DPPH
(Synonyms: 2,2-联苯基-1-苦基肼基,2,2-Diphenyl-1-Picrylhydrazyl DPPH radical) 目录号 : GC19475
A colorimetric probe for free radical scavengers
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, which is one of the best-known, frequently employed, and accurate methods. The delocalization causes 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 nonradical form of DPPH is obtained with 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].
DPPH（1,1-二苯基-2-丙酰肼基）是一种稳定的自由基，因为它在整个分子上具有多余的电子离域作用。DPPH测定法是最著名、常用且准确的方法之一。离域导致深紫色竹最大约520纳米。当DPPH的溶液与充当氢原子供体的底物混合时，获得稳定的非自由基形式的DPPH，同时紫色变为浅黄色[1]。
DPPH测定法已成功用于研究小麦颗粒和麸皮、蔬菜、共轭亚油酸、草药、食用籽油和面粉在几种不同溶剂体系中的抗氧化性能，包括乙醇、丙酮水溶液、甲醇、乙醇水溶液和苯[2，3]。DPPH测定法是对橄榄油、水果、果汁和葡萄酒中半胱氨酸、谷胱甘肽、抗坏血酸、生育酚和多羟基芳香族化合物[4]进行抗氧化测定的一种方便方法[5]。
References:
[1]. Szabo M, Idi?oiu 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.
DPPH assay [1]
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 | |
| 别名 | 2,2-联苯基-1-苦基肼基,2,2-Diphenyl-1-Picrylhydrazyl DPPH radical | ||
| 化学名 | 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-] | ||
| 分子式 | C18H12N5O6 | 分子量 | 394.3 |
| 溶解度 | 10mg/mL in DMF, 10mg/mL in Ethanol, 79mg/mL in DMSO | 储存条件 | Store at 2-8°C, protect from light |
| General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
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| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 2.5361 mL | 12.6807 mL | 25.3614 mL |
| 5 mM | 0.5072 mL | 2.5361 mL | 5.0723 mL |
| 10 mM | 0.2536 mL | 1.2681 mL | 2.5361 mL |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
| 第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方） | ||||||||||
| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
3. 以上所有助溶剂都可在 GlpBio 网站选购。
Quality Control & SDS
- View current batch:
- Purity: >98.00%
- 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