CN112321600A

CN112321600A - Method for extracting and separating rhodiola crenulata from rhodiola crenulata

Info

Publication number: CN112321600A
Application number: CN202011153655.4A
Authority: CN
Inventors: 李绪文; 张余; 金永日
Original assignee: Jilin University
Current assignee: Jilin University
Priority date: 2020-10-26
Filing date: 2020-10-26
Publication date: 2021-02-05

Abstract

A method for extracting and separating rhodiola crenulata from rhodiola crenulata comprises the steps of crushing rhodiola crenulata, adding 70-90% ethanol solution in volume ratio for reflux extraction, concentrating the extracting solution under reduced pressure, adding 100% ethanol for alcohol precipitation, concentrating the supernatant to obtain an extract, and separating and purifying the extract by column chromatography to obtain the rhodiola crenulata with the purity of more than 98%. The preparation method of the invention has high extraction efficiency and simple operation, is suitable for industrial production, and the obtained salidroside can be used for preparing pharmaceutical compositions and other products.

Description

Method for extracting and separating rhodiola crenulata from rhodiola crenulata

The technical field is as follows:

the invention belongs to the field of natural medicinal chemistry, and particularly relates to a method for extracting and separating rhodiola crenulata from rhodiola crenulata.

Background art:

plants of Rhodiola (Rhodiola L.) belonging to Crassulaceae (Crassulaceae) are perennial plants or subshrubular plants, grow in alpine regions of the northern hemisphere, and are mainly distributed in himalayas, western to northern asia, and are distributed in China, Korea, Japan to northern america. Chinese rhodiola plants have various types and abundant reserves, and are the main production area of rhodiola. There are more than 70 species, 2 subspecies and 7 varieties in the whole country, wherein 32 species (mainly concentrated in eastern part, southeast part and southern part of the Tibetan), 28 species (mainly concentrated in northwest part, western part and northern part of the Yunnan province) and 26 species of Sichuan (mainly distributed in plateau areas of the Sichuan west) are in the Tibetan region.

The medicinal history of rhodiola root is long. As early as 760 yuan, rhodiola rosea has been used by Tibetan medicine for treating various diseases, such as clearing lung and removing toxicity, nourishing vitality, treating edema of limbs, and the like. The medicinal value of rhodiola plant is also recorded in detail in medical books such as the fourth medical dictionary and the moon king medical treasure. Rhodiola rosea is listed as a 'herbal superior product' in 'herbal compendium', and has the effects of strengthening body resistance, consolidating constitution, benefiting qi, nourishing blood, clearing heat and moistening lung. Modern pharmacological research shows that rhodiola plants have multiple effects of resisting oxidation, reducing blood sugar, resisting fatigue, resisting stress, resisting depression, resisting inflammation, protecting nerves, resisting tumor, resisting altitude diseases, regulating immunity and the like.

To date, researchers at home and abroad have conducted studies on chemical compositions of more than 20 species of rhodiola plants, including rhodiola crenulata (r. crenulata), rhodiola sachalinensis (r. algida), rhodiola rosea (r. rosea), rhodiola rosea (r. wallichiana), rhodiola angusta (r. kirilowii), rhodiola elongata (r. fastiata), rhodiola tetrastigrina (r. drynariida), rhodiola yunnanensis (r. yunnanensis), rhodiola sacra (r.sacra s.h.fu), rhodiola sachalinensis (r.sachalinensis), rhodiola parvus (r. phariensis), and the like.

Research shows that rhodiola plants mainly contain tyrosol and glycosides, flavone and glycosides, phenylpropanoids, terpenes, cyanogenic glycosides or phenol glycosides and other glycosides, organic acids, volatile oil, polysaccharide, steroids and the like.

So far, no literature report on the separation of rhodiola crenulata from rhodiola crenulata is available.

The invention content is as follows:

the invention firstly separates the rhodiola crenulata from the rhodiola crenulata and further invents a separation method for extracting and separating the rhodiola crenulata from the rhodiola crenulata.

The specific technical scheme and research results of the invention are as follows:

pulverizing radix Rhodiolae, extracting with ethanol water under reflux for 1 hr for 3 times (12, 10, and 8 times of the total weight of the extract), filtering, mixing filtrates, concentrating, precipitating with ethanol, centrifuging, collecting supernatant, and recovering solvent under reduced pressure to obtain radix Rhodiolae extract.

The volume percentage of the extraction solvent ethanol-water solution is 30-100%, preferably 70-90%, the extraction method is reflux extraction, and the volume of the extraction solution is 7-15 times of the weight of the medicinal materials.

The alcohol precipitation is to concentrate the extracting solution to the density of 1.1-1.2, and then to add 95% or 100% ethanol 8-16 times of the extracting solution.

The column chromatography adopts silica gel column chromatography, mobile phases are ethyl acetate-methanol-water (17:2:2) and petroleum ether-ethyl acetate (25:1), the thin layer chromatography detects and combines the components containing the salidroside, and finally the salidroside monomer compound is obtained by recrystallization.

