JPS61137039A - Quantitative analysis of saponin of gynostemma pentaphyllum - Google Patents

Abstract

PURPOSE:To simply perform the quantitative analysis of total saponin, by contacting an aqueous extract of a saponin-containing specimen or an aqueous specimen with a polystyrene resin with a definite particle size to adsorb the same and subsequently washing the adsorbed resin with water, subsequently, dilute alcohol before eluting saponin by alcohol. CONSTITUTION:An aqueous extract or aqueous solution of Gynostemma pentaphyllum, tea thereof, an extract thereof, a drink thereof or a preparation of saponin thereof is adsorbed by a polystyrene resin with 50-200mesh. this resin is washed with water and 20-30% dilute alcohol and saponin is eluted by alcohol while the eluate is evaporated to dryness. The obtained residue is dissolved in alcohol to filter insolubles while the filtrate is evaporated to dryness and the wt. of the obtained residue is measured to perform the quantita tive analysis of saponin of Gynostemma pentaphyllum.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a simple method for quantifying Jiaogulan saponin.

Background of the invention Gymostemma pentaph
Yllum Maki no) is a perennial vine of the Cucurbitaceae family that grows wild in the mountains and fields of East Asia. Recently, Takemoto et al., one of the inventors of the present invention, isolated more than 50 types of saponins that are the same as medicinal ginseng or have structurally similar saponins from Jiaogulan and clarified their structures. On the other hand, interesting effects such as anti-bronchitis, anti-ulcer, anti-steroidal side effects, and anti-tumor effects have been found in Jiaogulan saponin, and it has come to be widely used as a health food for the treatment and prevention of these symptoms. Along with this, development as a pharmaceutical or cosmetic product was also considered.

However, since Jiaogulan is a natural product, the content of its active ingredient, saponin, varies significantly depending on the region of production, climate, natural features, time of collection, etc., and this makes it difficult to control the quality of Jiaogulan or the various products made from it. In order to solve this problem, there is a strong demand for a simple method for quantifying Jiaogulan saponin. As mentioned above, more than 50 types of saponins have been isolated from Jiaogulan, but not all saponins have been completely isolated yet. Therefore, it is extremely difficult and impractical to separate and quantify these individual saponins one by one. For quality control of actual raw materials and products, it is not necessary to quantify individual saponins; it is sufficient to know the total amount of saponin in the sample.

As a result of intensive research to establish a method for easily quantifying total saponin in a sample, the inventors of the present invention were able to easily quantify total saponin by adsorbing and separating saponin using polystyrene resin. The inventors have discovered that the invention can be obtained and completed the present invention.

Possible adsorbents for adsorbing and separating saponins include reverse-phase ODS resins or polystyrene resins, but ODS resins are expensive and are only used for high-performance liquid chromatography. It is not suitable for routine work such as quality control of Amachazol products. The present inventors have discovered that total saponin can be conveniently quantified by using a polystyrene-based resin that is inexpensive and strong, particularly a resin with an appropriate particle size.

That is, the method of the present invention consists of bringing an aqueous extract or an aqueous sample of a sample containing saponin into contact with a polystyrene resin of a certain particle size to adsorb it, washing with water and then diluted alcohol, and then eluting the saponin with alcohol. . The degree of saponin adsorption varies depending on the type of polystyrene resin used in the method of the present invention and its particle size, that is, the size of the surface area. When using Amberlite XAD-11, which is a typical commercially available polystyrene resin, 50 to 20
0 mesh is suitable; if the particle size is coarser than this, more saponin will pass through without being adsorbed.
If a resin finer than this is used, the passage of liquid will become difficult and analysis will take a long time.

The dilute alcohol used to wash the resin on which saponin has been adsorbed and the alcohol used to elute the saponin are preferably methanol or ethanol, and 20 to 30% of the dilute alcohol is suitable.

When saponin adsorbed on resin is eluted with alcohol, a small amount of substances other than saponin are also eluted.

Since this substance is poorly soluble in alcohol, this impurity substance and saponin can be separated by evaporating the eluate to dryness and dissolving the resulting residue in alcohol again. That is, the desired amount of saponin can be determined by removing impurities from this alcoholic solution by >Fi8, and then evaporating the Shido liquid again to dryness and measuring the weight of the resulting residue. Suitable alcohols for use in this step for separating impurities and saponins are ethanol and lobetanol.

In the method of the present invention, total saponin is determined by measuring the weight of the residue obtained by the above method. Methylaminohenzaldehyde, p-hydroquinobenzaldehyde, 4-
Ethoquino-3-hydroquinobenzaldehyde, 3.4-
It is also possible to develop color using dimethoquinobenozaldehyde, p-nitrohenzaldehyde, etc.) and a mineral acid for colorimetric determination.

As described above, the method of the present invention is very simple and does not require expensive measuring instruments, so it can be said to be a truly practical method.

Examples of the method of the present invention are shown below.

Example ! Determination of saponin in Jiaogulan herbal medicine Grind Jiaogulan herbal medicine, extract 20g of the powder with 50ml of water for 20 minutes, and shake. Repeat this operation three times and combine all the furnace liquids. On the other hand, polystyrene resin (
Amberlight XAD-1! , Organo Co., Ltd.) in a mortar, and collect the particles that pass through a No. 60 sieve but not a No. 100 sieve, and wash thoroughly with methanol.
1 and fill it into a 25ml bilette with a cotton plug at the bottom.

Next, 100ml of methanol and 50ml of water were passed through for washing, and then the above y solution was added, being careful not to drain the liquid! Pass at a flow rate of 1 ml per minute. It is then washed in a similar manner by passing 501 l of water and 50+ nl of 20% methanol. 50 ml of methanol are then passed through in the same manner, the eluate is collected, taken up in an evaporating dish and evaporated to dryness on a water bath. Add 2 liters of ethanol to this and dissolve by heating. treat Dilute the residue with ethanol 20
The furnace liquid and washing liquid were taken into an evaporating dish with a known weight, evaporated to dryness on a water bath, dried at 105°C for 1 hour, left to cool in a dencator (silica gel), and weighed. Measure and use as saponin nails.

Table 1 shows the results of quantifying the saponins of Jiaogulan grown in various places using the above method.

Table 1 The total saponins contained in Jiaogulan groups can also be quantified in exactly the same manner as above.

Example 2 Determination of saponin in Jiaogulan drink Jiaogulan 2. A Jiaogulan drink corresponding to Og is taken and treated in the same manner as the extraction furnace liquid in Example 1 to quantify the total saponin in the drink.

Aqueous preparations containing Jiaogulan saponin can also be quantified in a similar manner.

Claims (1)

[Claims] Jiaogulan, Jiaogulan tea, Jiaogulan extract,
An aqueous extract or aqueous solution of a preparation containing Jiaogulan link or Jiaogulan saponin is adsorbed onto a 50-200 mesh polystyrene resin, and the resin is mixed with water and 2
After washing with 0 to 30% diluted alcohol, eluate with alcohol, evaporate the eluate to dryness, dissolve the resulting residue in alcohol, filter off insoluble materials, and evaporate the filtrate to dryness. A method for quantifying Jiaogulan saponin comprising measuring the weight of the residue.