Folin ciocalteu s reagent

Manufactured by Thermo Fisher Scientific

Sourced in United Kingdom, United States

Folin-Ciocalteu's reagent is a colorimetric reagent used for the quantitative determination of phenolic and certain other easily oxidized compounds. It is commonly used in biochemistry and analytical chemistry laboratories for the analysis of total phenolic content in various samples.

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8 protocols using folin ciocalteu s reagent

Quantifying Total Phenolic Content

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Total phenolic content was determined using a modified protocol from Ainsworth and Gillespie (2007) (link). Briefly, 100 µL of methanolic sample extract was combined with 200 µL of Folin-Ciocalteu’s reagent (Thermo Fisher Scientific, MA, USA) and 800 µL of 700 mM sodium carbonate. The tubes were vortexed for 30 seconds then allowed to stand at room temperature for 2h. The absorbance was measured at 765 nm using a UV/VIS spectrophotometer. Total phenolic content was expressed as gallic acid equivalents.

Lanoue J., St. Louis S., Little C, & Hao X. (2022). Continuous lighting can improve yield and reduce energy costs while increasing or maintaining nutritional contents of microgreens. Frontiers in Plant Science, 13, 983222.

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Metabolite Extraction and Analysis

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The reagents used for the analysis, namely anhydrous methanol, 2,2′‐Azinobis‐(3ethylbenzothiazoline‐6‐sulfonic acid) diammonium salt (ABTS), methoxyamine hydrochloride and N‐methyl‐N‐trimethylsilyltrifuoroacetamide (MSTFA), cycloleucine, and heptadecanoic acid were obtained from Sigma‐Aldrich (Gillingham, UK). The anhydrous sodium carbonate, Folin–Ciocalteu's reagent and gallic acid were obtained from Fischer Scientific (Loughborough, UK).

de Falco B., Grauso L., Fiore A., Bochicchio R., Amato M, & Lanzotti V. (2021). Metabolomic analysis and antioxidant activity of wild type and mutant chia ( Salvia hispanica L.) stem and flower grown under different irrigation regimes. Journal of the Science of Food and Agriculture, 101(14), 6010-6019.

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Chemical Characterization of Extracts

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The reagents used for the extraction procedure and chemical characterization, namely anhydrous methanol (99.8%), anhydrous n-hexane (95%), 2,2`-Azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), derivatizating agents methoxyamine hydrochloride and N-methyl-N-trimethylsilyltrifuoroacetamide (MSTFA) were obtained from Sigma-Aldrich (Dorset, UK). N, O-Bis(trimethylsilyl)trifluoroacetamide, Trimethylchlorosilane (BSTFA with 1% TMCS) was purchased from Supelco Analytical (Bellefonte, PA). Pyridine, anhydrous sodium carbonate, Folin-Ciocalteu's reagent and gallic acid were obtained from Fischer Scientific (Loughborough, UK). Sugars, amino acids, organic acids and polyphenols used for identification and quantification were purchased from Sigma-Aldrich (Dorset, UK).

de Falco B., Fiore A., Bochicchio R., Amato M, & Lanzotti V. (2018). Metabolomic analysis by UAE-GC MS and antioxidant activity of Salvia hispanica (L.) seeds grown under different irrigation regimes. Industrial Crops & Products., 112.

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Comprehensive Chemical Characterization of Extracts

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The reagents used for the extraction procedure and chemical characterization, namely anhydrous methanol (99.8%), anhydrous n-hexane (95%), 2,2`-Azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), derivatizating agents methoxyamine hydrochloride and N-methyl-N-trimethylsilyltrifuoroacetamide (MSTFA), were obtained from Sigma-Aldrich (Dorset, UK). N, O-Bis(trimethylsilyl)trifluoroacetamide, Trimethylchlorosilane (BSTFA with 1% TMCS) was purchased from Supelco Analytical (Bellefonte, PA). Pyridine, anhydrous sodium carbonate, Folin-Ciocalteu's reagent and gallic acid were obtained from Fischer Scientific (Loughborough, UK). Sugars, amino acids, organic acids, polyphenols used for identification and quantification were purchased from Sigma-Aldrich (Dorset, UK).

de Falco B., Fiore A., Rossi R., Amato M, & Lanzotti V. (2018). Metabolomics driven analysis by UAEGC-MS and antioxidant activity of chia (Salvia hispanica L.) commercial and mutant seeds. Food chemistry, 254.

