CN1282654C - Process for preparing astraglus base total saponin - Google Patents

Process for preparing astraglus base total saponin Download PDF

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CN1282654C
CN1282654C CN 02155002 CN02155002A CN1282654C CN 1282654 C CN1282654 C CN 1282654C CN 02155002 CN02155002 CN 02155002 CN 02155002 A CN02155002 A CN 02155002A CN 1282654 C CN1282654 C CN 1282654C
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astragaloside
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extraction
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CN1425674A (en
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李慧梁
张卫东
邹豪
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上海博泰医药科技有限公司
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Abstract

本发明为一种黄芪总皂苷的制备方法。 The present invention is a method for preparing one kind of total astragalosides. 它以中药材黄芪为原料,经水热回流提取2次上,提取液过滤后直接上大孔吸附树脂柱,进行脱糖,脱色处理,再分离纯化得黄芪总皂苷混合物,其中黄芪总皂苷含量为80-98%,黄芪甲苷含量为50-80%。 Astragalus membranaceus after it as raw material, the water heat reflux extraction twice, the extract was filtered directly macroporous resin column for desugared, decolorized, and then isolated as a mixture of Astragalus total saponin, wherein the content of Radix 80 to 98%, astragaloside 50-80%. 本方法制得的黄芪甲苷和黄芪总皂苷颜色浅、纯度高,而且工艺简便规范,生产成本低,得率高。 Obtained by the method of the present Astragaloside and Radix light color, high purity, and the process is simple and standardized, low production cost, high yield.

Description

黄芪总皂苷的制备方法 Preparation of Radix

技术领域 FIELD

本发明属医药技术领域,具体涉及一种以中药材黄芪为原料制备黄芪总皂苷的方法。 The present invention belongs to the field of medical technology, in particular to a method for the preparation of starting materials Astragalus membranaceus Astragalus total saponins.

背景技术 Background technique

黄芪(Radix Astragali)是豆科植物蒙古黄芪Astragalus membranaceus(Fisch)Bge.Var.mongholicus(Bge.)Hsiao或膜荚黄芪Astragalus membranaceus(Fisch)Bge.的干燥根,为常用的补益药,味甘,性微温,传统医学认为其具有补气固表、利尿托毒、敛疮生肌之功效,药用历史悠久。 Astragalus (Radix Astragali) Mongolian legumes Astragalus Astragalus membranaceus (Fisch) Bge.Var.mongholicus (Bge.) Hsiao or Astragalus membranaceus Astragalus membranaceus (Fisch) Bge. Dry root, a commonly used tonics, sweet, tepid, traditional medicine with qi solid form, diuretic care drugs, sore myogenic effect, long history of medicinal. 现代研究表明,黄芪具有增强免疫、代谢、降压及利尿作用,其中黄芪皂苷,尤其是黄芪甲苷是其主要生理活性成分。 Modern research shows, Astragalus enhance immunity, metabolism, antihypertensive and diuretic effect, wherein astragaloside, especially astragaloside is the main bioactive component. 目前,黄芪皂苷的制备方法主要有水提醇沉法,醇溶液用于分离皂苷,采用这种方法,黄芪皂苷损失极大,得率低,生产成本很高;也可以用醇类有机溶剂提取,浓缩后,过大孔吸附树脂柱,以水和不同浓度的醇溶液梯度洗脱,采用这种方法,操作步骤复杂,杂质含量高,黄芪皂苷的颜色深,纯度较低。 Currently, the main method for preparing astragalosides water extraction and alcohol precipitation method, an alcohol solution for separating saponins, using this method, astragaloside great loss, low yield, high production cost; alcohol can be extracted with an organic solvent after concentration, too macroporous resin column with a gradient of solutions of different concentrations of alcohol and water elution, using this method, the steps are complex, high impurity content, Astragalin darker, less pure.

发明内容 SUMMARY

本发明的目的是提供一种自动化程度高、操作简便、产率高、无污染的黄芪总皂苷制备新方法。 Object of the present invention is to provide a high degree of automation, simple, high yield, pollution-free new method for preparation of Radix.

