WO2022178953A1 - 一种牵牛子提取物的制备方法 - Google Patents

一种牵牛子提取物的制备方法 Download PDF

Info

Publication number
WO2022178953A1
WO2022178953A1 PCT/CN2021/085215 CN2021085215W WO2022178953A1 WO 2022178953 A1 WO2022178953 A1 WO 2022178953A1 CN 2021085215 W CN2021085215 W CN 2021085215W WO 2022178953 A1 WO2022178953 A1 WO 2022178953A1
Authority
WO
WIPO (PCT)
Prior art keywords
morning glory
solution
extraction
soaking
extract
Prior art date
Application number
PCT/CN2021/085215
Other languages
English (en)
French (fr)
Inventor
刘景萍
刘全国
陈克领
王家
李党
吴伟贞
吴育强
Original Assignee
海南葫芦娃药业集团股份有限公司
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 海南葫芦娃药业集团股份有限公司 filed Critical 海南葫芦娃药业集团股份有限公司
Publication of WO2022178953A1 publication Critical patent/WO2022178953A1/zh

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/39—Convolvulaceae (Morning-glory family), e.g. bindweed
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/14—Prodigestives, e.g. acids, enzymes, appetite stimulants, antidyspeptics, tonics, antiflatulents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30—Extraction of the material
    • A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
    • A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30—Extraction of the material
    • A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/50—Methods involving additional extraction steps
    • A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/50—Methods involving additional extraction steps
    • A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/50—Methods involving additional extraction steps
    • A61K2236/55—Liquid-liquid separation; Phase separation
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

