WO2012081604A1 - Composition containing orotic acid, and process for production thereof - Google Patents

Abstract

The purpose of the present invention is to provide a process for producing a composition containing orotic acid and having a uric acid content of equal to or lower than a detection limit with high efficiency and at low cost by separating and removing uric acid from a liquid composition containing orotic acid and uric acid. The present invention provides a process for producing a composition containing orotic acid and having a uric acid content of equal to or lower than a detection limit, which comprises a step of bringing active carbon into contact with a liquid composition containing orotic acid and uric acid, a step of bringing the active carbon that has been subjected to the contacting procedure into contact with an alkaline solution to elute orotic acid, and a step of collecting the eluted alkaline solution containing orotic acid.

Description

Orotic acid-containing composition and method for producing the same

The present invention relates to a method for removing uric acid from a liquid composition containing orotic acid and uric acid, an orotic acid-containing composition from which uric acid has been removed, and a method for producing the same.

Orotic acid is a precursor of pyrimidine nucleotides, is biosynthesized from amino acids in vivo, and is frequently present in dairy products such as milk and whey in food (Non-patent Document 1).

Orotic acid has various effects that are effective in maintaining and promoting health, such as uric acid level lowering action, anti-inflammatory action, nourishing tonic action, liver function promoting action, etc. Therefore, alone or in combination with other ingredients The use for medicines, quasi drugs, health foods, etc. is under consideration.

¡Orotic acid is also included in by-products generated in the production of dairy products, but most of it is discarded as industrial waste. For example, cheese whey, which is a by-product of cheese manufacture, is partly pulverized and used as a food material, but at present, a lot of cheese whey is discarded worldwide. For this reason, if orotic acid can be recovered from the dairy by-product to be discarded, orotic acid can be provided at low cost.

However, milk by-products such as cheese whey contain a large amount of uric acid, which is a degradation product of purine nucleotides, in addition to orotic acid. Since an increase in uric acid level also causes gout, excessive intake of uric acid is not preferable for health, and it is desirable to remove uric acid as much as possible when orotic acid collected from milk by-products is used as a health food material. In addition, since orotic acid in cheese whey is a very small amount of about 0.1% per solid content, the solid content other than orotic acid is removed to increase the orotic acid content per solid content (hereinafter, orotic acid is concentrated) It is more desirable if it can be said.

On the other hand, as a method for separating nucleic acid-related substances, a solution containing nucleotides and nucleosides is adsorbed on activated carbon, and after washing with water, nucleotides and nucleobases are eluted with an aqueous alkali metal solution, and then nucleosides are eluted with an aqueous alkali metal solution containing ethanol. Thus, a method for separating nucleotides and nucleosides from each other has been reported (Patent Document 1). However, unlike nucleotides and nucleosides, nucleic acid-related substances that do not contain sugars or phosphate groups in the structures such as orotic acid and uric acid do not differ greatly in nature, so it is usually difficult to separate them using conventional methods. No method has been reported that can efficiently separate the two.

JP 59-104399 A

J. Dairy Sci., 67: 723-728 (1984)

An object of the present invention is to separate and remove uric acid from a liquid composition containing orotic acid and uric acid, and efficiently and inexpensively produce a composition containing orotic acid and having a uric acid content below the detection limit. It is to provide a method.

The present invention relates to the following [1] to [5].
[1] A liquid composition containing orotic acid and uric acid, comprising a step of bringing a liquid composition containing orotic acid and uric acid into contact with activated carbon, and a step of bringing an alkaline solution into contact with the activated carbon after the contact to elute orotic acid How to remove uric acid from things.
[2] A step of contacting activated carbon with a liquid composition containing orotic acid and uric acid, a step of contacting alkaline solution with activated carbon after the contact to elute orotic acid, and an alkali containing the eluted orotic acid A method for producing a composition containing orotic acid and having a uric acid content below the detection limit, from a liquid composition containing orotic acid and uric acid, comprising a step of recovering the solution.
[3] The method according to [1] or [2], further comprising a step of washing the activated carbon with water or acid following the step of contacting the activated carbon with a liquid composition containing orotic acid and uric acid.
[4] The method according to any one of [1] to [3], wherein the liquid composition containing orotic acid and uric acid is milk or a composition prepared from the milk.
[5] Milk or a composition prepared from the milk, which contains orotic acid and has a uric acid content below the detection limit.
This application claims the priority of the Japan patent application 2010-279296 for which it applied on December 15, 2010, and includes the content described in the specification of this patent application.

