CN102002090A

CN102002090A - Preparation method of Ombuoside H

Info

Publication number: CN102002090A
Application number: CN2010105541392A
Authority: CN
Inventors: 王峰; 王琳; 张发成
Original assignee: Suzhou Paiteng Biomedical Technology Co Ltd
Current assignee: Suzhou Paiteng Biomedical Technology Co Ltd
Priority date: 2010-11-23
Filing date: 2010-11-23
Publication date: 2011-04-06
Anticipated expiration: 2030-11-23
Also published as: CN102002090B

Abstract

The invention relates to a preparation method of Ombuoside H, which is convenience for operation and has less pollution and energy consumption. The preparation method comprises the processing steps of: adding coarse pokeberry root in a CO2 supercritical extractor and obtaining an extract with n-butyl alcohol as an entrainer; adding the extract on a macroporous adsorption resin column for adsorption; eluting by ethanol; collecting the eluent; decompressing to recover the solvent and drying the solvent; adding the dried solvent onto a silica chromatographic column to elute by methanol-chloroform mixed solvent, wherein the proportion of the methanol to the chloroform in the mixed solvent is 1: 1; dividing and collecting the eluent according to column volume; combining eluent at the 3-7 parts of column volume for concentration; adding acetone crystal; and separating, washing and drying to obtain the Ombuoside H. By adopting the preparation method of the invention, the obtained Ombuoside H has high purity and the industrial enlargement is easy to realize.

Description

The preparation method of a kind of ombuoside H

Technical field

The present invention relates to the preparation method of a kind of ombuoside H, especially a kind of preparation method who from plant, extracts ombuoside H.

Background technology

Ombuoside H (Esculentoside H, Phytolaccasaponin B), molecular formula: C ₄₈H ₇₆O ₂₁, molecular weight: 989.116, CAS accession number: 66656-92-6.Mainly be present in the Phytolaccaceae various plants, wherein content is higher in the Phytolaccaceae plant pokeweed Phytolacca acinosa Roxb. root, and raw material sources are abundant.Its molecular formula is as follows:

Modern study shows that ombuoside H has stronger anti-tumor activity, and it is also as the raw material that synthesizes other reactive derivative simultaneously.

The dry root of Phytolaccaceae plant pokeweed Phytolacca acinosa Roxb. is used as the Chinese medicine Phytolacca acinosa and uses, have the detumescence of relieving oedema or abdominal distension through diuresis or purgation, tonneau two just, the effect of detoxicating and resolving a mass.

In the prior art, still be not applicable to preparation technology's report of high purity ombuoside H industrialized production.

Summary of the invention

Technical problem to be solved by this invention provides a kind of preparation method who is beneficial to big production operation, ombuoside H that product purity is high.

For solving the problems of the technologies described above, the present invention adopts following technical proposal:

Get the Radix Phytolaccae meal, join CO ₂In the supercritical extraction device, propyl carbinol is as entrainment agent, and the volume percent that entrainment agent accounts for total extraction solvent is 2-4%, extracting pressure 20-40MPa, temperature 40-60 ℃, CO ₂Flow 1-3ml/g crude drug min, extraction time 80-150min gets extract, be added on the macroporous adsorptive resins and adsorb, the 60-80% ethanol elution is collected 3-8 and is doubly measured the column volume elutriant, decompression and solvent recovery is also dry, is added on the silica gel column chromatography, with methyl alcohol-chloroform mixed solvent wash-out, the ratio of methyl alcohol and chloroform is 1: 1 in this mixed solvent, and elutriant is pressed the column volume portioning and collected, and merges the elutriant of 3-7 part column volume, concentrate, add the acetone crystallization, separate, wash, be drying to obtain.

CO ₂The volume percent that the supercritical extraction entrainment agent accounts for total extraction solvent is preferably 3%.

CO ₂Supercritical extraction pressure is preferably 30MPa, and temperature is preferably 50 ℃, CO ₂Flow is preferably 2ml/g crude drug min.

CO ₂The supercritical extraction time is preferably 100min.

Macroporous adsorbent resin is selected from a kind of in D101 type, D102 type, the AB8 type macroporous adsorbent resin.

The macroporous adsorbent resin wash-out is preferably 70% with concentration of ethanol.

The macroporous adsorbent resin wash-out is preferably 5 times of amount column volumes with the alcoholic acid collecting amount.

Preparation gained ombuoside H can adopt following method to detect:

Test example 1 HPLC method is measured ombuoside H purity

Chromatographic condition

Chromatographic column: octadecylsilane bonding glue silica gel is weighting agent; Moving phase: methyl alcohol-0.4% glacial acetic acid solution (70: 30); Flow velocity: 1.0mL/min; Detect wavelength: 215nm; Column temperature: 30 ℃.

Measuring method

Precision takes by weighing ombuoside H2mg, places the 50mL measuring bottle, adds people's methyl alcohol 20mL, and sonic oscillation makes dissolving, and methanol constant volume is drawn 10 μ L to scale, injects high performance liquid chromatograph, adopts normalization method working sample purity.

Adopt the present invention to prepare ombuoside H, be beneficial to big production operation, energy consumption is little, pollutes little.

The present invention is further elaborated below in conjunction with embodiment, but the scope of protection of present invention is not limited to following embodiment.