  • the invention relates to the field of extracting effective components from morning glory seeds, in particular to a preparation method of a morning glory seed extract.
  • Morning glory belonging to the Convolvulaceae morning glory, alias, black ugly, white ugly, two ugly, morning glory, morning glory.
  • the colors of morning glory are blue, crimson, pink, purple, etc., and there are also mixed colors.
  • the fruit is ovoid and can be used as medicine.
  • the leaves of morning glory are three-lobed, the base is heart-shaped, the flowers are white, purple-red or purple-blue, funnel-shaped, and the whole plant is shaggy.
  • Divided-leaf morning glory is an annual entwined herbaceous vine, the whole plant is densely hirsute.
  • the top 5-lobed capsule is spherical, the seeds are ovate and triangular, the surface gray-black is black ugly, and the yellowish white is white ugly.
  • the present invention proposes a preparation method of morning glory seed extract to solve the above-mentioned technical problems.
  • a preparation method of a morning glory seed extract comprising the following steps:
  • Morning glory seeds are soaked in sodium bicarbonate solution after pulverizing, and then concentrated to obtain morning glory seeds powder;
  • step S2 morning glory powder is placed in 93-96% (v/v) ethanol solution and soaked by mass volume ratio g/mL1:5-15 to obtain soaking solution;
  • the soaking solution is concentrated under reduced pressure to obtain morning glory seed powder after soaking;
  • the first liquid residue is added to ethyl acetate for microwave extraction, the microwave power is 300-500W, the extraction time is 20-50min, the extraction temperature is 50-55°C, and the mass-volume ratio of the first liquid residue and ethyl acetate is g/ mL is 1:3-8, and the second extract and the second liquid residue are obtained;
  • the concentrate is added to deionized water to dissolve, and the mass-volume ratio g/mL of the concentrate and deionized water is 1:10-12, the obtained solution is slowly added to the ADS-17 macroporous resin for purification, and the solution is allowed to stand.
  • gradient elution was carried out with water:ethanol according to the volume ratio of 1:9, 3:7, and 5:5, and the fractions were collected, concentrated under reduced pressure, and vacuum-dried to obtain the morning glory extract.
  • the mass volume ratio g/mL of the morning glory seed and the sodium bicarbonate solution is 1:15-20, and the soaking time is 30-60min.
  • step S3 the soaking time of the ethanol solution is 1-3h, and the time is repeated 3-5 times.
  • the decompression concentration temperature is 50-60°C.
  • the application of the morning glory seed extract in the preparation of a purgative diuretic drug is preferable.
  • the preparation method of the morning glory seed extract of the present invention has the advantages of simple preparation process, short production period, ethanol and ethyl acetate can be recycled, and is environmentally friendly. Soak the morning glory seeds powder, part of the morning glory seeds is hydrolyzed into the morning glory acid, which can stimulate the intestinal mucosa, increase the peristalsis and lead to diarrhea, and at the same time reduce the toxicity of the morning glory glycosides, prevent injury to the righteousness, ease the medicinal properties, and ensure the safety and effectiveness of clinical medication.
  • microwave extraction has the advantages of simple operation, fast extraction speed, high yield, less damage to active ingredients, and less environmental pollution, and is suitable for large-scale industrial production.
  • the extraction method of the present invention has the advantages of simple process, short production period, green environmental protection, high extraction rate of the active components of the morning glory, suitable for industrial production, and provides a good way for the efficient utilization of the morning glory.
  • a preparation method of morning glory seed extract comprising the following steps:
  • Step S2 morning glory seed powder is placed in 95% (v/v) ethanol solution by mass volume ratio g/mL to be 1:5 and soaked for 1h to obtain soaking solution, repeated 3 times;
  • the soaking solution is concentrated under reduced pressure, and the concentration under reduced pressure temperature is 50° C. to obtain morning glory seed powder after soaking;
  • adding the first liquid residue to ethyl acetate is 1:3 by mass-volume ratio g/mL to carry out microwave extraction, microwave power 300W, extraction time 20min, extraction temperature 50°C, to obtain the second extraction solution and the second liquid residue;
  • the first extract and the second extract are mixed and filtered, the filtrate is concentrated under reduced pressure, and the temperature of concentration under reduced pressure is 50 ° C, and the extract is recovered from the solvent to a paste to obtain a concentrate;
  • a preparation method of morning glory seed extract comprising the following steps:
  • step S2 morning glory powder in 95% (v/v) ethanol solution by mass volume ratio of g/mL for 1:8 to soak for 2h, obtain soaking liquid, repeat 4 times;
  • the soaking solution is concentrated under reduced pressure, and the concentration under reduced pressure temperature is 50° C. to obtain morning glory seed powder after soaking;
  • adding the first liquid residue to ethyl acetate is 1:5 by mass-volume ratio g/mL to carry out microwave extraction, microwave power 400W, extraction time 30min, extraction temperature 55°C, to obtain the second extraction solution and the second liquid residue;
  • a preparation method of morning glory seed extract comprising the following steps:
  • Step S2 morning glory powder is placed in 95% (v/v) ethanol solution by mass to volume ratio of g/mL 1:10 and soaked for 1h to obtain soaking solution, repeated 5 times;
  • the soaking solution is concentrated under reduced pressure, and the concentration under reduced pressure temperature is 60° C. to obtain morning glory seed powder after soaking;
  • a preparation method of morning glory seed extract comprising the following steps:
  • Step S2 morning glory powder is placed in 95% (v/v) ethanol solution by mass-volume ratio g/mL to be 1:15 to soak for 3h, to obtain soaking liquid, repeat 3 times;
  • the soaking solution is concentrated under reduced pressure, and the concentration under reduced pressure temperature is 60° C. to obtain morning glory seed powder after soaking;
  • the first extract and the second extract are mixed and filtered, the filtrate is concentrated under reduced pressure, and the temperature of concentration under reduced pressure is 60 ° C, and the extract is recovered from the solvent to a paste to obtain a concentrate;
  • step S5 the soaked morning glory seed powder is added to ethyl acetate at a mass-to-volume ratio of g/mL of 1:20 for microwave extraction, the microwave power is 800W, the extraction time is 120min, and the extraction temperature 80°C to obtain the first extraction solution and the first liquid residue.
  • step S8 ADS-17 macroporous resin is not used, and the type of macroporous resin used is AB-7.
  • step S2 sodium bicarbonate solution is not used for soaking.
  • mice Male and female, weighing 18-20kg, and divide the mice into 9 groups, with 12 mice in each group.
  • the volume was continuously gavaged for 7 days, and the morning glory seed extracts prepared in Examples 1-4 and Comparative Examples 1-3 were continuously gavaged for 7 days at a capacity of 20 mL/kg, and then the mice were fasted for 16 hours without water, and the mice were observed.
  • Mice were given activated carbon mixture by gavage, mixed with 5 g of activated carbon and 10 mL of water, and after another 30 min, all mice were sacrificed by decapitation, and the small intestine propulsion rate was calculated.
  • A is the distance of activated carbon from the pylorus to the small intestine
  • B is the total length of the small intestine
  • the small intestine propulsion rate A/B ⁇ 100%.
  • the above results show that compared with the blank group, the small intestine propulsion rate of the atropine model group is significantly reduced, indicating that the atropine model induces small intestinal peristalsis in mice and has an inhibitory effect.
  • the propulsion rate of the small intestine by the bovine seed extract increased significantly, and the difference was statistically significant (P ⁇ 0.05), indicating that both the examples 1-4 and the comparative examples 1-3 had the small intestine propulsion function, and the examples 1-4 were prepared according to the method of the present invention.
  • the propelling effect of the morning glory seed extract is better, indicating that the morning glory seed extract prepared by the present invention can promote gastrointestinal motility.
  • mice with a body weight of 18-20 kg, and divide the mice into 8 groups on average, with 10 mice in each group.
  • the volume of kg was gavaged, and the blank group was directly gavaged with unextracted morning glory, and the mortality of mice was observed within 12 hours, as follows:
  • Example 1 Example 2 Example 3 Example 4 Comparative Example 1 Comparative Example 2 Comparative Example 3 blank group Number of mice (only) 10 10 10 10 10 10 10 10 Number of dead mice (only) 0 0 0 0 3 2 4 7
  • mice the drug toxicity of the morning glory seed extract prepared according to the method of the present invention is significantly reduced, and the mortality rate of mice in Comparative Examples 1-3 without the preparation of the morning glory seed extract according to the method of the present invention is higher, and the blank group
  • the mortality rate of mice directly gavaged with morning glory was the highest, mainly due to the hydrolysis of petrolidin into petronine acid during the preparation of petunia extract, thereby reducing the toxicity of petunia.