本发明提出的黄芪总皂苷的制备方法,以中药材黄芪为原料,包括提取、脱糖、脱色、纯化诸步骤,具体工艺如下:将黄芪根粉碎,在常压回流提取装置中,用水热回流提取二次以上(例如第一次,加6~8倍重量的水,回流提取1小时以上,第二次以后,各加4~6倍重量的水,每次回流提取30分钟以上);合并各部分的水提液,过滤,不需要进行任何其他处理,清液直接加入大孔吸附树脂柱上,以10倍以上体积的去离子蒸馏水洗脱糖类部分,再以5倍以上体积的0.05~0.5%的碱性水溶液进行洗脱,以5倍以上体积的10~40%的酒精洗脱进行脱色;再以70~90%的酒精洗脱黄芪皂苷,洗脱液浓缩到小体积,离心得到沉淀A部分,为高纯度的黄芪甲苷;向母液中加入酒精至酒精浓度为70~90%,离心,分得上清液,浓缩干燥后得到B部分;A部分和B部分混合后得到黄芪总 Radix preparation of the present invention proposed, as a raw material to Astragalus membranaceus, including extraction, desugared, bleaching, all purification steps, the following specific process: The pulverized astragalus root, reflux extraction apparatus at atmospheric pressure, heated under reflux with water extracting two or more times (e.g. the first time, plus 6 to 8 times by weight of water, reflux extraction for 1 hour or more after the second time, each plus 4 to 6 times by weight of water, reflux extraction for 30 minutes each); combined aqueous extract portions, filtered, does not require any additional treatment, the supernatant was added directly to a column of macroporous resin, in more than 10-fold volume of distilled deionized eluted carbohydrate moiety, then 5 volumes of more than 0.05 to 0.5% of an alkaline aqueous solution eluting with 10% alcohol to 40 volumes of elution than 5 decolorized; then 70 to 90% alcohol astrageloside elution, the eluate was concentrated to a small volume, centrifugation part a precipitate was obtained, high purity astragaloside; alcohol was added to the mother liquor to an ethanol concentration of 70 to 90%, by centrifugation, the supernatant share, dried and concentrated to give part B; obtained after mixing part a and part B Astragalus 苷。 Glycosides.

上述方法中,脱色采用的碱性水溶液有氢氧化钾、氢氧化钠或氢氧化钙水溶液。 In the above method, an aqueous alkaline solution bleaching employed are potassium hydroxide, sodium hydroxide or calcium hydroxide aqueous solution.

上述方法中,采用的大孔吸附树脂柱可以为多种,例如AB-8、D101、ZTC-1、DidaionHP20大孔树脂柱。 The above method, the macroporous adsorption resin column may be used for a variety of, for example, AB-8, D101, ZTC-1, DidaionHP20 macroporous resin column. 得到的A部分是高纯度的黄芪甲苷,其总皂苷含量为90~100%,黄芪甲苷含量为85~100%。 A part of the obtained high purity astragaloside, total saponin content of 90 to 100%, astragaloside 85 to 100%. A部分和B部分的混合物的总皂苷含量为80~98%,黄芪甲苷含量为50~80%。 Total saponin content of the mixture of the Part A and Part B is 80 to 98%, astragaloside content of 50 to 80%.

