Amberlite® XAD7HP (DuPont, Wilmington, DE, USA), chromatographic grade chemicals of hexane, water, acetic acid, sodium acetate trihydrate, methanol, ethanol, acetonitrile and analytical grade of chemicals of 2,2′-diphenyl-1-picrylhydrazyl radical (DPPH), Trolox®, 2,4,6-tripirydyl-Striazine (TPTZ) and ferric chloride were purchased from Fisher Scientific (Waltham, MA, USA). In addition, 96–98% (g/g) concentrated sulfuric acid, Folin–Ciocalteu reagents, sodium carbonate, phenolic compound standards of gallic acid, hydroxytyrosol, tyrosol, 4-hydroxyphenylacetic acid (4-HPA), vanillic acid, vanillin, o-coumaric acid, oleuropein, pinoresinol, cinnamic acid, caffeic acid, p-coumaric acid, ferulic acid, apigenin-7-glucoside, apigenin, luteolin-7-glucoside and luteolin were purchased from Sigma-Aldrich (St. Louis, MI, USA). Verbascoside was bought from the HWI group (Ruelzhelm, Germany). Rutin was bought from PhytoLab GmbH & Co. KG (Vestenbergsgreuth, Germany).
Phenolic compound standards
Manufactured by Merck Group
Sourced in United States, Italy, Germany
Phenolic compound standards are reference materials used to identify and quantify phenolic compounds in analytical procedures. They are designed to provide accurate and reproducible results for the evaluation of phenolic content in various samples.
Automatically generated - may contain errors
Lab products found in correlation
Folin ciocalteu reagent, by Merck Group (2 mentions)
Gallic acid, by Merck Group (2 mentions)
2 2 diphenyl 1 picrylhydrazyl radical dpph, by Thermo Fisher Scientific (1 mentions)
Sulfuric acid, by Merck Group (1 mentions)
Caffeic acid, by Merck Group (1 mentions)
Ferulic acid, by Merck Group (1 mentions)
Vanillic acid, by Merck Group (1 mentions)
P coumaric acid, by Merck Group (1 mentions)
Cinnamic acid, by Merck Group (1 mentions)
O coumaric acid, by Merck Group (1 mentions)
Ethanol, by Thermo Fisher Scientific (1 mentions)
Methanol, by Thermo Fisher Scientific (1 mentions)
Acetonitrile, by Thermo Fisher Scientific (1 mentions)
Sodium carbonate, by Merck Group (1 mentions)
Acetic acid, by Thermo Fisher Scientific (1 mentions)
Apigenin, by Merck Group (1 mentions)
Luteolin, by Merck Group (1 mentions)
Ferric chloride, by Thermo Fisher Scientific (1 mentions)
Vanillin, by Merck Group (1 mentions)
Sodium acetate trihydrate, by Thermo Fisher Scientific (1 mentions)
9 protocols using phenolic compound standards
1
Extraction and Analysis of Phenolic Compounds
2
Quantitative Analysis of Phenolic Compounds
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Phenolic compound standards were purchased from Sigma-Aldrich (St. Louis, MO, USA). Twenty milligrams of dried sample was extracted with 3 mL of 80% methanol (v/v). The crude mixture was ultrasonicated for 1 h at room temperature and then centrifuged at 12,000 rpm for 10 min. The final extract was filtered using a 0.45 μm Acrodisc syringe filter (Pall Corp.; Port Washington, NY, USA). The flow rate was maintained at 1 mL/min and a 280-nm wavelength was used for detection. Elution was performed using a binary gradient of mobile phase A (0.15% of acetic acid in H2O, v/v) and mobile phase B (methanol). The volume of the sample injected was 20 μL and column temperature was maintained at 30 °C for detection. The individual compounds were separated and identified in the Futecs model NS-4000 HPLC apparatus (Daejeon, Korea).
3
Procuring Chemical Standards for Analysis
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Phenolic compound standards were purchased from Sigma Aldrich (Milan, Italy). All the other chemicals and solvents were purchased from Fluka (Saint-Quentin Fallavier, France) at analytical or HPLC grade and were used as received.
4
Phytochemical Screening and Bioactivity Assays
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Acetonitrile 99.9% was of high-performance liquid chromatography (HPLC) grade, supplied by Lab-Scan (Lisbon, Portugal), and methanol was of analytical grade from Pronalab (Lisbon, Portugal). 2,2-Diphenyl-1-picrylhydrazyl (DPPH) was obtained from Alfa Aesar (Ward Hill, MA, USA). Dulbecco’s modified Eagle’s minimum essential medium (DMEM), fetal bovine serum (FBS), penicillin, streptomycin, Griess reagent system (Promega), dimethyl sulfoxide (DMSO), sulforhodamine B (SRB), lipopolysaccharide (LPS), 3,4-dihydroxy-L-phenylalanine (L-DOPA), dexamethasone, and mushroom tyrosinase enzyme were obtained from Sigma-Aldrich Co. (Saint Louis, MO, USA). Muller Hinton broth (MHB) and Tryptic Soy Broth (TSB) medium were purchased from Biomerieux (Marcy l’Etoile, France). Blood agar with 7% sheep blood and Mac Conkey agar plates were purchased from Biomerieux (Marcy l’Etoile, France). The p-iodonitrotetrazolium chloride (INT) dye and phenolic compound standards were purchased from Sigma-Aldrich (St. Louis, MO, USA).
