JP2011168649A - Pigment composition of high 13-cis-crocetin content and method for producing the same - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a pigment preparation of high 13-cis-crocetin content, and a method for producing the pigment preparation. <P>SOLUTION: This pigment preparation includes 13-cis-crocetin, and may comprise all-trans-crocetin or not. In a case of comprising all-trans-crocetin, when a peak area of all-trans-crocetin is 1 upon analysis by HPLC, the peak area of 13-cis-crocetin is ≥1. The method for producing pigment composition of a high 13-cis-crocetin content includes; a step of dissolving a mixture of 13-cis-crocetin and all-trans-crocetin in an alkaline condition to make a mixed solution which contains crocetin; a step of adjusting pH of the crocetin-containing mixed solution to 7.0-8.0 to deposit all-trans-crocetin; and a step of eliminating the deposited all-trans-crocetin to obtain the pigment composition of a high 13-cis-crocetin content. <P>COPYRIGHT: (C)2011,JPO&INPIT

Description

  The present invention relates to a pigment composition containing a large amount of crocetin, which is a kind of gardenia pigment, and a method for producing the same. The present invention particularly relates to a dye composition containing a large amount of highly soluble 13-cis-crocetin and a method for producing the same.

  Gardenia pigments derived from gardenia fruits have long been used as yellow pigments. Applications are mainly Chinese noodles, which are used for cup noodles, chilled yakisoba noodles, raw Chinese noodles used by ramen shops, and the like. Besides noodles, it is also used for confectionery (for example, cookies, candy, gum, pastry, confectionery bread, etc.) and chilled milk drinks. Since acidic solutions have low stability, they are not used for acidic beverages (such as lemon beverages). Gardenia pigment component is a kind of carotenoid, and is classified into crocin type and crocetin type from its structure (Table 1). The crocin-type pigment is water-soluble, has crocin as a main component, and exhibits a bright yellow color. The crocetin type pigment is insoluble in water but soluble in alkali and alcohol, and has crocetin as a main component and exhibits a greenish yellow color (lemon yellow). Crocin is a crocetin glycoside, and crocetin is obtained by desugaring it by alkaline hydrolysis or the like. Gardenia fruit exists in the form of crocin.

  Crocin and crocetin have a slightly different color tone, which is why there is a need for both. Especially for fried noodles, the color of crocetin may be preferred.

  Chinese noodles are alkaline because they use brine. Although crocin is water-soluble and crocetin is water-insoluble, since crocetin is also soluble in alkali, it can be used by dissolving crocetin in noodles using brine. However, since crocetin does not dissolve in milk beverages, crocetin cannot be used. Furthermore, when crocetin is placed in a neutral or acidic condition, the color changes from lemon yellow under an alkaline condition to an orangeish color, and therefore cannot be used as a pigment for coloring lemon yellow.

  A method for obtaining crocetin by hydrolyzing crocin derived from gardenia fruit has been known since ancient times. This crocetin is mainly composed of trans-crocetin (70% or more), and a patent application has been filed from a high purification method (for example, Patent Documents 1 to 3).

  The crocetin type gardenia pigment that is generally sold is mainly all-trans-crocetin.

  Furthermore, it was not known that crocetin type gardenia pigments include 13-cis-crocetin.

  Regarding crocetin, Patent Documents 1 to 6 are known. Patent Document 1 discloses a method for stably coloring noodles characterized by adding purified crocin or crocetin. Patent Document 1 discloses a gardenia fruit extract concentrate in Production Example 2, adjusted to pH 10 with caustic soda, heated at 95 ° C. for 1 hour, cooled, adjusted to pH 3 with hydrochloric acid, and the precipitated pigment was recovered by heating filtration. It describes that purified crocetin is obtained by washing with water, dissolving the pigment neutralized with caustic soda and recovering the precipitate, adding lactose as an excipient and spray drying.

  Patent document 2 is disclosing the manufacturing method of a powdery crocetin formulation. The crocetin described in Patent Document 2 is all-trans-crocetin and not 13-cis-crocetin. Furthermore, in the method described in Patent Document 2, after crocin is hydrolyzed, crocetin is precipitated by reducing the pH of the reaction solution to about 4.0 or lower, or by hydrolyzing with an acid, and centrifuged or separated. Crocetin is obtained by filtration. Therefore, crocetin obtained by this method is mainly all-trans-crocetin, and even if 13-cis-crocetin is mixed, it is less than 20%.

