1 1 diphenyl 2 picrylhydrazyl dpph

Manufactured by Maclin Biochemical Technology

Sourced in China

1,1-Diphenyl-2-picrylhydrazyl (DPPH) is a stable free radical used as a reagent in biochemical and chemical applications. It is commonly used to evaluate the antioxidant activity of various compounds.

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10 protocols using 1 1 diphenyl 2 picrylhydrazyl dpph

Evaluation of Guava Leaf Extracts

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Firstly, 1,1-Diphenyl-2-picrylhydrazyl (DPPH) and sodium alginate (product number: S817374; AR, purity ≥90%; viscosity (1%, 20 °C), 260 mPa·s) were purchased from Macklin Biochemical Co., Ltd. (Shanghai, China). Glycerol and Absolute ethanol (AR) were obtained from Tianjin Zhiyuan Chemical Reagent Co., Ltd. (Tianjin, China). Gallic acid, chlorogenic acid, p-hydroxybenzoic acid, caffeic acid, rutin, isoquercitrin, quercetin-3-O-α-L-arabinoside, ellagic acid, avicularin, quercetin, quercitrin, kaempferol were purchased from Aladdin Industrial Corporation (Shanghai, China).
Staphylococcus aureus (ATCC29215) and Escherichia coli (ATCC25922) were provided by the Microbiology Laboratory of South China University of Technology (Guangzhou, China).
Guava leaves were collected in October, 2018 and supplied by Jiangmen Nanyue Guava farmer cooperatives (Jiangmen, Guangdong, China). The guava leaves were naturally dried and stored in the dark (25 ± 2 °C). Dried guava leaves were grounded into powders and passed through an 80 mesh sieve size.

Luo Y., Liu H., Yang S., Zeng J, & Wu Z. (2019). Sodium Alginate-Based Green Packaging Films Functionalized by Guava Leaf Extracts and Their Bioactivities. Materials, 12(18), 2923.

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Antioxidant and Cytotoxicity Evaluation

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Phloretin (Phl, purity ≥ 98%), isoniazid (Inz, purity = 98%), 2-picolinic acid (99%), theophylline (99%), 4-aminobenzamide (98%), proline (99%), nicotinic acid (99%) and 4-picolinic acid (99%) were purchased from Shanghai Macklin Biochemical Co., Ltd (Shanghai, China). Phloretin (HPLC ≥ 98%) and maleic (99%) acid were purchased from Shanghai Yuanye Bio-Technology Co., Ltd. (Shanghai, China). 2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) was purchased from Sigma Life Science (St. Louis, MO, USA). 1,1-Diphenyl-2-picrylhydrazyl (DPPH) was purchased from Shanghai Macklin Biochemical Co., Ltd. (Shanghai, China). Methanol was obtained from Guangzhou Reagent Co., Ltd. (Guangzhou, China). The A549 cell line and HepG2 cell line were purchased from the Institute of Biochemistry and Cell Biology Sciences, Chinese Academy of Sciences (Shanghai, China).

Lu Z., Chen H., Mo J., Yuan X., Wang D., Zheng X, & Zhu W. (2023). Cocrystal of phloretin with isoniazid: preparation, characterization, and evaluation. RSC Advances, 13(16), 10914-10922.

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Bovine β-LG and Polyphenol Compounds

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Bovine β-LG (protein content ≥ 90%), and six compounds (phloretin, EGCG, naringenin, quercetin, kaempferol, and myricetin, HPLC ≥ 95%) were provided by Sigma Chemical Co., Ltd. (Shanghai, China). Reagents like 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2-azinobis(3-ethylbenzthiazoline)-6-sulfonic acid (ABTS) were obtained from Shanghai Macklin Biochemical Co., Ltd. (Shanghai, China). α-cyano-4-hydroxycinnamic acid (CHCA) was produced by Beijing Solarbio Technology Co., Ltd. (Beijing, China). Unless specifically mentioned, the remaining materials and chemicals met the criteria of being of analytical grade or the highest available grade.

Pu P., Deng Z., Chen L., Yang H, & Liang G. (2023). Reducing Antigenicity and Improving Antioxidant Capacity of β-Lactoglobulin through Covalent Interaction with Six Flavonoids. Foods, 12(15), 2913.

