WO2017185275A1 - 一种罗布麻叶提取物的制备方法 - Google Patents

一种罗布麻叶提取物的制备方法 Download PDF

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WO2017185275A1
WO2017185275A1 PCT/CN2016/080455 CN2016080455W WO2017185275A1 WO 2017185275 A1 WO2017185275 A1 WO 2017185275A1 CN 2016080455 W CN2016080455 W CN 2016080455W WO 2017185275 A1 WO2017185275 A1 WO 2017185275A1
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extract
apocynum venetum
extraction
residue
stage
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PCT/CN2016/080455
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French (fr)
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朱小涛
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朱小涛
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Priority to PCT/CN2016/080455 priority Critical patent/WO2017185275A1/zh
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    • 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)
    • A61K36/24—Apocynaceae (Dogbane family), e.g. plumeria or periwinkle

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  • the invention relates to the field of biological extraction technology, in particular to a preparation method of apocynum venetum leaf extract.
  • Apocynum venetum L. is a traditional Pinggan Xifeng drug. It has the functions of calming the liver and calming the nerves, nourishing blood and regulating menstruation, and relieving yin and antiperspirant. Modern pharmacological research has found that it also has a wide range of blood lipid lowering, anti-atherosclerosis and liver protection. Pharmacological effects. In recent years, "Apocynum venetum tea" and other commodities have appeared on the market in large quantities, and are often used clinically for the treatment of hypertension or blood pressure lowering, and the effective part thereof is a flavonoid compound.
  • quercetin-3-O- ⁇ -D-glucosyl- ⁇ -D-glucoside quercetin-3-O- ⁇ -D-glucuronide
  • the extraction methods of candidin, isoquercetin, quercetin and kaempferol have important research value and broad application prospects.
  • Apocynum venetum L. extract has strong anti-aging, anti-oxidation, ultraviolet protection, moisturizing, anti-bacterial and anti-inflammatory, ecchymosis and acne effects, and its skin protection effect is remarkable.
  • the present invention proposes a preparation method of apocynum venetum leaf extract, which has high extraction speed and extraction precision, low production cost and energy saving.
  • the preparation method of the Apocynum venetum leaf extract proposed by the invention comprises the following steps:
  • the Apocynum venetum leaves are dried at a low temperature, pulverized, sieved, and then degreased to obtain a filter cake;
  • the resin After mixing the extract and the extract, the resin is adsorbed on the macropores, and then eluted with water and an aqueous solution of ethanol, concentrated, and dried to obtain an extract of Apocynum venetum L. leaves.
  • the temperature for drying at a low temperature is 18 to 20 ° C
  • the pressure for drying at a low temperature is 3 to 6 Pa
  • the time for drying at a low temperature is 17 to 20 hours.
  • the ultrasonic extraction power is 620-650 w
  • the ultrasonic extraction frequency is 55-65 kHz
  • the ultrasonic extraction time is 3.5-4.5 h.
  • the specific operation of the multi-stage countercurrent pulse extraction is as follows: the residue is sent to the extractor, and the extractant of the Apocynum venetum L. extract is subjected to N-stage spray immersion extraction to obtain an extract and a wet residue;
  • the mass fraction of the extract of Apocynum venetum L. contained in the extractant used in the M grade is higher than the mass fraction of the extract of Apocynum venetum L. contained in the extractant used in the M-1 grade, 2 ⁇ M ⁇ N.
  • the advantage of the N-stage spray immersion extraction is that the pure extractant is used in each stage, and the amount of the extractant is very large, but the concentration of the extract of Apocynum venetum leaves is very low, and the above method can be used to reduce the extractant.
  • the amount is obtained, and a high concentration of Apocynum venetum L. leaf extract is obtained to reduce the load of the subsequent process.
  • the extractant is an aqueous solution of ethanol having a concentration of 40 to 50%.
  • the mass ratio of the extractant to the residue is 1.2 to 1.5:1.