  Patent Document 3 discloses a method for purifying crocetin. In the method described in Patent Document 3, crocetin obtained by hydrolyzing crocin in a plant extract is treated with a lower alcohol or a mixed solvent containing 50% by volume or more of a lower alcohol, and a step of removing the components of the solvent is performed. It is characterized by doing. Crocetin obtained by the method described in Patent Document 3 is mainly all-trans-crocetin, and even if 13-cis-crocetin is mixed, it is less than 20%.

  Patent Document 4 discloses a sleep improving agent containing crocetin or a pharmacologically acceptable salt thereof as an active ingredient. Patent Document 4 paragraph 0011 describes the formula for crocetin used, which is all-trans-crocetin and not 13-cis-crocetin. Furthermore, paragraph 0018 of Patent Document 4 describes that the purity of crocetin is preferably about 75% by mass or more. Furthermore, in the method described in Patent Document 4, after crocin is hydrolyzed, crocetin is precipitated by reducing the pH of the reaction solution to about 4.0 or lower, or by hydrolyzing with an acid, followed by centrifugation or Crocetin is obtained by filtration. Therefore, crocetin obtained by this method is mainly all-trans-crocetin, and even if 13-cis-crocetin is mixed, it is less than 20%.

  Patent Document 5 discloses an anti-fatigue agent and a food and drink characterized by containing crocetin or a pharmacologically acceptable salt thereof as an active ingredient. Patent Document 5 paragraph 0010 describes the formula of crocetin used, which is crocetin is all-trans-crocetin and not 13-cis-crocetin. Furthermore, paragraph 0018 describes that the purity of crocetin is preferably about 70% by mass or more. Furthermore, in the method described in Patent Document 5, after crocin is hydrolyzed, crocetin is precipitated by reducing the pH of the reaction solution to about 4.0 or lower, or by hydrolyzing with an acid, followed by centrifugation or Crocetin is obtained by filtration. Therefore, crocetin obtained by this method is mainly all-trans-crocetin, and even if 13-cis-crocetin is mixed, it is less than 20%.

  Patent Document 6 discloses an eye strain improving agent and a food and drink characterized by containing crocetin or a pharmacologically acceptable salt thereof as an active ingredient. Patent Document 6 paragraph 0010 describes the formula of crocetin used, which is crocetin is all-trans-crocetin and not 13-cis-crocetin. Furthermore, paragraph 0018 describes that the purity of crocetin is preferably about 70% by mass or more. Furthermore, in the method described in Patent Document 6, after crocin is hydrolyzed, crocetin is precipitated by reducing the pH of the reaction solution to about 4.0 or lower, or by hydrolyzing with an acid, followed by centrifugation or Crocetin is obtained by filtration. Therefore, crocetin obtained by this method is mainly all-trans-crocetin, and even if 13-cis-crocetin is mixed, it is less than 20%.

  Thus, 13-cis-crocetin is not described in the literature concerning the conventional crocetin, and even if it is mixed in the conventional crocetin, the amount thereof is small, less than 20% of the crocetin-type yellow pigment.

JP 54-064652 A JP 2006-335859 A JP 2004-269663 A JP 2008-273939 A WO2006 / 112283 pamphlet JP 2007-31426 A

  All-trans-crocetin, a crocetin that has been mainly used in the past, cannot give a lemon yellow color tone specific to crocetin except under alkaline conditions, so it can give a lemon yellow color tone specific to crocetin even if it is neutral or acidic. It has been desired to provide a dye preparation that can be used.

  Furthermore, all-trans-crocetin has a low solubility in water particularly in the neutral range and acidic range, and when used as a pigment, it was impossible to produce a liquid preparation. Therefore, it could not answer the needs of customers who prefer liquids. Furthermore, the powder preparation has problems such as dusting, and the liquid preparation may not be preferred at the manufacturing site. Therefore, it has been desired to provide a liquid formulation containing crocetin.

  The present invention is intended to solve these problems, and aims to provide a liquid formulation of crocetin and a method for producing the same.

  As a result of intensive studies to solve the above-mentioned problems, the present inventors have found that 13-cis-crocetin has higher water solubility than all-trans-crocetin, and 13-cis-crocetin and all-trans. -It discovered that it could isolate | separate easily from the difference in the solubility to the water of a specific pH range from crocetin, and completed this invention based on this.