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Antioxidant and Neuroprotective Effects of HA and Seleninic Acid

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High-molecular-weight HA (with a molecular weight of 2 × 10⁵ g mol⁻¹) and seleninic acid were procured from Huaxia Chemical Reagent Co., Ltd. (Chengdu, China). Sodium ascorbate was obtained from Aladdin Bio-Chem Technology Co., Ltd. (Shanghai, China). Dulbecco’s modified Eagle’s medium (DMEM) and fetal bovine serum were purchased from Gibco (Thermo Fisher Scientific, Waltham, MA, USA). 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), 2′,7′-dichlorodihydrofluorescein diacetate (DCFH-DA; D6883), and 4′,6-diamidino-2-phenylindole dihydrochloride (DAPI) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Hydrogen peroxide solution (30 wt% in water) was obtained from Sinopharm Chemical Reagent Co., Ltd. 1,1-Diphenyl-2-picrylhydrazyl (DPPH) was purchased from Shanghai Macklin Biochemical Co., Ltd. Anti-GFAP, anti-NF200, and anti-NeuN primary antibodies were obtained from Abcam (Cambridge, UK). Anti-CD44 and anti-caspase-3 antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). The dialysis membrane was obtained from Greenbird Technology Co., Ltd (Shanghai, China). The remaining compounds were obtained from commercial providers and used without any further manipulation.

Luo W., Li Y., Zhao J., Niu R., Xiang C., Zhang M., Xiao C., Liu W, & Gu R. (2024). CD44-targeting hyaluronic acid-selenium nanoparticles boost functional recovery following spinal cord injury. Journal of Nanobiotechnology, 22, 37.

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Antioxidant and Anti-inflammatory Properties of B. gymnorrhiza

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B. gymnorrhiza (L.) Lam. was provided by Nansha Wetland Park (Guangzhou, Guangdong, China), and was authenticated by one of our authors, Prof. Ziren Su of Guangzhou university of Chinese medicine, where a voucher specimen (Voucher 18-06-23) was deposited. HPLC grade methanol was purchased from Merck (Darmstadt, Germany). 1,1-Diphenyl-2-picryl-hydrazyl (DPPH, CAS:1898-66-4) was purchased from Shanghai Macklin Biochemical Co., Ltd. Dextran sulfate sodium (DSS) was bought from MP Biomedicals (molecular weight: 36,000~50,000, Canada). SASP was purchased from Shanghai Xinyi Tianping Pharmaceutical Co. Ltd (Shanghai, China). Myeloperoxidase (MPO) assay kit was obtained from Jiancheng Biotechnology Company (Nanjing, Jiangsu, China). MDA, SOD, and GSH assay kits were purchased from Jiancheng Biotechnology Company (Nanjing, Jiangsu, China). The enzyme-linked immunosorbent assay (ELISA) kits for TNF-α, IL-6, IL-1β, IFN-γ, IL-10, iNOS, and COX-2 were the products of Shanghai MLBIO Biotechnology Co. Ltd (Shanghai, China). The primary and secondary antibodies used in this study were purchased from Affinity Biosciences (OH, USA). The cDNAs for GCLC, GCLM, HO-1 and NQO1 were amplified by PCR with gene specific primers (Sangon Biotech Co. Ltd, Shanghai, China). Other chemicals used were of analytical grade or chromatographic grade.

Lin Y., Zheng X., Chen J., Luo D., Xie J., Su Z., Huang X., Yi X., Wei L., Cai J, & Sun Z. (2020). Protective Effect of Bruguiera gymnorrhiza (L.) Lam. Fruit on Dextran Sulfate Sodium-Induced Ulcerative Colitis in Mice: Role of Keap1/Nrf2 Pathway and Gut Microbiota. Frontiers in Pharmacology, 10, 1602.

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DPPH Antioxidant Assay Protocol

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1,1-Diphenyl-2-picrylhydrazyl (DPPH) was purchased from Shanghai Macklin Biochemical Co., Ltd. The ultrapure water (18.2 MΩ cm⁻¹) was used in this study, provided by a Simplicity-UV water purification system (Millipore, Bedford, USA).

Qiang Y., Si R., Tan S., Wei H., Huang B., Wu M., Shi M., Fang L., Fu J, & Zeng S. (2021). Spatial variation of volatile organic compounds and antioxidant activity of turmeric (Curcuma longa L.) essential oils harvested from four provinces of China. Current Research in Food Science, 4, 882-890.

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Enzymatic Assays for Antioxidant Analysis

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Papain was supplied by Yuanye Biotechnology Co., Ltd. (Shanghai, China), hydrogen peroxide (H₂O₂) was purchased from Chengdu Chron Chemicals Co., Ltd. (Chengdu, China) and Tris-HCl buffer was supplied by Nobleryder Technology Co. Ltd. (Beijing, China). Alcalase© 2.4 L, Chymotrypsin, potassium ferricyanide (C₆FeK₃N₆), ferrozine (C₂₀H₁₂N₄Na₂O₆S₂), sodium dodecyl sulphate (SDS), iron (II) chloride (FeCl₂), ferrous sulphate (FeSO₄), 1,10-phenanthroline (C₁₂H₈N₂), sodium hydrogen phosphate (Na₂HPO₄), sodium dihydrogen phosphate (NaH₂PO₄), trichloroacetic acid (C₂HCl₃O₂) and methyl red (C₁₅H₁₅N₃O₂) were acquired from Aladdin Industrial Co. (Shanghai, China). Hydrochloric acid (HCl) was obtained from Guangzhou Rongman Technology Co., Ltd. (Guangzhou, China), ethanol (CH₃CH₂OH) was purchased from Tianjin Fuyu Fine Chemical Co., Ltd. (Tianjin, China), bromocresol green (C₂₁H₁₄Br₄O₅S), iron (III) chloride (FeCl₃) and 1,1-diphenyl-2-picrylhydrazyl (DPPH) were bought from Shanghai Macklin Biochemical Co., Ltd. (Shanghai, China). Sodium hydroxide (NaOH) and boric acid (H₃BO₃) were procured from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China).