Embodiment

Embodiment 1

Get Radix Phytolaccae meal 10Kg, join CO ₂In the supercritical extraction device, propyl carbinol is as entrainment agent, and the volume percent that entrainment agent accounts for total extraction solvent is 2%, extracting pressure 20MPa, 40 ℃ of temperature, CO ₂Flow 1ml/g crude drug min, extraction time 80min, get extract, be added on the D101 type macroporous adsorptive resins and adsorb 60% ethanol elution, collect 3 times of amount column volume elutriants, decompression and solvent recovery is also dry, is added on the silica gel column chromatography, with methyl alcohol-chloroform mixed solvent wash-out, the ratio of methyl alcohol and chloroform is 1: 1 in this mixed solvent, elutriant is pressed the column volume portioning and is collected, and merges the elutriant of 3-7 part column volume, concentrates, add the acetone crystallization, separate washing, be drying to obtain white powder ombuoside H20.8g, detect through HPLC, purity is 94.0%, UV, IR, MS; ²HNMR, ¹³The data of its physical behavior of sign such as CNMR are consistent with prior art.

Embodiment 2

Get Radix Phytolaccae meal 10Kg, join CO ₂In the supercritical extraction device, propyl carbinol is as entrainment agent, and the volume percent that entrainment agent accounts for total extraction solvent is 4%, extracting pressure 40MPa, 60 ℃ of temperature, CO ₂Flow 3ml/g crude drug min, extraction time 150min, get extract, be added on the D102 type macroporous adsorptive resins and adsorb 80% ethanol elution, collect 8 times of amount column volume elutriants, decompression and solvent recovery is also dry, is added on the silica gel column chromatography, with methyl alcohol-chloroform mixed solvent wash-out, the ratio of methyl alcohol and chloroform is 1: 1 in this mixed solvent, elutriant is pressed the column volume portioning and is collected, and merges the elutriant of 3-7 part column volume, concentrates, add the acetone crystallization, separate washing, be drying to obtain white powder ombuoside H23.4g, detect through HPLC, purity is 93.6%, UV, IR, MS; ²HNMR, ¹³The data of its physical behavior of sign such as CNMR are consistent with prior art.

Embodiment 3

Get Radix Phytolaccae meal 10Kg, join CO ₂In the supercritical extraction device, propyl carbinol is as entrainment agent, and the volume percent that entrainment agent accounts for total extraction solvent is 3%, extracting pressure 30MPa, 50 ℃ of temperature, CO ₂Flow 2ml/g crude drug min, extraction time 100min, get extract, be added on the AB8 type macroporous adsorptive resins and adsorb 70% ethanol elution, collect 5 times of amount column volume elutriants, decompression and solvent recovery is also dry, is added on the silica gel column chromatography, with methyl alcohol-chloroform mixed solvent wash-out, the ratio of methyl alcohol and chloroform is 1: 1 in this mixed solvent, elutriant is pressed the column volume portioning and is collected, and merges the elutriant of 3-7 part column volume, concentrates, add the acetone crystallization, separate washing, be drying to obtain white powder ombuoside H22.7g, detect through HPLC, purity is 96.9%, UV, IR, MS; ²HNMR, ¹³The data of its physical behavior of sign such as CNMR are consistent with prior art.

Claims

1. the preparation method of an ombuoside H is characterized in that described method is made up of the following step: get the Radix Phytolaccae meal, join CO ₂In the supercritical extraction device, propyl carbinol is as entrainment agent, and the volume percent that entrainment agent accounts for total extraction solvent is 2-4%, extracting pressure 20-40MPa, temperature 40-60 ℃, CO ₂Flow 1-3ml/g crude drug min, extraction time 80-150min gets extract, be added on the macroporous adsorptive resins and adsorb, the 60-80% ethanol elution is collected 3-8 and is doubly measured the column volume elutriant, decompression and solvent recovery is also dry, is added on the silica gel column chromatography, with methyl alcohol-chloroform mixed solvent wash-out, the ratio of methyl alcohol and chloroform is 1: 1 in this mixed solvent, and elutriant is pressed the column volume portioning and collected, and merges the elutriant of 3-7 part column volume, concentrate, add the acetone crystallization, separate, wash, be drying to obtain.

2. according to the preparation method of the described a kind of ombuoside H of claim 1, it is characterized in that described CO ₂The volume percent that the supercritical extraction entrainment agent accounts for total extraction solvent is 3%.

3. according to the preparation method of the described a kind of ombuoside H of claim 1, it is characterized in that described CO ₂Supercritical extraction pressure 30MPa, 50 ℃ of temperature, CO ₂Flow 2ml/g crude drug min.

4. according to the preparation method of the described a kind of ombuoside H of claim 1, it is characterized in that described CO ₂The supercritical extraction time is 100min.

5. according to the preparation method of the described a kind of ombuoside H of claim 1, it is characterized in that described macroporous adsorbent resin is selected from a kind of in D101 type, D102 type, the AB8 type macroporous adsorbent resin.

6. according to the preparation method of the described a kind of ombuoside H of claim 1, it is characterized in that described macroporous adsorbent resin wash-out concentration of ethanol is 70%.

7. according to the preparation method of the described a kind of ombuoside H of claim 1, it is characterized in that described macroporous adsorbent resin wash-out is 5 times of amount column volumes with the alcoholic acid collecting amount.