藜麦麸皮不同极性部位的抑菌及酪氨酸酶抑制活性研究
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摘要
以抑菌圈大小及对酪氨酸酶活性的抑制率为指标,筛选出藜麦麸皮中具有抑菌活性以及抑制酪氨酸酶活性的极性部位。采用75%的乙醇对藜麦麸皮进行超声提取,三种不同极性溶剂石油醚、乙酸乙酯、正丁醇进行萃取,获取四个极性部位。纸片法观察不同的极性部位对于四种致病菌的体外抑制效果,同时探究对酪氨酸酶活性的抑制效果,利用液质推断其有效成分并用高效液相对高活性成分进行定量分析。结果表明:正丁醇萃取层的活性最高,且对革兰阳性菌较为敏感但对革兰阴性菌无明显效果。对金黄色葡萄球菌的最低抑菌浓度和最低杀菌浓度分别为1.04、2.08 mg/mL,对表皮葡萄球菌的最低抑菌浓度和最低杀菌浓度分别为0.52、1.04 mg/mL。比较可知藜麦麸皮极性部位的抑菌效果要优于某些常见的中药;在浓度为1 mg/mL时对酪氨酸酶活性的抑制率为55.86%,是同浓度VC抑制效果的58.96%;液质推断出正丁醇层主要物质为藜麦皂苷,定量分析藜麦皂苷的纯度为62.6%。综上,藜麦麸皮不同极性部位中,正丁醇萃取层的抑菌活性以及对于酪氨酸酶活性的抑制效果较好,且主要活性物质为藜麦皂苷。
The bacteriostatic activity and inhibition of tyrosinase activity of different polar sites in quinoa bran was studied with the target of the inhibition zone size and inhibition rate of tyrosinase activity.Different polar sites in quinoa bran were obtained by ultrasonic extraction method with 75% ethanol. In order to screen out the four polar sites,extracting sequentially with petroleum ether,ethyl acetate,n-butanol three different solvents.Paper disk method was used to observe the in vitro inhibitory of different polar sites on four pathogenic bacterias.And the inhibitory effect of tyrosinase was valued according to the inhibition rate of tyrosinase activity.Using LC-MS/MS to infer its active ingredients and using high performance liquid chromatography quantitative analysis of relatively high active ingredients.The results showed that the n-butyl alcohol extracts showed it had high bacteriostatic activity on Gram-positive bacterias,and no significant effect on gram-negative bacterias.The minimum inhibition concentration (MIC) on Staphylococcus aureus and Staphylococcus epidermidis were 1.04,0.52 mg/mL,respectively. And the minimum bactericidal concentration (MBC) were 2.08,1.04 mg/mL,respectively.By comparing the antibacterial effect,it could be concluded that the polar sites of quinoa bran was superior to the some common traditional Chinese medicine. When the concentration of n-butyl alcohol fraction was 1 mg/mL,its inhibition rate of tyrosinase activity was 55.86%,and its inhibition ability was 58.96% with that of vitamin C. Through full scan by HPLC,the main substance of n-butyl alcohol fraction was deducted as quinoa saponin,and the purity was 62.6%.In summary,N-butyl alcohol fraction has obvious antibacterial activity and better inhibition of tyrosinase activity in the different polar sites of quinoa bran.And the main active substance is quinoa saponin.
The bacteriostatic activity and inhibition of tyrosinase activity of different polar sites in quinoa bran was studied with the target of the inhibition zone size and inhibition rate of tyrosinase activity.Different polar sites in quinoa bran were obtained by ultrasonic extraction method with 75% ethanol. In order to screen out the four polar sites,extracting sequentially with petroleum ether,ethyl acetate,n-butanol three different solvents.Paper disk method was used to observe the in vitro inhibitory of different polar sites on four pathogenic bacterias.And the inhibitory effect of tyrosinase was valued according to the inhibition rate of tyrosinase activity.Using LC-MS/MS to infer its active ingredients and using high performance liquid chromatography quantitative analysis of relatively high active ingredients.The results showed that the n-butyl alcohol extracts showed it had high bacteriostatic activity on Gram-positive bacterias,and no significant effect on gram-negative bacterias.The minimum inhibition concentration (MIC) on Staphylococcus aureus and Staphylococcus epidermidis were 1.04,0.52 mg/mL,respectively. And the minimum bactericidal concentration (MBC) were 2.08,1.04 mg/mL,respectively.By comparing the antibacterial effect,it could be concluded that the polar sites of quinoa bran was superior to the some common traditional Chinese medicine. When the concentration of n-butyl alcohol fraction was 1 mg/mL,its inhibition rate of tyrosinase activity was 55.86%,and its inhibition ability was 58.96% with that of vitamin C. Through full scan by HPLC,the main substance of n-butyl alcohol fraction was deducted as quinoa saponin,and the purity was 62.6%.In summary,N-butyl alcohol fraction has obvious antibacterial activity and better inhibition of tyrosinase activity in the different polar sites of quinoa bran.And the main active substance is quinoa saponin.
引文
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[9]孙雪婷,袁俊杰,蒋玉蓉,等.藜麦种子总黄酮提取及其抗氧化性[J].江苏农业科学,2015,43(10):355-358.
