Deuterated methanol

Deuterated methanol, potassium dihydrogen phosphate, and 3-(trimethylsilyl)propionic acid sodium salt (TSP) were obtained from Merck Life Science (Pty) Ltd. (Modderfontein, South Africa). Pure standards of succinate (≥98% purity), lactose (≥98% purity), propylene glycol (99% purity), gallic acid (≥99% purity), and choline (≥99% purity) were also obtained from Merck Life Science (Pty) Ltd. (Modderfontein, South Africa).

UHPLC-MS Solvents, LC-MS formic acid, reagent grade lanzoprasole, formalin, ethanol, HCl, deuterated chloroform and deuterated methanol, and reagent grade chloroform were from Merck (Santiago, Chile). Ultrapure water was obtained from a Millipore water purification system (Milli-Q Merck Millipore, Santiago, Chile). UHPLC standards, (kaempferol, caffeic acid, isorhamnetin, hesperetin, eriodictyol, all standards with purity higher than 95% by HPLC) were purchased either from Sigma Aldrich (Saint Louis, Mo, USA), ChromaDex (Santa Ana, CA, USA), or Extrasynthèse (Genay, France).

Ethyl acetate (≥99.9%), cyclohexane (≥99.9%), methanol (≥99.9%), acetic acid (≥99.9%), dichloromethane (≥99.9%), formic acid (≥99.9%), dextrose (≥99.9%), agar (≥99.9%), silica gel, tetramethylsilane (TMS), deuterated methanol, deuterated pyridine and deuterated dimethyl sulfoxide were purchased from Sigma-Aldrich Co. LLC, Steinheim am Albuch, Germany. All solvents used were LC analytical grade. The mixture of alkanes standard from C10 to C40 was purchased from Merck KGaA, Darmstadt, Germany.

Deuterated methanol and water were purchased from Sigma-Aldrich (St. Louis, MO, USA). ¹H NMR spectra were recorded at 25 °C on a 600 MHz Bruker DMX-600 spectrometer (Bruker, Karlsruhe, Germany) operating at a proton NMR frequency of 600.13 MHz for ¹³C. CD₃OD was used as the internal control.
The ¹H NMR spectra were automatically reduced to ASCII files. Spectral intensities were scaled to total or internal standards (TMSP or TMS signals at δ 0.0) and reduced to integrated regions of equal width (δ 0.04) corresponding to the region of δ 0.0–10.0. The regions of δ 4.7–4.9 and δ 3.28–3.34 were excluded from the analysis because of the residual signals of D₂O and CD₃OD, respectively. Bucketing was performed using AMIX software (ver. 3.0 Bruker) with scaling on total intensity. Projections to latent structures (PLS) and orthogonal PLS (OPLS) with scaling based on unit variance were performed with SIMCA-P + software (v. 13.3, Umetrics, Umeå, Sweden).

p-Coumaric
acid (98%), sinapic acid (98%), ferulic acid (99%), N,N′-dicyclohexylcarbodiimide (DCC), sodium
bicarbonate (NaHCO₃), sodium hydroxide, 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic
acid) (ABTS), nitric acid, horseradish peroxidase type VI-A (HRP,
1.11.1.7, P6782), deuterated methanol, and 30% (w/w) hydrogen peroxide were purchased from Sigma-Aldrich (St. Louis,
MO, USA). Agmatine sulfate (97%) was purchased from Fisher Scientific
B.V. (Hampton, New Hampshire, USA), and 2,5-dihydroxybenzoic acid
was obtained from Bruker Daltonics (Bremen, Germany). Ammonium acetate,
potassium dihydrogenphosphate, and dipotassium hydrogen phosphate
were purchased from Merck KGaA (Darmstadt, Germany). Glacial acetic
acid was purchased from VWR International BV (Radnor, PA, USA). Ethyl
acetate, acetone, methanol (MeOH), hexane, HPLC grade acetonitrile
(ACN), UHPLC–MS grade ACN and water, and formic acid (FA) (99%
[v/v]) were purchased from Biosolve
(Valkenswaard, Netherlands). Water for purposes other than UHPLC was
purified using a Milli-Q purification system equipped with a 0.22
μm filter (Millipore, Molsheim, France).

Propylene carbonate, dioxane, sulfuric acid, hydrochloric acid, sodium hydroxide, salicylic aldehyde, diethylmalonate, piperidine, N, N-diethyl salicylic aldehyde, ethanol, glucose, fructose, sodium citrate, sodium acetate, ammonium chloride, ammonia, potassium dichromate, deuterated methanol were purchased from Sigma Aldrich (Poznań, Polska). Albumin was obtained from egg white. Lignin was derived from dried miscant plant via extraction. XTT (2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide) assay, DMEM without phenol red with high glucose, penicillin/streptomycin, trypsin, phosphate buffer, L929 mouse fibroblasts were purchased from Sigma Aldrich (Poznań, Polska). Sterile filters 0.22 µm were purchased from Bionovo (Legnica Poland) and regenerated cellulose membranes for dialysis (MWCO = 1000 Da) were obtained from VWR International (Gdańsk, Poland). Multiwhole plates (96 wholes) were purchased from Genoplast, Rokocin, Poland.

High pure CP was a gift sample from JW Pharmaceuticals, Seoul, Republic of Korea. HPLC grade methanol, chloroform, water and analytical reagent grade formic acid, ethyl acetate, triethylamine, phosphate buffer, deuterium oxide (D₂O, 100% atom ‘D’), deuterated methanol and _D-formic acid (98% atom ‘D’) used in the present study were purchased from Sigma Aldrich, Seoul, Republic of Korea and used without further purification.