Johnson Esua O., Sun D.W., Cheng J.H., Wang H, & Lv M. (2022). Functional and bioactive properties of Larimichthys polyactis protein hydrolysates as influenced by plasma functionalized water-ultrasound hybrid treatments and enzyme types. Ultrasonics Sonochemistry, 86, 106023.

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Biochemical Characterization of Oat Bran

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The OB was kindly provided by Xibeihuitong Agricultural Technology Co., Ltd.
α‐Amylase, pepsin, and pancreatin were purchased from Sigma‐Aldrich. 1,1‐diphenyl‐2‐picrylhydrazyl (DPPH), 2,2′‐azino‐bis(3‐ethylbenzothiazoline‐6‐sulfonic acid) (ABTS), cholesterol (>99%), bile acid, and O‐phthalaldehyde were purchased from Shanghai Macklin Biochemical Co., Ltd. The Megazyme β‐glucan mixed linkage assay kit was purchased from Megazyme International Ireland Ltd.. The total antioxidant capacity (T‐AOC) assay kit was purchased from Nanjing Jiancheng Bioengineering Institute. All other chemicals used in the experiments were of analytical grade.

Zhang Y., Zhang M., Guo X., Bai X., Zhang J., Huo R, & Zhang Y. (2022). Improving the adsorption characteristics and antioxidant activity of oat bran by superfine grinding. Food Science & Nutrition, 11(1), 216-227.

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Tannic Acid-Gold Nanoparticle Synthesis

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Tannic acid (TA) and HAuCl₄ were purchased from Aladdin Co., Shanghai, China. KH561 was received from Bolian Chemical Co., Zhengzhou, China. Trizma base (primary standard and buffer, ≥99.9% titration, crystalline) and DMSO were purchased from Sigma-Aldrich. AgNO₃ was purchased from Ourchem Co., Shanghai, China. 1,1-Diphenyl-2-picrylhydrazyl (DPPH) was purchased from Macklin Co., Shanghai, China. Minimum Essential Medium (MEM) was obtained from Hyclone (USA). 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and calcein/propidium iodide (PI) cell viability/cytotoxicity assay kits were purchased from Beyotime Biotechnology (Shanghai China). PBS solution was obtained from Sunshine Bio (Nanjing, China). Heat-inactivated horse serum was purchased from Gibco (USA). GlutaMAX, sodium pyruvate and non-essential amino acids were purchased from Invitrogen Corporation (USA). All other reagents were commercial chemicals and used as received, unless otherwise indicated.

Chen C., Yang X., Li S.J., Ma F.J., Yan X., Ma Y.N., Ma Y.X., Ma Q.H., Gao S.Z, & Huang X.J. (2021). Red wine-inspired tannic acid–KH561 copolymer: its adhesive properties and its application in wound healing. RSC Advances, 11(9), 5182-5191.

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Antioxidant and Bioactive Compound Assays

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Tannic acid was purchased from Aladdin Co., Shanghai, China. Tris base (primary standard and buffer, Z99.9% titration, crystalline) was purchased from Sigma-Aldrich, USA. Phytic acid (PA) was purchased from Zhejiang Dong Jin Biotechnology, Ningbo, China. Thioctic acid (a-lipoic acid) was purchased from Aladdin Co., Shanghai, China. 1,1-Diphenyl-2-picrylhydrazyl (DPPH) was purchased from Macklin Co., Shanghai, China. Dulbecco's Modified Eagle Medium (DMEM) was obtained from Invitrogen (USA). The PBS solution was obtained from Sunshine Bio (Nanjing, China). All other reagents were commercial chemicals and used as received except specially claimed. Ultrapure water (18 MO cm resistivity) was also used in the experiments.

Shao X.H., Yang X., Zhou Y., Xia Q.C., Lu Y.P., Yan X., Chen C., Zheng T.T., Zhang L.L., Ma Y.N., Ma Y.X, & Gao S.Z. (2022). Antibacterial, wearable, transparent tannic acid-thioctic acid-phytic acid hydrogel for adhesive bandages. Soft matter, 18(14).

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