According to the method of the present invention, it is possible to easily and efficiently produce a composition containing orotic acid from a liquid composition containing orotic acid and uric acid and having a uric acid content below the detection limit. In addition, it is possible to concentrate orotic acid to a high concentration by repeating the steps of the method of the present invention, and it is possible to continuously use activated carbon to be used. Therefore, the method of the present invention improves productivity and economy. Are better.

As the “orotic acid” in the present invention, in addition to orotic acid (uracil-4-carboxylic acid), an alkyl group having 1 to 6 carbon atoms such as methyl, ethyl, butyl, propyl, pentyl, hexyl, etc., in the 4-position carboxylic acid 1-, 2-, 3-, and 6-position hydrogens are alkyl groups having 1 to 6 carbon atoms such as methyl, ethyl, butyl, propyl, pentyl, hexyl, or the like, or methyl, Derivatives substituted with alkoxy groups having 1 to 6 carbon atoms such as ethyl, butyl, propyl, pentyl, hexyl, etc .; or alkali metal salts such as sodium salts and potassium salts thereof; magnesium salts, calcium salts, etc. Heavy metal salts such as alkaline earth metal salts and zinc salts of ammonium; ammonium salts such as ammonium and tetramethylammonium; morpholine, Pharmaceutically acceptable salts of organic amine addition salts obtained by adding such Perijin also encompassed.

The “uric acid” in the present invention includes, in addition to uric acid, salts such as sodium salt, potassium salt, ammonium salt, 3-N-methyluric acid, 3-N-lauryluric acid, 7-N-butyluric acid, 1 -N-alkyl-modified products such as -N-ethyluric acid, 9-N-lauryluric acid, 3,7-N-dimethyluric acid, uric acid glycosides such as 3-N-ribosyluric acid, 9-N-glycosyluric acid, etc. Are included, but uric acid or a salt thereof is preferred.

The “liquid composition containing orotic acid and uric acid” is not limited to an aqueous solution as long as it is a liquid composition containing orotic acid and uric acid. Product), a culture of microorganisms capable of producing orotic acid, a treated product of the culture, a supernatant of the culture, a beverage, etc., but milk and dairy products are preferred. It is done.

Examples of the “milk” include milk extracted from animals selected from cows, goats, sheep, camels, reindeer, yaks, horses, donkeys, etc., and milk derived from cows, goats, or sheep is particularly preferable. “Milk” in the present invention includes raw milk, cow milk, special milk, raw goat milk, pasteurized goat milk, raw noodle milk, partially skimmed milk, skimmed milk, and processed milk.

Compositions prepared from milk refer to so-called dairy products, cream, butter, butter oil, whey, concentrated whey, ice cream (ice cream, ice milk, lacto ice), concentrated milk, defatted concentrated milk, and sugar-free. Examples of the milk include sugar-free skim milk, sweetened milk, sweetened skim milk, yogurt (fermented milk), lactic acid bacteria beverages and milk beverages. In addition, powdered milk such as whole milk powder, skim milk powder, cream powder, whey powder, sweetened powdered milk, sweetened skim milk powder, and adjusted powdered milk may be dissolved or dispersed in a solvent such as milk or water. In the present invention, these milks and dairy products may be any one kind or a mixture of two or more kinds.

Further, as long as it is a liquid composition containing orotic acid and uric acid, it may be a composition obtained by roughly refining these compositions.

The contents of orotic acid and uric acid in the liquid composition containing orotic acid and uric acid and the ratio thereof are not particularly limited.

In addition, the contents of orotic acid and uric acid in the liquid composition can be quantified by a conventional method such as liquid chromatography described in JOURNAL OF AOAC INTERNATIONAL, 87, 1, 116-122 (2004).

The method for removing uric acid from a liquid composition containing orotic acid and uric acid according to the present invention includes a step of contacting activated carbon with a liquid composition containing orotic acid and uric acid (contacting step), and an alkaline solution on the activated carbon after the contact. A step of elution of orotic acid by contacting with (elution step).