Landscapes

  • Health & Medical Sciences (AREA)
  • Natural Medicines & Medicinal Plants (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Veterinary Medicine (AREA)
  • Chemical & Material Sciences (AREA)
  • Public Health (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • General Chemical & Material Sciences (AREA)
  • Biotechnology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Nutrition Science (AREA)
  • Alternative & Traditional Medicine (AREA)
  • Organic Chemistry (AREA)
  • Botany (AREA)
  • Medical Informatics (AREA)
  • Microbiology (AREA)
  • Mycology (AREA)
  • Epidemiology (AREA)
  • Medicines Containing Plant Substances (AREA)

Abstract

本发明提供一种牵牛子提取物的制备方法，包括以下步骤：S1.牵牛子中药材粉碎，过筛；S2.将粉碎后牵牛子浸泡于碳酸氢钠溶液，而后浓缩得牵牛子粉末；S3.将步骤S2牵牛子粉末放置乙醇溶液浸泡；S4.将浸泡液进行减压浓缩得到浸泡后牵牛子粉；S5.将浸泡后牵牛子粉加入乙酸乙酯进行微波提取，得到第一提取液和第一液渣；S6.将第一液渣加入乙酸乙酯进行微波提取得第二提取液和第二液渣；S7.将第一提取液和第二提取液混合后进行过滤，滤液减压浓缩，提取液回收溶剂至膏状；S8.加入去离子水溶解，将所得溶液缓慢加入大孔树脂中进行纯化，静置1-2h，用乙醇水溶液进行梯度洗脱，而后减压浓缩，真空干燥，得牵牛子提取物。

