CN110117231B - 一种从葫芦巴籽中提取4-羟基异亮氨酸及总皂苷的方法 - Google Patents
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Abstract
本发明公开了一种从葫芦巴籽中提取4‑羟基异亮氨酸及总皂苷的方法。葫芦巴籽中4‑羟基异亮氨酸及总皂苷具有降低血糖和调节血脂的功效，并且对治疗糖尿病并发症也有作用。采用本发明从葫芦巴种子中能高收率地得到提取物，且提取工艺简便可行，生产流程稳定可靠，具有较高的社会价值及经济价值。
Description
技术领域
本发明涉及一种植物中有效成分的提取方法，特别是涉及一种从葫芦巴籽中高效提取4-羟基异亮氨酸及总皂苷的方法，属于植物化学研究领域。
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
本发明的目的是提供一种以中药葫芦巴的种子为原料，工业提取天然4-羟基异亮氨酸及其总皂苷的方法。具体内容是：
取经粉碎机粉碎后的葫芦巴籽的粉末，在温度80℃下，用75%乙醇提取两小时，重复3次；将获得的提取液加入双效浓缩器中，在温度60℃下浓缩至无醇后，经过732阳离子交换树脂柱吸附；浓缩液经过无离子水水洗后，使用0.5mol/L氨水进行洗脱，同时吸附过柱流出液与水洗液的混合液A作为下一提取工序原液；洗脱液进入双效浓缩器，在温度60℃下浓缩；浓缩液经过喷雾干燥后，得到富含4-羟基异亮氨酸的葫芦巴提取物。
在提取葫芦巴的4-羟基异亮氨酸同时，将吸附过柱流出液与水洗液的混合液A，用SP207大孔吸附树脂进行吸附；经过无离子水水洗后，再用60%乙醇进行洗脱，同时水洗液循环进入下一次混合液A；洗脱液进入双效浓缩器在60℃温度下进行浓缩；浓缩液经过喷雾干燥，得到富含总皂苷的葫芦巴提取物。
附图说明
图1为葫芦巴提取物生产工艺流程图
具体实施方式
下面结合葫芦巴提取物生产工艺流程图和实施实例对本发明专利作进一步地说明。
实施实例
葫芦巴提取物生产工艺流程图如如图所示，其中中试实验过程及数据如下
1.取葫芦巴籽5kg，经粉碎机粉碎至1-2mm大小粉末；加入提取罐，料液比1:5，在温度80℃下，使用75%乙醇提取3次，每次2小时；合并提取液进入双效浓缩器，在温度60℃下浓缩至无醇，经过732阳离子交换树脂柱吸附，流速1BV/h，体积4BV；水洗2BV，再用0.5mol/L氨水进行洗脱，流速1BV/h，体积4BV；洗脱液进入双效浓缩器，在60℃温度下浓缩；浓缩液经过喷雾干燥后，得到含有20.4%的4-羟基异亮氨酸葫芦巴提取物105.44g，产品得率86%。
2.过柱流出液及水洗液的混合液用SP207大孔吸附树脂柱吸附，流速1BV/h，体积4BV；水洗2BV，再用60%乙醇进行洗脱，流速1BV/h，体积2BV；洗脱液进入双效浓缩器，在60℃温度下浓缩；浓缩液经过喷雾干燥，得到含有64%总皂苷的葫芦巴提取物152.76g，产品得率72%。
通过上述实例可以发现通过本发明获得提取物的工艺流程较为简单，便于实现工业化生产，而且产品获得率也较高，经济效益明显。
Claims (5)
1.一种从葫芦巴籽中提取4-羟基异亮氨酸及总皂苷提取物的方法，其特征是：取经粉碎机粉碎后的葫芦巴种子粉，加入提取罐，用体积分数为75％的乙醇在80℃温度下提取2小时，提取三次；提取液进入双效浓缩器，60℃浓缩至无醇，过732阳离子交换树脂柱吸附；经水洗树脂柱水洗后，再用氨水进行洗脱，其中吸附工艺的过柱流出液与水洗液的混合液A进入下一工序；洗脱液进入双效浓缩器，60℃浓缩；浓缩液喷雾干燥，得到富含4-羟基异亮氨酸的葫芦巴提取物；以及，将过柱流出液与水洗液的混合液A用SP207大孔吸附树脂进行吸附；经过水洗后，再用体积分数为60％的乙醇进行洗脱，同时水洗液进入下一次混合液；洗脱液进入双效浓缩器进行浓缩；浓缩液喷雾干燥，得到富含总皂苷的葫芦巴提取物。
2.根据权利要求1的提取方法，其特征在于，经过浓缩后，采用732阳离子交换树脂柱吸附，随后采用0.5mol/L的氨水进行洗脱，同时吸附过柱流出液与水洗液的混合液为下一工艺制备液。
3.根据权利要求1的提取方法，其特征在于，所述混合液使用SP207大孔树脂柱进行吸附，并循环采用水洗，水洗液再次进入混合液进行吸附，提高产品获得率。
4.根据权利要求1的提取方法，其特征在于，所述洗脱液进入双效浓缩器进行浓缩中的洗脱液采用甲醇。
5.根据权利要求1的提取方法，其特征在于，提取混合液、洗脱液都采用双效浓缩器在60℃下进行浓缩工艺，制成品前都对浓缩液采用喷雾干燥器进行干燥工艺获得成品。
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