P nitrophenyl α d glucopyranose

Manufactured by Merck Group

Sourced in United States, Germany

P-nitrophenyl-α-D-glucopyranose is a chemical compound used as a substrate for various enzymatic assays. It is a synthetic derivative of the natural disaccharide α-D-glucopyranose. The compound can be used to detect and quantify the activity of enzymes that cleave the glycosidic bond, such as α-glucosidases.

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6 protocols using p nitrophenyl α d glucopyranose

Phytochemical and Antioxidant Analysis Protocol

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Absolute ethanol, analytical grade methanol, dimethyl sulfoxide (DMSO), sodium carbonate, sodium nitroprusside, phosphate buffer saline, sulphanilamide, naphthylethylene diamine dihydrochloride, 85% phosphoric acid, monosodium phosphate, disodium phosphate, glycine, alpha-glucosidase enzyme, deuterated methanol-d4 (CD₃OD), nondeuterated potassium dihydrogen phosphate (KH₂PO₄), deuterium oxide (D₂O), trimethylsilyl propionic acid-d4 sodium salt (TSP) were supplied by Merck (Darmstadt, Germany) whereas gallic acid, Folin–Ciocalteau reagent, quercetin, 2,2-diphenyl-1-picrylhydrazyl (DPPH), p-nitrophenyl-α-D-glucopyranose (PNPG) were supplied by Sigma (Aldrich, Germany). Deionized water was purified by ultrapure water system from Sartorius (Sartorius AG, Göttingen, Germany).

Azizan A., Lee A.X., Abdul Hamid N.A., Maulidiani M., Mediani A., Abdul Ghafar S.Z., Zolkeflee N.K, & Abas F. (2020). Potentially Bioactive Metabolites from Pineapple Waste Extracts and Their Antioxidant and α-Glucosidase Inhibitory Activities by 1H NMR. Foods, 9(2), 173.

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Chromatographic Analysis of Phytochemicals

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Phosphate buffer, glycine, α-glucosidase enzyme, p-nitrophenyl-α-D-glucopyranose (PNPG) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) were purchased from Sigma-Aldrich (Hamburg, Germany). Analytical grade solvents including absolute ethanol, hexane, chloroform, ethyl acetate and methanol were supplied by Merck (Darmstadt, Germany). The NMR chemicals including deuterated dimethyl sulfoxide-d₆ (DMSO-d₆) and trimethylsilyl propionic acid-d4 sodium salt (TSP) were also supplied by Merck (Darmstadt, Germany). The LC/MS grade acetonitrile and formic acid were purchased from Merck (Darmstadt, Germany). Water was prepared using a Milli-Q purification system.

Lee S.Y., Mediani A., Ismail I.S., M.a.u.l.i.d.i.a.n.i, & Abas F. (2019). Antioxidants and α-glucosidase inhibitors from Neptunia oleracea fractions using 1H NMR-based metabolomics approach and UHPLC-MS/MS analysis. BMC Complementary and Alternative Medicine, 19, 7.

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Comprehensive Reagent Acquisition Protocol

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Ethanol, chloroform, methanol, ethyl acetate, hexane, formic acid, glacial acetic acid, 1-butanol, acetonitrile, sodium hydroxide (NaOH), hydrogen chloride (HCL), and dimethyl sulfoxide (DMSO) were purchased from Merck Millipore International (Darmstadt, Germany). The LCMS-grade acetonitrile, water, and formic acid were supplied by Fisher Scientific (Geel, Belgium). The sodium nitrate (NaNO₃), iron(III) chloride (FeCl₃), aluminum chloride hexahydrate (AlCl₃·6H₂O), ferrous sulfate heptahydrate (FeSO₄·7H₂O), gallic acid, sodium carbonate (Na₂CO₃), sodium acetate trihydrate, Folin-Ciocalteu reagent, 2, 2-diphenyl-1-picrylhydrazyl (DPPH), p-nitrophenyl-α-D-glucopyranose (PNPG), glycine, phosphate buffer, 2,4,6-tripyridyl-S-triazine (TPTZ) quercetin, and chlorohexidine were purchased from Sigma-Aldrich (St. Louis, MO, USA).