Description

一种牵牛子提取物的制备方法 技术领域
本发明涉及从牵牛子中提取有效成分领域，特别涉及一种牵牛子提取物的制备方法。
背景技术
牵牛，属旋花科牵牛属，别名，黑丑、白丑、二丑、喇叭花、牵牛。牵牛花的颜色有蓝、绯红、桃红、紫等，亦有混色的，花瓣边缘的变化较多，是常见的观赏植物。果实卵球形，可以入药，牵牛花叶子三裂，基部心形，花呈白色、紫红色或紫蓝色，漏斗状，全株有粗毛。裂叶牵牛为一年生缠绕性草质藤本，全株密被粗硬毛。顶端5浅裂蒴果球形，种子卵状三棱形，表面灰黑色者为黑丑，淡黄白色者为白丑。
现有文献报道了牵牛子包含苷类、生物碱类、黄酮类及蒽醌、酚酸类等多种化学成分，具有抗肿瘤作用、泻下利尿，兴奋离体兔肠、抑菌和兴奋离体大鼠子宫平滑肌等多种药理作用。传统提取牵牛子有效成分方法上包括振荡提取、溶剂提取、回流提取等，但传统方法具有温度高、时间长、提取效率率低，还会造成环境污染等问题，温度高可能会导致中药中组分被分解，现需一种方法来解决上述技术问题。
发明内容
鉴于此，本发明提出一种牵牛子提取物的制备方法，解决上述技术问题。
本发明的技术方案是这样实现的：一种牵牛子提取物的制备方法，包括以下步骤：
S1.取牵牛子，粉碎，过20-60目筛；
S2.将粉碎后牵牛子浸泡于碳酸氢钠溶液，而后浓缩得牵牛子粉末；
S3.将步骤S2牵牛子粉末放置93-96％(v/v)乙醇溶液按质量体积比g/mL1:5-15浸泡，得浸泡液；
S4.将浸泡液进行减压浓缩得到浸泡后牵牛子粉；
S5.将浸泡后牵牛子粉加入乙酸乙酯进行微波提取，微波功率300-500W，提取时间30-60min，提取温度60-70℃，所述牵牛子粉和乙酸乙酯的质量体积比g/mL为1:8-15，得到第一提取液和第一液渣；
S6.将第一液渣加入乙酸乙酯进行微波提取，微波功率300-500W，提取时间20-50min，提取温度50-55℃，所述第一液渣和乙酸乙酯的质量体积比g/mL为1:3-8，得第二提取液和第二液渣；
S7.将第一提取液和第二提取液混合后进行过滤，滤液减压浓缩，提取液回收溶剂至膏状，得到浓缩物；
S8.将浓缩物加入去离子水溶解，所述浓缩物和去离子水的质量体积比g/mL为1:10-12，将所得溶液缓慢加入ADS-17大孔树脂中进行纯化，静置1-2h，采用水：乙醇按照1:9、3:7、5:5体积比例进行梯度洗脱后收集流分而后减压浓缩，真空干燥得牵牛子提取物。
优选地，在步骤S2中，所述牵牛子和碳酸氢钠溶液的质量体积比g/mL为1:15-20，所述浸泡的时间为30-60min。
优选地，在步骤S3中，所述乙醇溶液浸泡时间为1-3h，重复3-5次。
优选地，所述减压浓缩温度为50-60℃。
优选地，所述牵牛子提取物在制备泻下利尿药物中的应用。
与现有技术相比，本发明的有益效果是：
本发明一种牵牛子提取物的制备方法，制备工艺简单，生产周期短，使用乙醇、乙酸乙酯都可以回收利用，对环境友好，本发明方法将牵牛子粉碎过筛，用碳酸氢钠溶液浸泡牵牛子粉末，牵牛子中部分牵牛子苷被水解成牵牛子酸，能刺激肠道黏膜，增加蠕动导致泻下，同时减弱牵牛子苷毒性，免伤正气，缓和药性，保证临床用药安全有效，采用乙酸乙酯作为溶剂进行微波提取，微波提取相对于沉淀提取、溶剂提取具有操作简单、提取速度快、收率高、有效成分破坏较少，不易造成环境污染等优点，适合大量工业生产。大孔树脂中进行纯化，用乙醇溶液进行梯度洗脱三次减压浓缩，制备牵牛子提取物过程中，设置优质配比，降低牵牛子提取物杂质含量，有效成分生物碱类、蒽醌和酚酸类提高。
而且，本发明的提取方法，工艺简单、生产周期短、绿色环保，牵牛子有效成分提取率高，适合于工业化生产，为高效利用牵牛子提供良好的途径。
具体实施方式
为了更好理解本发明技术内容，下面提供具体实施例，对本发明做进一步的说明。
本发明实施例所用的实验方法如无特殊说明，均为常规方法。
本发明实施例所用的材料、试剂等，如无特殊说明，均可从商业途径得到。
实施例1
一种牵牛子提取物的制备方法，包括以下步骤：
S1.取牵牛子，粉碎，过20目筛；
S2.将粉碎后牵牛子放置于碳酸氢钠溶液按质量体积比g/mL为1:15浸泡30min，后浓缩得牵牛子粉末；
S3.将步骤S2牵牛子粉末放置于95％(v/v)乙醇溶液按质量体积比g/mL为1:5浸泡1h，得浸泡液，重复3次；
S4.将浸泡液进行减压浓缩，减压浓缩温度为50℃，得到浸泡后牵牛子粉；
S5.将浸泡后牵牛子粉加入乙酸乙酯按质量体积比g/mL为1:8进行微波提取，微波功率300W，提取时间3min，提取温度60℃，得到第一提取液和第一液渣；
S6.将第一液渣加入乙酸乙酯按质量体积比g/mL为1:3进行微波提取，微波功率300W，提取时间20min，提取温度50℃，得第二提取液和第二液渣；
S7.将第一提取液和第二提取液混合后进行过滤，滤液减压浓缩，减压浓缩温度为50℃，提取液回收溶剂至膏状，得到浓缩物；
S8.将浓缩物加入去离子水按质量体积比g/mL为1:10进行溶解，将所得溶液缓慢加入ADS-17大孔树脂中进行纯化，静置1h，采用水：乙醇按照1:9、3:7、5:5体积比例进行梯度洗脱后收集流分而后减压浓缩，减压浓缩温度为50℃，真空干燥得牵牛子提取物。
实施例2
一种牵牛子提取物的制备方法，包括以下步骤：
S1.取牵牛子，粉碎，过30目筛；
S2.将粉碎后牵牛子放置碳酸氢钠溶液按质量体积比g/mL为1:18浸泡 45min，后浓缩得牵牛子粉末；
S3.将步骤S2牵牛子粉末放置95％(v/v)乙醇溶液按质量体积比为g/mL为1:8浸泡2h，得浸泡液，重复4次；
S4.将浸泡液进行减压浓缩，减压浓缩温度为50℃，得到浸泡后牵牛子粉；
S5.将浸泡后牵牛子粉加入乙酸乙酯按质量体积比g/mL为1:10进行微波提取，微波功率400W，提取时间50min，提取温度65℃，得到第一提取液和第一液渣；