  • the temperature of the multi-stage countercurrent pulse extraction is 45 to 50 ° C, and the extraction time of the N-stage spray immersion is 90 to 120 min.
  • the ratio of material to liquid is 1:10 to 12
  • the pH of the solution is 3 to 5.
  • the concentration of the aqueous ethanol solution in S4 is 50 to 60%.
  • the ultrasonic extraction of the invention can generate strong vibration, high acceleration, strong cavitation and agitation. When used for solvent-assisted extraction, it can accelerate the extraction of Apocynum venetum leaves into the solvent, thereby shortening the extraction time, saving the solvent, and avoiding the high temperature leading to Apocynum venetum.
  • the leaf extract is decomposed, but the extract of Apocynum venetum L. obtained by ultrasonic extraction is insufficient in purity and the residual rate in the residue is high.
  • the invention adopts ultrasonic extraction and multi-stage countercurrent pulse extraction for series connection, which not only improves the extraction speed and extraction precision, but also greatly reduces the production cost, and the extract of Apocynum venetum leaves extracted from Apocynum venetum leaves is developed for the prevention and treatment of related enterprises.
  • Medicines and health foods that play a role in atherosclerosis, hypolipidemic and liver-protecting gallbladder provide high-quality raw materials.
  • the preparation method of the Apocynum venetum leaf extract proposed by the invention comprises the following steps:
  • the Apocynum venetum leaves are dried at a low temperature, the temperature of low temperature drying is 18 ° C, the pressure of low temperature drying is 6 Pa, the drying time of low temperature is 17 h, pulverization, sieving, followed by degreasing to obtain a filter cake;
  • the ultrasonic extraction power is 650w
  • the ultrasonic extraction frequency is 55kHz
  • the ultrasonic extraction time is 4.5h
  • the extraction liquid and the residue are obtained by filtration
  • the residue is sent to the extractor, and the extracting agent of Apocynum venetum L. extract is used for 8-stage spray immersion extraction to obtain an extract and a wet residue ⁇ ; wherein the extract agent of the M-stage extract contains Apocynum venetum leaves.
  • the mass fraction is higher than the mass fraction of Apocynum venetum L. extract contained in the extractant used in the M-1 grade, 2 ⁇ M ⁇ 8; the extractant is a 40% ethanol aqueous solution; the mass ratio of the extractant to the residue is 1.5. :1; the temperature of multi-stage countercurrent pulse extraction is 45 °C, and the extraction time of N-stage spray immersion is 120 min; the mass fraction of Apocynum venetum L. extract in the extractant of 8 spray immersion extraction is 20wt%, 19wt%, 14 wt%, 11 wt%, 4 wt%, 3 wt%, 0.3 wt%, 0 wt%;
  • the macroporous adsorption resin After mixing the extract and the extract, the macroporous adsorption resin is used, the ratio of material to liquid is 1:12, the pH of the solution is 3, and then eluted with water and a 60% aqueous solution of ethanol, concentrated, and dried. , obtained apocynum venetum leaf extract.
  • the preparation method of the Apocynum venetum leaf extract proposed by the invention comprises the following steps:
  • the apocynum venetum leaves are dried at a low temperature, the temperature of low temperature drying is 20 ° C, the pressure of low temperature drying is 3 Pa, the time of low temperature drying is 20 h, pulverization, sieving, followed by degreasing to obtain a filter cake;
  • the ultrasonic extraction power is 620w
  • the frequency of ultrasonic extraction is 65kHz
  • the ultrasonic extraction time is 3.5h, and then the extract and the residue are filtered;
  • the residue is sent to the extractor, and the extracting agent of Apocynum venetum L. extract is used for 8-stage spray immersion extraction to obtain an extract and a wet residue ⁇ ; wherein the extract agent of the M-stage extract contains Apocynum venetum leaves.