5
Phytochemical Analysis of Non-THC Cannabis
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The plant material consists in female inflorescences of Cannabis sativa L cultivar “Futura 75”, a certified non-THC producing cultivar approved for industrial production, cultivated in Abruzzo Region (Italy), under controlled conditions, avoiding chemical additives. Female inflorescences were manually harvested from plants at full blooming state and then immediately dried in a ventilated oven (40 °C). Once they reached the constant weight, samples were clumsily chopped and stored in airtight plastic bags, in a dark and dry place at room temperature (22–24°C), before performing phytochemical and biological assays. Plant identity was confirmed botanically and morphologically by the co-author Prof. Luigi Menghini. Samples were kindly supplied by Hemp Farm Italia (Tortoreto (TE), Italy) during the cultivation season 2018. The product is sold by the company itself as industrial hemp, and is, therefore, certified for THC content results <0.2% w/w, according to the European Regulation EC no. 1124/2008—12 November 2008. TerpenoPhenolic compound standards were purchased from Focus Analytics Srl (Arcore (MB), Italy) and Comedical Srl (Trento, Italy). Phenolic compound standards were purchased from Sigma-Aldrich (Milano, Italy).
6
Folin-Ciocalteu Assay Reagents
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Sodium carbonate (≥99%), Folin's phenol reagent, and gallic acid (GA, ≥98%) were used in the Folin-Ciocalteu assay and were obtained from Sigma-Aldrich (Madrid, Spain). DPPH radical, TPTZ (2,4,6-Tris(2pyridyl)-s-triazine, 99%), aluminium chloride hexahydrate (99%) and 6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid (Trolox, 98%) were also from Sigma. Absolute anhydrous ethanol (p.a.), sodium nitrite (≥97%) and ascorbic acid (AA, 99.7%) were acquired from Carlo Erba (Peypin, France), Merck and Riedel-de Haën, respectively. Methanol and formic acid for HPLC analysis were gradient grade and got from Merck (Darmstadt, Germany). Phenolic compound standards were bought to Sigma-Aldrich (Madrid, Spain). Other chemicals were from Sigma (Madrid, Spain) and analytical grade.
7
Quantitative Analysis of Phytochemicals
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Phenolic compound standards were supplied by Sigma-Aldrich (Steinheim, Germany); Fluka (Buchs, Switzerland) and Extrasynthèse (Lyon, France). Polysaccharide standards were provided by Sigma-Aldrich (Steinheim, Germany).
Volatile compound standards were purchased from Fluka (Buchs, Switzerland), Sigma-Aldrich (Steinheim, Germany) and Lancaster (Strasbourg, France).
Ethanol (96%) was from Labkem (Barcelona, Spain). Acetonitrile and methanol for HPLC analyses were supplied by Carlo Erba (Sabadell, Spain), the remaining reagents-by Panreac (Madrid, Spain). Milli-Q water was obtained through a Millipore system (Bedford, MA, USA).
Volatile compound standards were purchased from Fluka (Buchs, Switzerland), Sigma-Aldrich (Steinheim, Germany) and Lancaster (Strasbourg, France).
Ethanol (96%) was from Labkem (Barcelona, Spain). Acetonitrile and methanol for HPLC analyses were supplied by Carlo Erba (Sabadell, Spain), the remaining reagents-by Panreac (Madrid, Spain). Milli-Q water was obtained through a Millipore system (Bedford, MA, USA).
8
Analytical Methods for Phenolic Compounds
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Acetonitrile, methanol and formic acid were High Performance Liquid Chromatography grade (HPLC) and purchased from Merck (Darmstadt, Germany). Ultra-pure water (MicroPure water purification system, 0.055 μS/cm; Thermofisher TKA, Germany) was used for solid phase extraction (SPE) and liquid chromatography/mass spectrometry (UPLC-MS/MS) analyses. The Oasis HLB bcc/200 µm SPE cartridges (Waters, Milford, MA, USA), were used for SPE of samples. Phenolic compound standards were purchased from Sigma-Aldrich (Steinheim, Germany) and Fluka (Buch, Switzerland). Trolox (6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid), 2,2-azino-bis(3ethylbenzothiazoline-6-sulfonic acid) (ABTS), 2,2diphenyl-1-picrylhydrazyl (DPPH), and potassium persulfate were purchased from Sigma-Aldrich (Steinheim, Germany). Folin-Ciocalteu (FC) reagent was obtained from Merck (Darmstadt, Germany). TPTZ (2,4,6-trypyridyl-s-triazine) was purchased from Fluka Analytical (Switzerland).
9
Antioxidant Capacity Determination Protocol
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Phenolic compound standards, 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (trolox), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), 2,4,6-tri(2-pyridyl)-S-triazine (TPTZ), Folin-Ciocalteau phenol reagent were purchased from Sigma (Milan, Italy). Methanol and formic acid were obtained from Carlo Erba (Milan, Italy). All MS/MS reagents were from Bio-Rad (Hercules, CA, U.S.A.). Chemicals and enzymes for the digestion procedure were purchased from Sigma-Aldrich (Milan, Italy). All the materials and chemicals for cell culture were from Euroclone (Milan, Italy). MTS cell proliferation assay kit was purchased from Promega (Milan, Italy). Solid phase extraction (SPE) column (C18, 50 μm, 60 Å, 500 mg) were supplied by Waters (Milan, Italy).
The absorbance was read using a Jasco V-550 UV/Vis spectrophotometer.
The absorbance was read using a Jasco V-550 UV/Vis spectrophotometer.
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