According to a preferred embodiment of the present invention, the following inventions are provided:
(Item 1) A dye preparation containing 13-cis-crocetin, which contains or does not contain all-trans-crocetin, and contains all-trans-crocetin, and all-trans-crocetin as analyzed by HPLC A dye preparation in which the peak area of 13-cis-crocetin is 1 or more when the peak area of 1 is 1.

  (Item 2) The dye preparation according to Item 1, wherein the peak area of 13-cis-crocetin when the peak area of all-trans-crocetin when analyzed by HPLC is 1 is 1 or more and 10,000 or less.

  (Item 3) The pigment preparation according to Item 1 or 2, wherein the color value is 100 or more and 32000 or less.

  (Item 4) The pigment preparation according to any one of Items 1 to 3, wherein the content of the 13-cis-crocetin is 0.05% by weight or more.

  (Item 5) The pigment preparation according to any one of Items 1 to 4, wherein a content of the 13-cis-crocetin is 0.1 wt% or more and 75 wt% or less.

  (Item 6) The pigment preparation according to any one of Items 1 to 5, which is a powder preparation.

  (Item 7) The pigment preparation according to any one of Items 1 to 5, which is a liquid preparation.

  (Item 8) The pigment preparation according to item 7, comprising water or alcohol as a solvent.

(Item 9) A method for producing a dye composition having a high content of 13-cis-crocetin,
Dissolving a mixture containing 13-cis-crocetin and all-trans-crocetin under alkaline conditions to obtain a mixed solution containing crocetin;
A step of precipitating all-trans-crocetin by adjusting the pH of the crocetin-containing mixed solution to pH 7.0-8.0; and removing the precipitated all-trans-crocetin to contain a high content of 13-cis-crocetin A method comprising the step of obtaining a dye composition.

(Item 10) A mixture containing the 13-cis-crocetin and all-trans-crocetin,
Extracting a crocin-containing raw material with a solvent to obtain a crocin extract; and hydrolyzing crocin in the crocin extract or a concentrate thereof to obtain a mixture containing 13-cis-crocetin and all-trans-crocetin 10. The method according to item 9, obtained by a process.

  (Item 11) The method of item 9 or 10 whose pH of the said precipitation process is 7.2-7.8.

  (Item 12) The manufacturing method of the colored food / beverage products including the process of mixing the pigment | dye formulation of any one of the items 1-9 with food / beverage products or food-drinks raw materials, and coloring.

  (Item 13) The manufacturing method of the item 12 whose said food / beverage products are neutral food / beverage products.

  (Item 14) The manufacturing method of the item 13 whose said neutral food / beverage products are milk drinks or a flower paste.

  (Item 15) Food / beverage products containing 13-cis-crocetin, with or without all-trans-crocetin and all-trans-crocetin, when all-trans-crocetin is analyzed by HPLC Food / beverage products whose peak area of 13-cis-crocetin when the peak area of 1 is set to 1 is 1 or more.

  (Item 16) Food / beverage products of item 15 whose said food / beverage products are neutral food / beverage products.

  (Item 17) Food / beverage products of item 16 whose said neutral food / beverage products are milk drinks or flower paste.

  In a particular embodiment, the production method of the present invention is characterized in that the originally present trans-crocetin component and 13-cis-crocetin component are separated at a specific pH.

  For example, crocin is produced by hydrolyzing crocin derived from gardenia fruit. This crocetin generally contains about 10 to 20% of 13-cis-crocetin. By adjusting the pH of the crocetin-containing material to a specific value, 13-cis-crocetin and all-trans-crocetin can be converted to 13-cis- by utilizing the difference in water solubility between neutral and weakly acidic regions. Crocetin can be isolated. In this way, for example, a mixture of about 13-cis-crocetin: all-trans-crocetin = 8: 2 can be obtained. A pigment powder can also be provided by adding an auxiliary agent such as dextrin to the mixture and pulverizing the mixture.

  13-cis-crocetin exhibits a higher solubility in water in the neutral range than a pigment preparation based on all-trans-crocetin.

  According to the present invention, it is possible to provide a pigment preparation that can be colored to lemon yellow peculiar to crocetin even under conditions other than alkaline.