S6.将第一液渣加入乙酸乙酯按质量体积比g/mL为1:5进行微波提取，微波功率400W，提取时间30min，提取温度55℃，得第二提取液和第二液渣；
S7.将第一提取液和第二提取液混合后进行过滤，滤液减压浓缩，减压浓缩温度为50℃，提取液回收溶剂至膏状得到浓缩物；
S8.将浓缩物加入去离子水按质量体积比g/mL为1:10进行溶解，将所得溶液缓慢加入ADS-17大孔树脂中进行纯化，静置1h，采用水：乙醇按照1:9、3:7、5:5体积比例进行梯度洗脱后收集流分而后减压浓缩，减压浓缩温度为50℃，真空干燥得牵牛子提取物。
实施例3
一种牵牛子提取物的制备方法，包括以下步骤：
S1.取牵牛子，粉碎，过50目筛；
S2.将粉碎后牵牛子放置于碳酸氢钠溶液按质量体积比g/mL为1:15浸泡60min，后浓缩得牵牛子粉末；
S3.将步骤S2牵牛子粉末放置95％(v/v)乙醇溶液按质量体积比为g/mL1:10浸泡1h，得浸泡液，重复5次；
S4.将浸泡液进行减压浓缩，减压浓缩温度为60℃，得到浸泡后牵牛子粉；
S5.将浸泡后牵牛子粉加入乙酸乙酯按质量体积比g/mL为1:10进行微波提取，微波功率300W，提取时间60min，提取温度65℃，得到第一提取液和第一液渣；
S6.将第一液渣加入乙酸乙酯按质量体积比g/mL1:6进行微波提取，微波功率300W，提取时间30min，提取温度55℃，得第二提取液和第二液渣；
S7.将第一提取液和第二提取液混合后进行过滤，滤液减压浓缩，减压浓缩 温度为60℃，提取液回收溶剂至膏状，得浓缩物；
S8.将浓缩物加入去离子水按质量体积比g/mL为1:10进行溶解，将所得溶液缓慢加入ADS-17大孔树脂中进行纯化，静置1h，采用水：乙醇按照1:9、3:7、5:5体积比例进行梯度洗脱后收集流分而后减压浓缩，减压浓缩温度为60℃，真空干燥得牵牛子提取物。
实施例4
一种牵牛子提取物的制备方法，包括以下步骤：
S1.取牵牛子，粉碎，过60目筛；
S2.将粉碎后牵牛子放置碳酸氢钠溶液按质量体积比g/mL为1:20浸泡60min，后浓缩得牵牛子粉末；
S3.将步骤S2牵牛子粉末放置95％(v/v)乙醇溶液按质量体积比g/mL为1:15浸泡3h，得浸泡液，重复3次；
S4.将浸泡液进行减压浓缩，减压浓缩温度为60℃，得到浸泡后牵牛子粉；
S5.将浸泡后牵牛子粉加入乙酸乙酯按质量体积比g/mL为1:15进行微波提取，微波功率500W，提取时间60min，提取温度70℃，得到第一提取液和第一液渣；
S6.将第一液渣加入乙酸乙酯按质量体积比g/mL为1:8进行微波提取，微波功率300W，提取时间20min，提取温度50℃，得第二提取液和第二液渣；
S7.将第一提取液和第二提取液混合后进行过滤，滤液减压浓缩，减压浓缩温度为60℃，提取液回收溶剂至膏状，得浓缩物；
S8.将浓缩物加入去离子水按质量体积比g/mL为1:12进行溶解，将所得溶液缓慢加入ADS-17大孔树脂中进行纯化，静置1h，采用水：乙醇按照1:9、3:7、5:5体积比例进行梯度洗脱后收集流分而后减压浓缩，减压浓缩温度为60℃，真空干燥得牵牛子提取物。
对比例1
本对比例与实施例3区别在于，在步骤S5中，将浸泡后牵牛子粉加入乙酸乙酯按质量体积比g/mL为1:20进行微波提取，微波功率800W，提取时间120min，提取温度80℃，得到第一提取液和第一液渣。
对比例2
本对比例与实施例3区别在于，在步骤S8中，没有使用ADS-17大孔树脂，使用的大孔树脂的型号为AB-7。
对比例3
本对比例与实施例3区别在于，在步骤S2中，没有使用碳酸氢钠溶液浸泡。
一、牵牛子提取物对小肠推进影响试验
选取昆明种小鼠108只，雌雄不分，体重为18-20kg，将小鼠平均分为9组，每组12只，空白组给予纯水灌胃7天，阿托品模型组按20mL/kg的容量连续灌胃7天，实施例1-4和对比例1-3制备牵牛子提取物均按20mL/kg的容量连续灌胃7天，而后16h禁食不禁水，观察小鼠，30min后，小鼠给予活性炭混合物灌胃，活性炭5g，水10mL混合，再30min后，所有小鼠脱颈处死，计算小肠推进率。A为活性炭从幽门到小肠中的距离，B为小肠的总长度，小肠推进率＝A/B×100％。
Figure PCTCN2021085215-appb-000001
上述结果表明，与空白组比较，阿托品模型组小肠推进率显著降低，说明阿托品模诱导小鼠小肠蠕动起到抑制效果，与阿托品模型组比较，实施例1-4和对比例1-3制备牵牛子提取物对小肠推进率均显著上升，差异有统计学意义(P＜0.05)，说明实施例1-4和对比例1-3都具有小肠推进功能，实施例1-4按照本发明方法制备牵牛子提取物推进效果更佳，说明本发明制备牵牛子提取物可促进胃肠运动。
二、毒性试验
选择小白鼠雄鼠80只，体重为18-20kg，将小鼠平均分为8组，每组10只，分别将实施例1-4和对比例1-3制备出牵牛子提取物按20mL/kg的容量灌胃，空白组是直接将未提取牵牛子灌胃，12h内观察小鼠死亡率，如下：
实施例1 实施例2 实施例3 实施例4 对比例1 对比例2 对比例3 空白组
小鼠数量(只) 10 10 10 10 10 10 10 10
小鼠死亡数量(只) 0 0 0 0 3 2 4 7
上述结果表明，从小鼠死亡率可以看出，按照本发明方法制备牵牛子提取物药物毒性明显降低，对比例1-3没有按照本发明方法制备牵牛子提取物小鼠死亡率较高，空白组直接将牵牛子灌胃小鼠死亡率最高，主要是由于制备牵牛子提取物过程中牵牛子苷被水解成牵牛子酸，从而降低牵牛子毒性。
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 (5)