Moreover, the method for producing a composition containing orotic acid of the present invention and having a uric acid content below the detection limit is a step of contacting a liquid composition containing orotic acid and uric acid with activated carbon (contact step). And a step of bringing the alkaline solution into contact with the activated carbon after the contact to elute orotic acid (elution step), and a step of collecting the alkaline solution containing the eluted orotic acid (recovery step).

In the orotic acid-containing composition after removing uric acid, the content of uric acid is below the detection limit so that the orotic acid concentration is 10 mg / L by suspending the composition in water or the like as necessary. When prepared, it can be confirmed by the fact that no ultraviolet absorption of uric acid is detected at 280 nm. Usually, when calculating based on the detection limit value of uric acid, the removal rate of uric acid in the raw material is calculated to be 99% or more.

Hereinafter, each step will be described.
First, in the contacting step, activated carbon and uric acid in the liquid composition are adsorbed on the activated carbon by bringing activated carbon into contact with the liquid composition containing orotic acid and uric acid.

The activated carbon is not particularly limited, and any raw material (woody raw material, mineral raw material, etc.) and shape (granular, powdery, granular, etc.) may be used. For example, granular white leopard LH2c or granular white leopard KL manufactured by Nippon Enviro Chemicals can be used.

The liquid composition containing orotic acid and uric acid may be contacted with activated carbon, and any method such as a contact filtration method, a fixed bed adsorption method, a moving bed adsorption method, or a fluidized bed adsorption method may be used. A bed adsorption method, for example, a method in which activated carbon is packed in a column and the liquid composition is passed from the upper part or the lower part of the column is preferable because it is simple and has high efficiency of adsorbing orotic acid and uric acid.

The amount of activated carbon used varies depending on the type, but is usually about 1 to 200 times, preferably about 10 to 100 times that of orotic acid and uric acid contained in the liquid composition to be contacted.

The pH of the liquid composition containing orotic acid and uric acid to be brought into contact with the activated carbon is 1 to 7, preferably 2 to 7, from the viewpoint of recovery of orotic acid and removal of uric acid. For pH adjustment, an acid such as hydrochloric acid, sulfuric acid, nitric acid, citric acid, lactic acid, and acetic acid, and an alkali such as sodium hydroxide can be used.

Further, after the contacting step, before the elution step, a step of washing the activated carbon with acid or water (washing step) may be performed. In the washing step, the activated carbon that has been contacted with the liquid composition containing orotic acid and uric acid in the previous step is washed with water or acid. Examples of the acid used for washing include hydrochloric acid, sulfuric acid, nitric acid, citric acid, lactic acid, and acetic acid. The amount of water or acid used is not particularly limited, but is usually about 0.1 to 5 times, preferably about 1 to 3 times the amount of the liquid composition brought into contact with activated carbon.

Next, in the elution step, orotic acid adsorbed on the activated carbon can be eluted while contacting uric acid on the activated carbon by contacting the activated carbon after contact with an alkaline solution. As the alkaline solution, an aqueous sodium hydroxide solution, an aqueous potassium hydroxide solution, aqueous ammonia, or the like can be used. When using a sodium hydroxide aqueous solution or a potassium hydroxide aqueous solution, the concentration is preferably from 0.1 mol / L to 2 mol / L. When ammonia water is used, the concentration is preferably 1% or more and 10% or less. The amount of the alkaline solution to be used is usually about 0.01 to 1 times, preferably about 0.1 to 0.2 times that of the liquid composition brought into contact with the activated carbon in the contact step.

Finally, in the recovery step, an alkaline solution containing the eluted orotic acid is recovered. This alkaline solution contains orotic acid as it is, but can be used as a composition in which the content of uric acid is below the detection limit (hereinafter also referred to as “the composition of the present invention”). Further, when a volatile alkali such as ammonia water is used as an alkaline solution for eluting orotic acid in the elution step, the alkali is removed by distillation operation combined with heating or reduced pressure, and this is used as the composition of the present invention. Alternatively, when a non-volatile alkali is used, the alkali may be removed by combining neutralization with an acid and electrodialysis desalting, and this may be used as the composition of the present invention. Orotic acid can also be recovered from the alkaline solution using a synthetic resin or the like.

By the above method, the composition of the present invention can be easily produced using a liquid composition containing orotic acid and uric acid as raw materials.

It is possible to increase the adsorption amount of orotic acid per activated carbon by repeating the contact step and the washing step, or the contact step, the washing step and the elution step as necessary. The number of repetitions is not particularly limited, but is, for example, about 1 to 5 times.