[10]任卓伟,倪文杰,刘森.藜麦皂甙的测定研究[J].山西农业科学,2015,43(8):932-935.
[11]刘伟锐,张霞,姜蕊,等.紫外可见分光光度发测定苣荬菜根部总三萜皂苷的含量[J].西北药学杂志,2014,29(6):554-557.
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[13]董飞,郭晓农.藜麦种子总黄酮的提取及体外抑菌作用[J].甘肃农业科技,2018(4):14-18.
[14]梁霞,周柏玲,刘森,等.响应面法优化藜麦总皂苷提取工艺研究[J].中国粮油学报,2017,32(11):40-46.
[15]熊成文,李晓伟,徐得娟,等.分光光度法测定藜麦中总皂苷的含量[J].安徽农业科学,2017,45(26):96-98.
[16]Filho A M,Pirozi M R,Borges J T,et al.Quinoa:Nutritional,functional,and antinutritional aspects[J].C R C Critical Reviews in Food Technology,2017,57(8):1618-1630.
[17]魏爱春,杨修仕,么杨,等.藜麦营养功能成分及生物活性研究进展[J].食品科学,2015,36(15):272-276.
[18]许效群,赵文婷,苗玲香,等.藜麦麸皮总皂苷的提取纯化工艺研究[J].食品工业科技,2017,38(18):215-220.
[19]杨洁,高凤祥,杨敏,等.藜麦皮总皂苷微波辅助提取工艺及其抗氧化活性研究[J].食品与机械,2017(12).148-153.
[20]李溯,丁劲松.黑色素生物合成与酪氨酸酶抑制剂的研究进展[J].中南药学,2013,11(4):278-282.
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[22]Tiwatt K,Piyanut T,Wa Lee C,et al.Triterpene saponins from Chenpodium quinoa Willd[J].Phytochemistry,2008(69):1919-1926.
[23]Xue P,Yao Y,Yang X S,et al.Improved antimicrobial effect of ginseng extract by heat transformation[J].Journal of Ginseng Research,2017,41(2):180-187.
[24]Nicke L J,Spanier L P,Bote Lho F T,et al.Effect of different types of processing on the total phenolic compound content,antioxidant capacity,and saponin content of Chenopodium quinoa Willd grains[J].Food Chemistry,2016,209:139-143.
[25]高立新.HPLC-UV法测定商陆中商陆皂苷甲的含量[J].中国临床药理学杂志,2015(20):2052-2054.
[26]魏娜,王琪,官杰,等.黎药角花胡颓子提取物不同极性部位抗菌活性研究[J].中国医药导报,2012,09(14):35-36.
[27]安惠霞,斯拉甫·艾白,古力娜·达吾提,等.地锦草不同萃取部位体外抗真菌作用研究[J].中国中医药信息杂志,2010,17(1):40-42.
[28]刘计权,谢树莲,李洋洋,等.绵马贯众不同萃取部位的抑菌活性[J].中国实验方剂学杂志,2016(3):137-142.
[29]韩毅丽,张娟娟,张淑蓉.米口袋不同萃取部位体外抑菌活性研究[J].山西中医学院学报,2015,16(6):21-23.
[30]黄梦姣,卢添林,王瑶,等.粗壮女贞不同萃取部位抑菌活性成分研究[J].现代预防医学,2016,43(5):864-866,902
[31]杜静婷.藜麦种皮皂苷的提取、纯化、抗氧化、抑菌及皂苷元成分鉴定[D].太原:山西大学,2017.
[32]徐晓敏.藜麦皂苷的提取、分离纯化以及生物活性研究[D].呼和浩特:内蒙古农业大学,2015.
[33]王建华,雷帆.20种中药对酪氨酸酶抑制作用的研究[J].中国药学杂志,2000,35(4):232-234.
[34]李俊强,李星辰,刘晓静.甘草总皂苷生物转化与抑制酪氨酸酶活性研究[J].中国药事,2018(2):234-241.
[35]Madl T,Sterk H,Mittelbach M,et al.Tandem mass spectrometric analysis of a complex triterpene saponin mixture o Chenopodium quinoa[J].Journal of the American Society for Mass Spectrometry,2006,17(6):795-806.
[2]赵文婷.藜麦麸皮总皂苷的提取纯化及其抗氧化和免疫增强作用[D].太原:山西农业大学,2015.
[3]兀浩.表面活性剂提取薯蓣皂苷及薯蓣皂苷元水解工艺的研究[D].西安:陕西科技大学,2013.
[4]Jarvis,David E,Ho Y S,et al.The genome of Chenopodium quinoa[J].Nature,2017,542(7641):307-312.
[5]Oelke E A,Putnam D H,Teynor T M,et al.Quinoa Alternative Field Crops Manual[J].University of Wisconsin-Extension accessed,2012,21:4-16.
[6]申瑞玲,张文杰,董吉林,等.藜麦的主要营养成分、矿物元素及植物化学物质含量测定[J].郑州轻工业学院学报:自然科学版,2015,30(5):17-21.