  According to the present invention, a liquid formulation of crocetin has become possible. Since 13-cis-crocetin has high solubility in water in the neutral range, it is easy to use in the preparation of dye preparations and use as a dye.

  The production method of the present invention is a method that cannot be obtained by a normal method and is a unique special production method. In particular, the method of separating 13-cis-crocetin and all-trans-crocetin using the difference in solubility in water at a specific pH is inexpensive and easy to implement on an industrial scale, and is useful for industrial production. Is suitable.

  13-cis-crocetin was an important substance characterizing the individuality (physical properties, etc.) of gardenia pigments. Specifically, compared with all-trans-crocetin, the solubility in water is high and the color tone is different.

  The 13-cis-crocetin high-content crocetin pigment preparation of the present invention has a high 13-cis-crocetin content compared to the all-trans-crocetin content, a high solubility in water in the neutral range, It is characteristic that it can be dissolved and colored to lemon yellow characteristic of crocetin. The marketed crocetin pigment preparation is based on trans-crocetin.

FIG. 1 shows an HPLC chart of 13-cis-crocetin-rich crocetin powder produced in Example 1 at pH 7.5 in stage a. FIG. 2 shows an HPLC chart when the peak 2 compound fractionated by HPLC is measured again by HPLC. FIG. 3 is a photograph showing the results of coloring test 2. (1) Yellow only: Marigold dye preparation; (2) Yellow only: Crocin dye preparation; (3) Yellow only: 13-cis-crocetin high content crocetin powder; (4) Yellow only: Safflower dye preparation; (5) Blue addition: Marigold dye preparation; (6) Blue addition: Crocin dye preparation; (7) Blue addition: 13-cis-crocetin dye preparation; (8) Blue addition: safflower dye preparation; (9) Yellow only: All- Trans-crocetin high pigment preparation; (10) Yellow only: 13-cis-crocetin high pigment preparation; (11) Milk only. FIG. 4 is a photograph showing the results of a flour coloring test. (A) shows the coloration when there is powdered water, and (B) shows the color when there is no powdered water. (1) All-trans-crocetin high content dye preparation, (2) 13-cis-crocetin high content dye preparation, (3) Crocin dye preparation. FIG. 5 is a photograph showing the results of a flour paste coloring test. (1) All-trans-crocetin high pigment preparation, (2) 13-cis-crocetin high pigment preparation.

  Hereinafter, the present invention will be described in detail.

(Definition)
As used herein, the term “crocin” refers to a compound in which R ₁ and R ₂ are gentiobiose in formula 1 below:

  As used herein, the term “all-trans-crocetin” refers to a compound represented by Formula 2 below:

  As used herein, the term “13-cis-crocetin” refers to a compound represented by Formula 3 below:

In the present specification, the term “color value” is a unit that expresses the color intensity of a dye, and a Kolthoff buffer solution (50 mM Na ₂ CO ₃ -50 mM Na) is used in a reliable concentration range in an absorptiometer. ₂ B ₄ O ₇ , pH 10.0) means a value obtained by converting the absorbance at the maximum absorption wavelength when measured using a 1 cm cell into a value in a 10 wt% solution. This color value can be indicated by the following symbols:

In the present specification, this color value is indicated as “color value E (10%, 1 cm)” or “color value”.

  As used herein, the term “cake” as used in connection with a purification procedure refers to a residue that remains on a filter paper, filter, etc. when a solution containing insoluble matter is filtered. The filtration may be filtration without reducing pressure, or may be filtration under reduced pressure.

(1. Raw material for crocetin)
Crocetin is usually contained in plants as its glycoside crocin in the natural state. As the crocin-containing raw material, any plant containing crocin can be used. Preferably, the crocin-containing raw material is Rubiaceae gardenia (also called Gardenia jasminoides Ellis, Gardenia augusta Merril) and its related species, saffron stigmas, and the like. Gardenia fruit, saffron stigma, etc. are usually marketed as dried products. The dried fruit of gardenia is easily available and inexpensive, and it is preferable to use the dried fruit of gardenia as the crocin-containing material. Since extraction is easy, it is preferable that the crocin containing raw material is fragmented, ground or powdered.

