JPS6112679B2 - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for separating ubiquinone.

Ubiquinones are widely present in plants and animals ranging from higher animals and plants to microorganisms, and in particular, one of their homologues, ubiquinone Q ₁₀ , has come to be recognized for its importance as a pharmaceutical agent. However, the content of ubiquinone in animals and plants is extremely small, and conventional extraction methods simultaneously extract a large amount of other various substances, making it difficult to separate and purify ubiquinone alone.

The present invention was made for the purpose of providing a method for easily and efficiently separating ubiquinone from extracts from animal and plant tissues, and the gist thereof is to separate ubiquinone from extracts containing ubiquinone from animal and plant or microbial tissues. When separating,
The lipids are eluted from the hydrophilic polymer material on which the extract has been adsorbed using a mixed solvent of an organic solvent and water that is soluble in water, and then adsorbed onto the hydrophilic polymer material using the organic solvent that does not contain water. The present invention relates to a method for separating ubiquinone, which is characterized by eluting ubiquinone.

In the present invention, the hydrophilic polymer substance has one or more types of functional groups such as -OH group, -CONH- group, or -O- group in its molecule, and has a molecular weight of 10
Refers to polymeric substances that have the property of becoming wet with water. In order to produce the polymeric substance, known aqueous suspension polymerization techniques are suitably employed. for example,
Methyl acrylate, methyl methacrylate, ethylene glycol diacrylate (or methacrylate), tetraethylene glycol diacrylate (or methacrylate), hydroxyethyl acrylate (or methacrylate), acrylamide, methoxymethyl acrylamide, tetramethylolmethane triacrylate (or methacrylate) Select one or two or more monomers from the following and polymerize them in an aqueous suspension containing a polymerization catalyst and a suspension stabilizer to form small polymer particles,
Among these, those that satisfy the above conditions can be employed as the hydrophilic polymer material in the present invention.

Additionally, if an organic solvent is added to the polymerization system that dissolves the monomer but does not dissolve the polymer, the resulting spherical polymer becomes porous, increasing its surface area, and the present invention It is effective in improving the separation efficiency in the method.

Further, it is effective to crosslink the new aqueous polymeric substance with a suitable crosslinking agent or the like in order to improve the mechanical strength when used as a filler.

Note that the hydrophilic polymer substance is preferably used in the form of particles with a diameter of 5 to 200 microns, and is usually used in a state packed in a column for liquid chromatography.

Next, in order to separate ubiquinone by the method of the present invention, first, an extract containing ubiquinone extracted from animal, plant, or microbial tissue is injected into a column packed with the hydrophilic polymer substance, and the extract is It is adsorbed onto a filler made of molecular substances. The above extract contains ubiquinone (trans-2,3-dimethoxy-5-methyl-6-multiprenyl-1,4-benzoquinone), most of which is contained in plants, animals, and microorganisms. The problem lies in the removal of this lipid.

Next, the lipids are eluted by passing a mixed solvent of a water-soluble organic solvent and water through the filler on which the extract has been adsorbed as described above, but at this time, ubiquinone is not eluted and is absorbed into the filler. It remains absorbed.

It was discovered for the first time in the present invention that lipids and ubiquinone, which make up the majority of the extract, can be separated using the above mixed solvent.The reason why such separation is possible is that both lipids and ubiquinone are hydrophobic. This is thought to be due to the fact that lipids have higher polarity, and their behavior towards the above mixed solvents is different. The water-soluble organic solvents used in the above mixed solvent include methyl alcohol, ethyl alcohol, propyl alcohol, isopropyl alcohol, butyl alcohol, isobutyl alcohol, tertiary butyl alcohol, acetone, methyl ethyl ketone, acetonitrile, dimethyl sulfoxide, and diethyl alcohol. Ether etc. are used alone or in combination of two or more,
As for the mixing ratio of the organic solvent and water, the organic solvent is usually used in a larger amount than the water, but a mixing ratio suitable for eluting only lipids may be selected as appropriate.

Next, the organic solvent containing no water is passed through the packing material from which the lipids have been eluted as described above, whereby the ubiquinone adsorbed on the hydrophilic polymer substance is eluted. By concentrating the solvent solution, a concentrate of ubiquinone can be obtained. The organic solvent used to elute ubiquinone has the property of being soluble in water, but it may be the same or different from the organic solvent used for lipid elution as described above. An appropriate organic solvent may be selected from the organic solvents already exemplified.

Since the method for separating ubiquinone of the present invention is as described above, ubiquinone can be efficiently separated from extracts of animals, plants, and microorganisms with simple operations.

The present invention will be explained below based on examples.

Example 1 Rhodotorula cultured using waste molasses
3N hydrochloric acid was added to 100 g of dried cells of mucilaginosa (yeast), centrifuged, washed with water, suspended in ethyl alcohol, and extracted by stirring for about 3 hours while maintaining the temperature at 70°C in a nitrogen stream. After filtering and washing with ethyl alcohol, 2.2 g of a lipid-based extract containing 38 mg of ubiquinone Q ₁₀ was obtained.

Meanwhile, 4% by weight was placed in a 2L separable flask equipped with a condenser, stirrer, thermometer and dropping funnel.
A mixed solution consisting of 400 ml of an aqueous polyvinyl alcohol solution, 100 g of tetramethylolmethane triacrylate, and 1.5 g of benzoyl peroxide was supplied.

Next, the temperature was raised to 80° C. while stirring at a stirring speed of 400 rpm, reaction was carried out for 10 hours, and the mixture was cooled. After cooling, the polymerization product was separated from the mother liquor and washed with hot water and acetone to obtain a spherical polymer having a particle size of 5 to 20 microns.

The spherical polymer thus obtained had a molecular weight of 100,000 or more and was wettable with water.
Particles of 8 to 12 microns, obtained by removing fine particles and coarse particles, were dispersed in 80 ml of ion-exchanged water and placed in a stainless steel column (diameter 28 mm, length 25 cm).
ion exchange water by high pressure constant flow pump to 1.6
A column packed with a hydrophilic polymer substance was prepared by packing by force feeding at a rate of ml/min.

Next, the extract obtained above was dissolved in 10 ml of isopropyl alcohol and injected into the column packed with the above-mentioned hydrophilic polymer substance to allow the extract components to be adsorbed to the column packing.
Using 1000 ml of the mixture with volume part as the eluent,
₁₀ ml of 100% isopropyl alcohol.
Components mainly consisting of were eluted and flowed out. Concentration resulted in 162 mg, which contained 36 mg of ubiquinone Q ₁₀ .

Example 2 10 g of concentrated fish oil (bonito) containing 4 mg of ubiquinone Q ₁₀ in 1 g was treated in the same manner as in Example 1 to obtain 130 mg of a concentrate containing 25 mg of ubiquinone Q ₁₀ .

Claims (1)

[Claims] 1. When separating ubiquinone from extracts containing ubiquinone from animal, plant or microbial tissue, lipids are eluted from the hydrophilic polymeric substance on which the extract has been adsorbed using a mixed solvent of water and an organic solvent that is soluble in water; A method for separating ubiquinone, which comprises then eluting the ubiquinone adsorbed onto the hydrophilic polymer substance with the organic solvent that does not contain water.