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Organic Solvents and Chemicals for Analysis

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The organic solvents used in this study were Acetone, hexane, methanol, ethyl acetate, and diethyl ether were obtained from Fisher Scientific Ltd. (Ottawa, ON, Canada). The following chemicals, anhydrous sodium sulfate, sodium hydroxide, sodium chloride, dibasic and monobasic sodium phosphates, sodium nitrite, HC l, AlCl₃, potassium ferricyanide, ferrous chloride, Folin-Ciocalteu's reagent, ferric chloride, and sodium carbonate were purchased from Fisher Scientific Ltd. (Ottawa, ON, Canada) and Sigma-Aldrich Canada Ltd. (Oakville, ON, Canada). Gallic acid, catechin, Trolox, DPPH radical, and a number of phenolic acid standards were purchased from Sigma-Aldrich Canada Ltd. (Oakville, ON, Canada).

Rahman M.J., Malunga L.N., Eskin M., Eck P., Thandapilly S.J, & Thiyam-Hollander U. (2021). Valorization of Heat-Treated Brewers' Spent Grain Through the Identification of Bioactive Phenolics by UPLC-PDA and Evaluation of Their Antioxidant Activities. Frontiers in Nutrition, 8, 634519.

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Pharmacological Evaluation of Diazepam and Piracetam

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Diazepam (Valium) and Piracetam (Nootropil) injection were purchased from local pharmacy. Ethanol, sulphuric acid, carboxymethyl cellulose, chloroform, iodine, sodium hydroxide, folin-ciocalteu’s reagent, potassium acetate, mEthanol, formalin, ferric chloride, n-hexane, sodium carbonate, HCL, ninhydrin reagent, ferric cyanide, and 2,2-diphenyl-1-picrylhydrazyl (DPPH) were purchased from Fisher chemicals (Loughborough, UK), Ascorbic Acid, Gallic acid, Rutin and Trichloroacetic acid were obtained from Sigma Aldrich chemicals (Darmstadt, Germany).

Mir N.T., Saleem U., Anwar F., Ahmad B., Ullah I., Hira S., Ismail T., Ali T, & Ayaz M. (2019). Lawsonia Inermis Markedly Improves Cognitive Functions in Animal Models and Modulate Oxidative Stress Markers in the Brain. Medicina, 55(5), 192.

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Detailed Reagents for Antioxidant Assays

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Acetic acid (>99%), m ethanol (>99%), ethanol (99.8%), sodium carbonate (Na₂CO₃) (99.5%) and Folin-Ciocalteu’s reagent (2.0 N) were purchased from Fisher Chemical (Fair Lawn, NJ, USA). Acetic acid potassium (99%), aluminum chloride hexahydrate (AlCl₃·6H₂O) (99%), EDTA (99%), ferrozine, gallic acid (98%), iron (II) chloride tetrahydrate (FeCl₂·4H₂O) and iron (III) chloride hexahydrate (FeCl₃·6H₂O) were supplied by Acros (Geel, Belgium). DPPH (2,2-diphenyl-1-picrylhydrazyl) (95%) and p-anisidine (99%) were purchased from Alfa Aesar (Haverhill, MA, USA) and Sigma-Aldrich (St. Louis, MO, USA), respectively.

Chuang K.J., Chen Z.J., Cheng C.L, & Hong G.B. (2018). Investigation of the Antioxidant Capacity, Insecticidal Ability and Oxidation Stability of Chenopodium formosanum Seed Extract. International Journal of Molecular Sciences, 19(9), 2726.

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Comprehensive Analysis of Polyphenolic Compounds

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Citric acid (CTA), sucrose, ascorbic acid (AA), glucose, fructose, malic acid (MA), trans-p-coumaric acid ( p-CA), 3-hydroxybenzoic acid (3-HBA), 4-hydroxybenzoic acid (4-HBA), 3, 4-dihydroxybenzoic acid (3, 4-diHBA), caffeic acid (CFA), gallic acid (GA), trolox standards were purchased from Sigma-Aldrich (St. Louis, MO, USA); ferulic acid (FA) was from ChromaDex Inc. (Irvine, CA, USA). Chemical reagents (all HPLC grade) including ammonium formate, ammonium acetate, ammonium hydroxide, formic acid, acetonitrile (ACN) and methanol were from Fisher Scientific (Fair Lawn, NJ, USA). Reagents (all ACS grade) including ethyl-enediaminetetraacetic acid (EDTA), Folin Ciocalteu's reagent, sodium carbonate, 2, 3, 5-triphenyltetrazolium chloride (TPTZ), 2, 2′-azinobis( 3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), potassium persulfate, sodium acetate trihydrate, glacial acetic acid, hydrochloric acid were from Fisher Scientific (Fair Lawn, NJ, USA). Pierce™ LC-MS water was from Thermo Fisher Scientific (Waltham, MA, USA).

Lyu W., Yuan B., Liu S., Simon J.E, & Wu Q. (2021). Assessment of lemon juice quality and adulteration by ultra-high performance liquid chromatography/triple quadrupole mass spectrometry with interactive and interpretable machine learning. Journal of Food and Drug Analysis, 29(2), 275-286.

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