  1. 一种牵牛子提取物的制备方法，其特征在于，包括以下步骤：
    S1.取牵牛子中药材，粉碎，过20-60目筛；
    S2.将粉碎后牵牛子浸泡于碳酸氢钠溶液，而后浓缩得牵牛子粉末；
    S3.将步骤S2牵牛子粉末放置93-96％v/v乙醇溶液按质量体积比g/mL为1:5-15浸泡，得浸泡液；
    S4.将浸泡液进行减压浓缩得到浸泡后牵牛子粉；
    S5.将浸泡后牵牛子粉加入乙酸乙酯进行微波提取，微波功率300-500W，提取时间30-60min，提取温度60-70℃，所述牵牛子粉和乙酸乙酯的质量体积比g/mL为1:8-15，得到第一提取液和第一液渣；
    S6.将第一液渣加入乙酸乙酯进行微波提取，微波功率300-500W，提取时间20-50min，提取温度50-55℃，所述第一液渣和乙酸乙酯的质量体积比g/mL为1:3-8，得第二提取液和第二液渣；
    S7.将第一提取液和第二提取液混合后进行过滤，滤液减压浓缩，提取液回收溶剂至膏状，得浓缩物；
    S8.将浓缩物加入去离子水溶解，所述浓缩物和去离子水的质量体积比g/mL为1:10-12，将所得溶液加入ADS-17大孔树脂中进行纯化，静置1-2h，采用水：乙醇按照1:9、3:7、5:5体积比例进行梯度洗脱后收集流分而后减压浓缩，真空干燥，得牵牛子提取物。
  2. 根据权利要求1所述的牵牛子提取物的制备方法，其特征在于，在步骤S2中，所述牵牛子和碳酸氢钠溶液的质量体积比g/mL为1:15-20，所述浸泡的时间为30-60min。
  3. 根据权利要求1所述的牵牛子提取物的制备方法，其特征在于，在步骤S3中，所述浸泡的时间为1-3h，重复3-5次。
  4. 根据权利要求1所述的牵牛子提取物的制备方法，其特征在于，所述减压浓缩温度为50-60℃。
  5. 根据权利要求1至4任一项所述的牵牛子提取物的制备方法制得牵牛子提取物应用于制备泻下利尿药物。
PCT/CN2021/085215 2021-02-26 2021-04-02 一种牵牛子提取物的制备方法 WO2022178953A1 (zh)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN202110215606.7A CN112972519B (zh) 2021-02-26 2021-02-26 一种牵牛子提取物的制备方法
CN202110215606.7 2021-02-26

