CN116270779B - 一种三七提取物的制备方法 - Google Patents
一种三七提取物的制备方法 Download PDFInfo
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- CN116270779B CN116270779B CN202310242527.4A CN202310242527A CN116270779B CN 116270779 B CN116270779 B CN 116270779B CN 202310242527 A CN202310242527 A CN 202310242527A CN 116270779 B CN116270779 B CN 116270779B
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Abstract
本发明涉及中药提取技术领域,具体涉及一种三七提取物的制备方法,所述的制备方法包括使用提取溶剂对三七进行提取,所述提取溶剂按照重量份数计包括柠檬酸水溶液10‑30份、丙二醇1‑12份和海藻酸钠固定木聚糖酶0.1‑5份。本发明通过控制提取溶剂的组成和溶剂中各成分的含量和三七与提取溶剂的质量比,提高了对三七提取物中活性成分的提取效率,充分的发挥了三七提取物的药效,可用于制备药物及功能食品。
Description
技术领域
本发明涉及中药提取技术领域,具体涉及一种三七提取物的制备方法。
背景技术
三七,性温,味甘、微苦,归肝经、胃经,具有散瘀止血、消肿定痛功效。三七可用于治疗吐血、便血、外伤出血、胸腹刺痛、跌扑肿痛等。三七花为五加科人参属植物三七的干燥花蕾(Flowerbuds of PanaxNotoginseng(Burk)
F.H.Chen)。三七花及其茎叶具有增强免疫力、抗肿瘤等作用。据研究报道,从三七中分离出超过200种化学成分,主要成分是三七总皂苷(Panax notoginseng saponins,PNS),分为四种类型:原七二醇、原七三醇、雌黄型和齐墩果酸。其他成分还包括黄酮类、环肽类、甾醇类、糖类、挥发油类、氨基酸类及有机酸类等成分。
皂苷,是苷元为三萜或螺旋甾烷类化合物的一类糖苷,从三七中共分离鉴定出人参皂苷、三七皂苷、绞股蓝皂苷等100多种皂苷类化合物。从三七中分离出的皂苷主要为三七总皂苷,三七总皂苷是三七的主要活性成分之一,也是主要的活性化合物。共鉴定出27中皂苷类化合物,其中9种为主要成分,分别为三七皂苷R1、人参皂苷Rb1、Rb2、Rb3、Rc、Rd、Re、Rf、Rg1。人参皂甙Rb1、Rg1和三七皂苷R1具有维持血液循环、抗氧化等作用,人参皂苷Rd可缓解动脉粥样硬化,人参皂苷Rb1、Re可降低血糖。
中国专利CN114231357A中公开了一种高海拔地区三七花纯露的提取方法,包括如下步骤:(1)将三七花清洗后放入在蒸馏釜中,加入去离子水粉碎得到三七花浆;(2)向蒸馏釜中的三七花浆加入含氯化镁和乙二胺四乙酸二钠的去离子水混合;(3)加热蒸馏釜,保持沸腾状态蒸馏3-5h;(4)待蒸馏釜温度降低到55℃时打开冷凝器,经过冷凝分离得到蒸馏液,静置过滤得到三七花纯露。该发明制备的三七花纯露无任何添加组分,保留了三七花中皂苷及挥发油成分,具有抗炎、保肝、降血脂、降血压、抗肿瘤、平喘等功效。
