采用超声波辅助法提取杜仲叶茯砖茶中的绿原酸,利用高效液相色谱法(HPLC)测定绿原酸含量,优化提取工艺,并对其体外降血糖抗氧化活性进行了研究。结果表明,杜仲叶茯砖茶绿原酸最佳提取工艺为:超声功率90 W,乙醇体积分数39%,提取时间50 min,液料比25∶1(mL∶g),提取温度60 ℃,在该最佳条件下,杜仲叶茯砖茶绿原酸得率为2.446 1%,与模型预测值基本吻合,表明该提取工艺稳定可行。体外降血糖实验发现,1.4 mg/mL杜仲叶茯砖茶绿原酸对α-胰淀粉酶的抑制率为83.26%,0.18 mg/mL杜仲叶茯砖茶绿原酸对α-葡萄糖苷酶的抑制率为96.80%,体现了较好的降血糖活性。抗氧化结果表明,当杜仲叶茯砖茶绿原酸质量浓度为24 μg/mL时,对DPPH自由基的清除率可达97.94%,相同质量浓度Vc对DPPH自由基的清除率为96.90%,略低于绿原酸,当杜仲叶茯砖茶绿原酸质量浓度为0.28 mg/mL时,其还原力为1.75,显著高于Vc,说明杜仲叶茯砖茶绿原酸抗氧化活性较好。
The eucommia ulmoides leaves Fu brick tea was used to extracted chlorogenic acid by ultrasonic-assisted method,the content of chlorogenic acid was determined by HPLC. On the basis of the single factor experiment,response surface methodology was used to optimize the extraction process,and evaluate the hypoglycemic and antioxidant activity in vitro. The results showed that the optimum condition were as follows:ultrasonic power 90 W,ethanol concentration 39%,extraction time 50 min, ratio of liquid to solid 25∶1 (mL∶g),and extraction temperature 60 ℃, under the best condition, the yield of eucommia ulmoides leaves Fu brick tea chlorogenic acid was 2.446 1%,and which was consistent with the predicted results,it showed that the extraction process is stable and feasible.When the eucommia ulmoides leaves Fu brick tea chlorogenic acid concentration was 1.4 mg/mL,the inhibition rate to α-pancreatic amylase was 83.26%, and when the eucommia ulmoides leaves fu brick tea chlorogenic acid concentration was 0.18 mg/mL, the inhibition rate to α-glucosidase was 96.80%.When the eucommia ulmoides leaves fu brick tea chlorogenic acid concentration was 24 μg/mL,the rate of scavenging DPPH radicals was 97.94% ,while the same concentration of Vc on the scavenging DPPH radicals was 96.90%,slightly lower than chlorogenic acid.and when the eucommia ulmoides leaves fu brick tea chlorogenic acid concentration was 0.28 mg/mL, the reducing power reached to 1.75,significantly higher than Vc.The results shows eucommia ulmoides leaves fu brick tea chlorogenic acid has good effect on hypoglycemic and antioxidant activity.
[1] 李旭,刘停,陈时建,等.杜仲叶绿原酸提取工艺优化及对自发性高血压大鼠的降压作用[J].食品科学,2013,34(14):30-34.
[2] 国家药典委员会,中华人民共和国药典[M].北京:中国医药科技出版社, 2015,166-166.
[3] MOCAN A,VLASE L,ARSENE A L,et al. HPLC/MS analysis of caffeic and chlorogenic acids from three romanian veronica species and their antioxidant and antimicrobial properties[J].Farmacia,2015,63(6):890-896.
[4] KABIR F,KATAYAMA S,TANJI N,et al.Antimicrobial effects of chlorogenic acid and related compounds[J].J Korean Soc Appl Biol Chem,2014,57(3):359-365.
[5] FUENTES E,CABALLERO J,ALARCON M,et al.Chlorogenic acid inhibits human platelet activation and thrombus formation[J]. Plos One,2014,9(3):e90699.
[6] STEFANELLO N,SCHMATZ R,PEREIRA B.L,et al.Effects of chlorogenic acid, caffeine, and coffee on behavioraland biochemical parameters of diabetic rats[J].Mol Cell Biochem,2014,388(1/2):277-286.
[7] CHO A S,JEON S M,KIM M J,et al.Chlorogenic acid exhibits anti-obesity property and improves lipid metabolism in high-fat diet-induced-obese mice[J].Food and Chemical Toxicology.2010,48(3):937-943.
[8] HAO B J,WU Y H,WANG J G,et al.Hepatoprotective and antiviral properties of isochlorogenic acid A from Laggera alata against hepatitis B virus infection[J].Journal of Ethnopharmacology,2012,144(1):190-194.
[9] LIU Y J,ZHOU C Y,QIU C H,et al.Chlorogenic acid induced apoptosis and inhibition of proliferation in human acute promyelocytic leukemia HL-60 cells[J].Molecular Medicine Reports,2013,8(4):1 106-1 110.
[10] YAN Y,LI J,HAN J,et al.Chlorogenic acid enhances the effects of 5-fluorouracil in human hepatocellular carcinoma cells through the inhibition of extracellular signal-regulated kinases[J].Anti-Cancer Drugs,2015,26(5):540-546.
[11] 艾光丽,曾桢,王维,等.金银花提取物的质量分析方法评价[J].中国实验方剂学杂志,2017,23(5):22-26.
[12] 杨秀芳,翁昆,祝雅松,等.加工中添加天然物质对改善茯砖茶感官品质的研究初报[J].中国茶叶加工,2007(3):24-25.
[13] 曹永,赵谋明,赵甜甜,等.不同黑茶提取物功能性成分分析及活性评价[J].食品科学,2017,38(18):54-59.
[14] 侯爱香,李宗军.茯砖茶的化学成分及其功能特性研究进展[J].中国食物与营养,2016,22(6):63-67.
[15] 陈媛,唐铁鑫,陈金龙,等.高效率高纯度绿原酸制备工艺研究[J].天然产物研究与开发,2014,26:1 503-1 509.
[16] 叶琼仙,刘静,苗爱清,等.白叶单枞黑茶抗氧化及体外降血糖活性研究[J].食品工业科技,2014,35(16):153-157.
[17] 赖小燕,姜泽东,倪辉,等.茶花粉黄酮对α-葡萄糖苷酶抑制作用的研究[J].食品工业科技,2016,37(5):353-357.
[18] 展锐,邵金辉.大枣多糖抗氧化及抗炎活性的研究[J].现代食品科技,2017,33(12):1-6.
[19] 熊春华,周苏果,沈忱,等.响应面法优化提取菊米黄酮及抗氧化活性研究[J].中国食品学报,2014,14(7):118-123.
[20] 康娜,梁永锋. 茵陈中绿原酸提取工艺及抗氧化活性研究[J].中国中医药科技,2017,24(5):587-598.
[21] 宋佳敏,王鸿飞,孙朦,等.响应面法优化金蝉花多糖提取工艺及抗氧化活性分析[J].食品科学,2018,39(4):275-281.