  • the mass fraction is higher than the mass fraction of Apocynum venetum L. extract contained in the extractant used in the M-1 grade, 2 ⁇ M ⁇ 8; the extractant is an aqueous ethanol solution having a concentration of 50%; the mass ratio of the extractant to the residue is 1.2. :1; the temperature of multi-stage countercurrent pulse extraction is 50 °C, the time of N-stage spray immersion extraction is 90 min; the mass fraction of Apocynum venetum L. extract in the extractant of 8 spray immersion extraction is 24wt%, 17wt%, 16 wt%, 9 wt%, 6 wt%, 1.5 wt%, 0.8 wt%, 0 wt%;
  • the macroporous adsorption resin is used, the ratio of material to liquid is 1:10, the pH of the solution is 5, and then eluted with water and a 50% aqueous solution of ethanol, concentrated, and dried. , obtained apocynum venetum leaf extract.
  • the preparation method of the Apocynum venetum leaf extract proposed by the invention comprises the following steps:
  • the apocynum venetum leaves are dried at a low temperature, the temperature of low temperature drying is 19 ° C, the pressure of low temperature drying is 5 Pa, the drying time of low temperature is 18 h, pulverization, sieving, followed by degreasing to obtain a filter cake;
  • the ultrasonic extraction power is 630w
  • the ultrasonic extraction frequency is 60kHz
  • the ultrasonic extraction time is 4h
  • the extract and the residue are obtained by filtration
  • the residue is sent to the extractor, and the extracting agent of Apocynum venetum L. extract is used for 8-stage spray immersion extraction to obtain an extract and a wet residue ⁇ ; wherein the extract agent of the M-stage extract contains Apocynum venetum leaves.
