KR880000537B1 - Method of preparing extract containing salivin from cardus maliarus - Google Patents

Abstract

The salivin (A)-cotg. ext. is prepd. from cardus maliarus seeds. Thus, 813g defatted seed is extd. with AcOEt. The ext. is vacuum- distld. The resulting residue is treated with cyclohexane, filtered, dried, suspended in 23% MeOH soln, stirred, filtered and dried to give solid (I) (33.3g). Solid (I) is dissolved in AcOEt and eluted with 50% MeOH soln.. The organic phase is pooled and dried to give 28.9g solid (II), which is dissolved in AcOEt, treated with charcoal and filtered. The filtrate is vacuumdistld. and stand for 5-20days at 4-5≰C to give 19.4g crystalline material contg. salivin. compd. (A) has effect on the treatment of liver disorders.

Description

Process for the preparation of high concentrations of silybin-containing extracts from seeds of Cardus marianus

The present invention relates to a process for the preparation of high concentrations of silinine-containing extracts from seeds of Cardus marianus. More specifically, degreased Kartus marianus was extracted with ethyl acetate, followed by removing the residual oil and removing the polar substance to prepare a medicinal extract of Cardus marianus with a total flavonoid content of 69-86% silybin and 20-57%. It is about how to.

Cardus marianus is a plant belonging to the Asteraceae family and is planted in southern Europe and North Africa. It is native to southern Europe and North Africa. In Europe, the plant has been widely used for the treatment of liver disease, and its efficacy has been demonstrated by a series of recent studies. Wagner and Hansel have separated the active ingredient from this seed and confirmed its chemical formula. In an earlier study, a group of substances with pharmacological action was isolated and called flagliignan, called silymarin, whose structural formula (1) is

The silymarin contains flavonoids, such as silydianin and silycristine, in addition to the above-mentioned silbin.

Conventionally, a method for producing a medicinal extract from seeds of Cardus marianus is disclosed in Patent Publication Nos. 76-461 and 83-2765, and the like. As 2765 shows various shortcomings of the manufacturing method, it is a multi-step, very complicated method, and it is not economical because it uses an expensive mixed solvent, and the method disclosed in the specification of Patent Publication No. 83-2765 is Although slightly simpler, this method also has the disadvantage of using an expensive mixed solvent and a flammable solvent. Also, it is impossible to obtain an extract of 35-57% of the silicide content, and the silymarin content (total flavone content) is 69. It was not possible to get an extract of -85%. Moreover, it is important to note that the above known methods do not have an oil removal process, so the oil is contained in the extract as it is, so that the pharmacological effect may not be reduced, but a high concentration of extract could not be prepared. In the known method, a precipitation method using a mixed solvent is used to remove the polar substance, but this method causes a complicated manufacturing method and a disadvantage of increasing the manufacturing cost.

The present inventors earnestly researched and examined to remove the various drawbacks, and after removing the oil from the seed of Cardus marianus by primary extraction, cyclohexane, petroleum ether, and n-hexane were removed at least one selected from the group consisting of The present invention was completed by discovering that a high concentration of silane extract, which is the object of the present invention, can be prepared by removing the polar substance with an aqueous methanol solution.

Hereinafter, the present invention will be described in detail.

The seed of Cardus marianus contains 20-30% oil. This oil is removed by a normal compression method. For example, even using a pressurized milking machine used for the production of sesame oil can remove more than 90% of the oil. Seeds of crushed Cardus Maianus which have passed through the milking machine may be used for the following operations without further grinding. The skim seeds thus obtained are placed in an extractor, and ethyl acetate is added in an amount of about 3-7 times the weight of skim seeds, and then extracted three times or more at about 20-30 ° C. for about 1-5 hours. The combined ethyl acetate solvents are depressurized to remove ether acetate to give a residue containing a significant amount of oil. If you do not remove the oil at this stage, it will be a huge obstacle in the next stage. Solvent fractionation was used to remove this fraction. Solvents used are n-hexane, cyclohexane, petroleum ether, benzene, toluene.

