薄层生物自显影技术比较新疆2种洋甘菊抗氧化活性

Comparison of antioxidant activity between two species of chamomiles produced in Xinjiang by TLC-bioautography

目的:比较新疆2种洋甘菊——母菊和罗马洋甘菊抗氧化活性成分差异。方法:采用薄层-生物自显影技术,以1,1-二苯基苦基苯肼(1,1-diphenyl-2-picrylhydrazyl,DPPH)自由基为实验模型,经薄层扫描获得各抗氧化成分的峰面积,从而对2种洋甘菊的挥发油提取物、黄酮提取物进行抗氧化活性成分的分析,以总峰面积大小为指标比较不同提取物的抗氧化能力,并与2种洋甘菊黄酮提取物总的抗氧化活性进行比较。结果:母菊挥发油提取物显现烯炔双环醚等4个白色抗氧化斑点,罗马洋甘菊挥发油提取物显现1个白色抗氧化斑点,薄层扫描结果表明母菊挥发油提取物抗氧化斑点峰面积大于罗马洋甘菊挥发油提取物;2种洋甘菊黄酮提取物紫外-可见分光光度法抗氧化活性测定结果表明,母菊清除50%DPPH自由基的质量浓度为0.66 g·L-1,罗马洋甘菊清除50%DPPH自由基的质量浓度为0.33 g·L-1;2种洋甘菊黄酮提取物薄层生物自显影实验结果表明,母菊黄酮提取物显现芹菜素、芹苷元-7-葡萄糖苷等7个淡黄色抗氧化斑点,罗马洋甘菊黄酮提取物显现芹菜素、芹苷元-7-葡萄糖苷等8个淡黄色抗氧化斑点,薄层扫描结果表明罗马洋甘菊黄酮提取物抗氧化斑点峰面积大于母菊黄酮提取物。结论:2种洋甘菊挥发油提取物抗氧化活性TLC差异显著,母菊挥发油提取物抗氧化活性强于罗马洋甘菊挥发油提取物;母菊挥发油提取物中已知抗氧化活性成分为烯炔双环醚,其他3个抗氧化活性成分以及罗马洋甘菊挥发油提取物中1个抗氧化活性成分均为未知化合物,有待进一步确定;鉴于2种洋甘菊挥发油提取物抗氧化活性斑点数差异显著,可应用于2种洋甘菊的鉴别区分。2种洋甘菊黄酮提取物抗氧化活性测定结果、薄层生物自显影实验结果一致,母菊、罗马洋甘菊黄酮提取物都具有较强的抗氧化活性,且罗马洋甘菊黄酮提取物的抗氧化活性强于母菊黄酮提取物;2种洋甘菊黄酮提取物中均含已知抗氧化活性成分芹菜素、芹苷元-7-葡萄糖苷,母菊黄酮提取物中其他5个抗氧化活性成分以及罗马洋甘菊中其他6个抗氧化活性成分均为未知成分,有待进一步确定。该方法为进一步鉴定洋甘菊抗氧化活性成分奠定基础。

Objective： To compare the antioxidant active components from two species of ehamomile-matricaria and Roman chamomile produced in Xinjiang. Method： The TLC-bioautography was used, with 1,1-Diphenyl-2-picrylhydrazyl （DPPH） radical as the experimental model. The peak areas of various antioxidant components were obtained by TLC-scanning for analyzing antioxidant ac- tive components contained in volatile oil extracts and flavone extracts from the two species of chamomiles. The total peak area was taken as the indicator for comparing the antioxidant capacities of the two types of extracts, and comparing them with the total antioxidant ac- tivity of flavone extracts of the two species of chamomiles. Result ： According to the result of TLC-bioautography in volatile oil extracts from the two species of chamomiles, volatile oil extracts from chamomile showed four white antioxidant spots, including en-yne-dieycl- oether, and volatile oil extracts from Roman chamomile showed only one white antioxidant spot. The TLC-scanning result showed that the peak area of antioxidant spots of volatile oil extracts from chamomile was significantly larger than that of volatile oil extracts from Ro- man chamomile. According to the test on the antioxidant activity of the two species of chamomiles with ultraviolet-visible spectrophotom- etry, the concentration of chamomile after scavenging 50% of DPPH radicals was 0.66 g · L^- 1 , whereas the figure for Roman chamo- mile was 0.33 g · L^- 1. According to the result of TLC-bioautography in flavone extracts from the two species of chamomiles, fiavone extracts from chamonfile showed seven yellowish antioxidant spots, including apigenin and apigenin-7-glueoside, and flavone extracts of Roman chamomile showed eight yellowish antioxidant spots, including apigenin and apigenin-7-glueoside. The TLC-seanning results showed that the peak area of antioxidant spots of flavone extracts from Roman chamomile was significantly larger than that of flavone ex- tracts from chamomile. Conclusion： Volatile oil extracts from the two species of chamomiles have significant difference in the antioxi- dant activity in TLC-bioautography. Specifically, the antioxidant activity of volatile oil extracts from chamomile is stronger than volatile oil extracts from Roman chamomile; the known antioxidant active components in volatile oil extracts from chamomile is en-yne-dicycl- oether, while all of the other three antioxidant active components as well as antioxidant active components in volatile oil extracts from Roman chamomile are unknown components and remain to be further determined. Considering the significant difference in the number of antioxidant active spots in volatile oil extracts from the two species of chamomiles, the result can be applied to distinguish the two species of chamomiles. The antioxidant activity determination result for flavone extracts from two species of chamomiles was consistent with the result of TLC-bioautography, showing that flavone extracts from chamomile and Roman chamomile are more antioxidant active, while that of Roman chamomile is stronger than chamomile. Flavone extracts from both of the two species of chamomiles contain apige- nin and pigenin-7-glucoside, which are known, while all of the other five antioxidant active components contained in flavone extracts from chamomile and the other six antioxidant active components contained in flavone extracts from Roman chamomile are unknown and remain to be further identified. The method lays a foundation for further identification of antioxidant active components contained in chamomile.