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Radix bupleuri total saponins as well as extraction process and application thereof

An extraction process, a technology of total saponins, applied in the field of total saponins and its extraction process, and the preparation of asthma medicines, to achieve the effects of inhibiting the increase of inflammatory factors, safety and side effects, and good clinical application prospects

Inactive Publication Date: 2021-03-16
CHINA PHARM UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The therapeutic effect of Bupleurum saponins on asthma has not yet been studied, so we optimized and established the extraction and refining process of Bupleurum saponins under the concept of environmental protection, and further studied the chemical composition and pharmacological effects of the refined products

Method used

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  • Radix bupleuri total saponins as well as extraction process and application thereof
  • Radix bupleuri total saponins as well as extraction process and application thereof
  • Radix bupleuri total saponins as well as extraction process and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1: Extraction and Refining Process of Bupleurum Total Saponins

[0035] Weigh the Bupleurum radix powder that has been crushed and passed through a 60-mesh sieve into a round bottom flask, add 12 times the volume of 65% ethanol, adjust the pH of the solution to 10 with NaOH, soak for 1.5 hours, reflux extraction at 90°C for 1.5 hours, and filter with suction The obtained filtrate was concentrated under reduced pressure at 50°C until it had no alcohol smell, and was appropriately diluted with distilled water as a sample solution for later use. Soak the AB-8 macroporous resin in 95% ethanol for more than 24 hours, wash with distilled water until there is no alcohol smell, wet-pack the column, add eight times the column volume of distilled water to balance, load the sample, and let it stand overnight. Next, 10 column volumes of distilled water and 10% ethanol solution were used to remove impurities in sequence, and 10 column volumes of 70% ethanol solution were use...

Embodiment 2

[0042] Example 2: Characterization of the chemical components of the refined product of Bupleurum saponins

[0043] 1: Sample pretreatment

[0044] Weigh an appropriate amount of refined Bupleurum saponins prepared in Example 1, add 50% methanol to make the final concentration 500 μg / mL, ultrasonically extract at room temperature for 30 minutes, centrifuge at 12,000 r / min for 10 minutes, and transfer the supernatant to a sample injection vial In UPLC-QTOF / MS analysis.

[0045] Two: Liquid chromatography / mass spectrometry (LC / MS) analysis conditions

[0046] Chromatographic conditions: Waters ACQUITY UPLC TM BEH chromatographic column (2.1mm×150mm, 1.7μm), mobile phase: A (0.1% formic acid aqueous solution, v / v), B (acetonitrile solution). The positive and negative ion gradient elution procedures are the same, see Table 3 for specific information, the flow rate is 0.3mL / min, the column temperature is 40°C, the sample tray temperature is 4°C, and the injection volume is 2μL....

Embodiment 3

[0060] Example 3: Study on the Activity of Bupleurum Saponins on Asthma

[0061] One: Experimental materials

[0062] 1.1 Experimental Instruments

[0063] Microplate reader, high-speed low-temperature centrifuge, upright biological microscope, digital pathological slide scanner, magnetic stirrer, pipette gun, etc.

[0064] 1.2 Experimental animals

[0065] Sixty clean-grade C57BL / 6J female mice, aged 6-8 weeks and weighing 18-20 g, were raised in the Experimental Animal Center of China Pharmaceutical University. The experimental mice were raised in accordance with the "Ministry of Health of the People's Republic of China Laboratory Animal Environment and Facilities Standards". During the experiment, common feed and common drinking water were fed, day and night alternated for 12 hours, and the temperature and humidity were suitable.

[0066] 1.3 Preparation of related reagents

[0067] (1) Sensitizer: The freeze-dried powder of house dust mite was dissolved in physiologic...

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Abstract

The invention discloses a radix bupleuri total saponin extraction process, which avoids the traditional n-butyl alcohol extraction process, adopts a macroporous resin combined method, and optimizes aseries of conditions to enable the saponin content in the final refined product to reach more than 75%. Component characterization is carried out by using high-resolution mass spectrometry, and 33 saponin compounds are characterized in total in a positive and negative ion mode. Meanwhile, animal experiments prove that the radix bupleuri total saponins can obviously inhibit increase of inflammatoryfactors of house dust mite sensitized asthma mice and improve pathological changes of lung tissues, are safe and free of toxic and side effects, can be used for preparing the medicine for preventingor treating asthma, and have good clinical application prospects.

Description

technical field [0001] The invention belongs to the technology of natural medicines, and in particular relates to a total saponin of Bupleurum bupleuri and its extraction process, as well as its use in the preparation of medicines for treating asthma. Background technique [0002] Bronchial asthma (Asthma) is a chronic airway inflammation involving a variety of inflammatory cells (such as eosinophils, neutrophils, airway epithelial cells, etc.) and their components, and is recognized by the world medical community as one of the four chronic diseases one. The pathological changes are characterized by reversible airway obstruction, airway hyperresponsiveness, eosinophil infiltration, and chronic airway inflammation supplemented by mast cells, lymphocytes, and macrophages. [0003] Traditional Chinese medicine believes that bronchial asthma belongs to the scope of "wheezing disease". The etiology and pathogenesis of the patient's spleen, lung, and kidney are weak, and they can...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K36/233A61P11/06A61P11/00A61K125/00
CPCA61K36/233A61K2236/15A61K2236/333A61K2236/39A61K2236/51A61K2236/53A61K2236/55A61P11/00A61P11/06
Inventor 辛贵忠翟思琦刘丽芳
Owner CHINA PHARM UNIV
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