荒漠肉苁蓉多糖提取及纯化工艺优化
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摘要
该研究以新疆荒漠肉苁蓉为实验材料,通过单因素、正交实验探究水浸提法提取荒漠肉苁蓉多糖的最优工艺;通过Sevag法和酶法及两者联用探究其对蛋白去除和对多糖保留的效果,确定荒漠肉苁蓉多糖最适去蛋白法;通过AB-8大孔树脂法和活性炭法探究脱色和纯化方法,确定肉苁蓉多糖纯化的最佳工艺。结果表明,水浸提法对荒漠肉苁蓉多糖提取的最佳工艺为提取温度75℃,提取时间165 min,料液比1∶55,在此工艺下,荒漠肉苁蓉多糖提取率为18.40%;最适去蛋白工艺为温度50℃,时间为2 h,木瓜蛋白酶用量为0.2%,在此脱蛋白体系下,脱蛋白率可达31.40%,多糖的保留率可达96.00%;在荒漠肉苁蓉多糖脱色方面,活性炭脱色率为80.37%,多糖的回收率为77.05%,AB-8大孔树脂脱色率为86.43%,多糖的回收率为91.93%,表明宜用大孔树脂进行脱色;AB-8大孔树脂纯化工艺为:上样浓度取4 g·L~(-1),洗脱液的浓度为60%乙醇,流速1 mL·min~(-1),多糖粗提取液的纯度从67.70%升高至84.80%,纯化效果明显。
In this study, taking Cistanche deserticola in Xinjiang as the experimental material, the optimal process for extracting polysaccharides from C. deserticola with water extraction was studied by using single factor and orthogonal experiment. Its effects on protein removal and polysaccharides retaining were investigated by using Sevag, enzymatic method or combination of these two methods, so as to determine the optimal method for protein removal from polysaccharides of C. deserticola; the decolorization and purification methods such as macroporous resin of AB-8 and activated Carbon were used to determine the optimal process. The results showed that the extraction rate of polysaccharides from C. deserticola was 18.40% during the optimal process of the water extraction as follows: extraction temperature 75 ℃, extraction time 165 min and solid-liquid ratio 1∶55. The protein removal rate can reach 31.40% and polysaccharide retention rate can reach 96.00% under the optimal protein removal process: temperature 50 ℃, time 2 h, and papain dosage 0.2%. The decolorization rate of activated Carbon and macroporous resin called AB-8 was 80.37% and 86.43%, and the recovery rate of polysaccharides was 77.05% and 91.93%, respectively, suggesting that macroporous resin was more suitable for decoloration. Macroporous resin named AB-8 increased the purity of the polysaccharide crude extract from 67.70% to 84.80% under the following conditions: concentration of the sample 4 g·L~(-1), concentration of the eluent 60% ethanol, and the flow rate 1 mL·min~(-1), showing significant purification effect.
In this study, taking Cistanche deserticola in Xinjiang as the experimental material, the optimal process for extracting polysaccharides from C. deserticola with water extraction was studied by using single factor and orthogonal experiment. Its effects on protein removal and polysaccharides retaining were investigated by using Sevag, enzymatic method or combination of these two methods, so as to determine the optimal method for protein removal from polysaccharides of C. deserticola; the decolorization and purification methods such as macroporous resin of AB-8 and activated Carbon were used to determine the optimal process. The results showed that the extraction rate of polysaccharides from C. deserticola was 18.40% during the optimal process of the water extraction as follows: extraction temperature 75 ℃, extraction time 165 min and solid-liquid ratio 1∶55. The protein removal rate can reach 31.40% and polysaccharide retention rate can reach 96.00% under the optimal protein removal process: temperature 50 ℃, time 2 h, and papain dosage 0.2%. The decolorization rate of activated Carbon and macroporous resin called AB-8 was 80.37% and 86.43%, and the recovery rate of polysaccharides was 77.05% and 91.93%, respectively, suggesting that macroporous resin was more suitable for decoloration. Macroporous resin named AB-8 increased the purity of the polysaccharide crude extract from 67.70% to 84.80% under the following conditions: concentration of the sample 4 g·L~(-1), concentration of the eluent 60% ethanol, and the flow rate 1 mL·min~(-1), showing significant purification effect.
引文
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[23] 王奇志,王娜,龙瑞敏,等.费菜多糖脱蛋白方法及工艺研究[J].热带作物学报,2013,34(7):1384.
[24] 方积年,丁侃.天然药物——多糖的主要生物活性及分离纯化方法[J].中国天然药物,2007(5):338.
[25] 赵瑞希. 海藻多糖提取、纯化及其补体结合活性的研究[D].哈尔滨:哈尔滨工业大学,2015.
[26] 瞿明. 芦笋多糖提取分离、纯化、初步结构鉴定及其抗氧化活性研究[D].武汉:华中农业大学,2011.
[27] 芦金清, 刘尚桃. 吸附树脂法提取绞股蓝皂甙[J]. 中成药, 1992(4):2.
[28] 马振山. 大孔吸附树脂在药学领域中的研究应用[J]. 中成药, 1997(12):40.
[29] 杨波, 韩凤波. AB-8和D-101大孔吸附树脂分离纯化玉竹多糖[J]. 食品研究与开发, 2014(23):109.