The suitability of the solvent was determined by quantifying the total amount of flavonoids eluted in the supernatant obtained by adding 200 ml of solvent to 100 g of dehydrated seed powder, stirring at 20 ° C. for 2 hours, and then standing. Determination of total flavonoids was followed by the method of the eighth edition of the German Pharmacopoeia, and the more solvents that elute flavonoids, the more unsuitable for use.

According to the above method, 0.08 g in n-hexane, 0.045 g in cyclohexane, 0.067 g in fiber ether, 0.15 g in benzene, and 0.133 g in toluene were eluted. This result is the average of three replicates.

According to the above results, the degreasing solvent is preferably cyclohexane, petroleum ether and n-hexane, benzene and toluene are inadequate, and solvents having higher polarity than benzene or toluene are more theoretically obvious.

Cyclohexane and n-hexane are most preferred in consideration of stability and recovery rate among the three solvents determined as suitable as described above.

The solid remaining after degreasing was suspended in 10-30% aqueous methanol solution, stirred at 20-30 ° C. for 1 hour, then stopped, and the insolubles were filtered off and dried. The yield of the dried product was 2.8-3.3% by weight of the seed used. The dry matter contains 69-78% of the total flavonoids, depending on the conditions of seed production, harvest time, storage condition and harvesting year. (Building (1))

After dissolving the above dried product (1) in 5-15 times of ethyl acetate, the same amount of solution and the same amount of 10-50% methanol aqueous solution was added, shaken for 10 minutes and left for 1 hour. After separating the conflict and repeating the same operation once more combined and dry the solution is obtained in the yield of 2.2-2.8% by weight of the seed used. This dry matter contains 75-86% of the total flavonoids, depending on the conditions of seed production, harvesting time, storage condition, and harvesting year. (Dry matter (2))

After dissolving the dried product (2) in 5-15 times of ethyl acetate again, decolorized in activated charcoal, distilled under reduced pressure and left for 5-20 days at 4-5 ℃ to obtain a crystalline material. The yield of the combined crystalline material up to the second crystal was 1.46-1.94% by weight of the seed used.

This crystalline material contains 35-57% of silybin.

The present invention is explained in more detail by the following examples.

Example 1

1000 g of seeds are degreased by passing through a milking machine. 813 g of crushed seeds obtained by degreasing were added to the extractor with 31 ethyl acetate and extracted for 3 hours at room temperature. After filtration, the filtrate is stored and the plant residue is extracted twice more under the same conditions. The combined ethyl acetate extracts were added to a vacuum distillation and the ethyl acetate was distilled off. 200 ml of cyclohexane was added to the remaining oily semi-solid material, followed by shaking for 15 minutes. The solids remaining on the filter are washed twice with 100 ml of cyclohexane. The solid thus obtained was dried, suspended in 23% aqueous methanol solution, stirred at room temperature for 1 hour, filtered, and dried. The dry weight is 33.30 g.

Example 2

After dissolving 33.3 g of the solid obtained in Example 1 in 170 ml of ether acetate, 170 ml of 50% aqueous methanol solution was added thereto, shaken for 10 minutes, and left for 1 hour. The organic layer (supernatant) is separated separately and this operation is repeated once more. Drying the organic layer yields 28.9 g.

Example 3

28.9 g of the solid obtained in Example 2 was dissolved in 300 ml of ethyl acetate, 8 g of activated carbon was added thereto, stirred at room temperature for 2 hours, filtered, and the filtrate was added to a still, and ethyl acetate was evaporated until insoluble matter began to form. This concentrate is left to stand at 4-5 ° C for 15 days. The resulting primary crystals are dried and stored, the filtrate is placed in a concentrator and again subjected to the above operation to obtain secondary crystals. The combined dry crystals weighed 19.4 g.

Claims (1)

After extracting Kardus Marianus degreased with a milking machine with methyl acetate to make an ethyl acetate extract, the residual oil contained in the extract is removed with cyclohexane, and the polar substance contained in the remaining solid is the solid itself or its A method of preparing Cardus Marianus extract, wherein the ethyl acetate solution is eluted with an aqueous 10-15% methanol solution, and the total flavonoid content is 69-86%, and the silybin content is 20-57%.