In particular, since the composition of the present invention obtained by repeating the above contact step and washing step has a higher concentration than the orotic acid concentration in the raw material, the above method concentrates orotic acid simultaneously with the removal of uric acid. Also excellent in that it can.

In the above method, the activated carbon after the elution step can be used repeatedly. Even if it is used repeatedly, the recovered amount of orotic acid is hardly lowered and uric acid can be removed, which is economically advantageous.

Hereinafter, the present invention will be described more specifically with reference to examples. However, the present invention is not limited to these examples. An example of the batch method is shown, but the column method can be carried out without any problem.

(Example 1) Recovery of orotic acid from orotic acid and uric acid-containing liquid 1 g of commercially available whey permeate powder (containing 1.8 mg of orotic acid and 0.55 mg of uric acid) in 50 ml of water (pH 6. 5) and 1 g of whey permeate powder were dissolved in 50 ml of water, and then an aqueous solution having a pH of 2.8 with 1 mol / l hydrochloric acid was prepared.

0.1 g of activated carbon was added to each whey permeate aqueous solution, and orotic acid was adsorbed while stirring the solution for 30 minutes. Thereafter, the activated carbon was washed with 50 ml of water, orotic acid was eluted with 50 ml of 5% aqueous ammonia, and the eluted orotic acid was separated and quantified by high performance liquid chromatography (HPLC) (detection wavelength 280 nm).

As a result, when using a whey permeate aqueous solution adjusted to pH 6.5, about 0.4 mg of orotic acid can be recovered, and when using a whey permeate aqueous solution adjusted to pH 2.8 which is an acidic condition, About 1.2 mg of orotic acid could be recovered. Uric acid was not detected in the orotic acid eluate after any treatment.

(Example 2) Study on continuous use of activated carbon and recovery of orotic acid from milk raw material 1 g of commercially available whey permeate powder (containing 1.8 mg of orotic acid and 0.55 mg of uric acid) in 50 ml of water, 1 mol An aqueous solution having a pH of 2.8 with / l hydrochloric acid was prepared. 0.1 g of activated carbon was added to the aqueous whey permeate solution, and orotic acid was adsorbed while stirring the solution for 30 minutes. Thereafter, the activated carbon was washed with 50 ml of water, and orotic acid was eluted with 50 ml of 5% ammonia water.

Subsequently, a new whey permeate aqueous solution was prepared each time using the same activated carbon, and the above operation was repeated a total of 6 times. The orotic acid in each eluate was separated and quantified using HPLC under the same conditions as in Example 1.

The obtained eluate was dried under reduced pressure, and the purity of orotic acid per solid content was measured. As a result, the recovered amount of orotic acid was about 0.84 mg on average, and the orotic acid content averaged 7.5% per solid (w / w).

In addition, the recovery amount of orotic acid in 6 operations was 0.7 mg or more in any recovery, and no tendency to decrease was observed even after repeated operations. In addition, no uric acid was detected in these 6 eluates.

INDUSTRIAL APPLICABILITY The present invention can be used in the field of manufacturing foods and drinks such as functional foods and supplements that use orotic acid effective for maintaining and promoting health and pharmaceuticals.
All publications, patents and patent applications cited herein are incorporated herein by reference in their entirety.

Claims (5)

Uric acid from a liquid composition containing orotic acid and uric acid, comprising a step of contacting activated carbon with a liquid composition containing orotic acid and uric acid, and a step of contacting the activated carbon after the contact with an alkaline solution to elute orotic acid How to remove. A step of contacting activated carbon with a liquid composition containing orotic acid and uric acid, a step of bringing alkaline solution into contact with activated carbon after the contact and eluting orotic acid, and collecting an alkaline solution containing the eluted orotic acid A method for producing a composition containing orotic acid and having a uric acid content below the detection limit from a liquid composition containing orotic acid and uric acid. The method according to claim 1, further comprising a step of washing the activated carbon with an acid or water following the step of bringing the activated carbon into contact with a liquid composition containing orotic acid and uric acid. The method according to any one of claims 1 to 3, wherein the liquid composition containing orotic acid and uric acid is milk or a composition prepared from the milk. Milk or a composition prepared from milk containing orotic acid and having a uric acid content below the detection limit.