CN110117232A - 葫芦巴籽中有效成分的提取方法 - Google Patents
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Abstract
本发明涉及葫芦巴籽中有效成分的提取方法，充分考虑了原料葫芦巴籽的特性和目标产物4‑羟基异亮氨酸的特性，经过系统的筛选，对提取工艺进行优化。本发明方法步骤简单，水使用量少，不使用有机溶剂，避免后续除杂工艺，降低了生产成本，在提高产品纯度的同时还提升了提取率，工艺中水可以循环使用，增加了资源利用率。
Description
技术领域
本发明涉及一种提取方法，更具体地，涉及葫芦巴籽中有效成分的提取方法。
背景技术
葫芦巴（拉丁学名：Trigonella foenum-graecum L.），是豆科胡卢巴属植物胡卢巴的习称，也作胡芦巴。一年生草本植物。高30-80厘米，根系发达，茎直立，羽状三出复叶，叶柄平展。花无梗，萼筒状，花冠黄白色或淡黄色，子房线形，荚果圆筒状，种子长圆状卵形。花期4-7月，果期7-9月。
分布于中国、地中海东岸、中东、伊朗高原以至喜马拉雅地区。中国南北各地均有栽培，在西南、西北各地呈半野生状态。多生于田间、路旁。
葫芦巴，种子入药，可补肾壮阳、祛痰除湿；嫩茎、叶可作蔬菜食用；茎、叶或种子晒干磨粉掺入面粉中蒸食作增香剂；干全草可驱除害虫；可作饲料。
一年生草本，高30-80厘米。主根深达土中80厘米，根系发达。茎直立，圆柱形，多分枝，微被柔毛。
羽状三出复叶；托叶全缘，膜质，基部与叶柄相连，先端渐尖，被毛；叶柄平展，长6-12毫米；小叶长倒卵形、卵形至长圆状披针形，近等大，长15-40毫米，宽4-15毫米，先端钝，基部楔形，边缘上半部具三角形尖齿，上面无毛，下面疏被柔毛，或秃净，侧脉5-6对，不明显；顶生小叶具较长的小叶柄。
花无梗，1-2朵着生叶腋，长13-18毫米；萼筒状，长7-8毫米，被长柔毛，萼齿披针形，锥尖，与萼等长；花冠黄白色或淡黄色，基部稍呈堇青色，旗瓣长倒卵形，先端深凹，明显地比冀瓣和龙骨瓣长；子房线形，微被柔毛，花柱短，柱头头状，胚珠多数。荚果圆筒状，长7-12厘米，径4-5毫米，直或稍弯曲，无毛或微被柔毛，先端具细长喙，喙长约2厘米（包括子房上部不育部分），背缝增厚，表面有明显的纵长网纹，有种子10-20粒。种子长圆状卵形，长3-5毫米，宽2-3毫米，棕褐色，表面凹凸不平。花期4-7月，果期7-9月。
适应性强，抗寒，生长迅速。葫芦巴喜冷凉干旱气候，对土壤、气候的适应性很强，在中国“三北”地区和中南等地均有栽培。中国葫芦巴的适生地区是宁夏、甘肃、青海、新疆、内蒙古等省区。葫芦巴提取物为豆科植物戎芦巴Trigonella foenum-graecum L.的种子提取物，具有温肾、祛寒、止痛等功效，用于肾脏虚冷、小腹冷痛、小肠疝气、寒湿脚气等症状。
作为治疗上述病症外，近年来发现并公认了葫芦巴种子的提取物具有降低血糖和调节血脂的功效，并且治疗糖尿病并发症也有作用。结合前期药理学研究，葫芦巴总皂苷和4-羟基异亮氨酸可能是通过促胰岛素分泌作用、延缓葡萄糖在肠道的吸收，改善外周的葡萄糖利用，对胰岛素受体发挥其对抗作用，从而取到降糖作用，同时减少肝脏中沉积的脂质，减轻脂质过氧化，葫芦巴提取物所具有的降糖、降脂和抗氧化作用，均有益于延缓糖尿病并发症的进展。
发明内容
本发明提供了葫芦巴籽中有效成分的提取方法，包括以下步骤：
（1）将干燥的葫芦巴籽粉碎，以70%-80%乙醇水溶液加热提取3次，提取温度80℃，每次提取时间2h，合并提取液，得混合液A；
（2）将混合液A在55-65℃温度下浓缩至无醇，得到浓缩液B；
（3）浓缩液B过层析柱Ⅰ，层析柱Ⅰ吸附剂为732阳离子交换树脂,收集过柱流出液C；水洗层析柱Ⅰ，收集水洗液D；将过柱流出液C和水洗液D混合，得到混合液H；混合液H过层析柱Ⅱ，层析柱Ⅱ吸附剂为SP207大孔吸附树脂；水洗层析柱Ⅱ，收集水洗液J，水洗液J进入下一次混合液H；
（4）使用0.4-0.7mol/L氨水洗脱层析柱Ⅰ，得到洗脱液E；将洗脱液E在55-60℃温度下浓缩，得到浓缩液F；浓缩液F进行喷雾干燥，得到产品G；