(2. Method for producing 13-cis-crocetin of the present invention)
In a preferred embodiment, the production method of the present invention is a method for producing a dye composition having a high content of 13-cis-crocetin, wherein a mixture containing 13-cis-crocetin and all-trans-crocetin is dissolved under alkaline conditions. To obtain a crocetin-containing mixed solution; a step of precipitating all-trans-crocetin by adjusting the pH of the crocetin-containing mixed solution to pH 7.0 to 8.0; and the precipitated all-trans-crocetin A step of removing to obtain a dye composition having a high content of 13-cis-crocetin.

  In another embodiment, the production method of the present invention is a method for producing a 13-cis-crocetin-containing dye preparation, the step of extracting a crocin-containing raw material with a solvent to obtain a crocin extract; A step of hydrolyzing crocin in the concentrate to obtain a crocetin-containing mixed solution; and a step of obtaining a 13-cis-crocetin-containing pigment preparation by removing all-trans-crocetin from the crocetin-containing mixed solution.

  In a preferred embodiment, in this method, the removal of all-trans-crocetin from the crocetin-containing mixed solution is carried out by precipitating all-trans-crocetin from the crocetin-containing mixed solution at pH 7.2 to 7.8. Is called.

  In a further preferred embodiment, the removal of all-trans-crocetin from the crocetin-containing mixed solution brings the pH of the liquid containing the crocetin-containing mixture to 12.0 or higher, and then the crocetin-containing mixed solution and the acid are mixed. Then, the pH of the crocetin-containing mixed solution is set to 7.2 to 7.8 to precipitate all-trans-crocetin, and then the precipitated all-trans-crocetin is removed from the crocetin-containing mixed solution.

  In another embodiment, the method of the present invention is a method for producing a dye preparation containing 13-cis-crocetin, the method comprising extracting a crocin-containing raw material with a solvent to obtain a crocin extract; the crocin extract or crocin thereof A step of mixing the concentrated solution and an alkali to obtain an alkaline mixed solution having a pH of 12.0 to 12.5; stirring the alkaline mixed solution for 2 to 40 hours to hydrolyze crocin in the alkaline mixed solution to crocetin A step of obtaining a first crocetin-containing alkaline mixture; and mixing the first crocetin-containing alkaline mixture with an acid to adjust the pH of the mixture to 2.8 to 3.2 to precipitate crocetin. A step of filtering out the crocetin to obtain a crocetin cake; a mixture of the crocetin cake, water and an alkali to form a second clostin having a pH of 12.0 or more. A step of obtaining a tin-containing alkaline mixed solution; an acid is added to the second crocetin-containing alkaline mixed solution to adjust the pH to 7.2 to 7.8 to precipitate all-trans-crocetin to form a crocetin-containing weak alkaline mixed solution. Obtaining a 13-cis-crocetin-rich filtrate by filtering the weakly alkaline mixture containing crocetin and collecting the filtrate; and drying the 13-cis-crocetin-rich filtrate 13- A step of obtaining a cis-crocetin-containing dye composition.

  Each step will be described in more detail.

(2.1 Extraction of crocin)
First, crocin is extracted from the crocin-containing raw material. Crocin can be extracted from the crocin-containing raw material according to any method known in the art. Preferably, the crocin-containing raw material is extracted with a solvent. As the solvent, water, alcohol or a mixture thereof can be used. Examples of alcohol include ethanol. The solvent is preferably water or a hydrous alcohol. The proportion of alcohol in the hydrous alcohol can be arbitrarily set according to the production equipment, production conditions, and the like. For example, it may be about 10% by volume or more, about 30% by volume or more, about 50% by volume or more. The ratio of the alcohol in the hydrous alcohol may be, for example, about 95% by volume or less, about 80% by volume or less, about 60% by volume or less. The hydrous alcohol is preferably hydrous ethanol (ethanol aqueous solution).

  The crocin-containing raw material may be a plant in a raw state, or a plant that has been partially or almost completely dried. The plant material used for extraction is preferably fragmented, ground or pulverized.

