Ding Li, Zhang Xiaomin, Zhang Jiajia
Zhejiang Pharmaceutical College, Ningbo, Zhejiang, China.
Int J Anal Chem. 2021 Nov 23;2021:7195125. doi: 10.1155/2021/7195125. eCollection 2021.
This study aimed to investigate the extract with high antioxidant activity of Diels et Gilg and identify the antioxidant components in vitro. , -Diphenyl--picrylhydrazyl (DPPH) radical assay, Trolox equivalent antioxidant capacity (TEAC) assay, ferric reducing antioxidant power (FRAP), and hydroxyl radical scavenging method were used to screen the extract with high antioxidant activity. The antioxidant capacity of the extracts was evaluated by the free radical scavenging ability of DPPH. The ability of extracts to scavenge 2, 2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) free radical was evaluated by TEAC assay. The FRAP method was used to evaluate the ability of extracts to reduce Fe. The ability to scavenge hydroxyl radicals produced by the interaction of hydrogen peroxide and Fe was measured by monitoring the change in the absorbance of the reaction mixture at 536 nm. Then, high-performance liquid chromatography-DPPH (HPLC-DPPH) and HPLC-hydroxyl radical scavenging methods were used to screen the antioxidant components in the extract. The molecular weight of the above antioxidant components was investigated using the qualitative analytical method of high-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (HPLC-Q-TOF LC/MS). Based on the concentrations of the samples (0.2-4 mg/mL), the DPPH free radical scavenging ability, ABTS+ free radical scavenging ability, hydroxyl free radical scavenging ability, and Fe reducing ability of the ethyl acetate extract (EAE) were stronger than that of the crude extract (CE), petroleum ether extract (PEE), and n-butanol extract (BE). The EAE has higher antioxidant activity than CE, PEE, and BE. Six antioxidant components, rutin, quercetin, isoquercetin, astragalin, kaempferol, and kaempferol-3-o-rutoside, were identified in the EAE.
本研究旨在对藏药川西獐牙菜提取物进行体外抗氧化活性筛选及抗氧化成分的鉴定。采用1,1-二苯基-2-三硝基苯肼（DPPH）自由基清除法、Trolox当量抗氧化能力（TEAC）法、铁离子还原抗氧化能力（FRAP）法、羟自由基清除法对提取物进行抗氧化活性筛选。以DPPH自由基清除能力评价提取物的抗氧化能力，以TEAC法评价提取物清除2,2'-联氮-双-3-乙基苯并噻唑啉-6-磺酸（ABTS）自由基的能力，采用FRAP法评价提取物还原Fe的能力，通过监测反应混合物在536nm处吸光度的变化测定提取物对过氧化氢与Fe相互作用产生的羟自由基的清除能力。然后采用高效液相色谱-DPPH（HPLC-DPPH）法和HPLC-羟自由基清除法对提取物中的抗氧化成分进行筛选，运用高效液相色谱-四极杆飞行时间串联质谱联用（HPLC-Q-TOF LC/MS）定性分析方法对上述抗氧化成分的分子量进行研究。基于样品浓度（0.2～4mg/mL），乙酸乙酯提取物（EAE）的DPPH自由基清除能力、ABTS+自由基清除能力、羟自由基清除能力和Fe还原能力均强于粗提物（CE）、石油醚提取物（PEE）和正丁醇提取物（BE）。EAE的抗氧化活性高于CE、PEE和BE。从EAE中鉴定出6种抗氧化成分，分别为芦丁、槲皮素、异槲皮素、黄芪甲苷、山柰酚和山柰酚-3-O-芸香糖苷。