中国专利CN113713002A中公开了一种三七提取物的提取方法。所述三七提取物的提取方法包括将三七根粉碎,按三七根重量的4-5倍加入70-80%的乙醇溶液中浸渍12-20h,期间每半小时搅拌1-2次,得三七浸渍液;按三七根重量的8-10倍加入水对三七浸渍液进行溶解后超声5-10min,得到三七水溶液;经循环树脂吸附,在循环流动过程,树脂上不断吸附三七提取物,循环流动20-30min;排出吸附余液,并采用乙醇的碱性水溶液作为解析液进行洗脱;浓缩去除水,按照三七根重量的2-3倍加入90-95%的酒精,搅拌均匀,醇沉过夜,过滤减压浓缩即可。该发明提高了三七提取物的提取效率。
但是从三七中提取得到有效成分的纯度更高和提取效率更高的制备方法仍然是令人期待的。
发明内容
针对上述存在的问题,本发明旨在提供一种三七提取物的制备方法,所述的制备方法得到的三七提取物的有效成分纯度和提取效率较高。
为实现上述发明目的,本发明的技术方案如下:
一方面,本发明提供了一种三七提取物的制备方法,包括使用提取溶剂对三七进行提取,所述提取溶剂按照重量份数计包括柠檬酸水溶液10-30份、丙二醇1-12份和海藻酸钠固定木聚糖酶0.1-5份。
优选地,所述的提取溶剂按照重量份数计包括柠檬酸水溶液12-25份、丙二醇3-9份和海藻酸钠固定木聚糖酶1-4份;
进一步优选地,所述的提取溶剂按照重量份数计包括柠檬酸水溶液18份、丙二醇7份和海藻酸钠固定木聚糖酶3份。
进一步优选地,所述的柠檬酸水溶液的浓度为1-5%;
再进一步优选地,所述的柠檬酸水溶液的浓度为3%。
优选地,所述的三七与提取溶剂的质量体积比为1:(5-20);
进一步优选地,所述的三七与提取溶剂的质量体积比为1:(12-18);
最优选地,所述的三七与提取溶剂的质量体积比为1:14。
具体的,所述的丙二醇的化学式为C3H8O2,CAS号为57-55-6。
具体的,所述的海藻酸钠的化学式为C6H9NaO7,CAS号为9005-38-3。
具体的,所述的海藻酸钠固定木聚糖酶的制备方法包括但不限于直接包埋法、交联包埋法、包埋交联法。
进一步具体的,所述的海藻酸钠固定木聚糖酶的制备方法为直接包埋法。
进一步具体的,所述的直接包埋法为将25mL酶液与15.5mL质量分数为3.5%海藻酸钠溶液混合,冷却,吸取10mL混合液注入质量份数为2%氯化钙溶液中,形成光滑凝胶小球,滤出凝胶小球,在4℃条件下硬化,用氯化钠洗涤备用。
在一些实施例中,所述的三七提取物的制备方法,包括以下步骤:
S1、酸洗:取三七放入酸溶液中浸泡;
S2、碱洗:酸洗后的三七碱溶液中浸泡;
S3、提取:将S2所得三七用提取溶剂提取,合并提取液,过滤;
S4、将过滤后的溶液减压浓缩至浸膏,加入乙醇,混匀,过滤,回收乙醇,浓缩,干燥,即得。
优选地,上述S1-S4中所述的三七为三七根,所述的三七根包括三七根茎、三七主根和三七须根。
优选地,所述的S1中酸溶液选自盐酸、硝酸、硫酸中的至少一种。
进一步优选地,所述的S1中酸溶液选自盐酸、硝酸中的至少一种。
进一步优选地,所述的酸溶液为盐酸。
进一步优选地,所述的酸溶液的浓度为3-5%;
再进一步优选地,所述的酸溶液的浓度为4%。
优选地,所述的S1中将三七在酸溶液中浸泡后还包括水洗步骤;
进一步优选地,所述的水洗所用水的pH值为6.5-9;
再进一步优选地,所述的水洗所用水的pH值为7.5。
优选地,所述的S2中碱溶液选自氢氧化钾、氨水、氢氧化钙、甲醇钠、碳酸氢钠中的一种或多种。
进一步优选地,所述的S2中碱溶液选自氢氧化钾、氢氧化钙、碳酸氢钠中的一种或多种。
进一步优选地,所述的碱溶液为氢氧化钾。
进一步优选地,所述的碱溶液的浓度为3-5%;
再进一步优选地,所述的碱溶液的浓度为4%。
