Reagents for antioxidant activities and cholinesterase inhibition assays were from Sigma-Aldrich (St. Louis, MO, USA and Steinheim, Germany), except magnesium chloride hexahydrate, which was bought from VWR (Leuven, Belgium), sodium chloride from Fisher Scientific (Fair Lawn, NJ, USA), sodium nitroprusside dihydrate from Fluka, and sodium carbonate from Merck. Hydrogen peroxide 35.5% w/w was purchased from Labchem (Zelienople, PA, USA). Methanol Chromasolv for HPLC was from Riedel-de Haën (Seelze, Germany), and formic acid was from Carlo Erba (Val de Reuil, France). The standards used for HPLC analyses were acquired from Sigma-Aldrich (4-O-caffeoylquinic acid, quercetin-3-O-rutinoside hydrate, protocatechuic acid, ferulic acid, and p-coumaric acid), Fluka (caffeic acid), Alfa Aesar (5-O-caffeoylquinic acid), and Extrasynthèse (Genay, France): 3-O-caffeoylquinic acid, apigenin-8-C-glucoside, apigenin-6-C-glucoside, quercetin-3-O-glucoside, kaempferol-3-O-glucoside, kaempferol-3-O-rutinoside, and tiliroside. Ultrapure water (with resistivities of 18.2 MΩ/cm) was obtained using a Milli-Q water purification system from Millipore (Molsheim, France). Standard solutions of Pb and Cd, Atomic Absorption standard with 4% HNO3, were purchased from SPC Science (Quebec, Canada). Suprapur nitric acid 65% (v/v) and NH4H2PO4 (Suprapur) were from Merck (Darmstad, Germany).
Apigenin 6 c glucoside
Manufactured by Extrasynthese
Sourced in France
Apigenin-6-C-glucoside is a flavone glycoside compound. It is a naturally occurring substance that can be isolated and purified for use in various applications.
Automatically generated - may contain errors
Lab products found in correlation
Quercetin 3 o rutinoside, by Extrasynthese (6 mentions)
Quercetin 3 o glucoside, by Extrasynthese (5 mentions)
Caffeic acid, by Extrasynthese (5 mentions)
Luteolin 7 o glucoside, by Extrasynthese (5 mentions)
Rosmarinic acid, by Extrasynthese (5 mentions)
Naringenin, by Extrasynthese (4 mentions)
Chlorogenic acid, by Extrasynthese (4 mentions)
Kaempferol 3 o glucoside, by Extrasynthese (3 mentions)
Formic acid, by Merck Group (3 mentions)
Hesperetin, by Extrasynthese (3 mentions)
Acetonitrile 99.9, by Thermo Fisher Scientific (3 mentions)
Apigenin 8 c glucoside, by Extrasynthese (2 mentions)
Kaempferol 3 o rutinoside, by Extrasynthese (2 mentions)
Acetonitrile, by Thermo Fisher Scientific (2 mentions)
Gallic acid, by Extrasynthese (2 mentions)
Luteolin 6 c glucoside, by Extrasynthese (2 mentions)
Apigenin 7 o glucoside, by Extrasynthese (2 mentions)
6 hydroxy 2 5 7 8 tetramethylchroman 2 carboxylic acid trolox, by Merck Group (2 mentions)
2 2 diphenyl 1 picrylhydrazyl (dpph), by Thermo Fisher Scientific (2 mentions)
Milli q water purification system, by Merck Group (2 mentions)
10 protocols using apigenin 6 c glucoside
1
Antioxidant and Cholinesterase Inhibition Assays
2
Phytochemical and Enzymatic Analysis
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Acetonitrile (99.9%) was of HPLC grade from Fisher Scientific (Lisbon, Portugal). Phenolic compound standards (apigenin-6-C-glucoside, quercetin-3-O-glucoside, quercetin-3-O-rutinoside, peonidin-3-O-glucoside) were from Extrasynthèse (Genay, France). Formic acid, DPPH· (2,2-diphenyl-1-picrylhydrazyl), TPTZ (2,4,6-tris(2-pyridyl)-s-triazine), ATCI (acetylthiocholine iodide), acetylcholinesterase (AChE), monoamine oxidase A (MAO A), clorgiline, tris–HCl and pyrogallol were purchased from Sigma–Aldrich (St. Louis, MO, USA). DNTB (5,5′-dithiobis (2-nitrobenzoic acid)) and juglone (5-hydroxy-1,4-naphthoquinone) were from Alfa Aesar (Ward Hill, MA, USA), Folin-Ciocalteu reagent was purchased from Chem-lab (Zeldelgem, Belgium). Water was treated in a Milli-Q water purification system (TGI Pure Water Systems, Greenville, SC, USA).