Publications (1)

Publication Number Publication Date
WO2022178953A1 true WO2022178953A1 (zh) 2022-09-01

Family

ID=76350967

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2021/085215 WO2022178953A1 (zh) 2021-02-26 2021-04-02 一种牵牛子提取物的制备方法

Country Status (2)

Country Link
CN (1) CN112972519B (zh)
WO (1) WO2022178953A1 (zh)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112972519B (zh) * 2021-02-26 2022-05-17 海南葫芦娃药业集团股份有限公司 一种牵牛子提取物的制备方法

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102283971A (zh) * 2011-07-27 2011-12-21 孔维巍 一种治疗各类胃炎的中成药
CN104558221A (zh) * 2013-10-25 2015-04-29 匡海学 一种牵牛子及其炮制品多糖提取物的制备方法及用途
CN112972519A (zh) * 2021-02-26 2021-06-18 海南葫芦娃药业集团股份有限公司 一种牵牛子提取物的制备方法

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107467264A (zh) * 2017-08-21 2017-12-15 佛山市三水区嘉信农业技术研究院（普通合伙） 一种改善便秘的黑茶饮

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102283971A (zh) * 2011-07-27 2011-12-21 孔维巍 一种治疗各类胃炎的中成药
CN104558221A (zh) * 2013-10-25 2015-04-29 匡海学 一种牵牛子及其炮制品多糖提取物的制备方法及用途
CN112972519A (zh) * 2021-02-26 2021-06-18 海南葫芦娃药业集团股份有限公司 一种牵牛子提取物的制备方法

Non-Patent Citations (7)