优选地,所述的S2中将三七在碱溶液中浸泡后还包括水洗步骤;
进一步优选地,所述的水洗所用水的pH值为6.5-9;
再进一步优选地,所述的水洗所用水的pH值为7.5。
优选地,所述的S3中加热提取的温度为40-60℃;
进一步优选地,所述的S3中加热提取的温度为50℃;
优选地,所述的S3中提取的次数为1-4次,提取的时间为1-5h;
进一步优选地,所述的提取的次数为2次,提取的时间为1-3h。
优选地,所述的提取次数为2次时,第一次提取的时间为2h,第二次提取的时间为1h。
优选地,所述的S4中浸膏的相对比重为1.05-1.15,浸膏的温度为50-60℃;
进一步优选地,所述的S4中浸膏的相对比重为1.10,浸膏的温度为55℃。
优选地,所述的S4中料液的温度为10-30℃;进一步优选地,所述料液的温度为20℃。
优选地,所述的S4中乙醇的体积分数为50-70%;进一步优选地,所述的乙醇的体积分数为50-55%。
再进一步优选地,所述的乙醇的体积分数为55%。
优选地,所述的S4中加入乙醇后料液的含醇量为40-50%;
进一步优选地,所述的S4中加入乙醇后料液的含醇量为40-45%;
再进一步优选地,所述的S4中加入乙醇后料液的含醇量为45%。
优选地,所述的S4中混匀的时间为10-20分钟;所述混匀的方式包括但不限于搅拌。
进一步优选地,所述的S4中混匀的时间为15分钟。
优选地,所述的S4中混匀后还包括静置步骤,所述的S4中静置的时间为12-18h;
进一步优选地,所述的静置的时间为14-16h;
进一步优选地,所述的静置的时间为15h。
优选地,所述的S4中浓缩至相对比重为1.15-1.25,浓缩时的温度为50-60℃;
进一步优选地,所述的S4中浓缩至相对比重为1.20,浓缩时的温度为55℃。
优选地,所述的S4中干燥的温度为65±5℃。
优选地,所述的粉碎的方式包括但不限于球磨、粉碎机粉碎、研磨、高压均质。
在一些实施例中,所述S1中还包括将三七粉碎步骤;所述的三七为三七根。
进一步优选地,所述的S1中粉碎的后过筛的目数为15-30目;
再进一步优选地,所述的S1中粉碎的后过筛的目数为20目。
又一方面,本发明提供了按照上述的制备方法制备的三七提取物。
又一方面,本发明提供了上述的制备方法制备的三七提取物或上述的三七提取物在制备药物及功能食品中的应用。
具体的,所述的药物剂型选自片剂、粉剂、胶囊剂、软胶囊剂、滴丸剂、颗粒剂、口服液中的至少一种。
进一步具体的,所述的药物剂型选自胶囊剂、软胶囊剂、滴丸剂、片剂中的至少一种。
具体的,所述的功能食品包括但不限于固体饮料、液体饮料、酒。
与现有技术相比,本发明的有益效果为:
本发明公开了一种三七提取物的制备方法,通过控制提取溶剂的组成和溶剂中各成分的含量和三七与提取溶剂的质量比,最大限度的浓缩和保留有效成分,并且提高了对三七提取物中活性成分的提取效率,三七总皂苷含量不低于84.2%,有效成分中人参皂甙R1不少于8.1%,人参皂苷Rg1不少于34.6%,人参皂苷Rb1不少于37.2%,人参皂苷Rd不少于5.5%,可用于制备药物及功能食品。
具体实施方式
为了使本发明实现的技术手段、创作特征、达成目的与功效易于明白了解,下面结合具体实施例,进一步阐明本发明,但下述实施例仅为本发明的优选实施例,并非全部。基于实施方式中的实施例,本领域技术人员在没有做出创造性劳动的前提下所获得其它实施例,都属于本发明的保护范围。下述实施例中,若无特殊说明,所用的操作方法均为常规操作方法,所用设备均为常规设备,各个实施例所用设备材料均相同。