3
Phenolic Profile and Antioxidant Capacity
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Total Phenols (TP) was measured using the Folin-Ciocalteu method as described by Martín-Diana et al. (2017) . Results were expressed as μmol gallic acid equivalents (GAE) g−1 of sample using a calibration curve with gallic acid as standard (9.8–70 mM).
Phenolic compound profile was determined using HPLC-DAD-ESI/MS chromatography, as described in Dueñas et al. (2015) . The standards, gallic acid, quercetin 3-O-glucoside, kaempferol 3-O-glucoside, isorhamnetin 3-O-glucoside, myricetin 3-O-glucoside, apigenin 6-C-glucoside, were obtained from Extra synthese (Genay, France). Galloyl glucose derivatives were quantified using gallic acid standard curve; derivatives of apigenin, quercetin, kaempferol, isorhamnetin and myricetin were quantified using apigenin 6-C-glucoside, quercetin 3-O-glucoside, kaempferol 3-O-glucoside, isorhamnetin 3-O-glucoside and myricetin 3-O-glucoside standard curves, respectively. Results were expressed as μg g−1 sample (d.w.b.).
Phenolic compound profile was determined using HPLC-DAD-ESI/MS chromatography, as described in Dueñas et al. (2015) . The standards, gallic acid, quercetin 3-O-glucoside, kaempferol 3-O-glucoside, isorhamnetin 3-O-glucoside, myricetin 3-O-glucoside, apigenin 6-C-glucoside, were obtained from Extra synthese (Genay, France). Galloyl glucose derivatives were quantified using gallic acid standard curve; derivatives of apigenin, quercetin, kaempferol, isorhamnetin and myricetin were quantified using apigenin 6-C-glucoside, quercetin 3-O-glucoside, kaempferol 3-O-glucoside, isorhamnetin 3-O-glucoside and myricetin 3-O-glucoside standard curves, respectively. Results were expressed as μg g−1 sample (d.w.b.).
4
Flavonoid Extraction and Quantification
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All reagents and solvents for extraction, as well as UPLC and HPLC-MS analyses (methanol, acetonitrile, formic acid, dimethylsulfoxide, polyamide and chloroform), were from Sigma-Aldrich (Poznań, Poland), and ultrapure water was obtained from a Millipore Direct Q3 device. Standards of flavonoids (apigenin, apigenin-7-O-glucoside, apigenin-6-C-glucoside, apigenin-8-C-glucoside, isorhamnetin, luteolin, luteolin 7-O-glucoside, luteolin 4′-O-glucoside, luteolin-8-C-glucoside, luteolin-6-C-glucoside, quercetin 3-O-galactoside, quercetin, rutin, isorhamnetin, isorhamnetin 7-O-glucoside) were purchased from Extrasynthese (Genay, France).
Extracts were prepared according to [21 (link)]. Weighted samples of frozen leaves (about 100 mg) were placed in Eppendorf tubes with 1.4 mL of 80% methanol and the internal standard, homogenized using a ball mill (MM 400, Retsch, Haan, Germany) and subsequently placed in an ultrasonic bath for 30 min and centrifuged (11,000× g) for 10 min. apigenin (5 µL of 1 mg/mL solution in dimethylsulfoxide) was used as the internal standard in extracts prepared for the UPLC analyses. Samples of the secondary metabolite extract were directly subjected to HPLC-MS analysis and, prior to the UPLC analysis, 200 µL were diluted with 80% methanol (800 µL).
