响应面法优化黄芪总黄酮提取工艺及初步纯化
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摘要
黄芪总黄酮具有抗氧化、增强机体免疫等功能,常作为免疫增强剂添加在食品和饲料中。通过单因素试验研究乙醇体积分数、料液比和提取时间对黄芪总黄酮提取量的影响,进一步用Box–Behnken法优化黄芪总黄酮最佳提取工艺,并利用大孔吸附树脂–聚酰胺联用技术对其进行富集纯化。结果表明:在乙醇体积分数70%,料液比1∶17,提取时间70 min的条件下提取效果最佳,总黄酮提取量为2.78 mg·g~(–1),相对标准偏差为1.41%。经HPD722大孔吸附树脂–聚酰胺联用富集纯化后,黄芪总黄酮提取量为310.63 mg·g~(–1),增加112倍,其中毛蕊异黄酮含量由0.13 mg·g~(–1)增至42.56 mg·g~(–1),增加327倍;芒柄花素含量由0.06 mg·g~(–1)增至7.89 mg·g~(–1),增加132倍。
Flavonoids in astragali radix exert biological health functions actions such as antioxidant and immune-enhancing effects. They are often added to food and feed as immunopotentiators. The effects of ethanol concentration, the solid/liquid ratio, and extraction time on the extraction content of total flavonoids(TFA) from Astragalus membranaceus were evaluated using single factor experiments. Furthermore, the optimum beneficiation and depuration technology of TFA was investigated with HPD722 macroporous resin-polyamide combination method. The results showed that the extraction content of TFA and relative standard deviation(RSD) were 2.78 mg·g~(–1) and 1.41%, respectively, under the conditions of 70% ethanol concentration, solid/liquid ratio of 1∶17, and an extraction time of 70 min. When TFA were enriched and purified using HPD722 macroporous resin-polyamide, the extraction content of TFA was 311 mg·g~(–1) and its purity was increased 112 times. As the main component of TFA, the content of calycosin was increased 327 times from 0.13 mg·g~(–1) to 42.56 mg·g~(–1) and formononetin was increased 132 times from 0.06 mg·g~(–1) to 7.89 mg·g~(–1).
Flavonoids in astragali radix exert biological health functions actions such as antioxidant and immune-enhancing effects. They are often added to food and feed as immunopotentiators. The effects of ethanol concentration, the solid/liquid ratio, and extraction time on the extraction content of total flavonoids(TFA) from Astragalus membranaceus were evaluated using single factor experiments. Furthermore, the optimum beneficiation and depuration technology of TFA was investigated with HPD722 macroporous resin-polyamide combination method. The results showed that the extraction content of TFA and relative standard deviation(RSD) were 2.78 mg·g~(–1) and 1.41%, respectively, under the conditions of 70% ethanol concentration, solid/liquid ratio of 1∶17, and an extraction time of 70 min. When TFA were enriched and purified using HPD722 macroporous resin-polyamide, the extraction content of TFA was 311 mg·g~(–1) and its purity was increased 112 times. As the main component of TFA, the content of calycosin was increased 327 times from 0.13 mg·g~(–1) to 42.56 mg·g~(–1) and formononetin was increased 132 times from 0.06 mg·g~(–1) to 7.89 mg·g~(–1).
引文
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[20]陈红梅,谢翎.响应面法优化半枝莲黄酮提取工艺及体外抗氧化性分析.食品科学,2016,37(2):45-50.CHEN H M,XIE L.Optimization of extraction process for flavonoid from Soutellaria barbata by response surface methodology and evaluation of its antioxidant activity.Food Science,2016,37(2):45-50.
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[2]刘月涛,贾璐,秦雪梅.多效黄芪物质基础的研究进展.中草药,2018,49(6):1476-1480.LIU Y T,JIA L,QIN X M.Research advances of material basis of multi-effect Astragali radix.Chinese Traditional and Herbal Drugs,2018,49(6):1476-1480.
[3]游庭活,刘凡,温露,邹宇晓,廖森泰.黄酮类化合物抑菌作用研究进展.中国中药杂志,2013,38(21):3645-3649.YOU T H,LIU F,WEN L,ZOU Y X,LIAO S T.Advance in studies on antibacterial effect of flavonoids.China Journal of Chinese Materia Medica,2013,38(21):3645-3649.
[4]刘少静,盛耀光,马晓娇,万凤凤,杨黎彬.黄芪中总黄酮的提取纯化工艺研究.化学工程师,2018(5):9-12.LIU S J,SHENG Y G,MA X J,WAN F F,YANG L B.Optimization of the extraction and purification technologies of total flavonoids from Radix astragali.Chemical Engineer,2018(5):9-12.
[5]余小翠,刘高峰.响应面分析法在中药提取和制备工艺中的应用.中药材,2010,33(10):1651-1655.YU X C,LIU G F.Application of response surface analysis in extraction and preparation of traditional Chinese medicine.Journal of Chinese Medicinal Materials,2010,33(10):1651-1655.
[6]陈况况,帕塔尔·尼牙孜,章宏慧,刘东红,叶兴乾.响应面法优化水芹黄酮提取工艺及其成分研究.中国食品学报,2014,14(11):83-89.CHEN K K,Pataer·Niyazi,ZHANG H H,LIU D H,YE X Q.Optimization of flavonoids extration from Oenanthe javanica by response surface methodology and the study on chemical compositions.Journal of Chinese Institute of Food Science and Technology,2014,14(11):83-89.