实施例1-7和对比例1-3中的海藻酸钠固定木聚糖酶的制备方法为:将25mL酶液与15.5mL质量分数为3.5%海藻酸钠溶液混合,冷却,吸取10mL混合液注入质量份数为2%氯化钙溶液中,形成光滑凝胶小球,滤出凝胶小球,在4℃条件下硬化,用氯化钠洗涤备用;所述的木聚糖酶的酶活为5000U/mL。
实施例1
一种三七提取物的制备方法,包括以下步骤:
S1、酸洗:取三七根茎放入3%的盐酸溶液中浸泡,用pH值为7.5的水进行水洗;
S2、碱洗:酸洗后的三七根茎放入3%的氢氧化钾溶液中浸泡,用pH值为7.5的水进行水洗;
S3、提取:将S2所得三七根茎用提取溶剂提取,三七根茎与提取溶剂的质量体积比为1:5,提取溶剂包括10份柠檬酸水溶液,3份丙二醇、0.5份海藻酸钠固定木聚糖酶,提取的温度为50℃,提取2次,第一次提取1h,第二次提取2h,合并提取液,过滤;
S4、将过滤后的溶液减压浓缩至浸膏的相对比重为1.05,加入50%乙醇,加入乙醇后料液的含醇量为40%,搅拌混匀10分钟,静置12小时,过滤,浓缩至相对比重为1.15,65℃进行干燥,即得。
实施例2
一种三七提取物的制备方法,包括以下步骤:
S1、酸洗:取三七根茎放入4%的盐酸溶液中浸泡,用pH值为7.5的水进行水洗;
S2、碱洗:酸洗后的三七根茎放入4%的氢氧化钾溶液中浸泡,用pH值为7.5的水进行水洗;
S3、提取:将S2所得三七根茎用提取溶剂提取,三七根茎与提取溶剂的质量体积比为1:14,提取溶剂包括18份柠檬酸水溶液,7份丙二醇、3份海藻酸钠固定木聚糖酶,提取的温度为60℃,提取2次,第一次提取2h,第二次提取1h,合并提取液,过滤;
S4、将过滤后的溶液减压浓缩至浸膏的相对比重为1.10,加入55%乙醇,加入乙醇后料液的含醇量为45%,搅拌混匀15分钟,静置15小时,过滤,浓缩至相对比重为1.20,65℃进行干燥,即得。
实施例3
一种三七提取物的制备方法,包括以下步骤:
S1、酸洗:取三七根茎放入4%的盐酸溶液中浸泡,用pH值为7.5的水进行水洗;
S2、碱洗:酸洗后的三七根茎放入4%的氢氧化钾溶液中浸泡,用pH值为7.5的水进行水洗;
S3、提取:将S2所得三七根茎用提取溶剂提取,三七与提取溶剂的质量体积比为1:18,提取溶剂包括30份柠檬酸水溶液,12份丙二醇、5份海藻酸钠固定木聚糖酶,提取的温度为60℃,提取2次,第一次提取2h,第二次提取1h,合并提取液,过滤;
S4、将过滤后的溶液减压浓缩至浸膏的相对比重为1.10,加入55%乙醇,加入乙醇后料液的含醇量为45%,搅拌混匀15分钟,静置14小时,过滤,浓缩至相对比重为1.20,55℃进行干燥,即得。
实施例4
一种三七提取物的制备方法,包括以下步骤:
S1、酸洗:取三七根茎放入4%的盐酸溶液中浸泡,用pH值为7.5的水进行水洗;
S2、碱洗:酸洗后的三七根茎放入4%的氢氧化钾溶液中浸泡,用pH值为7.5的水进行水洗;
S3、提取:将S2所得三七用提取溶剂提取,三七与提取溶剂的质量体积比为1:14,提取溶剂包括18份柠檬酸水溶液,7份丙二醇、3份海藻酸钠固定木聚糖酶,提取的温度为60℃,提取2次,第一次提取2h,第二次提取1h,合并提取液,过滤;
S4、将过滤后的溶液减压浓缩至浸膏的相对比重为1.10,加入55%乙醇,加入乙醇后料液的含醇量为45%,搅拌混匀20分钟,静置16小时,过滤,浓缩至相对比重为1.20,70℃进行干燥,即得。
实施例5
一种三七提取物的制备方法,包括以下步骤:
S1、将三七主根粉碎过20目筛,放入4%的盐酸溶液中浸泡,用pH值为6.5的水进行水洗;
S2、酸洗后的三七主根放入4%的氢氧化钾溶液中浸泡,用pH值为6.5的水进行水洗;
S3、提取:将S2所得三七主根用提取溶剂提取,三七与提取溶剂的质量体积比为1:14,提取溶剂包括18份柠檬酸水溶液,7份丙二醇、3份海藻酸钠固定木聚糖酶,提取的温度为50℃,提取2次,第一次回流提取2h,第二次回流提取1h,合并提取液,过滤;
S4、将过滤后的溶液减压浓缩至浸膏的相对比重为1.05,加入50%乙醇,加入乙醇后料液的含醇量为40%,搅拌混匀15分钟,静置14小时,过滤,浓缩至相对比重为1.15,60℃进行干燥,即得。
实施例6
一种三七提取物的制备方法,包括以下步骤:
S1、将三七须根粉碎过20目筛,放入4%的盐酸溶液中浸泡,用pH值为7.5的水进行水洗;
S2、酸洗后的三七须根放入4%的氢氧化钾溶液中浸泡,用pH值为7.5的水进行水洗;
S3、提取:将S2所得三七须根用提取溶剂提取,三七与提取溶剂的质量体积比为1:14,提取溶剂包括18份柠檬酸水溶液,7份丙二醇、3份海藻酸钠固定木聚糖酶,提取的温度为60℃,提取2次,第一次回流提取2h,第二次回流提取1h,合并提取液,过滤;
S4、将过滤后的溶液减压浓缩至浸膏的相对比重为1.10,加入55%乙醇,加入乙醇后料液的含醇量为45%,搅拌混匀18分钟,静置15小时,过滤,浓缩至相对比重为1.20,65℃进行干燥,即得。
实施例7
一种三七提取物的制备方法,包括以下步骤:
S1、将三七须根粉碎过20目筛,放入4%的盐酸溶液中浸泡,用pH值为7.5的水进行水洗;
S2、酸洗后的三七须根放入4%的氢氧化钾溶液中浸泡,用pH值为7.5的水进行水洗;
S3、提取:将S2所得三七须根用提取溶剂提取,三七与提取溶剂的质量体积比为1:18,提取溶剂包括25份柠檬酸水溶液,9份丙二醇、4份海藻酸钠固定木聚糖酶,提取的温度为50℃,提取2次,第一次回流提取2h,第二次回流提取1h,合并提取液,过滤;
S4、将过滤后的溶液减压浓缩至浸膏的相对比重为1.15,加入60%乙醇,加入乙醇后料液的含醇量为50%,搅拌混匀20分钟,静置16小时,过滤,浓缩至相对比重为1.20,65℃进行干燥,即得。
对比例1
与实施例2的区别在于,提取溶剂中用乙醇替换柠檬酸水溶液,其余皆相同。
对比例2
与实施例2的区别在于,三七与提取溶剂的质量体积比为1:25,其余皆相同。
对比例3
与实施例2的区别在于,提取溶剂中缺少丙二醇,提取溶剂具体包括18份柠檬酸水溶液、3份海藻酸钠固定木聚糖酶,其余皆相同。
对比例4
与实施例2的区别在于,提取溶剂中缺少海藻酸钠固定木聚糖酶,提取溶剂按照重量份数计包括柠檬酸水溶液18份、丙二醇7份,其余皆相同。
实验例1
对以上实施例1-7以及对比例1-4得到的三七提取物进行纯度的检测。
纯度检测方法:照高效液相色谱法(《中国药典》通则0512)测定。
色谱条件与系统适用性试验:以十八烷基硅烷键合硅胶为填充剂;以乙腈为流动相A,以0.2%磷酸溶液为流动相B,按下表1中的规定进行梯度洗脱;检测波长为203nm。
对照品溶液的制备:取人参皂苷Rb3对照品适量,精密称定,加乙醇制成每1ml含0.5mg的溶液,即得。
供试品溶液的制备:取本品50mg,精密称定,置10ml量瓶中,用乙醇溶解并稀释至刻度,摇匀,滤过,取续滤液,即得。
测定法分别精密吸取对照品溶液与供试品溶液各10μL,注入液相色谱仪,测定,即得。
表1色谱条件与系统适用性试验梯度洗脱方案
检测结果如下表2。
表2.
