Milli q gradient system

Most native and deuterated standards for analysis of oxylipins, endocannabinoids, and endocannabinoid-related lipids, including 12-[[(cyclohexylamino)carbonyl]amino]-dodecanoic acid (CUDA) used for recovery calculation purposes, were obtained from Cayman Chemical (Ann Arbor, MI, USA). Only 9,10,13-trihydroxyoctadecenoic acid (TriHOME) and 9,12,13-TriHOME were obtained from Larodan (Malmö, Sweden). Acetonitrile (ACN) and methanol (MeOH) were from Merck (Darmstadt, Germany). Isopropanol was from VWR PROLABO (Fontenay-sous-Bois, France). Acetic acid was purchased from Aldrich Chemical Company, Inc. (Milwaukee, WI, USA). Butylhydroxytoluene (BHT) was from Cayman Chemical (Ann Arbor, MI, USA) and ethylenediaminetetraacetic acid (EDTA) from Fluka Analytical, Sigma-Aldrich (Buchs, Switzerland). Glycerol was from Fischer Scientific (Loughborough, UK). All solvents and chemicals were of HPLC grade or higher and ultrapure water was used (Milli-Q Gradient system, Millipore, Milford, MA, USA).

All reagents and solvents used in these studies were commercial and high purity of analytical grade or ultra-gradient LC-MS-grade purchased from MilliporeSigma (St. Louis, MO, USA), J.T. Baker (Deventer, The Netherlands), Merck (Darmstadt, Germany), Riedel-de Haën (Seelze, Germany) or Thermo Fisher Scientific (Waltham, MA, USA), unless otherwise stated. Water was purified using a Milli-Q Gradient system (Millipore, Milford, MA, USA). The following reagents were used for basic coagulology screening, Bio-Ksel System APTTs reagent and calcium chloride (Bio-Ksel, Grudziądz, Poland), Bio-Ksel PT plus reagent (thromboplastin and solvent, Bio-Ksel, Grudziądz, Poland), and thrombin (3.0 UNIH/mL, Bio-Ksel, Grudziądz, Poland). Coefficients of variability were calculated using Bio-Ksel normal plasma (Bio-Ksel, Poland), and water for injection (Polpharma, Gdańsk, Poland). Triton X-100 used in the erythrotoxicity test was obtained from Polish Chemical Reagents (Gliwice, Poland).

Methanol for HPLC and formic acid were purchased from RCI Labscan Limited (Bangkok, Thailand) and Sigma-Aldrich (St. Louis, MO, USA), respectively. Ultrapure water was obtained using a Milli-Q Gradient system, coupled with an Elix Advantage 10, both from Millipore (Merck Millipore, Darmstadt, Germany).

Norcantharidin (NCTD) was obtained from Alfa Aesar (> 98%, Ward Hill, MA). Soybean oil, olive oil, ethyl oleate, isopropyl myristate, triacetin, tributyrin, Tween 80, Tween 20, ethanol, ethylene glycol, 1-propanol, isopropanol, propylene glycol, 1-butanol, glycerol and PEG 400 were purchased from Aladdin (> 98%, Shanghai, China). Medium chain triglyceride (MCT), Cremophor EL and Cremophor RH 40 were obtained from BASF (Ludwigshafen, Germany). All other chemicals and solvent were of analytical grade and used without further purification.
Deionized water was generated from the Milli-Q gradient system of Millipore (Synergy, Millipore SAS, Molsheim, France).

Methanol (MeOH), LC–MS grade, was obtained from Biosolve (Valkenswaard, Netherlands), and HPLC-grade MeOH was obtained from VWR International (Zaventem, Belgium). Ethyl acetate (EtOAc), dichloromethane (DCM) and acetone [dimethyl ketone (DMK)] were purchased from Acros Organics (Geel, Belgium). Sigma-Aldrich (Bornem, Belgium) supplied ammonium formate (HCOONH₄). Formic acid (HCOOH, Merck, Darmstadt, Germany) was used. Ultrapure H₂O, used during these analyses was produced in-house with a Milli-Q Gradient System (Millipore, Brussels, Belgium). Ultrafree^®-MC centrifugal filter units (0.22 μm) from Millipore (Bedford, MA, United States) were used for filtration and reconstitution of fungal extracts. Sigma-Aldrich was our source for fungal growth media, such as agar, corn steep solids, dextrose, peptone, sucrose, yeast extract, dipotassium hydrogen phosphate trihydrate (K₂HPO₄⋅3H₂O), magnesium sulfate heptahydrate (MgSO₄⋅7H₂O), and iron(II) sulfate heptahydrate (FeSO₄⋅7H₂O). Other compounds necessary for this study included Triton X-100, potassium chloride (KCl), and sodium nitrate (NaNO₃), which were obtained from Merck. Standards of AFB₁, AFB₂, AFG₁, AFG₂, AFM₁, and ST (sterigmatocystin) were purchased from Oskar Tropitzsch (Marktredwitz, Germany).

The material used as the dispersant phase was washed sea sand, 200–300 μm Scharlau (Chemie S.A., Barcelona, Spain), the extraction solvent was LC-MS grade methanol supplied by Scharlau (Chemie S.A., Barcelona, Spain), the ultrapure water was produced in the laboratory with a Milli-Q gradient system (Millipore, Bedford, MA, USA) and the formic acid was supplied by Merck (Darmstadt, Germany). Folin and Ciocalteau phenol reagent obtained from Sigma–Aldrich and sodium carbonate (Na₂CO₃, Panreac, Barcelona, Spain) were used for the determination of the total polyphenolic index. The 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox^®) were purchased from Sigma–Aldrich (Chemie Check GmbH, Steinheim, Germany) and used for the determination of the scavenging activity of the apple peel extracts.
Individual standard stock solutions (2000–8000 µg/mL) were prepared in methanol from pure polyphenols, procyanidin A2, B1, B2, catechin, epicatechin, chlorogenic acid, gallic acid monohydrate (99%), epicatechin gallate, quercetin, quercetin-3-glucoside and quercetin-3-rutinoside, which were all supplied by Sigma–Aldrich (Chemie Check GmbH, Steinheim, Germany). The prepared solutions were stored at −20 °C and protected from light.

N-acetylcarnosine was commercially purchased from Toronto Research Chemicals (Toronto, Ontario Canada) with >98% purity. The molecular weight of the NAC was 268.27 g mol⁻¹. PLGA (lactide:glicolide = 50:50, M_w ∼ 38–54 kDa) and poly(vinyl alcohol) (PVA) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Dichloromethane (DCM) was also purchased from Merck Millipore (>99.5%) (Darmstadt, Germany). All the chemicals and solvents were of analytical grade. Ultrapure water was obtained using the Millipore MilliQ Gradient System. Molecular dynamics (MD) simulations were performed using GROMACS code (Van Der Spoel et al., 2005 (link)) for 5 ns.