  • the mass fraction is higher than the mass fraction of Apocynum venetum L. extract contained in the extractant used in the M-1 grade, 2 ⁇ M ⁇ 8; the extractant is a 45% ethanol aqueous solution; the mass ratio of the extractant to the residue is 1.3. :1; the temperature of multi-stage countercurrent pulse extraction is 48 °C, and the extraction time of N-stage spray immersion is 105 min;
  • the mass fraction of Apocynum venetum L. extract in the extractant of 8 spray immersion extraction is 22wt%, 18wt%, 15 wt%, 10 wt%, 5 wt%, 2 wt%, 0.5 wt%, 0 wt%;
  • the macroporous adsorption resin After mixing the extract and the extract, the macroporous adsorption resin is used, the ratio of material to liquid is 1:11, the pH of the solution is 4, and then eluted with water and a 55% aqueous solution of ethanol, concentrated, and dried. , obtained apocynum venetum leaf extract.

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Abstract

一种罗布麻叶提取物的制备方法，包括：将罗布麻叶低温干燥后，粉碎，过筛，脱脂得到滤饼；将滤饼干燥后，加入无水乙醇中进行超声提取，过滤得到提取液和残渣；将残渣进行多级逆流脉冲萃取得到萃取液；将萃取液和提取液混合后，上大孔吸附树脂，然后依次采用水和乙醇水溶液洗脱，浓缩，干燥，得到罗布麻叶提取物。多级逆流脉冲萃取的具体操作如下：将残渣送入萃取器中，用罗布麻叶提取物的萃取剂进行N级喷淋浸泡萃取，得到萃取液和湿残渣粕；第M级所用萃取剂中所含罗布麻叶提取物质量分数高于第M-1级所用萃取剂中所含罗布麻叶提取物质量分数，2≤M≤N。该方法不仅提取速度和提取精度高，而且生产成本低，节约能耗。

Description

一种罗布麻叶提取物的制备方法 技术领域
本发明涉及生物提取技术领域，尤其涉及一种罗布麻叶提取物的制备方法。
背景技术
中药罗布麻叶是一味传统的平肝息风药，具有平肝安神、养血调经、敛阴止汗的作用，现代药理研究发现其还具有降血脂、抗动脉粥样硬化和肝保护等广泛的药理作用。近年来，“罗布麻茶”等商品在市场上大量出现，临床上常用于治疗高血压或辅助降血压等，其有效部位为黄酮类化合物。由于常用提取方法诸如“醇提-水沉”或“水提-醇沉”等提取罗布麻叶时，由于受到植物细胞壁的阻挡，细胞内的化学成分较难被水或乙醇溶液提取出，此外，由于常用提取方法中芦丁的含量很高，而槲皮素-3-O-β-D-葡萄糖基-β-D-葡萄糖苷、萹蓄苷、槲皮素-3-O-β-D-葡萄糖醛酸苷、金丝桃苷、异槲皮素、槲皮素和山奈酚等成分含量很低。因此，找到一种方便、且能提高槲皮素-3-O-β-D-葡萄糖基-β-D-葡萄糖苷、槲皮素-3-O-β-D-葡萄糖醛酸苷、金丝桃苷、异槲皮素、槲皮素和山奈酚的提取方法具有重要研究价值和广泛的应用前景。
而且罗布麻叶提取物作为化妆品添加剂使用时具有较强的抗衰老、抗氧化、紫外线防护、保湿、抑菌消炎、祛斑祛痘等功效，其保护皮肤的效果显著。
发明内容
基于背景技术存在的技术问题，本发明提出了一种罗布麻叶提取物的制备方法，不仅提取速度和提取精度高，而且生产成本低，节约能耗。
本发明提出的一种罗布麻叶提取物的制备方法，包括如下步骤：
S1、将罗布麻叶低温干燥后，粉碎，过筛，接着脱脂得到滤饼；
S2、将滤饼干燥后，加入无水乙醇中进行超声提取，接着过滤得到提取液和残渣；
S3、将残渣进行多级逆流脉冲萃取得到萃取液；