根据上表2的检测的数据可以看出,本发明实施例1-7提供的制备方法得到的三七提取物的纯度均较高,尤其是实施例2中得到的三七提取物的纯度为99.1%。对比例1中用乙醇替换柠檬酸水溶液,对比例2中改变了三七和提取溶剂的质量比,均会在一定程度上影响三七提取物的纯度。对比例3-4改变了提取溶剂的组成,也会在一定程度上影响三七提取物的纯度。
同时,从上表3中可以看出,实施例1-7的三七总皂苷含量大于对比例1-4,本申请实施例1-7的方法提取得到的三七提取物中三七总皂苷含量为84.2-89.0%,与对比例1-4相比较,提取效率更高。从上表可知,人参皂甙R1、人参皂苷Rg1、人参皂苷Rb1和人参皂苷Rd的含量比与对比例1-4相比较均有所提高。对比例1中用乙醇替换柠檬酸水溶液,对比例2中改变了三七和提取溶剂的质量比,使得人参皂甙R1、人参皂苷Rg1、人参皂苷Rb1和人参皂苷Rd的含量降低。对比例3的提取溶剂中缺少丙二醇,对比例4的提取溶剂中缺少海藻酸钠固定木聚糖酶,该提取溶剂提取三七时的提取效率较实施例1-7差。由此可知,本申请的实施例1-7中通过控制提取溶剂的组成和溶剂中各成分的含量和三七与提取溶剂的质量比,提高了对三七提取物中活性成分的提取效率,充分的发挥了三七提取物的功效。
以上所述仅为本发明的较佳实施例而已,并不用以限制本发明,凡在本发明的精神和原则之内,所作的任何修改、等同替换、改进等,均应包含在本发明的保护范围之内。
Claims (9)
1.一种三七提取物的制备方法,其特征在于,包括以下步骤:
S1、酸洗:取三七放入3%或4%酸溶液中浸泡;
S2、碱洗:酸洗后的三七放入3%或4%碱溶液中浸泡;
S3、提取:将S2所得三七用提取溶剂提取,提取的温度为50℃或60℃,合并提取液,过滤;
S4、将过滤后的溶液减压浓缩至浸膏的相对比重为1.05或1.10或1.15,加入50%或55%或60%乙醇,混匀,过滤,回收乙醇,浓缩至相对比重为1.15或1.20,干燥,即得;
步骤S3中所述的提取溶剂按照重量份数计由柠檬酸水溶液10-30份、丙二醇1-12份和海藻酸钠固定木聚糖酶0.1-5份组成。
2.根据权利要求1所述的制备方法,其特征在于,所述的提取溶剂按照重量份数计由柠檬酸水溶液18份、丙二醇7份和海藻酸钠固定木聚糖酶3份组成。
3.根据权利要求1所述的制备方法,其特征在于,所述的三七与提取溶剂的质量体积比为1:(5-20)。
4.根据权利要求3所述的制备方法,其特征在于,所述的三七与提取溶剂的质量体积比为1:14。
5.根据权利要求1所述的制备方法,其特征在于,S1-S3中所述的三七为三七根,所述的三七根包括三七根茎、三七主根和三七须根。
6.根据权利要求1所述的制备方法,其特征在于,所述的S3中提取的次数为1-3次,提取的时间为1-5h。
7.根据权利要求1所述的制备方法,其特征在于,所述的S4中加入乙醇后料液的含醇量为40-50%。
8.根据权利要求1-7任一项所述的制备方法制备的三七提取物。
9.根据权利要求1-7任一项所述的制备方法制备的三七提取物或权利要求8所述的三七提取物在制备药物及功能食品中的应用。
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