  By contacting the crocin-containing raw material with a solvent, crocin is extracted into the solvent. The extraction conditions can be appropriately selected depending on the solvent used. For example, when hydrous alcohol is used as the extraction solvent, the temperature of the solvent can be arbitrarily set according to the production equipment, production conditions, and the like. For example, it may be about 0 ° C. or higher, about 10 ° C. or higher, about 20 ° C. or higher. For example, when hydrous alcohol is used as the extraction solvent, the temperature of the solvent can be, for example, about 100 ° C. or lower, about 80 ° C. or lower, and the like. When water is used as the solvent, it is preferably hot water. In general, the higher the temperature of the solvent, the faster the extraction, unless crocin does not decompose and the solvent volatilizes rapidly. Therefore, the temperature of the solvent is preferably as high as possible. During the crocin extraction, it is preferable to continuously or intermittently stir the mixture of the crocin-containing raw material and the solvent.

  The extraction time can be arbitrarily set according to manufacturing equipment, manufacturing conditions, and the like. The extraction time can be, for example, about 30 minutes or longer, about 1 hour or longer. The extraction time can be, for example, about 24 hours or less, about 12 hours or less, about 3 hours or less, and the like. The suitable extraction time can vary with the extraction temperature. For example, when the extraction temperature is about 50-70 ° C., the extraction time can be, for example, about 30 minutes or more, about 1 hour or more, etc .; the extraction time is, for example, about 12 hours or less, about 3 hours or less, about 2 It can be less than hours. The extraction operation may be performed once or a plurality of times. It is preferable to perform extraction several times using a new solvent. When the extraction is performed a plurality of times, the obtained plurality of extracts can be combined and used in the next step.

  In the present invention, the term “extract” refers to a liquid obtained by extraction from a crocin-containing raw material or a dried product thereof. When the extract is a liquid, it is also called an extract. The extract is obtained, for example, by bringing a crocin-containing raw material into contact with an arbitrary liquid solvent. Usually, in the extract, the ratio of one or more extracted components in the crocin-containing material to non-extracted components is higher than the ratio of the extracted components in the natural crocin-containing material to non-extracted components. The extract may be a solution obtained by transferring the extraction components into a liquid solvent, or a concentrate or dry solid obtained by partially or almost completely evaporating the solvent of this solution. There may be. The extract may be liquid or solid.

  The crocin-containing extract thus obtained may be used in the next step as it is, may be used after being concentrated, or may be concentrated to dryness and dissolved in a suitable solvent at the time of use. Good. As a solvent for dissolution, the same solvent as that which can be used for extraction of crocin (for example, water, alcohol or a mixture thereof) can be used. The crocin-containing extract is preferably concentrated under reduced pressure.

(2.2 Hydrolysis of crocin)
Next, crocin in the crocin-containing extract is hydrolyzed. Hydrolysis can be performed according to any method known in the art. The hydrolysis of crocin can be performed by using, for example, alkali, acid, hydrolase and the like. Any conventionally known alkali, acid, hydrolase and the like can be used, and those usable for food or food production are preferably used. Here, the acid refers to a substance from which an acidic aqueous solution can be obtained when dissolved in pure water. Examples of acids that can be used for the hydrolysis of crocin include hydrochloric acid, sulfuric acid, phosphoric acid and the like. Examples of alkalis that can be used for the hydrolysis of crocin include potassium hydroxide, sodium hydroxide, sodium carbonate, potassium carbonate and the like. Here, the alkali refers to a substance from which an alkaline aqueous solution can be obtained when dissolved in pure water. The alkali used for the hydrolysis of crocin is preferably a substance in which the aqueous solution obtained when dissolved in pure water has a pH suitable for the hydrolysis of crocin (for example, pH 9.5 or higher). The acid, alkali or hydrolase may be mixed with the crocin-containing extract as it is, or may be mixed after being dissolved or dispersed in an appropriate solvent (for example, water, alcohol or a mixture thereof). Since it is cheap, it is preferable to use an alkali or an acid, and it is more preferable to use an alkali. It is preferable to stir during the hydrolysis.