[7]李静,张朝辉,段筱杉,应锐.聚酰胺-大孔树脂联用纯化大叶藻黄酮.食品科技,2016,41(3):203-207.LI J,ZHANG Z H,DUAN X S,YING R.Purifcation of flavonoids from Zostera marina L.with polyamide-macroporous resin.Food Science and Technology,2016,41(3):203-207.
[8]王冬,关宏峰,刘晓秋.中药中重金属和残留农药去除方法研究进展.沈阳药科大学学报,2009,26(2):152-155.WANG D,GUAN H F,LIU X Q.Research progress on techniques of removing heavy metals and pesticides from traditional Chinese medicine.Journal of Shenyang Pharmaceutical University,2009,26(2):152-155.
[9]张蕾,廖琼峰,谢智勇,冯志强,曾元儿.聚酰胺-大孔树脂联用富集苦参总黄酮.中草药,2011,42(10):1968-1972.ZHANG L,LIAO Q F,XIE Z Y,FENG Z Q,ZENG Y E.Enrichment of total flavonoids from Sophora flavescens with polyamidemacroporous resin.Chinese Traditional and Herbal Drugs,2011,42(10):1968-1972.
[10]刘鄂湖,鄢丹,蔡光明,肖小河,夏新华.大孔吸附树脂技术在中药中应用现存问题分析与探讨.中草药,2007,38(5):I0003-I0005.LIU E H,YAN D,CAI G M,XIAO X H,XIA X H.Issues analysis and discussion on application of macroporous adsorbing resins in traditonal Chinese medicine.Chinese Traditional and Herbal Drugs,2007,38(5):I0003-I0005.
[11]张俊,李进,吕海英,马雪.响应面法优化扁桃果皮熊果酸的纯化工艺.食品与发酵工业,2017,43(10):170-176.ZHANG J,LI J,LYU H Y,MA X.Optimization of purification process for ursolic acid from Amygdalus communis L.peel by response surface methodology.Food and Fermentation Industries,2017,43(10):170-176.
[12]ZHANG J,HAYAT K,ZHANG X M,ZHANG X M,TONG J M,XIA S Q.Separation and purification of flavonoid from ginkgo extract by polyamide resin.Separation Science and Technology,2010,45(16):2413-2419.
[13]金慧鸣,郭红英,谭兴和,刘梦浩,王锋,蔡文,张喻,邓洁红.大孔吸附树脂-聚酰胺联用纯化茅岩莓总黄酮工艺优化.食品科学,2016,37(12):13-18.JIN H M,GUO H Y,TAN X H,LIU M H,WANG F,CAI W,ZHANG Y,DENG J H.Purification of total flavonoids from Ampelopsis grossedentata by combined use of macroporous adsorption resin and polyamide.Food Science,2016,37(12):13-18.
[14]林丽,杨扶德,李硕,张毅.甘肃7个不同产地蒙古黄芪质量的比较研究.湖南中医药大学学报,2009,29(4):24-26.LIN L,YANG F D,LI S,ZHANG Y.Quality comparison of Radix astragali from seven regions of Gansu.Journal of Traditional Chinese Medicine University of Hunan,2009,29(4):24-26.
[15]包强,刘丽梅,王瑞海,叶迎,柏冬,刘卫东,田生华.甘肃不同产地黄芪和红芪药材中总黄酮含量对比研究.中药材,2016(10):2281-2284.BAO Q,LIU L M,WANG R H,YE Y,BAI D,LIU W D,TIAN S H.Comparative study on total flavonoid content between Radix astragali and Radix hedysari from different regions of Gansu.Journal of Chinese Medicinal Materials,2016(10):2281-2284.
[16]白生文,汤超,田京,闫宏斐,许晓莎,范惠玲.沙棘果渣总黄酮提取工艺及抗氧化活性分析.食品科学,2015,36(10):59-64.BAI S W,TANG C,TIAN J,YAN H F,XU X S,FAN H L.Extraction and antioxidant activity of total flavonoids from sea buckthorn pomace.Food Science,2015,36(10):59-64.
[17]吴博,刘春梅,刘冰,赵旭,毕开顺,贾英.益智总黄酮大孔吸附树脂纯化工艺研究.中草药,2015,46(9):1321-1325.WU B,LIU C M,LIU B,ZHAO X,BI K S,JIA Y.Purification technology for flavone from Alpiniae oxyphyllae Fructus with macroporous resins.Chinese Traditional and Herbal Drugs,2015,46(9):1321-1325.
[18]林春梅.聚酰胺层析法分离纯化牛蒡叶中的总黄酮.食品与发酵工业,2014,40(2):236-238.LIN C M.Separation and purification of total flavones in burdock leaves by polyamide column.Food and Fermentation Industries,2014,40(2):236-238.
[19]李涛,刘晓璠,杨小兰.啤酒花总黄酮测定方法的比较研究.食品科学,2014,35(18):89-92.LI T,LIU X F,YANG X L.Comparative study on assays for determination of flavonoids in hops(Humulus lupulus L.).Food Science,2014,35(18):89-92.
[20]陈红梅,谢翎.响应面法优化半枝莲黄酮提取工艺及体外抗氧化性分析.食品科学,2016,37(2):45-50.CHEN H M,XIE L.Optimization of extraction process for flavonoid from Soutellaria barbata by response surface methodology and evaluation of its antioxidant activity.Food Science,2016,37(2):45-50.
[21]刘玉芬,夏海涛.响应面法优化碱蒿黄酮提取工艺及其体外抗氧化作用.食品科学,2012,33(12):63-68.LIU Y F,XIA H T.Optimization of extraction process for flavonoids from Artemisia anethifolia Web.ex Stechm by response surface methodology and evaluation of their antioxidant activity.Food Science,2012,33(12):63-68.
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