为了更好的理解本发明,下面对本发明的原理说明如下:大孔吸附树脂是20世纪60年代发展起来的一类有机高聚物吸附剂,具有良好的吸附性能,近十余年来逐渐被应用于中草药化学成分的分离和中药新药的开发研制。 For a better understanding of the invention, following the principles of the present invention is described below: macroreticular resin is a polymer-based organic adsorbents developed in the 1960s, and has a good adsorption performance, the past decade gradually It applied to the development of Chinese medicine and the separation of the chemical constituents of herbs. 大孔吸附树脂是一类以吸附为特点,对有机物有浓缩分离作用的高分子聚合物。 Macroporous resins are a class of adsorption characteristics of the polymer concentration and separation of organic matter effects. 在水溶液中,大孔吸附树脂对有机物具有良好的吸附选择性,可以固相萃取的方法,在吸附黄芪皂苷的同时将糖类成分去除。 In an aqueous solution, macroporous adsorption resin having good selectivity for organic compounds, solid phase extraction method can, while adsorption astragalosides saccharide component removed. 大孔吸附树脂的理化性质稳定,不溶于酸、碱及有机溶剂,对有机物的选择性较好,不受无机盐类及强离子低分子化合物的影响。 Macroporous resin physicochemical properties stable, insoluble in acids, alkalis and organic solvents, good selectivity of organic matter, is not affected by strong ionic inorganic salts and low-molecular compound. 一般而言,酸性化合物在碱性溶液中被解吸附,碱性化合物在酸性溶液中被解吸附。 In general, acidic compounds are desorbed in an alkaline solution, the alkaline compounds are desorbed in the acidic solution. 黄芪皂苷为中性化合物,而植物提取物中的色素成分多为酸性化合物。 Astragaloside neutral compound, and pigment components in the plant extract is more acidic compounds. 因此,本发明采用稀碱溶液对大孔吸附树脂柱洗脱,可最大限度的将色素杂质洗脱,而且黄芪皂苷不受影响,并且了提高产品的纯度。 Accordingly, the present invention employs dilute alkaline solution of macroporous resin column elution, the eluted dye to maximize the impurities, and astragaloside affected, and improved product purity. 大孔吸附树脂的吸附量一般与上样溶液浓度成反比,通常以较低浓度进行吸附较为有利。 Macroporous resin adsorption amount is generally inversely proportional to the concentration of the sample solution, the adsorption is generally carried out at low concentrations advantageous. 本发明采取水提液不经浓缩处理,直接上大孔吸附树脂柱的方法,使黄芪皂苷最大限度的被吸附在树脂柱上,极大的提高了黄芪皂苷的转移率和产品得率。 The present invention takes the concentrated aqueous extract without treatment, direct adsorption on macroporous resin column, so that the maximum astrageloside is adsorbed on the resin column, which greatly improves the transfer rate and the product yield astragalosides. 黄芪皂苷在大孔吸附树脂柱的70~90%酒精洗脱部位,浓缩后,黄芪甲苷由于在低浓度醇溶液中溶解度较小而析出。 Astragaloside 70-90% alcohol macroporous resin column elution fraction, concentrated and astragaloside since the low concentration alcohol solution is less soluble precipitates. 本发明经过离心得到高纯度的黄芪甲苷(A部分),含量达到85~100%,由于离心后的母液中含有少量黄芪皂苷和一些醇不溶性成分,本发明采用醇沉法对母液进行处理,可最大限度的减少黄芪皂苷的损失,进一步提高产品得率。 The present invention is centrifuged to obtain a high purity of astragaloside (A part), content of 85 to 100%, since the mother liquor after centrifugation containing insoluble components small amounts astragalosides and a number of alcohol, the present invention mother liquid treatment using alcohol precipitation, You can minimize the loss of Astragalus saponin, to further improve product yield.

以本发明提供的方法对五批黄芪进行了中试生产,测定了药材甲苷含量、总皂苷得率、总皂苷含量、黄芪甲苷含量。 The method of the present invention to provide a batch of five Astragalus pilot production were measured medicine ASIV content, total yield saponins, the content of total saponins, astragaloside. 其中,黄芪甲苷的含量应用HPLC-ELSD检测法测定,药材样品按国家药典2000版规定的方法进行处理,测定药材中甲甙含量;产品用甲醇溶解超滤后作为测试样品,测定产品中的甲苷含量。 Wherein the content of the application A of AST measured HPLC-ELSD detection, medicinal sample is processed according to the method specified in Pharmacopoeia 2000, A determination of glucosinolate content in medicine; product after ultrafiltration was dissolved in methanol as the test sample, the product A glucosinolate content. 总皂苷的含量以黄芪甲苷做标准品,选用香草醛-高氯酸发色体系,测定波长560nm,分别测定不同浓度的标准品的吸收度,绘制标准曲线。 The total content of saponin to make Astragaloside standards, selection vanillin - perchloric acid chromophoric system, measurement wavelength of 560 nm, the absorbance of standards were measured at different concentrations of the standard curve. 用甲醇溶解制备的黄芪总皂苷产品,测定其吸收度,由标准曲线得到甲苷含量,再计算出总皂苷含量。 Dissolved in methanol prepared Radix product, the absorbance was measured by a standard curve A glucosinolate content, and then to calculate the total saponin content. 结果如表1所示。 The results are shown in Table 1.