（5）使用55-65%乙醇水溶液洗脱层析柱Ⅱ，得到洗脱液K；将洗脱液K在55-60℃温度下浓缩，得到浓缩液L；浓缩液L进行喷雾干燥，得到产品M。
其中，步骤（1）中乙醇水溶液优选75%。
其中，步骤（1）中每次提取时，优选葫芦巴籽与乙醇水溶液料液比1:3至1：6，更优选葫芦巴籽与乙醇水溶液料液比1:5。
其中，步骤（3）中，层析柱Ⅰ吸附流速0.8-1.2BV/h，体积4BV；层析柱Ⅱ吸附流速0.8-2.2BV/h，体积4BV。
其中，步骤（4）中，氨水浓度为0.5mol/L；洗脱流速0.8-1.5BV/h，体积4BV。
其中，步骤（5）中，乙醇水溶液浓度为60%；流速0.8-1BV/h，体积2BV。
本发明方法步骤简单，水使用量少，不使用有机溶剂，避免后续除杂工艺，降低了生产成本，在提高产品纯度的同时还提升了提取率，工艺中水可以循环使用，增加了资源利用率。
具体实施方式
下面结合以下具体实施方式对本发明作更为详细地说明。
将干燥的葫芦巴籽5kg粉碎至直径1-2mm，加入提取罐，以75%乙醇水溶液加热提取3次，料液比1:5，提取温度80℃，每次提取时间2h，合并提取液，得混合液A；使用双效浓缩器对混合液A进行浓缩，在60℃温度下浓缩至无醇，得到浓缩液B；浓缩液B过层析柱Ⅰ，层析柱Ⅰ吸附剂为732阳离子交换树脂,吸附流速1BV/h，体积4BV，并收集过柱流出液C；水洗层析柱Ⅰ，体积2BV，收集水洗液D；将过柱流出液C和水洗液D混合，得到混合液H；混合液H过层析柱Ⅱ，层析柱Ⅱ吸附剂为SP207大孔吸附树脂，吸附流速1BV/h，体积4BV；水洗层析柱Ⅱ，体积2BV，收集水洗液J，水洗液J进入下一次混合液H。
使用0.5mol/L氨水洗脱层析柱Ⅰ,洗脱流速1BV/h，体积4BV，得到洗脱液E；使用双效浓缩器将洗脱液E在60℃温度下浓缩，得到浓缩液F；浓缩液F进行喷雾干燥，得到产品G；产品G重量105.44g，其中4-羟基异亮氨酸含量20.4%，得率86%。
使用60%乙醇水溶液洗脱层析柱Ⅱ，流速1BV/h，体积2BV,得到洗脱液K；使用双效浓缩器将洗脱液K在60℃温度下浓缩，得到浓缩液L；浓缩液L进行喷雾干燥，得到产品M；产品M重量152.76g，其中葫芦巴总皂苷含量64%，得率72%。
Claims (6)
1.葫芦巴籽中有效成分的提取方法，其特征在于：
（1）将干燥的葫芦巴籽粉碎，以70%-80%乙醇水溶液加热提取3次，提取温度80℃，每次提取时间2h，合并提取液，得混合液A；
（2）将混合液A在55-65℃温度下浓缩至无醇，得到浓缩液B；
（3）浓缩液B过层析柱Ⅰ，层析柱Ⅰ吸附剂为732阳离子交换树脂,收集过柱流出液C；水洗层析柱Ⅰ，收集水洗液D；将过柱流出液C和水洗液D混合，得到混合液H；混合液H过层析柱Ⅱ，层析柱Ⅱ吸附剂为SP207大孔吸附树脂；水洗层析柱Ⅱ，收集水洗液J，水洗液J进入下一次混合液H；
（4）使用0.4-0.7mol/L氨水洗脱层析柱Ⅰ，得到洗脱液E；将洗脱液E在55-60℃温度下浓缩，得到浓缩液F；浓缩液F进行喷雾干燥，得到产品G；
（5）使用55-65%乙醇水溶液洗脱层析柱Ⅱ，得到洗脱液K；将洗脱液K在55-60℃温度下浓缩，得到浓缩液L；浓缩液L进行喷雾干燥，得到产品M。
2.如权利要求1所述的制备方法，步骤（1）中乙醇水溶液优选75%。
3.如权利要求1所述的制备方法，步骤（1）中每次提取时，优选葫芦巴籽与乙醇水溶液料液比1:3至1：6，更优选葫芦巴籽与乙醇水溶液料液比1:5。
4.如权利要求1所述的制备方法，步骤（3）中，层析柱Ⅰ吸附流速0.8-1.2BV/h，体积4BV；层析柱Ⅱ吸附流速0.8-2.2BV/h，体积4BV。
5.如权利要求1所述的制备方法，步骤（4）中，氨水浓度为0.5mol/L；洗脱流速0.8-1.5BV/h，体积4BV。
6.如权利要求1所述的制备方法，步骤（5）中，乙醇水溶液浓度为60%；流速0.8-1BV/h，体积2BV。
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