丹参茎叶提取物抗氧化活性物质基础与量效关系研究

目的以7月份和12月份采收丹参茎叶的水提物和醇提物为研究对象,评价丹参茎叶提取物的抗氧化效应,并探讨丹参茎叶提取物抗氧化活性的物质基础。方法采用超高效液相-三重四级杆质谱联用技术(UPLC-TQ/MS)对各提取物中化学成分进行定性定量分析,明确丹参茎叶提取物中主要丹酚酸类化合物(丹参素、咖啡酸、迷迭香酸和丹酚酸B),基于1,1-二苯基-2-三硝基苦肼(DPPH)、2,2-联氮-二(3-乙基-苯并噻唑-6-磺酸)二铵盐(ABTS)自由基清除法和铁还原/抗氧化能力(FRAP)法对丹参茎叶提取物进行抗氧化活性评价,同时以市售丹参(根及根茎)样品作对照。结果 7月份采收丹参茎叶水提物(SY-7)抗氧化活性最强,且总酚酸的量最高(75.663 mg/g),其次为7月份采收丹参茎叶醇提物(CY-7),市售丹参提取物抗氧化活性及总酚酸的量均低于7月份采收丹参茎叶提取物。丹酚酸单体化合物丹参素、咖啡酸、迷迭香酸和丹酚酸B均具有明显抗氧化、清除自由基能力,且表现出明显的量效关系。结论丹参茎叶水提物和醇提物具有较强的体外抗氧化活性,且含有丰富的丹酚酸类化合物,其中含有的丹参素、咖啡酸、迷迭香酸和丹酚酸B具有明显抗氧化活性。

Study on chemical constituents and dose-effect correlation of anti-oxidative activities among extracts of stems and leaves of Salvia miltiorrhiza

Objective To evaluate the anti-oxidative effect and chemical constituents of stems and leaves of Salvia miltiorrhiza (SMSL) collected in July and December, water extracts and alcohol extracts of SMSL collected in July and December were taken as the subject, therefore provide scientific basis for the comprehensive development and utilization of SMSL. Methods The chemical constituents in the extracts were identified and determined by Ultra-high performance liquid chromatography combined with triple quadrupole electrospray tandem mass spectrometry (UPLC-TQ/MS), then confirmed the main salvianolic acids (danshensu, caffeic acid, rosmarinic acid, and salvianolic acid B) as the material basis of anti-oxidant activity of SMSL. Moreover, based on the anti-oxidant activity evaluation index:1,1-diphenyl-2-picryl-hydrazyl (DPPH), 2,2''-azinobis-(3-ethylbenzthiazoline-6-sulphonate) (ABTS) free radical scavenging and iron reduction/anti-oxidant capacity (FRAP), anti-oxidant activity of SMSL was evaluated. Meanwhile Salvia Miltiorrhiza Radix et Rhizoma (SM) from market was used to be control. Results It showed that the water extracts of SMSL in July possessed strong anti-oxidant activities, and the total salvianolic acids with the content of 75.663 mg/g was the highest; Followed by the alcohol extracts of SMSL in July, anti-oxidant activity and total phenolic acid contents of SM extracts were all lower than that of SMSL in July. Danshensu, caffeic acid, rosmarinic acid, and salvianolic acid B showed obvious anti-oxidative activities and significant dose-dependent effect in scavenging free radicals. Conclusion It revealed that SMSL possessed strong in vitro anti-oxidant activity. Additonally, it is shown that SMSL was rich in salvianolic acids, in which danshensu, caffeic acid, rosmarinic acid, and salvianolic acid B also had obvious anti-oxidant activity.