The preparation method disclosed by the invention has the advantages of high efficiency, high speed, high product purity, less organic solvent consumption, environmental friendliness and the like.

Drawings

FIG. 1 mass spectrum of rhodiola crenulata

FIG. 2. rhodiola crenulata¹H-NMR chart

FIG. 3 is a view of rhodiola crenulata¹³C-NMR chart

The specific implementation mode is as follows:

the present invention will be further described with reference to specific examples.

Example 1:

taking 15kg of dried rhodiola crenulata, crushing, carrying out reflux extraction for 3 times by using 70% ethanol-water solution, wherein the extraction time is respectively 2.0,1.5 and 1.0 hours, the liquid-material ratio is respectively 12,10 and 8 times, filtering, combining filtrates, concentrating until no alcohol smell exists, adding absolute ethanol for alcohol precipitation under stirring, centrifuging, recovering a solvent from a supernatant under reduced pressure to obtain an extract, carrying out silica gel column chromatography on the extract by using ethyl acetate-methanol-water (17:2:2) as a mobile phase, carrying out second silica gel column chromatography by using petroleum ether-ethyl acetate (25:1) as a mobile phase to obtain a component containing the salidroside, and finally carrying out methanol recrystallization to obtain a salidroside monomeric compound. The obtained salidroside has purity of 99.5% by HPLC-ELSD detection. Comparing the nuclear magnetic data of the separated salidroside monomer compound with the nuclear magnetic data of the salidroside reported in the literature, and finally determining that the salidroside is the salidroside. The developing solvent for thin layer chromatography is chloroform-methanol-water (15: 5:1) and chloroform-methanol-water (180: 20: 1).

Measured rhodiola crenulata¹³C NMR (DMSO, 150MHz) nuclear magnetic data were as follows:

δc：146.98(C-2)，136.08(C-3)，176.11(C-4)，152.24(C-5)，98.03(C-6)，148.71(C-7)，124.36(C-8)，143.53(C-9)，104.14(C-10)，121.63(C-1’)，129.56(C-2’)，115.5(C-3’)，159.36(C-4’)，115.5(C-5’)，129.56(C-6’)，126.76(C-1”)，111.79(C-2”)，147.61(C-3”)，147.18(C-4”)，115.34(C-5”)，120.56(C-6”)，77.04(C-7”)，77.61(C-8”)，60.04(C-9”)，55.7(O-CH3)。

HPLC-ELSD chromatographic conditions were as follows:

chromatograph: agilent Technologies 1200series

A chromatographic column: COSMOSIL 5C 18-MS-II (4.6ID X250 mm)

Mobile phase: acetonitrile-water (40: 60);

flow rate: 1L/min;

column temperature: 30 deg.C

Sample introduction amount: 20 mu L of the solution;

ELSD detector conditions: the drift tube temperature is 99 ℃; the state of the impactor: and closing. Carrier gas pressure: 0.45 Mpa;

flow rate of the atomizer: 2.8L/min; magnification: 1.0 times.

Example 2:

taking 15kg of dried rhodiola crenulata, crushing, extracting with methanol under reflux for 3 times, wherein the extraction time is 2.0,1.5 and 1.0 hours respectively, the liquid-material ratio is 12,10 and 8 times respectively, filtering, combining filtrates, concentrating until no alcohol smell exists, adding 95% ethanol for alcohol precipitation under stirring, centrifuging, recovering the solvent from the supernatant under reduced pressure to obtain an extract, performing silica gel column chromatography on the extract by using chloroform-methanol-water (15: 5:1) as a mobile phase, performing second silica gel column chromatography by using chloroform-methanol-water (180: 20: 1) as a mobile phase to obtain a component containing the rhodiola crenulata, and finally recrystallizing by using methanol to obtain a monomer compound of the rhodiola crenulata, wherein the purity of the rhodiola crenulata is 99.0% by HPLC-ELSD detection.

Claims

1. A method for extracting and separating the rhodiola crenulata is characterized in that the raw material (medicinal material) for preparing the rhodiola crenulata is rhodiola crenulata.

2. The method of claim 1, wherein the extracting and purifying step comprises pulverizing radix Rhodiolae, extracting with ethanol, filtering, mixing filtrates, concentrating, precipitating with ethanol, recovering ethanol from the supernatant under reduced pressure to obtain extract, and separating by column chromatography to obtain rhodiola crenulata.

3. The method as claimed in claim 1, wherein the volume percentage concentration of the ethanol is 70-90%, the extraction method is reflux extraction, and the dosage is 7-15 times of the medicinal materials.

4. The method according to claim 1, characterized in that the column chromatography stationary phase is silica gel and the mobile phase systems are ethyl acetate-methanol-water (17:2:2) and petroleum ether-ethyl acetate (25: 1).