* Cited by examiner, † Cited by third party
Title
"Explanations of 8404 Types of Natural Medicines", 30 June 2014, PEOPLE'S MILITARY MEDICAL PRESS, CN, ISBN: 978-7-5091-7256-8, article HUANG, CHUANGUI: "Semen Pharbitidis", pages: 1622 - 1623, XP009539322 *
AO DONGMEI , WEI QUN: "Research progress of morning glory", CHINESE JOURNAL OF INFORMATION ON TRADITIONAL CHINESE MEDICINE, 15 April 2003 (2003-04-15), pages 77 - 80, XP055961785 *
CHEN LI-NA,LI PING: "Study on Chemical Constituents of Semen Pharbitidis (II)", CHEMISTRY AND INDUSTRY OF FOREST PRODUCTS, vol. 27, no. 6, 15 December 2007 (2007-12-15), pages 105 - 108, XP055961794, ISSN: 0253-2417 *
CHEN LI-NA,LI PING: "Study on Chemical Constituents of Semen Pharbitidis", CHINESE JOURNAL OF NATURAL MEDICINES, no. 03, 1 January 2004 (2004-01-01), CN , pages 146 - 148, XP009539308, ISSN: 2095-6975 *
SUN YANPING;WANG YANHONG;YANG BINGYOU;GE GUANGBO;YANG LING;WANG QIUHONG;KUANG HAIXUE: "Evaluation on the Property and Flavor Pharmacology of Chemical Split Fractions from Semen Pharbitidis and Study on Its Flavor Ascription", WORLD CHINESE MEDICINE, 15 December 2015 (2015-12-15), pages 1837 - 1846+1853, XP055961775 *
ZHANG DANYU; JI YUBIN; XU XUDONG; MA GUOXU: "Research progress on chemical constituents and pharmacological effects of morning glory", SCIENTIFIC AND TECHNOLOGICAL INNOVATION, 25 October 2018 (2018-10-25), pages 13 - 14, XP055961780 *
ZHANG SONG; CHEN ZHAOWEN; QIANG MEIYU; CHEN SUFEN; ZHOU HAOBING: "Study on Morning Glory", JOURNAL OF CHINA PHARMACEUTICAL UNIVERSITY, 10 June 1959 (1959-06-10), pages 36 - 40, XP055961790 *

Also Published As

Publication number Publication date
CN112972519B (zh) 2022-05-17
CN112972519A (zh) 2021-06-18

Similar Documents

Publication Publication Date Title
CN101721452B (zh) 一种提高紫草利用率的新工艺
CN109810201A (zh) 一种蛹虫草中虫草多糖和虫草素的超声波复合酸性水提取方法
WO2020073976A1 (zh) 一种叶黄素和万寿菊黄酮的工业化制备方法
CN102002108A (zh) 一种拐枣多糖的制备方法
CN103113195A (zh) 一种快速制备羟基酪醇的新方法
CN102180940A (zh) 一种化合物文冠果壳苷的制备方法
CN111732622A (zh) 从枳实中提取橙皮苷的方法
WO2022178953A1 (zh) 一种牵牛子提取物的制备方法
CN104623095A (zh) 枸杞活性成分的提取方法
CN114425029A (zh) 具有抗炎舒敏的植物提取组合物、制备方法及其应用
CN114774493A (zh) 一种桑黄活性物质多糖的提取及纯化方法
CN108383885B (zh) 从槐角中提取槐角苷的工艺及其在抗炎症中的用途
CN112760163A (zh) 一种山桐子油脂制备工艺
CN107793492A (zh) 一种火麻仁多糖的提取方法
CN111777656A (zh) 一种从新鲜黄芪中提取黄芪甲苷的方法
CN103073599B (zh) 一种皂素的浓缩提取方法
CN106117284B (zh) 一种同时提取富集金银花中六种环烯醚萜苷类成分的方法
CN110841004A (zh) 一种从叠鞘石斛中热回流低温提取叠鞘石斛提取物的工艺
CN101623310B (zh) 从萝芙木中提取总碱、利血平和芦丁的方法
CN106635394B (zh) 一种油茶籽或茶枯的综合利用方法
CN112717009A (zh) 一种栀子提取物的制备方法
CN111329888A (zh) 一种三七花片及其生产制备工艺
CN112592413B (zh) 一种金荞麦多糖的制备方法
CN116622002B (zh) 一种辣木叶提取物的制备方法
CN108929350A (zh) 一种从醒脑静注射液蒸馏提取废液中回收栀子苷的方法

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 21927387

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 21927387

Country of ref document: EP

Kind code of ref document: A1