表1黄芪总皂苷含量及甲苷含量 Table 1 Content and Radix Astragali content

以上的所有产品颜色均为白色至淡黄色,其中生产的沉淀A部分黄芪甲苷的含量以HPLC-ELSD检测,都在95%以上。 All the above products are white to pale yellow color, wherein the content of astragaloside A part of the production of the precipitate to HPLC-ELSD detected in more than 95%.

本发明方法制备的黄芪甲苷和黄芪总苷颜色浅、纯度高,制备方法具有得率高、生产成本低、工艺简便规范的特点。 Astragaloside astragalus and method of the present invention produced total glycosides light color, high purity, having a high yield production method, low production cost, simple process specification features.

经临床应用证明,由本发明方法得到的A部分以及A部分和B部分的混合物具有降压、抗炎、稳定红细胞膜、提高血浆cAMP含量、促进小鼠再生肝DNA合成和增强免疫功能的作用。 Proven clinical application, Part A and Part B and Part A mixture was obtained by the method of the present invention have antihypertensive, anti-inflammatory, red cell membrane stability, increased plasma cAMP levels, promote liver regeneration in mice DNA synthesis and enhanced immune function.

具体实施方式 Detailed ways

下面结合实施例来进一步说明本发明的内容:实施例1:黄芪根10kg粉碎后,在加热回流提取罐中用水回流提取三次。 The following Examples further illustrate embodiments in conjunction with the present invention: Example 1: Astragalus Root 10kg pulverized, extracted three times with water heated to reflux reflux extraction tank. 合并提取液,加入到AB-8大孔吸附树脂柱上,以100升的去离子蒸馏水洗脱糖类,再以50升0.1%的氢氧化钠水溶液进行洗脱,继以50升20%的酒精洗脱进行脱色。 The combined extracts were added to the AB-8 macroporous resin column and eluted with 100 liters of deionized, distilled water elution sugars, then 50 liters of 0.1% aqueous sodium hydroxide was eluted, followed by 50 liters of 20% alcohol eluting bleaching. 再以25升75%的酒精洗脱黄芪皂苷,洗脱液浓缩到小体积,离心得到沉淀A部分5.435克。 Then to 25 liters of 75% ethanol elution astragaloside, the eluate was concentrated to a small volume, the precipitate obtained by centrifugation Part A 5.435 g. 向母液中加入酒精至酒精浓度为70%,离心,分得上清夜,浓缩干燥后得到B部分。 Alcohol is added to the mother liquor to the alcohol concentration of 70%, centrifuged, the supernatant share, dried and concentrated to give Part B. A部分和B部分混合后得到黄芪总皂苷8.637克,得率0.08637%。 Part A and Part B to give a total mixed astragalosides 8.637 g, 0.08637% yield.

