Chen Xu, Zhou Tao, Huang Zhi-Fang, Chen Yan, Liu Yu-Hong, Liu Yun-Hua, Yi Jin-Hai
Sichuan Provincial Key Laboratory of Quality and Innovation Research of Chinese Materia,Sichuan Academy of Chinese Medicine Sciences Chengdu 610041,China School of Pharmacy of Chengdu University of Traditional Chinese Medicine Chengdu 611130,China.
Sichuan Provincial Key Laboratory of Quality and Innovation Research of Chinese Materia,Sichuan Academy of Chinese Medicine Sciences Chengdu 610041,China Department of Pharmacy,Children's Hospital Affiliated to Chongqing Medical University Chongqing 400014,China.
Zhongguo Zhong Yao Za Zhi. 2021 Mar;46(5):1148-1154. doi: 10.19540/j.cnki.cjcmm.20201210.301.
There is no consensus on the content, accumulation, transformation and content determination methods of phenolic acids in fresh Salvia miltiorrhiza. In order to find out the true content of phenolic acids in fresh S. miltiorrhiza, a variety of treatment me-thods were used in this study to prepare sample solution. The content changes of phenolic acids in S. miltiorrhiza samples with different dehydration rates were investigated during drying and shade drying processes. Polyphenol oxidase(PPO) of S. miltiorrhiza was extracted and purified by ammonium sulfate precipitation and dialysis to investigate the enzymatic properties. The content of rosmarinic acid, lithosperic acid and S. nolic acid B in S. miltiorrhiza was determined by UPLC. The results showed that the content of phenolic acids in fresh S. miltiorrhiza was highest when it was homogenized with 1 mol·L(-1) HCl solution or 1 mol·L(-1) HCl methanol solution. There was no significant difference in the content of phenolic acids in S. miltiorrhiza with different dehydration rates, indicating that there was no correlation between phenolic acid content and dehydration rate. The optimum pH of S. miltiorrhiza PPO was 7.6 and the optimum temperature was 40 ℃. With catechol as substrate, S. miltiorrhiza PPO had the enzymatic browning reaction which was in compliance with Michaelis equation, with Michaelis constant K_m of 0.12 mol·L(-1) and V_(max) of 588.23 U·min(-1). The inhibitory effect of citric acid, disodium ethylenediamine tetraacetate, ascorbic acid and sodium sulfite on S. miltiorrhiza PPO increased with the increase of inhibitor concentration, and sodium sulfite showed the strongest inhibitory effect. The present study proved that there were a large number of phenolic acids in fresh S. miltiorrhiza, which were the secondary metabolite of primitive accumulation during the growth of S. miltiorrhiza, rather than the induced product of postharvest drying and dehydration stress. This study has reference value and significance for the cultivation, harvest and processing of S. miltiorrhiza.
对于新鲜丹参中酚酸类成分的含量、积累、转化及含量测定方法，目前尚无统一认识。为明确新鲜丹参中酚酸类成分的真实含量，本研究采用多种处理方法制备样品溶液，考察了不同干燥速率的丹参样品在烘干和阴干过程中酚酸类成分的含量变化；通过硫酸铵沉淀和透析法提取并纯化丹参多酚氧化酶（PPO），研究其酶学性质；采用超高效液相色谱法（UPLC）测定丹参中迷迭香酸、紫草酸和丹酚酸B的含量。结果表明，新鲜丹参用1 mol·L⁻¹ HCl溶液或1 mol·L⁻¹ HCl甲醇溶液匀浆处理时，酚酸类成分含量最高；不同干燥速率的丹参中酚酸类成分含量差异不显著，表明酚酸类成分含量与干燥速率无关。丹参PPO的最适pH为7.6，最适温度为40℃；以邻苯二酚为底物时，丹参PPO的酶促褐变反应符合米氏方程，米氏常数K_m为0.12 mol·L⁻¹，最大反应速度V_(max)为588.23 U·min⁻¹。柠檬酸、乙二胺四乙酸二钠、抗坏血酸和亚硫酸钠对丹参PPO的抑制作用随抑制剂浓度的增加而增强，其中亚硫酸钠的抑制作用最强。本研究证明新鲜丹参中存在大量酚酸类成分，它们是丹参生长过程中原始积累的次生代谢产物，而非采后干燥和脱水胁迫诱导产生的。本研究对丹参的栽培、采收及加工具有参考价值和意义。
