CN101665401A - Method for separating alkene effective constituents in ginger essential oil - Google Patents

Abstract

The invention relates to a method for separating a natural active product, in particular to a method for separating various alkene effective constituents in ginger essential oil by column chromatography, which comprises the following steps: quantitatively weighing the ginger essential oil, adding absolute ethyl alcohol to dissolve the ginger essential oil, adding chromatographic silica gel with the granularity of 48-75 mu m, and evenly stirring to remove the solvent so as to obtain the silica gel absorbed with the ginger essential oil; filling some other chromatographic silica gel into a chromatographic column, putting the silica gel absorbed with the ginger essential oil on the top end of the chromatographic column, eluting with a mixed eluent of aether and n-hexane or sherwood oil in volume ratio of 2-5:98-95, and orderly collecting three eluents respectively containing beta-phellandrene, zingiberene and alpha-curcumene in three stages; and concentrating the three eluents in three stages to obtain three concentrating products respectively mainly containing beta-phellandrene, zingiberene, alpha-curcumene and beta-bisabolene. The method has the advantages of simple technique, shortperiod and easy realization of industrialization.

Description

A method for isolating effective components of alkenes in ginger essential oil

1. Technical field

The invention relates to a method for separating natural active products, in particular to a method for separating various alkene active components in ginger essential oil by column chromatography.

2. Technical background

Ginger essential oil is a volatile component obtained from the fresh rhizome of Zingiber officinale Rosc. by means of steam distillation, pressing, supercritical carbon dioxide extraction, etc. It is a brown oily liquid with a strong aroma and spicy taste of ginger. In the early days, it was mainly used as a flavoring agent for food and beverages, and it was also an important flavor raw material. Now its medicinal value is gradually being paid attention to.

Ginger essential oil mainly contains two major components of alkenes and phenols. Alkenes include gingerene, α-curcumene, β-bisabolene, β-phellandrene, β-sesquiphellandrene and other components. Also includes multiple ingredients. Each component has certain biological activity and application value. For example, zingiberene is one of the main components of ginger essential oil, which has various activities such as anti-virus, anti-ulcer, and anti-fertility; both α-curcumene and β-bisabolene have good anti-fertility activity; β-phellandrene is a kind of spice and spice intermediate, and also a natural pesticide; gingerol has multiple uses such as anti-oxidation, anti-tumor, and prevention and treatment of cardiovascular diseases. In view of the fact that various chemical components in ginger essential oil have different medicinal or other effects, it is a research trend in this field to finely separate them, and the obtained isolates have broad application prospects.

At present, the research on ginger essential oil at home and abroad mainly focuses on the composition analysis of ginger essential oil and the separation of gingerols, while there are few studies on the separation of olefins in ginger essential oil. Chinese patent CN1251367A discloses a method for supercritical carbon dioxide extraction and separation of active ingredients in ginger, which is a complex mixture of ginger oleoresin and ginger essential oil (there are more than 30 identified components in ginger essential oil) , there are more than 40 identified components in ginger oleoresin). Chinese patent CN1762956A discloses a method for preparing total gingerols by supercritical column chromatography, but does not mention the alkene components in ginger essential oil. Jocelyn G.Millar of the U.S. published "Rapid and Simple Isolation of Zingiberene from Ginger Essential Oil" (Jocelyn G.Millar. Rapid and Simple Isolation of Zingiberene from Ginger Essential Oil[J].J.Nat.Prod.1998,61,1025-1026 ) paper, narration in the paper: at first ginger essential oil is carried out Kugelrohr distillation (trace rotary distillation), get the cut that mainly contains sesquiterpene, add dienophile material, make it and contain the zingiberene of homocyclic conjugated diene Diels-Alder addition reaction occurs, the adduct is concentrated and purified by flash chromatography, and then zingiberene is reduced in KOH ethanol solution, and traces of aniline are removed by a silica gel G column, and finally the purity of 99% can be obtained. Gingerene (gas analysis). This method has many steps and high cost, and more chemical reagents are added in the reaction and subsequent treatment process. This method has strong selectivity, and the process route is only designed for zingiberene, which is suitable for the preparation of zingiberene standard samples in laboratories. But the standard sample of zingiberene has not been commercialized so far. Summarizing the existing literature, there is no report on the separation of various olefinic substances in ginger essential oil or ginger oleoresin, nor any report on the separation process of olefinic components with industrialization prospects. Therefore, it is necessary to do further research on the separation of effective components of olefins in ginger essential oil.

Chromatography is the use of differences in the physical and chemical properties of the components in the mixture (such as adsorption, molecular shape and size, molecular affinity, distribution coefficient, etc.) to make each component in two phases (one phase is fixed, called fixed phase). phase; the other phase flows through the stationary phase, known as the mobile phase) in different degrees of distribution, so that the components move at different speeds to achieve the purpose of separation. Silica gel column chromatography is based on the different adsorption and dissolution properties of different substances, and the distribution coefficients in the silica gel stationary phase and eluent mobile phase are different, so as to achieve the purpose of separation and purification. It has fast separation speed, high efficiency and good separation effect. Obvious and other characteristics, can be applied to the separation of biologically active substances.

3. Contents of the invention

The object of the present invention is to provide a process for obtaining β-phellandrene, zingiberene, α-curcumene and other alkenes active ingredients through one-time separation of ginger essential oil.

