Method for preparing secondary platycodins capable of improving permeability of cell membranes

A technology of permeability and cell membrane, which is applied in the field of preparation of Platycodon grandiflora secondary saponins, can solve problems that do not involve the preparation process and method of Platycodon grandiflorum secondary saponins, the active ingredient of Platycodon grandiflora

Active Publication Date: 2019-07-19
NANKAI UNIV
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  • Abstract
  • Description
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Problems solved by technology

Although the total saponins of Platycodon grandiflora have a strong hemolytic effect in vitro [Sun H et al. 420mg / kg, while the safe dose of saponin D exceeds 2g / kg [Lee WH et al. Toxicological Research.2011,27:217-224]
However, there is no report concerning the medicinal components in platycodon saponins, especially the preparation process and method of platycodon secondary saponins

Method used

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  • Method for preparing secondary platycodins capable of improving permeability of cell membranes
  • Method for preparing secondary platycodins capable of improving permeability of cell membranes
  • Method for preparing secondary platycodins capable of improving permeability of cell membranes

Examples

Experimental program
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Effect test

Embodiment 1

[0029] Embodiment 1: Separation and preparation of Platycodon grandiflorum total saponins

[0030] Take 2.5 kg of Platycodon grandiflora decoction pieces (purchased from Xi'an Tianrui Biotechnology Co., Ltd., batch number TR-170810), add 25 liters of 10 times the amount of distilled water, and heat and reflux for extraction for 2 hours. Filtrate while hot, and concentrate the filtrate under reduced pressure to about 500 ml. After cooling, add absolute ethanol to 1500 ml to make the ethanol ratio reach 70%, mix well and let stand overnight. The supernatant was collected by filtration, concentrated under reduced pressure to obtain 210 grams of platycodon saponin crude extract. Take 200 grams of it and add 20 liters of distilled water to dissolve it, configure it into a 1% solution, and load it on AB-8 macroporous adsorption resin (diameter: 5.5 cm; column height: 30 cm), and use ethanol solutions of different concentrations (0 % → 30% → 70% → 100% ethanol) gradient elution, ea...

Embodiment 2

[0033] Embodiment 2: Analysis of Platycodon grandiflorum total saponins

[0034] Platycodon saponins are mainly pentacyclic triterpenoid saponins, which mainly include platycodon saponin and platycodon acid saponin. Adopt UPLC-Q / TOF to analyze the platycodon total saponins of embodiment 1 and find that it mainly contains 10 platycodon saponins (see figure 2 ). It is speculated that the secondary saponins platycodon saponin 682 and platycodon saponin 696 that enter the blood are derived from the in vivo metabolites of platycodon saponins and platycodon acid saponins, respectively.

[0035] UPLC-Q / TOF experimental conditions:

[0036] 1) Liquid chromatography conditions:

[0037] ColumnWaters ACQUITY BEH C18 chromatographic column (2.1×100mm, 1.7μm); flow rate 0.4ml / min; PDA detector, detection wavelength 210nm; sample injection volume: 5.0μl; column temperature 30℃; mobile phase: A is 0.1% formic acid- Aqueous solution, B is acetonitrile; binary gradient elution: 0~2 min...

Embodiment 3

[0044] Embodiment 3: the investigation of the hydrolysis condition of Platycodon grandiflora secondary saponin

[0045] Using platycodon saponin D (Shanghai Boka Chemical Technology Co., Ltd., batch number: J-013-161216) as the reaction substrate (1 mg / ml), the hydrolysis conditions were investigated in a water bath at 85°C. The effects of acid (pH=1) and alkali (pH=14) on the hydrolysis efficiency were discussed respectively. And by the HPLC method of Example 1, the formation of Platycodon grandiflora secondary saponins 682 and 696 was investigated. The result is as image 3 As shown, the yield of secondary saponin 682 by alkali hydrolysis is significantly better than the effect of acid hydrolysis, so it is determined to use the method of alkali hydrolysis to optimize the hydrolysis conditions in the next step.

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Abstract

The invention discloses a method for preparing secondary platycodins capable of improving permeability of cell membranes. The method comprises the following steps: using total platycodin as a raw material; hydrolyzing by alkali to obtain a main secondary platycodin metabolized in platycodin; separating the obtained secondary platycodin with middle-pressure preparation liquid chromatography to obtain two main secondary platycodins: and secondary platycodin 682 and secondary platycodin 696. The secondary platycodin 682 and thsecondary platycodin 696 can effectively improve the permeability of cell membranes, assist drugs to enter cells, and can be used for assisting drug delivery.

Description

technical field [0001] The invention belongs to the technical field of chemical extraction, and relates to a preparation technology of platycodon grandiflorum secondary saponins capable of improving cell membrane permeability. Background technique [0002] Platycodon grandiflorum (Jacq.) A.DC. is the root of Platycodon grandiflorum (Jacq.) A.DC. It was first recorded in "Shen Nong's Materia Medica". Platycodon grandiflorum is bitter, pungent and flat in nature, and belongs to the lung meridian. It has anti-inflammatory, expectorant, antitussive, anti-ulcer, lowering blood pressure, dilating blood vessels, antipyretic, analgesic and sedative, lowering blood sugar, anticholinergic, and promoting bile acid secretion, etc. Wide range of pharmacological effects. Platycodon grandiflorum is a commonly used traditional Chinese medicine for treating the lungs clinically, and is known as a medicine for boats and boats. According to "Materia Medica Seeking Truth" records: "Pianidospe...

Claims

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Application Information

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IPC IPC(8): C07J63/00C07H1/08C07H15/256A61K47/28
CPCA61K47/28C07H1/08C07H15/256C07J63/008
Inventor 白钢姜志宏侯媛媛王静荣吴文博唐仲尧申福葵张曼姜民高洁
Owner NANKAI UNIV
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