实施例2:黄芪根10kg粉碎后,在加热回流提取罐中用水回流提取三次。 Example 2: 10kg pulverized astragalus root, reflux extraction three times with water heated to reflux extraction tank. 合并提取液,加入到D101大孔吸附树脂柱上,以90升的去离子蒸馏水洗脱糖类,再以50升0.1%的氢氧化钾水溶液进行洗脱,继以50升20%的酒精洗脱进行脱色。 The combined extracts were added to D101 macroporous resin column with 90 liters of deionized, distilled water elution sugars, then 50 l of 0.1% potassium hydroxide aqueous solution was eluted followed by 50 liters of 20% ethanol wash off decolorization. 再以25升80%的酒精洗脱黄芪皂苷,洗脱液浓缩到小体积,离心得到沉淀A部分5.754克。 Then to 25 liters of 80% ethanol elution astragaloside, the eluate was concentrated to a small volume, the precipitate obtained by centrifugation Part A 5.754 g. 向母液中加入酒精至酒精浓度为70%,离心,分得上清夜,浓缩干燥后得到B部分,A部分和B部分混合后得到黄芪总皂苷9.036克,得率0.09036%。 Alcohol is added to the mother liquor to the alcohol concentration of 70%, centrifuged, the supernatant share, dried and concentrated to give Part B, A and Part B to give partial mixing astragalosides the total 9.036 g, 0.09036% yield.

实施例3:黄芪根10kg粉碎后,在加热回流提取罐中用水回流提2次。 Example 3: astragalus root 10kg crushed, heated to reflux extraction tank reflux water extract 2 times. 合并提取液,加入到ZTC-1大孔吸附树脂柱上,以110升的去离子蒸馏水洗脱糖类,再以50升0.1%的氢氧化钠水溶液进行洗脱,继以50升20%的酒精洗脱进行脱色。 The combined extracts were added to the ZTC-1 macroporous resin column with 110 liters of deionized, distilled water elution sugars, then 50 liters of 0.1% aqueous sodium hydroxide was eluted, followed by 50 liters of 20% alcohol eluting bleaching. 再以25升80%的酒精洗脱黄芪皂苷,洗脱液浓缩到小体积,离心得到沉淀A部分5.872克。 Then to 25 liters of 80% ethanol elution astragaloside, the eluate was concentrated to a small volume, the precipitate obtained by centrifugation Part A 5.872 g. 向母液中加入酒精至酒精浓度为80%,离心,分得上清夜,浓缩干燥后得到B部分,A部分和B部分混合后得到黄芪总皂苷9.263克,得率0.09263%。 Alcohol is added to the alcohol concentration in the mother liquor to 80%, by centrifugation, the supernatant share, dried and concentrated to give Part B, A and Part B to give partial mixing astragalosides the total 9.263 g, 0.09263% yield.

实施例4:黄芪根30kg粉碎后,在加热回流提取罐中用水回流提取三次。 Example 4: 30kg pulverized astragalus root, reflux extraction three times with water heated to reflux extraction tank. 合并提取液,加入到AB-8大孔吸附树脂柱上,以150升的去离子蒸馏水洗脱糖类,再以100升0.1%的氢氧化钠水溶液进行洗脱,继以50升20%的酒精洗脱进行脱色。 The combined extracts were added to the AB-8 macroporous resin column with 150 liters of deionized, distilled water elution sugars, and then 100 liters of 0.1% aqueous sodium hydroxide was eluted, followed by 50 liters of 20% alcohol eluting bleaching. 再以50升90%的酒精洗脱黄芪皂苷,洗脱液浓缩到小体积,离心得到沉淀A部分15.872克。 Again with 50 liters of 90% ethanol elution astragaloside, the eluate was concentrated to a small volume, the precipitate obtained by centrifugation Part A 15.872 g. 向母液中加入酒精至酒精浓度为80%,离心,分得上清夜,浓缩干燥后得到B部分,A部分和B部分混合后得到黄芪总皂苷27.895克,得率0.09238%。 Alcohol is added to the alcohol concentration in the mother liquor to 80%, by centrifugation, the supernatant share, dried and concentrated to give Part B, A and Part B to give partial mixing astragalosides After 27.895 g total, yield 0.09238%.