[30] 舒祝明,蒋剑平,王建平,等.大孔树脂纯化胡柚皮总黄酮的工艺研究[J].中国现代应用药学,2017,34(5):649.
[31] 黄海波,刘华春,金朱明,等.银杏叶原花青素的制备及其抗氧化活性研究[J].中国现代应用药学,2016,33(6):686.
[32] 张瑜,杨健,秦振娴,等.大孔吸附树脂富集金银花中环烯醚萜苷类成分的工艺优选[J].中国实验方剂学杂志,2016,22(24):18.
[2] 陈飞,陈卓,邢雪飞,等.肉苁蓉的研究进展[J].药物评价研究,2013,36(6):469.
[3] 中国药典.一部[S]. 2010.
[4] 谭西,周欣,陈华国.植物多糖构效关系研究进展[J].中国中药杂志,2017,42(21):4104.
[5] 李媛,宋媛媛,张洪泉.肉苁蓉的化学成分及药理作用研究进展[J].中国野生植物资源,2010,29(1):7.
[6] 王丽波,程龙,徐雅琴,等.南瓜籽多糖热水提取工艺优化及其抗氧化活性[J].农业工程学报,2016,32(9):284.
[7] 赵芷芊,王敏,张志清.植物多糖的提取及抗氧化功效的研究进展[J].食品工业科技,2018,39(13):337.
[8] 徐灿,刁嘉茵,王淑美.银耳多糖的药理研究进展[J].今日药学,2018,28(3):207.
[9] 许春平,杨琛琛,郑坚强,等.植物叶多糖的提取和生物活性综述[J].食品研究与开发,2014,35(14):111.
[10] 肖道安. 肉苁蓉多糖脱蛋白工艺的优化[J]. 江苏农业科学, 2015, 43(7):314.
[11] 屈贺幂. 茯苓多糖提取纯化及指纹图谱研究[D]. 长沙:湖南大学, 2007.
[12] 高建德, 刘雄, 范凌云,等. 肉苁蓉酶解提取工艺及抗氧化活性研究[J]. 中成药, 2014, 36(12):2621.
[13] 张丽红,谢三都,徐芳,等.紫苏叶多糖活性炭脱色工艺优化[J].食品与机械,2015,31(3):224.
[14] 杨波,韩凤波,杨波.AB-8和D-101大孔吸附树脂分离纯化玉竹多糖[J].食品研究与开发,2014,35(23):67.
[15] 张锐, 曾冬云, 龚兴国,等. 羊栖菜多糖的提取工艺研究[J]. 中国食品学报, 2006, 6(3):14.
[16] 姜军. 山药多糖的分离纯化及其化学结构的初步研究[D].扬州:扬州大学,2007.
[17] 夏锦锦,王睿,祝缘,等.利用AB-8大孔树脂纯化‘黑宝石’李果实花色苷的研究[J].食品工业科技,2017,38(6):266.
[18] 成娟. 山药多糖提取工艺及质量控制研究[D].成都:西南交通大学,2014.
[19] 王媛媛,许兰淑,樊玮,等.山药多糖得率-活性综合优化提取工艺[J].食品安全质量检测学报,2015,6(6):1980.
[20] 龙菊,孙翠英,张红霞,等.正交设计与响应面法优化山药多糖提取工艺[J].食品研究与开发,2017,38(4):54.
[21] 郭咪咪. 紫薯多糖的分离纯化及抗氧化性研究[D].洛阳:河南科技大学,2013.
[22] 吕志琴. 沙棘多糖的提取、分离纯化工艺研究[D].石河子:石河子大学,2013.
[23] 王奇志,王娜,龙瑞敏,等.费菜多糖脱蛋白方法及工艺研究[J].热带作物学报,2013,34(7):1384.
[24] 方积年,丁侃.天然药物——多糖的主要生物活性及分离纯化方法[J].中国天然药物,2007(5):338.
[25] 赵瑞希. 海藻多糖提取、纯化及其补体结合活性的研究[D].哈尔滨:哈尔滨工业大学,2015.
[26] 瞿明. 芦笋多糖提取分离、纯化、初步结构鉴定及其抗氧化活性研究[D].武汉:华中农业大学,2011.
[27] 芦金清, 刘尚桃. 吸附树脂法提取绞股蓝皂甙[J]. 中成药, 1992(4):2.
[28] 马振山. 大孔吸附树脂在药学领域中的研究应用[J]. 中成药, 1997(12):40.
[29] 杨波, 韩凤波. AB-8和D-101大孔吸附树脂分离纯化玉竹多糖[J]. 食品研究与开发, 2014(23):109.
[30] 舒祝明,蒋剑平,王建平,等.大孔树脂纯化胡柚皮总黄酮的工艺研究[J].中国现代应用药学,2017,34(5):649.
[31] 黄海波,刘华春,金朱明,等.银杏叶原花青素的制备及其抗氧化活性研究[J].中国现代应用药学,2016,33(6):686.
[32] 张瑜,杨健,秦振娴,等.大孔吸附树脂富集金银花中环烯醚萜苷类成分的工艺优选[J].中国实验方剂学杂志,2016,22(24):18.