Medium (BB), de Man Rogosa Sharpe (MRS) broth, glycerol, human salivary α-amylase, porcine pepsin, rabbit gastric extract for gastric lipase, bovine bile, and porcine pancreatin were purchased from Sigma-Aldrich (St. Louis, MO, United States). Modified Gifu anaerobic medium (mGAM) broth was purchased from HyServe GmbH and Co., KG (Germany). Ultrapure water used throughout the study was prepared by a Milli-Q water purification system (Millipore, Molsheim, France).

Ethanol, acetone, 2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), Folin–Ciocalteu reagent, KCl, NaHCO₃, KH₂PO₄, (NH4)₂CO₃, NaCl, HCl, MgCl₂·(H₂O)₆ and thin-layer chromatography (TLC) plates were purchased from Merck (Darmstadt, Germany). α-amylase, pepsin from porcine gastric mucosa, pancreatin from porcine pancreas and bile salts were purchased from Alfa Aesar (Karslruhe, Germany). All reagents used in the experimental part were of analytical purity.

1,5-Carboxy-seminaphtorhodafluor
acetoxymethylester (SNARF-1-AM) and Dulbecco’s modified Eagle’s
medium GlutaMAX (DMEM) were purchased from Thermo Fisher Scientific.
Fetal bovine serum (FBS Supreme), trypsin/ethylenediaminetetraacetic
acid, and penicillin–streptomycin were obtained from PAN-Biotech
GmbH (Aidenbach, Germany). Phosphate buffered saline was bought from
Biozym Scientific GmbH (Hessisch Oldendorf, Germany). Dimethyl sulfoxide
(DMSO) was purchased from Carl Roth (Karlsruhe, Germany). CaCl₂, casein from bovine milk, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium
bromide (MTT), d-glucose, formic acid, HCl, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic
acid (HEPES), KCl, KH₂PO₄, KOH, MeCN, MgCl₂(H₂O)₆, MgSO₄, NaCl, NaHCO₃, (NH₄)₂CO₃, and pepsin from
porcine gastric mucosa were ordered from Merck KGaA (Darmstadt, Germany).
Custom peptides (PVVVPPFLQPEVM, VAPFPEVF, YFYPEL, YQEPVLGPVRGPFPIIV,
and YYVPLGTQ) were synthesized by Genscript Biotech with a purity
of >95% (New Jersey, USA). Double-distilled water (ddH₂O) from Elga Purelab Classic (Veolia Water Solutions & Technologies,
France) was used for all experiments. Krebs-Ringer–HEPES buffer
(KRHB) contains 130 mM NaCl, 4.7 mM KCl, 1.3 mM CaCl₂,
1.2 mM MgSO₄, 1.2 mM KH₂PO_4, 11.7
mM d-glucose, and 10 mM HEPES; the pH was adjusted to 7.4
with KOH.

Ethanol 96%, 2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), 2,4,6-tripyridyl-s-triazine (TPTZ), FeCl₃·6H₂O,Folin–Ciocalteu reagent, ascorbic acid, CH₃COONa, 36.5% HCl, glacial acetic acid, H₂SO₄ (0.1 N), KMnO₄, KCl, NaHCO₃, KH₂PO₄, (NH₄)₂CO₃, NaCl, MgCl₂⋦(H₂O)₆ were purchased from Merck. α-Amylase, pepsin from porcine gastric mucosa, pancreatin from porcine pancreas, bile salts were purchased from Alfa Aesar (Karslruhe, Germany). All reagents used in the experimental part were of analytical purity. Sweeteners used in this study were purchased from a health food store and local market.

All materials were standard analytical grade. α-amylase was obtained from Bacillus licheniformis (E-BLAAM EC.3.2.1.1; Megazyme International Ireland Ltd., Wicklow, Ireland) and was supplied at a concentration of 10000 U/ml on soluble starch at pH 6.5 and 40°C. Glucoamylase was obtained from Aspergillus niger (AMG Conc. BG; Novozymes, Bagsværd, Denmark) and had an activity of 1300 AGU/g. Pepsin (P6887; Sigma-Aldrich S.r.l. Milan, Italy) was obtained from porcine gastric mucosa and had an activity between 3200 and 4500 U/mg protein. Creon® 10000 U.Ph.Eur. (Abbott Products S.p.A; Sesto San Giovanni (Mi), Italy), which is normally used in the treatment of pancreatic exocrine insufficiency, was available as brown/clear capsules. Each capsule contained lipase (10000 Ph.Eur. units), amylase (8000 Ph.Eur. units), and protease (600 Ph.Eur. units). CaCl₂, KCl, KH₂PO₄, NaHCO₃, NaCl, MgCl₂(H₂O)₆, (NH₄)₂CO₃ and HCl were purchased from Sigma-Aldrich (Sigma-Aldrich, Sigma-Aldrich S.r.l. Milan, Italy).

Acetic acid (≥99.5%), 2Prop-GLS (≥99%), l-ascorbic acid (≥99%), benzonitrile (≥99.9%), d/l-dithiothreitol, FeSO₄(H₂O)₇ (≥99%), myrosinase (thioglucosidase from Sinapis alba seeds; ≥100 units/g) and NaHCO₃ (≥99.5%) were purchased from Sigma-Aldrich Chemie GmbH, (Steinheim, Germany). 4-Hydroxybenzyl GLS (≥99%) was purchased from Carl Roth GmbH, Karlsruhe, Germany and NaCH₃COO(H₂O)₃ (≥99.5%) was purchased from Merck (Darmstadt, Germany), respectively. Simulated salivary fluid (saliva buffer) was prepared according to60 (link) (pH 7) using chemicals from Merck (Darmstadt, Germany) (KCl, KH₂PO₄, MgCl₂(H₂O)₆, CaCl₂(H₂O)₂) and Sigma-Aldrich Chemie GmbH, (Steinheim, Germany) (α-amylase. NaHCO₃, HCl (37%), (NH₄)₂CO₃ (≥30%, NH₃ basis)) with a purity of ≥99% if not stated otherwise. All solvents were of LC-MS grade and water was of Milli-Q quality.

The simulated digestive fluids to conduct in vitro digestions were prepared with KCl, KH₂PO₄, NaCl, NaHCO₃, MgCl₂ (H₂O)₆, (NH₄)₂CO₃, CaCl₂, human α-amylase (1000–3000 U/mg protein), pepsin from porcine gastric mucosa ( ≥ 2500 U/mg protein) and bovine bile extract, all of which were obtained from Sigma-Aldrich Chemical Company (St Louis, MO, USA). The pancreatic enzyme supplements came from Kreon 10,000 LU (Mylan, Canonsburg, PA, USA). For the analytical determinations, Triton-X 100%, trichloroacetic acid (TCA), hexane, Folin-Ciocalteu reagent, Na₂CO₃, gallic acid (GA), trolox (TX) and the analytical standards were acquired from Sigma-Aldrich Chemical Company (St Louis, MO, USA). Ethanol (95% v/v for analysis), NaOH and HCl, were from AppliChem Panreac (Barcelona, Spain).