Extracts were prepared according to [21 (link)]. Weighted samples of frozen leaves (about 100 mg) were placed in Eppendorf tubes with 1.4 mL of 80% methanol and the internal standard, homogenized using a ball mill (MM 400, Retsch, Haan, Germany) and subsequently placed in an ultrasonic bath for 30 min and centrifuged (11,000× g) for 10 min. apigenin (5 µL of 1 mg/mL solution in dimethylsulfoxide) was used as the internal standard in extracts prepared for the UPLC analyses. Samples of the secondary metabolite extract were directly subjected to HPLC-MS analysis and, prior to the UPLC analysis, 200 µL were diluted with 80% methanol (800 µL).
5
Quantitative Analysis of Phenolic Compounds
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The extracts at a concentration of 10 mg mL -1 were analyzed using a Dionex Ultimate 3000 UPLC chromatographic system (Thermo Scientific, San Jose, CA, USA). This system consists of a diode array detector (using 280, 330 and 370 nm as wavelengths) coupled to an electrospray ionization mass detector working in negative mode (Linear Ion Trap LTQ XL mass spectrometer, Thermo Finnigan, San Jose, CA, USA), following a procedure previously performed by the authors. 17 Calibration curves of the available phenolic standards (apigenin-6-C-glucoside, caffeic acid, chlorogenic acid, hesperetin, luteolin-7-O-glucoside, naringenin, quercetin-3-O-rutinoside and rosmarinic acid, Extrasynthese, Genay, France) were constructed based on the UV signal to perform quantitative analysis. In the case of the unavailable commercial standards, the compounds were quantified via the calibration curve of the most similar standard available. The results are expressed as mg per g of dry extract.
6
Quantification of Phenolic Compounds
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Composition of phenolic compounds of the aerial plant parts was determined by HPLC-DAD-ESI/MS (Dionex Ultimate 3000, Thermo Scientific, San Jose, CA) according to the method previously described by Bessada, Barreira, Barros, Ferreira, and Oliveira (2016) (link). Calibration curves for each available phenolic standard (chlorogenic acid, apigenin-6-C-glucoside, apigenin-7-O-glucoside, rosmarinic acid, quercetin-3-Oglucoside, Extrasynthèse, Genay, France) were prepared. The results were expressed as mg per g of extract.
7
Characterization of Trixis lignosa Extracts
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Samples of T. lignosa, available as dried rosettes of leaves and inflorescences, were obtained from a local herbal shop in Bragança (North-eastern Portugal). Botanical identification of all plant materials used was previously confirmed. Amber Perspex routine dosimeters, Batch V, were purchased from Harwell Company (Oxfordshire, UK). Organic acids (oxalic, quinic, shikimic and succinic acids) and trolox (6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid) were purchased from Sigma (St. Louis, MO, USA). Acetonitrile (99.9%, HPLC grade) was purchased from Fisher Scientific (Lisbon, Portugal). Formic acid was purchased from Prolabo (VWR International, France). The phenolic compound standards (apigenin-6-C-glucoside, p-coumaric acid, ellagic acid, gallic acid, kaempferol-3-O-glucoside, kaempferol-3-O-rutinoside, luteolin-6-C-glucoside, quercetin-3-O-glucoside and quercetin-3-O-rutinoside) were purchased from Extrasynthese (Genay, France). 2,2-Diphenyl-1-picrylhydrazyl (DPPH) was obtained from Alfa Aesar (Ward Hill, MA, USA). All other chemicals and solvents were of analytical grade and purchased from common sources. Water was treated in a Milli-Q water purification system (Merck Millipore, model A10, Billerica, MA, USA).