The basic idea of the present invention is to adopt column chromatography, the stationary phase is chromatographic silica gel, the mobile phase is a mixed eluent, ginger essential oil is placed on the top of the chromatography column, and the eluent is eluted, and the eluent is collected successively and then concentrated, Concentrated products containing different active ingredients are obtained. The method steps are:

1. Take ginger essential oil, add absolute ethanol to dissolve, then add chromatographic silica gel with a particle size of 48-75 μm, stir evenly and remove the solvent to obtain silica gel with ginger essential oil adsorbed, the mass ratio of ginger essential oil, absolute ethanol and chromatographic silica gel 1:2-5:1-5;

2. Take another chromatographic silica gel and put it into the chromatographic column. The mass ratio of chromatographic silica gel and ginger essential oil is 20-50:1, and then put the silica gel adsorbed with ginger essential oil on the top of the chromatographic column, and use diethyl ether and n-hexane or a mixed eluent of petroleum ether for elution, the volume ratio of the mixed eluent is 2-5:98-95, and the β-phellandrene, gingerene, α-curcumene and β-bisabolene are collected successively The three-stage eluent;

3. Concentrate the three eluents respectively to obtain concentrated products.

Among the three concentrated products in step 3, the first is β-phellandrene concentrate, the second is zingiberene concentrate, and the third is α-curcumene and β-bisabolene concentrate.

Application effect of the present invention:

1. The present invention can directly separate a variety of active ingredients with higher content from ginger essential oil through silica gel column chromatography only once, and the process is simple and the cycle is short.

2. The technology adopted in the present invention is very easy to realize large-scale, as long as a simple scale-up test is carried out, industrialized production can be realized.

3. Among the products obtained in the present invention, β-phellandrene is a kind of spice and spice intermediate, and is also a natural insecticide. Zingiberene has multiple activities such as anti-virus, anti-ulcer, and anti-fertility. α-Curcuma longa The efficacy of β-bisabolene and β-bisabolene is similar, and a good separation according to the efficacy of the active ingredients has been achieved. At the same time, it is guaranteed that there is no gingerol in the isolate. The active ingredient content of the isolated product can meet the general requirements for its efficacy.

4. Description of drawings

Figure 1: It is the first concentrated product, containing 79.76% of β-phellandrene, a three-dimensional high-performance liquid chromatogram;

Figure 2: is the second concentrated product, containing 94.91% zingiberene three-dimensional high-performance liquid chromatography;

Fig. 3: It is the three-dimensional high-performance liquid chromatogram of the third concentrated product, containing 75.85% of α-curcumene and 14.29% of β-bisabolene.

5. Specific implementation

In conjunction with the accompanying drawings, the implementation process of the method will be described in detail.

Example 1

Weigh 5g of ginger essential oil, add 10g of absolute ethanol to dissolve, then add 8g of chromatographic silica gel with a particle size of 48-75μm, remove the solvent after stirring evenly, and obtain silica gel with ginger essential oil absorbed; another 180g of chromatographic silica gel is loaded into the chromatography In the column, the silica gel adsorbed with ginger essential oil is placed on the top of the chromatography column, and the volume ratio of the mixed eluent ether and n-hexane is 2:98, and the β-phellandrene, zingiberene and α-curcumene are collected successively. The three-stage eluate; the three-stage eluate is concentrated respectively to obtain three products: the relative contents of the main components in the product are respectively: the first concentrated product contains β-phellandrene 79.76%, and the second concentrated The product contains 94.91% zingiberene, and the third concentrated product contains 75.85% α-curcumene and 14.29% β-bisabolene (see accompanying drawings 1, 2, 3 for details).

Example 2

Weigh 7g of ginger essential oil, add 30g of absolute ethanol to dissolve, then add 30g of chromatographic silica gel with a particle size of 48-75μm, stir evenly and remove the solvent to obtain silica gel with ginger essential oil absorbed; another 180g of chromatographic silica gel is loaded into the chromatography column In the process, the silica gel adsorbed with ginger essential oil was placed on the top of the chromatographic column, and the volume ratio of the mixed eluent diethyl ether and petroleum ether was 5:95. The three-stage eluate; the three-stage eluate is concentrated respectively to obtain three products: the first product contains β-phellandrene 78.37%, the second product contains zingiberene 93.41%, and the third product contains α-phellandrene Curcumene 66.22%, β-bisabolene 20.92%.

Claims (1)

1, a kind of method of separating alkene effective constituents in the ginger essential oil, adopt column chromatography, stationary phase is a chromatographic silica gel, moving phase is mixtures of eluents, ginger essential oil places the chromatography column top, and the eluent wash-out concentrates after collecting elutriant successively, obtain containing the enriched product of different effective constituents, it is as follows to it is characterized by method steps:

1. get ginger essential oil, add anhydrous alcohol solution, add the chromatographic silica gel of granularity 48-75 μ m again, solvent is removed in the back that stirs, and obtains being adsorbed with the silica gel of ginger essential oil, and the mass ratio of ginger essential oil, dehydrated alcohol and chromatographic silica gel is 1: 2-5: 1-5;

2. getting chromatographic silica gel in addition packs in the chromatography column, the mass ratio of chromatographic silica gel and ginger essential oil is 20-50: 1, the silica gel that will be adsorbed with ginger essential oil again places the chromatography column top, adopt the mixtures of eluents of ether and normal hexane or sherwood oil to carry out wash-out, the volume ratio of mixtures of eluents is 2-5: 98-95, collects three sections elutriants that contain β-phellandrene, zingiberene, α-curcumene and β-heerabolene successively;

3. three sections elutriants are concentrated, obtain three kinds of enriched products.

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