S4、将萃取液和提取液混合后，上大孔吸附树脂，然后依次采用水和乙醇水溶液洗脱，浓缩，干燥，得到罗布麻叶提取物。
进一步地，S1中，低温干燥的温度为18～20℃，低温干燥的压力为3～6Pa，低温干燥的时间为17～20h。
进一步地，S2中，超声提取的功率为620～650w，超声提取的频率为55～65kHz，超声提取的时间为3.5～4.5h。
进一步地，S3中，多级逆流脉冲萃取的具体操作如下：将残渣送入萃取器中，用罗布麻叶提取物的萃取剂进行N级喷淋浸泡萃取，得到萃取液和湿残渣粕；其中，第M级所用萃取剂中所含罗布麻叶提取物质量分数高于第M-1级所用萃取剂中所含罗布麻叶提取物质量分数，2≤M≤N。
N级喷淋浸泡萃取的优点为：相对于每一级都采用纯的萃取剂，其萃取剂的用量非常大，但罗布麻叶提取物的浓度很低，采用上述方式，可以减少萃取剂的用量，并得到具有较高浓度的罗布麻叶提取物，减少后续工序的负荷。
进一步地，S3的多级逆流脉冲萃取过程中，萃取剂为浓度为40～50％的乙醇水溶液。
进一步地，S3的多级逆流脉冲萃取过程中，萃取剂与残渣的质量比为1.2～1.5：1。
进一步地，S3的多级逆流脉冲萃取过程中，多级逆流脉冲萃取的温度为45～50℃，N级喷淋浸泡萃取的时间为90～120min。
进一步地，S3的多级逆流脉冲萃取过程中，N＝8，八级喷淋浸泡萃取的萃取剂中罗布麻叶提取物质量分数依次为20～24wt％、17～19wt％、14～16wt％、9～11wt％、4～6wt％、1.5～3wt％、0.3～0.8wt％、0wt％。
进一步地，S4的上大孔吸附树脂过程中，料液比为1：10～12，溶液的pH值为3～5。
进一步地，S4中的乙醇水溶液浓度为50～60％。
本发明超声提取可以产生强烈振动、高加速度、强烈空化作用及搅拌作用，用于溶剂辅助提取时，可以加速罗布麻叶提取物进入溶剂，从而缩短提取时间、节约溶剂，避免高温导致罗布麻叶提取物分解，但超声提取得到的罗布麻叶提取物纯度不足，残渣中残留率高。
本发明采用超声提取和多级逆流脉冲萃取进行串联，不仅提高了提取速度和提取精度，而且大大降低了生产成本，从罗布麻叶中提取的罗布麻叶提取物为相关企业开发生产预防和治疗动脉粥样硬化、降血脂和保肝利胆等作用的药品和保健食品提供了优质生产原料。
具体实施方式
下面，通过具体实施例对本发明的技术方案进行详细说明。
实施例1
本发明提出的一种罗布麻叶提取物的制备方法，包括如下步骤：
S1、将罗布麻叶低温干燥，低温干燥的温度为18℃，低温干燥的压力为6Pa，低温干燥的时间为17h，粉碎，过筛，接着脱脂得到滤饼；
S2、将滤饼干燥后，加入无水乙醇中进行超声提取，超声提取的功率为650w，超声提取的频率为55kHz，超声提取的时间为4.5h，接着过滤得到提取液和残渣；
S3、将残渣送入萃取器中，用罗布麻叶提取物的萃取剂进行8级喷淋浸泡萃取，得到萃取液和湿残渣粕；其中，第M级所用萃取剂中所含罗布麻叶提取物质量分数高于第M-1级所用萃取剂中所含罗布麻叶提取物质量分数，2≤M≤8；萃取剂为浓度为40％的乙醇水溶液；萃取剂与残渣的质量比为1.5：1；多级逆流脉冲萃取的温度为45℃，N级喷淋浸泡萃取的时间为120min；8级喷淋浸泡萃取的萃取剂中罗布麻叶提取物质量分数依次为20wt％、19wt％、14wt％、11wt％、4wt％、3wt％、0.3wt％、0wt％；
S4、将萃取液和提取液混合后，上大孔吸附树脂，料液比为1：12，溶液的pH值为3，然后依次采用水和浓度为60％的乙醇水溶液洗脱，浓缩，干燥，得到罗布麻叶提取物。
实施例2
本发明提出的一种罗布麻叶提取物的制备方法，包括如下步骤：
S1、将罗布麻叶低温干燥，低温干燥的温度为20℃，低温干燥的压力为3Pa，低温干燥的时间为20h，粉碎，过筛，接着脱脂得到滤饼；
S2、将滤饼干燥后，加入无水乙醇中进行超声提取，超声提取的功率为620w，超声提取的频率为65kHz，超声提取的时间为3.5h，接着过滤得到提取液和残渣；
S3、将残渣送入萃取器中，用罗布麻叶提取物的萃取剂进行8级喷淋浸泡萃取，得到萃取液和湿残渣粕；其中，第M级所用萃取剂中所含罗布麻叶提取物质量分数高于第M-1级所用萃取剂中所含罗布麻叶提取物质量分数，2≤M≤8；萃取剂为浓度为50％的乙醇水溶液；萃取剂与残渣的质量比为1.2：1；多级逆流脉冲萃取的温度为50℃，N级喷淋浸泡萃取的时间为90min；8级喷淋浸泡萃取的萃取剂中罗布麻叶提取物质量分数依次为24wt％、17wt％、16wt％、9wt％、6wt％、1.5wt％、0.8wt％、0wt％；
