[以夏枯草抗氧化活性与其总酚酸含量的相关性研究为例探讨引入生物测定法进行中药质量评价的可行性]
[Exploration on feasibility of introducing bioassay method into quality evaluation of Chinese herbal medicines by studying on the correlation between antioxidant activity of Prunella vulgaris and its total phenolic acids content for example].
作者信息
Feng Wei-Hong, Li Chun, Xin Wei-Mei, Lin Li-Mei, Xia Bo-Hou, Rong Li-Xin, Yang Li-Xin, Yi Hong, Zhang Yong-Xin, Chen Liang-Mian, Wang Zhi-Min
机构信息
Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China.
National Engineering Laboratory for Quality Control Technology of Chinese Herbal Medicines, Beijing 100700, China.
出版信息
Zhongguo Zhong Yao Za Zhi. 2016 Jul;41(14):2660-2668. doi: 10.4268/cjcmm20161416.
This paper aims to investigate the correlation between the antioxidant activity of Prunella vulgaris and its total phenolic acids content by measuring the antioxidant activity of different sources and different organs of P. vulgaris and the total contents of protocatechuic acid, protocatechuic aldehyde, caffeic acid, salviaflaside and rosmarinic acid in these samples. Using the 50% methanol extract of P. vulgaris samples as the research object, DPPH method and HPLC method were used respectively to determine the antioxidant activities and the total contents of the above-mentioned five analytes in P. vulgaris samples. 0.5 mL of 50% methanol extract of P. vulgaris reacts with 0.1 mmol•L⁻¹ DPPH ethanol solution for 60 min, then the absorbance of the reaction solution was measured at 517 nm, scavenging rate and IC₅₀ values were calculated by the absorbance and the sample concentration for evaluating the antioxidant activity. HPLC analysis was made on a C₁₈ Epic column, with acetonitrile-0.1% formic acid aqueous solution as mobile phase (gradient elution), and the detection wavelength was set at 280 nm. The correlation between the antioxidant capacity of different habitats and different organs of P. vulgaris and the total contents of five kinds of phenolic acids was analyzed by partial least squares method. The reaction dose-response range of 50% methanol extract of P. vulgaris with 0.1 mmol•L⁻¹ DPPH ethanol solution was 0.300-1.65 g•L⁻¹. When the quantities of potocatechuic acid, protocatechuic aldehyde, caffeic acid, salviaflaside and rosmarinic acid were respectively in 0.007 84-0.980, 0.011 5-1.44, 0.008 64-1.08, 0.080 0-1.00 and 0.079 8-0.998 μg range, their quantities were in good linear relationship with the corresponding peak areas. The average recovery of 5 components were 97.76%, 96.88%, 100.3%, 102.1%, 104.5%, with RSD of 1.8%, 1.6%, 1.7%, 1.6% and 1.7%, respectively. In a certain range of crude drug quantity, the antioxidant activity of each organ of P. vulgaris and total phenolic acids content inside has a good linear correlation. Therefore, in certain quality range of crude drug, DPPH bioassay combined with HPLC content determination can be used for the quality control of P. vulgaris, as is a new method for the quality control of P. vulgaris.
本文旨在通过测定夏枯草不同来源、不同器官的抗氧化活性以及其中原儿茶酸、原儿茶醛、咖啡酸、丹参素和迷迭香酸的总含量，研究夏枯草抗氧化活性与其总酚酸含量之间的相关性。以夏枯草样品的50%甲醇提取物为研究对象，分别采用DPPH法和HPLC法测定夏枯草样品的抗氧化活性及上述5种分析物的总含量。取0.5 mL夏枯草50%甲醇提取物与0.1 mmol•L⁻¹ DPPH乙醇溶液反应60 min，然后在517 nm处测定反应溶液的吸光度，根据吸光度和样品浓度计算清除率和IC₅₀值，以评价抗氧化活性。采用C₁₈ 艾彼克柱进行HPLC分析，以乙腈 - 0.1%甲酸水溶液为流动相（梯度洗脱），检测波长设定为280 nm。采用偏最小二乘法分析夏枯草不同产地、不同器官的抗氧化能力与5种酚酸总含量之间的相关性。夏枯草50%甲醇提取物与0.1 mmol•L⁻¹ DPPH乙醇溶液的反应剂量 - 响应范围为0.300 - 1.65 g•L⁻¹。当原儿茶酸、原儿茶醛、咖啡酸、丹参素和迷迭香酸的量分别在0.007 84 - 0.980、0.011 5 - 1.44、0.008 64 - 1.08、0.080 0 - 1.00和0.079 8 - 0.998 μg范围内时，它们的量与相应峰面积呈良好的线性关系。5种成分的平均回收率分别为97.76%、96.88%、100.3%、102.1%、104.5%，相对标准偏差分别为1.8%、1.6%、1.7%、1.6%和1.7%。在一定的药材量范围内，夏枯草各器官的抗氧化活性与其中总酚酸含量具有良好的线性相关性。因此，在一定的药材质量范围内，DPPH生物测定法结合HPLC含量测定可用于夏枯草的质量控制，是一种新的夏枯草质量控制方法。
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