Claims (3)

1.一种黄芪总皂苷的制备方法,以中药材黄芪为原料,包括提取、脱糖、脱色、纯化诸步骤,其特征是将黄芪根粉碎,在常压提取装置中,用水热回流提取2次以上;合并各部分提取液,过滤,清液加入大孔吸附树脂柱上,以10倍以上体积的去离子蒸馏水洗脱糖类部分,再以5倍以上体积的0.05~0.5%的氢氧化钠、氢氧化钾或氢氧化钙水溶液进行洗脱,继以5倍以上体积的10~40%的酒精洗脱进行脱色;再以70~90%的酒精洗脱黄芪皂甙,洗脱液浓缩到小体积,离心得到沉淀A部分,为高纯度的黄芪甲苷;向母液中加入酒精至酒精浓度为70~90%,离心,分得上清液,浓缩干燥后得到B部分;A部分和B部分混合后得到黄芪总皂苷,总皂苷含量为80-98%,其中黄芪甲苷含量为50-80%。 1. A method for preparing total astragalosides to Astragalus membranaceus as raw material, including extraction, desugared, bleaching, various purification steps, wherein the pulverized astragalus root, extraction apparatus at atmospheric pressure, hot water reflux extraction 2 or more times; portions were combined extracts were filtered, and the supernatant was added macroporous adsorption resin column with 10 times volume of distilled deionized eluted carbohydrate moiety, then 5 volumes of more than 0.05 to 0.5% by hydroxide sodium, potassium or calcium hydroxide aqueous solution elution, followed alcoholic eluting with 5 more volumes of 10 to 40% by decolorization; then 70 to 90% ethanol elution astragaloside, the eluate was concentrated small volume, the precipitate obtained by centrifugation part a, a high purity of astragaloside; alcohol was added to the mother liquor to an ethanol concentration of 70 to 90%, by centrifugation, the supernatant share, dried and concentrated to give part B; a and part B part astragalosides mixed to give a total, total saponin content of 80-98%, wherein the content of astragaloside 50-80%.
2.一种黄芪甲苷的制备方法,以中药材黄芪为原料,包括提取、脱糖、脱色、纯化诸步骤,其特征是将黄芪根粉碎,在常压提取装置中,用水热回流提取2次以上;合并各部分提取液,过滤,上清液加入大孔吸附树脂柱上,以10倍以上体积的去离子蒸馏水洗脱糖类部分,再以5倍以上体积的0.05~0.5%氢氧化钠、氢氧化钾或氢氧化钙水溶液进行洗脱,继以5倍以上体积的10~40%的酒精洗脱进行脱色;再以70~90%的酒精洗脱黄芪皂甙,洗脱液浓缩到小体积,离心得到沉淀,为高纯度的黄芪甲苷,含量为85~100%。 2. A process for preparing Astragaloside to Astragalus membranaceus as raw material, including extraction, desugared, bleaching, various purification steps, wherein the pulverized astragalus root, extraction apparatus at atmospheric pressure, hot water reflux extraction 2 or more times; portions were combined extracts were filtered, and the supernatant was added macroporous adsorption resin column with 10 times volume of distilled deionized eluted carbohydrate moiety, then 5 volumes of more than 0.05 to 0.5% sodium hydroxide sodium, potassium or calcium hydroxide aqueous solution elution, followed alcoholic eluting with 5 more volumes of 10 to 40% by decolorization; then 70 to 90% ethanol elution astragaloside, the eluate was concentrated small volume, the precipitate obtained by centrifugation, a high purity of astragaloside content of 85% to 100%.
3.根据权利要求1所述的黄芪总皂苷的制备方法,其特征在于黄芪药材水热回流提取二次以上,第一次加6-8倍重量的水,回流提取1小时以上,第二次后,各加4-6倍重量的水,回流30分钟以上。 3. Preparation method of Radix according to claim 1, characterized in that the hydrothermal Radix Astragali reflux extraction twice more, plus the first 6-8 times the weight of water, reflux extraction for 1 hour or more, the second after each addition of 4-6 times the weight of water, was refluxed for 30 minutes.
CN 02155002 2002-12-19 2002-12-19 Process for preparing astraglus base total saponin CN1282654C (en)

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