[7]董晶,张焱,曹赵茹,等.藜麦总黄酮的超声波法提取及抗氧化活性[J].江苏农业科学,2015,43(4):267-269.
[8]董施彬,宁亚萍,杨喆,等.藜麦总黄酮提取及大孔树脂纯化工艺的研究[J].食品工业科技,2015,36(16):272-278.
[9]孙雪婷,袁俊杰,蒋玉蓉,等.藜麦种子总黄酮提取及其抗氧化性[J].江苏农业科学,2015,43(10):355-358.
[10]任卓伟,倪文杰,刘森.藜麦皂甙的测定研究[J].山西农业科学,2015,43(8):932-935.
[11]刘伟锐,张霞,姜蕊,等.紫外可见分光光度发测定苣荬菜根部总三萜皂苷的含量[J].西北药学杂志,2014,29(6):554-557.
[12]Brown D G,Lister T,May-dracka T L,et al.New natural products as new Leads for antibacterial drug discovery[J].Bioorg Med Chem Lett,2014,24(2):413-418.
[13]董飞,郭晓农.藜麦种子总黄酮的提取及体外抑菌作用[J].甘肃农业科技,2018(4):14-18.
[14]梁霞,周柏玲,刘森,等.响应面法优化藜麦总皂苷提取工艺研究[J].中国粮油学报,2017,32(11):40-46.
[15]熊成文,李晓伟,徐得娟,等.分光光度法测定藜麦中总皂苷的含量[J].安徽农业科学,2017,45(26):96-98.
[16]Filho A M,Pirozi M R,Borges J T,et al.Quinoa:Nutritional,functional,and antinutritional aspects[J].C R C Critical Reviews in Food Technology,2017,57(8):1618-1630.
[17]魏爱春,杨修仕,么杨,等.藜麦营养功能成分及生物活性研究进展[J].食品科学,2015,36(15):272-276.
[18]许效群,赵文婷,苗玲香,等.藜麦麸皮总皂苷的提取纯化工艺研究[J].食品工业科技,2017,38(18):215-220.
[19]杨洁,高凤祥,杨敏,等.藜麦皮总皂苷微波辅助提取工艺及其抗氧化活性研究[J].食品与机械,2017(12).148-153.
[20]李溯,丁劲松.黑色素生物合成与酪氨酸酶抑制剂的研究进展[J].中南药学,2013,11(4):278-282.
[21]Medina-Meza I G,Aluwi N A,Saunders S R,et al.Profiling of Triterpenoid Saponins from 28 Quinoa Varieties(Chenopodium quinoa Willd)grown in Washington State by GC-MS[J].Journal Agricultural Food Chemistry,2016,64(45):8583-8591.
[22]Tiwatt K,Piyanut T,Wa Lee C,et al.Triterpene saponins from Chenpodium quinoa Willd[J].Phytochemistry,2008(69):1919-1926.
[23]Xue P,Yao Y,Yang X S,et al.Improved antimicrobial effect of ginseng extract by heat transformation[J].Journal of Ginseng Research,2017,41(2):180-187.
[24]Nicke L J,Spanier L P,Bote Lho F T,et al.Effect of different types of processing on the total phenolic compound content,antioxidant capacity,and saponin content of Chenopodium quinoa Willd grains[J].Food Chemistry,2016,209:139-143.
[25]高立新.HPLC-UV法测定商陆中商陆皂苷甲的含量[J].中国临床药理学杂志,2015(20):2052-2054.
[26]魏娜,王琪,官杰,等.黎药角花胡颓子提取物不同极性部位抗菌活性研究[J].中国医药导报,2012,09(14):35-36.
[27]安惠霞,斯拉甫·艾白,古力娜·达吾提,等.地锦草不同萃取部位体外抗真菌作用研究[J].中国中医药信息杂志,2010,17(1):40-42.
[28]刘计权,谢树莲,李洋洋,等.绵马贯众不同萃取部位的抑菌活性[J].中国实验方剂学杂志,2016(3):137-142.
[29]韩毅丽,张娟娟,张淑蓉.米口袋不同萃取部位体外抑菌活性研究[J].山西中医学院学报,2015,16(6):21-23.
[30]黄梦姣,卢添林,王瑶,等.粗壮女贞不同萃取部位抑菌活性成分研究[J].现代预防医学,2016,43(5):864-866,902
[31]杜静婷.藜麦种皮皂苷的提取、纯化、抗氧化、抑菌及皂苷元成分鉴定[D].太原:山西大学,2017.
[32]徐晓敏.藜麦皂苷的提取、分离纯化以及生物活性研究[D].呼和浩特:内蒙古农业大学,2015.
[33]王建华,雷帆.20种中药对酪氨酸酶抑制作用的研究[J].中国药学杂志,2000,35(4):232-234.
[34]李俊强,李星辰,刘晓静.甘草总皂苷生物转化与抑制酪氨酸酶活性研究[J].中国药事,2018(2):234-241.
[35]Madl T,Sterk H,Mittelbach M,et al.Tandem mass spectrometric analysis of a complex triterpene saponin mixture o Chenopodium quinoa[J].Journal of the American Society for Mass Spectrometry,2006,17(6):795-806.