DPPH (Synonyms: 2,2-Diphenyl-1-Picrylhydrazyl DPPH radical) |
Catalog No.GC19475
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DPPH (1,1-diphenyl2-picrylhydrazyl) is a stable free radical because of its spare electron delocalization over the whole molecule. The DPPH assay, which is one of the best-known, frequently employed, and accurate methods.
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Cas No.: 1898-66-4
Sample solution is provided at 25 µL, 10mM.
DPPH (1,1-diphenyl2-picrylhydrazyl) is a stable free radical because of its spare electron delocalization over the whole molecule. The DPPH assay, is one of the best-known, frequently employed, and accurate methods. The delocalization of DPPH results in a deep violet color with λmax around 520 nm. When a solution of DPPH is mixed with a substrate acting as a hydrogen atom donor, a stable non-radical form of DPPH is obtained with a simultaneous change of the violet color to pale yellow[1].
DPPH assay has been successfully utilized for investigating antioxidant properties of wheat grain and bran, vegetables, conjugated linoleic acids, herbs, edible seed oils, and flours in several different solvent systems including ethanol, aqueous acetone, methanol, aqueous alcohol, and benzene [2,3]. DPPH assay is a convenient method for the antioxidant assay of cysteine, glutathione, ascorbic acid, tocopherol, and polyhydroxy aromatic compounds [4], for olive oil, fruits, juices and wines [5].
References:
[1]. Szabo M, Idioiu C, Chambre D, et al. Improved DPPH determination for antioxidant activity spectrophotometric assay[J]. Chemical Papers, 2007, 61(3): 214-216.
[2]. Yu L. Free radical scavenging properties of conjugated linoleic acids[J]. Journal of Agricultural and Food Chemistry, 2001, 49(7): 3452-3456.
[3]. Parry J, Su L, Luther M, et al. Fatty acid composition and antioxidant properties of cold-pressed marionberry, boysenberry, red raspberry, and blueberry seed oils[J]. Journal of agricultural and food chemistry, 2005, 53(3): 566-573.
[4]. Nishizawa M, Kohno M, Nishimura M, et al. Non-reductive scavenging of 1, 1-diphenyl-2-picrylhydrazyl (DPPH) by peroxyradical: a useful method for quantitative analysis of peroxyradical[J]. Chemical and Pharmaceutical bulletin, 2005, 53(6): 714-716.
[5]. Sanchez-Moreno C. Methods used to evaluate the free radical scavenging activity in foods and biological systems[J]. Food science and technology international, 2002, 8(3): 121-137.
The DPPH free radical scavenging activities of the extracts were analyzed. DPPH solution (40 μg mL-1) was prepared in methanol. To each well on a microtiter plate was added 20 μL of sample (in triplicate) of appropriate concentration (1000, 500, 100, 50, and 10 μg mL-1) and 180 μL of DPPH solution. As a negative control, 20 μL of methanol was used instead of a sample. BHA, BHT, and ascorbic acid (AA) were used as positive controls. The reaction plates were kept in the dark at 37℃ for 30 minutes, after which the absorbance was measured at 517 nm using Multiskan Sky Thermo Scientific Microtiter plate reader. The inhibition of DPPH radicals in the test sample was calculated using the following formula and expressed as a percentage (%):
Where Ab is the absorbance of negative control (without sample) and As is the absorbance of the sample at different concentration and the positive controls, as well. The results are presented as the mean of percentage of DPPH radicals inhibition ± standard error.
This protocol only provides a guideline, and should be modified according to your specific needs
References:
[1]. Mandić M R, Oalđe M M, Lunić T M, et al. Chemical characterization and in vitro immunomodulatory effects of different extracts of moss Hedwigia ciliata (Hedw.) P. Beauv. from the Vršačke Planine Mts., Serbia[J]. PloS one, 2021, 16(2): e0246810.
| Cas No. | 1898-66-4 | SDF | |
| Synonyms | 2,2-Diphenyl-1-Picrylhydrazyl DPPH radical | ||
| Chemical Name | 2,2-diphenyl-1-(2,4,6-trinitrophenyl)-hydrazinyl | ||
| Canonical SMILES | O=[N+](C1=C(C([N+]([O-])=O)=CC([N+]([O-])=O)=C1)[N]N(C2=CC=CC=C2)C3=CC=CC=C3)[O-] | ||
| Formula | C18H12N5O6 | M.Wt | 394.3 |
| Solubility | 10mg/mL in DMF, 10mg/mL in Ethanol, 79mg/mL in DMSO | Storage | Store at 2-8°C, protect from light |
| General tips | Please select the appropriate solvent to prepare the stock solution according to the
solubility of the product in different solvents; once the solution is prepared, please store it in
separate packages to avoid product failure caused by repeated freezing and thawing.Storage method
and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored
at -20°C, please use it within 1 month. To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time. |
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| Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. | ||
| Prepare stock solution | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 2.5361 mL | 12.6807 mL | 25.3614 mL |
| 5 mM | 507.2 μL | 2.5361 mL | 5.0723 mL |
| 10 mM | 253.6 μL | 1.2681 mL | 2.5361 mL |
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
g
μL
Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)
Calculation results:
Working concentration: mg/ml;
Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )
Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL saline, mix and clarify.
Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.
Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.
Quality Control & SDS
- View current batch:
- Purity: >98.00% Appearance: A solid
- COA (Certificate of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
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Related Biological Data
D Free-radical scavengabilities of ascorbic acid, PADM, and AgNP–PADM hydrogels in range 50–250μg mL^−1. E IC50 values of ascorbic acid, PADM, and AgNP–PADM hydrogel groups IC50: the 50% inhibiting concentration
Then, the ascorbic acid, the PADM hydrogel, and the AgNP–PADM hydrogel prepared using the 80-μg mL^-1 AgNP solution were added to a 0.4 mM DPPH(GlpBio) anhydrous ethanol solution, and the solution was stored in the dark for 10 min.
Biomater Res 26.1 (2022): 36. PMID: 35879746 IF: 11.2997 -
Related Biological Data
The free radical scavenging ratio was determined by the DPPH method. CNMs at different MIC levels were mixed with DPPH solution. The mixtures were shaken well and put in the dark at room temperature for 30 min.
CNMs at different MIC levels were mixed with DPPH(GlpBio) solution. The mixtures were shaken well and put in the dark at room temperature for 30 min.
Front Immunol 12 (2022): 5955. PMID: 35126369 IF: 8.7864
Average Rating: 5 (Based on Reviews and 6 reference(s) in Google Scholar.)