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通过电子顺磁共振光谱法评估草药的抗氧化活性。

Antioxidant activity of herbal medicine evaluated by electron paramagnetic resonance spectroscopy.

作者信息

Quan Li, Zhou En-Chao, Guo Xue-Wen, Yin Gui

机构信息

The First School of Clinical Medicine, Jiangsu University Key Laboratory of Tonifying Kidney and Anti-senescence, Nanjing University of Chinese Medicine, Nanjing, 210023, China.

Department of Nephrology, Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing, 210029, China.

出版信息

Anal Methods. 2025 Jan 16;17(3):476-484. doi: 10.1039/d4ay02045f.

Abstract

The hydroxyl radical (˙OH) is one of the most reactive and harmful reactive oxygen species in living organisms. In this study, we used electron paramagnetic resonance (EPR) spectroscopy to evaluate the antioxidant activity of herbal medicine (PL) by measuring its scavenging activity against ˙OH radicals. Using the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical as a scavenging model, the PL extract (0.6 mg mL) was able to scavenge 95.1% of 0.5 mmol L DPPH radicals, showing a quantitative linear relationship between the DPPH radical concentration and the double integral area under the characteristic peaks of the EPR spectra, with a coefficient of determination = 0.9986. Additionally, we also studied the scavenging activity of the PL extract towards hyperactive ˙OH radicals through the reduction of the intensity of the DMPO-OH˙ adducts. The PL extract (0.5 mg mL) showed a high scavenging ability against extremely short-lived ˙OH radicals, with a scavenging rate of 93.6% for ˙OH radicals generated by continuous ultraviolet-irradiation of a 5 mM HO aqueous solution for 30 minutes. This study developed a method based on EPR spectroscopy measurements to detect ˙OH radicals captured by herbal medicine, providing a reference for the application of PL in antioxidant and clinical disease therapies.

摘要

羟基自由基(˙OH)是生物体内最具活性和危害性的活性氧物种之一。在本研究中,我们使用电子顺磁共振(EPR)光谱法,通过测量草药(PL)对˙OH自由基的清除活性来评估其抗氧化活性。以1,1-二苯基-2-苦基肼(DPPH)自由基作为清除模型,PL提取物(0.6 mg/mL)能够清除95.1%的0.5 mmol/L DPPH自由基,DPPH自由基浓度与EPR光谱特征峰下的二重积分面积之间呈现定量线性关系,决定系数R² = 0.9986。此外,我们还通过降低DMPO-OH˙加合物的强度,研究了PL提取物对高活性˙OH自由基的清除活性。PL提取物(0.5 mg/mL)对极短寿命的˙OH自由基表现出较高的清除能力,对5 mM H₂O₂水溶液连续紫外照射30分钟产生的˙OH自由基的清除率为93.6%。本研究建立了一种基于EPR光谱测量的方法来检测草药捕获的˙OH自由基,为PL在抗氧化和临床疾病治疗中的应用提供了参考。

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