S4、将萃取液和提取液混合后，上大孔吸附树脂，料液比为1：10，溶液的pH值为5，然后依次采用水和浓度为50％的乙醇水溶液洗脱，浓缩，干燥，得到罗布麻叶提取物。
实施例3
本发明提出的一种罗布麻叶提取物的制备方法，包括如下步骤：
S1、将罗布麻叶低温干燥，低温干燥的温度为19℃，低温干燥的压力为5Pa，低温干燥的时间为18h，粉碎，过筛，接着脱脂得到滤饼；
S2、将滤饼干燥后，加入无水乙醇中进行超声提取，超声提取的功率为630w，超声提取的频率为60kHz，超声提取的时间为4h，接着过滤得到提取液和残渣；
S3、将残渣送入萃取器中，用罗布麻叶提取物的萃取剂进行8级喷淋浸泡萃取，得到萃取液和湿残渣粕；其中，第M级所用萃取剂中所含罗布麻叶提取物质量分数高于第M-1级所用萃取剂中所含罗布麻叶提取物质量分数，2≤M≤8；萃取剂为浓度为45％的乙醇水溶液；萃取剂与残渣的质量比为1.3：1；多级逆流脉冲萃取的温度为48℃，N级喷淋浸泡萃取的时间为105min；8级喷淋浸泡萃取的萃取剂中罗布麻叶提取物质量分数依次为22wt％、18wt％、15wt％、10wt％、5wt％、2wt％、0.5wt％、0wt％；
S4、将萃取液和提取液混合后，上大孔吸附树脂，料液比为1：11，溶液的pH值为4，然后依次采用水和浓度为55％的乙醇水溶液洗脱，浓缩，干燥，得到罗布麻叶提取物。
以上所述，仅为本发明较佳的具体实施方式，但本发明的保护范围并不局限于此，任何熟悉本技术领域的技术人员在本发明揭露的技术范围内，根据本发明的技术方案及其发明构思加以等同替换或改变，都应涵盖在本发明的保护范围之内。

Claims (10)

  1. 一种罗布麻叶提取物的制备方法，其中，包括如下步骤：
    S1、将罗布麻叶低温干燥后，粉碎，过筛，接着脱脂得到滤饼；
    S2、将滤饼干燥后，加入无水乙醇中进行超声提取，接着过滤得到提取液和残渣；
    S3、将残渣进行多级逆流脉冲萃取得到萃取液；
    S4、将萃取液和提取液混合后，上大孔吸附树脂，然后依次采用水和乙醇水溶液洗脱，浓缩，干燥，得到罗布麻叶提取物。
  2. 根据权利要求1所述罗布麻叶提取物的制备方法，其中，S1中，低温干燥的温度为18～20℃，低温干燥的压力为3～6Pa，低温干燥的时间为17～20h。
  3. 根据权利要求1所述罗布麻叶提取物的制备方法，其中，S2中，超声提取的功率为620～650w，超声提取的频率为55～65kHz，超声提取的时间为3.5～4.5h。
  4. 根据权利要求1所述罗布麻叶提取物的制备方法，其中，S3中，多级逆流脉冲萃取的具体操作如下：将残渣送入萃取器中，用罗布麻叶提取物的萃取剂进行N级喷淋浸泡萃取，得到萃取液和湿残渣粕；其中，第M级所用萃取剂中所含罗布麻叶提取物质量分数高于第M-1级所用萃取剂中所含罗布麻叶提取物质量分数，2≤M≤N。
  5. 根据权利要求4所述罗布麻叶提取物的制备方法，其中，S3的多级逆流脉冲萃取过程中，萃取剂为浓度为40～50％的乙醇水溶液。
  6. 根据权利要求4所述罗布麻叶提取物的制备方法，其中，S3的多级逆流脉冲萃取过程中，萃取剂与残渣的质量比为1.2～1.5：1。
  7. 根据权利要求4所述罗布麻叶提取物的制备方法，其中，S3的多级逆流脉冲萃取过程中，多级逆流脉冲萃取的温度为45～50℃，N级喷淋浸泡萃取的时间为90～120min。
  8. 根据权利要求4所述罗布麻叶提取物的制备方法，其中，S3的多级逆流脉冲萃取过程中，N＝8，八级喷淋浸泡萃取的萃取剂中罗布麻叶提取物质量分数依次为20～24wt％、17～19wt％、14～16wt％、9～11wt％、4～6wt％、1.5～3wt％、 0.3～0.8wt％、0wt％。
  9. 根据权利要求1所述罗布麻叶提取物的制备方法，其中，S4的上大孔吸附树脂过程中，料液比为1：10～12，溶液的pH值为3～5。
  10. 根据权利要求1所述罗布麻叶提取物的制备方法，其中，S4中的乙醇水溶液浓度为50～60％。
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