8
Phenolic Compound Extraction and Analysis
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Acetonitrile 99.9% was of HPLC grade from Fisher Scientific (Lisbon, Portugal). Phenolic compound standards (apigenin-6-Cglucoside, caffeic acid, chlorogenic acid, hesperetin, luteolin-7-Oglucoside, naringenin, quercetin-3-O-rutinoside and rosmarinic acid) were from Extrasynthese (Genay, France). Fetal bovine serum (FBS), L-glutamine, Hank's balanced salt solution (HBSS), trypsin-EDTA (ethylenediaminetetraAcetic acid), penicillin/streptomycin solution (100 U/mL and 100 mg/mL, respectively), RPMI-1640 and DMEM media were from Hyclone (Logan, UT, USA). Acetic acid, formic acid, ellipticine, sulforhodamine B (SRB), trypan blue, trichloroAcetic acid (TCA) and Tris were from Sigma Chemical Co. (St. Louis, MO, USA). Water was treated in Milli-Q water purification system (TGI Pure Water Systems, Greenville, SC, USA). Ferrous ammonium sulfate(II) hexahydrate, sodium chloride and sulfuric acid, all with PA purity, were purchased from Panreac S.A. (Barcelona, Spain) (proanalysis), in order to prepare the acid aqueous Fricke dosimeter solution.
9
Quantification of Phenolic Compounds
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Acetonitrile 99.9% was of HPLC grade from Fisher Scientific (Lisbon, Portugal). Phenolic compound standards: apigenin-6-Cglucoside, caffeic acid, chlorogenic acid, hesperetin, luteolin-7-O-glucoside, naringenin, quercetin-3-O-rutinoside and rosmarinic acid were purchased from Extrasynthese (Genay, France).
10
Preparation of Phenolic Compound Standards
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Acetonitrile 99.9% was of HPLC grade from Fisher Scientific (Lisbon, Portugal).
Phenolic compound standards (apigenin-6-C-glucoside, caffeic acid, luteolin-7-Oglucoside, naringenin, quercetin-3-O-rutinoside and rosmarinic acid) were from Extrasynthese (Genay, France). Trolox (6-hydroxy-2,5,7,8-tetramethylchroman-2carboxylic acid) was purchased from Sigma (St. Louis, MO, USA) and 2,2-diphenyl-1picrylhydrazyl (DPPH•) was obtained from Alfa Aesar (Ward Hill, MA, USA). β-Carotene and linoleic acid were acquired from Sigma-Aldrich (St. Louis, Missouri, USA) and Tween 80 from Panreac (Barcelona, Spain). All other solvents and reagents were acquired from scientific retailers. Ferrous ammonium sulfate(II) hexahydrate, sodium chloride and sulfuric acid, all with PA purity, were purchased from Panreac S.A. (Barcelona, Spain), in order to prepare the acid aqueous Fricke dosimeter solution.
Water was treated in a Milli-Q water purification system (TGI Pure Water Systems, Greenville, SC, USA).
Phenolic compound standards (apigenin-6-C-glucoside, caffeic acid, luteolin-7-Oglucoside, naringenin, quercetin-3-O-rutinoside and rosmarinic acid) were from Extrasynthese (Genay, France). Trolox (6-hydroxy-2,5,7,8-tetramethylchroman-2carboxylic acid) was purchased from Sigma (St. Louis, MO, USA) and 2,2-diphenyl-1picrylhydrazyl (DPPH•) was obtained from Alfa Aesar (Ward Hill, MA, USA). β-Carotene and linoleic acid were acquired from Sigma-Aldrich (St. Louis, Missouri, USA) and Tween 80 from Panreac (Barcelona, Spain). All other solvents and reagents were acquired from scientific retailers. Ferrous ammonium sulfate(II) hexahydrate, sodium chloride and sulfuric acid, all with PA purity, were purchased from Panreac S.A. (Barcelona, Spain), in order to prepare the acid aqueous Fricke dosimeter solution.
Water was treated in a Milli-Q water purification system (TGI Pure Water Systems, Greenville, SC, USA).
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