Buzgaia N., Lee S.Y., Rukayadi Y., Abas F, & Shaari K. (2021). Antioxidant Activity, α-Glucosidase Inhibition and UHPLC–ESI–MS/MS Profile of Shmar (Arbutus pavarii Pamp). Plants, 10(8), 1659.

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Phytochemical Extraction and Analysis

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Folin–Ciocalteu reagent and absolute ethanol were purchased from Merck (Darmstadt, Germany). Sodium carbonate, DPPH, α-glucosidase enzyme, p-nitrophenyl-α-d-glucopyranose (PNPG), glycine, phosphate-buffered saline, sodium nitroprusside (SNP), and other standard compounds used, including gallic acid and quercetin, were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA). LC-MS-grade methanol, purified water, formic acid (FA), and dimethyl sulfoxide (DMSO) were supplied by Fisher Scientific (Geel, Belgium).

Wong P.L., Fauzi N.A., Mohamed Yunus S.N., Abdul Hamid N.A., Abd Ghafar S.Z., Azizan A., Zolkeflee N.K, & Abas F. (2020). Biological Activities of Selected Plants and Detection of Bioactive Compounds from Ardisia elliptica Using UHPLC-Q-Exactive Orbitrap Mass Spectrometry. Molecules, 25(13), 3067.

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Inhibition of Digestive Enzymes

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Intestinal α-glucosidase, pancreatic α-amylase, p-nitrophenyl-α-D-glucopyranose (PNPG), 1, 1-diphenyl-2-picrylhydrazyl (DPPH), acetylcholine iodide, butyrylcholine iodide, acarbose, Ellman’s reagent (5,5’-dithiobis (2-nitrobenzoic acid), DTNB), 2,2- azinobis (3-ethyl-benzothiazoline-6-sulfonic acid) (ABTS), butylated hydroxytoluene (BHT), gallic acid, quercetin and were procured from Sigma-Aldrich, Inc., (Saint Louis, MO), and other chemicals used were of analytical grades and prepared in all-glass apparatus using sterilized distilled water.

Afolabi O.B., Olasehinde O.R., Olanipon D.G., Mabayoje S.O., Familua O.M., Jaiyesimi K.F., Agboola E.K., Idowu T.O., Obafemi O.T., Olaoye O.A, & Oloyede O.I. (2023). Antioxidant evaluation and computational prediction of prospective drug-like compounds from polyphenolic-rich extract of Hibiscus cannabinus L. seed as antidiabetic and neuroprotective targets: assessment through in vitro and in silico studies. BMC Complementary Medicine and Therapies, 23, 203.

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LCMS-based Biochemical Assay Protocol

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LCMS grade formic acid, methanol, acetonitrile, water, dimethyl sulfoxide (DMSO), sodium phosphate buffer, and absolute ethanol were supplied by Thermo Fisher Scientific (Waltham, MA, USA). Other chemicals including p-nitrophenyl-α-D-glucopyranose (PNPG), α-glucosidase enzyme, α-amylase enzyme, glycine, 3,5-dinitrosalicylic acid (DNS), potato starch, and sodium phosphate buffered saline (PBS) were purchased from Sigma Aldrich (St. Louis, MO, USA). All the standard compounds including acarbose, quercetin, daidzein, quercitrin, kaempferol, formononetin, 6-hydroxyflavanone, and geniposide were purchased from Acros Organics (Geel, Belgium) with >98% purity.

Zolkeflee N.K., Ramli N.S., Azlan A, & Abas F. (2022). In Vitro Anti-Diabetic Activities and UHPLC-ESI-MS/MS Profile of Muntingia calabura Leaves Extract. Molecules, 27(1), 287.

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