  When hydrolyzing crocin with an alkali, an alkaline mixed liquid is obtained by mixing the crocin extract and the alkali. The pH of the alkaline mixed solution can be, for example, about 11.0 or more, about 12.0 or more, and the like. The pH of the alkaline mixture may be, for example, about 14.0 or less, about 13.0 or less. Hydrolysis of crocin proceeds in the alkaline mixture. The temperature and time for performing the hydrolysis can be arbitrarily set. For example, the hydrolysis temperature (that is, the temperature of the alkaline mixture) can be arbitrarily set according to the production equipment, production conditions, and the like. The temperature of hydrolysis can be, for example, about 10 ° C. or higher, about 20 ° C. or higher. The hydrolysis temperature (that is, the temperature of the alkaline mixture) can be, for example, about 70 ° C. or less, about 50 ° C. or less, and the like. In general, unless the conditions are such that crocin does not decompose and the solvent volatilizes rapidly, the higher the temperature of the alkaline mixture, the faster the decomposition. Therefore, the temperature of the solvent is preferably as high as possible. The time for performing hydrolysis with an alkali can be arbitrarily set according to the production equipment, production conditions, and the like. The time for hydrolysis may be, for example, about 1 hour or more, about 3 hours or more, about 10 hours or more. The time for performing the hydrolysis with alkali may be, for example, about 48 hours or less, about 24 hours or less, and the like. By stirring / standing the alkaline mixed solution for a predetermined time, crocin in the alkaline mixed solution is hydrolyzed to become crocetin, and a first crocetin-containing alkaline mixed solution is obtained.

  When crocin is hydrolyzed with an acid, an acidic mixture is obtained by mixing the crocin extract and the acid. The pH of the acidic mixture can be, for example, about 1.0 or more, about 2.0 or more, about 3.0 or more, and the like. The pH of the acidic mixed solution can be, for example, about 4.0 or less, about 3.0 or less, and the like. Hydrolysis of crocin proceeds in the acidic mixture. The temperature and time for performing the hydrolysis can be arbitrarily set. For example, the hydrolysis temperature (that is, the temperature of the acidic mixed solution) can be arbitrarily set in accordance with production equipment, production conditions, and the like. The temperature of hydrolysis can be, for example, about 10 ° C. or higher, about 20 ° C. or higher. The hydrolysis temperature (that is, the temperature of the acidic mixture) can be, for example, about 70 ° C. or less, about 50 ° C. or less, and the like. Generally, unless the conditions are such that crocin does not decompose and the solvent volatilizes rapidly, the higher the temperature of the acidic mixture, the faster the decomposition. Therefore, the temperature of the solvent is preferably as high as possible. The time for performing hydrolysis with an acid can be arbitrarily set according to the production equipment, production conditions, and the like. The time for hydrolysis may be, for example, about 1 hour or more, about 3 hours or more, about 10 hours or more. The time for performing the acid hydrolysis generally can be, for example, about 48 hours or less, about 24 hours or less, and the like. By stirring / standing the acidic mixture for a predetermined time, crocin in the acidic mixture is hydrolyzed to crocetin, and a crocetin-containing acidic mixture is obtained.

  When crocin is hydrolyzed with a hydrolase, a crocin-hydrolase mixture is obtained by mixing the crocin extract and the hydrolase. The pH of the crocin-hydrolase mixed solution is preferably set according to the optimum pH of the hydrolase. As the hydrolase, any hydrolase can be used as long as it is a hydrolase capable of degrading the sugar moiety of crocin. An example of such a hydrolase is β-glucosidase. Any commercially available hydrolase that can degrade the sugar portion of crocin may be used. It is preferable to use a hydrolase having an optimum pH in the alkaline region. For example, when β-glucosidase is used as a hydrolase, the pH is, for example, within the range of optimum pH ± 1.0 of the β-glucosidase used, within the range of optimum pH ± 0.5 of the reaction, It may be in the range of optimum pH ± 0.1, optimum reaction pH, and the like. The temperature of the crocin-hydrolase mixed solution is preferably set in accordance with the optimum reaction temperature of the hydrolase. The temperature of the crocin-hydrolase mixture can be arbitrarily set according to the production equipment, production conditions, and the like. The temperature of the crocin-hydrolase mixed solution may be, for example, a reaction optimum temperature range of ± 10 ° C., a reaction optimum temperature range of ± 5 ° C., a reaction optimum temperature, or the like. The time for performing the hydrolysis can be arbitrarily set. The time for performing the hydrolysis by the hydrolase is until the hydrolysis is completed, and may be, for example, about 30 minutes or more, about 45 minutes or more, about 1 hour or more, for example, about 3 hours or less, about It may be 2.5 hours or less, about 2 hours or less, and the like. By stirring / standing the crocin-hydrolase mixture for a predetermined time, crocin in the crocin-hydrolase mixture is hydrolyzed to crocetin, and a crocetin-containing mixture is obtained.

(2.3 Removal of all-trans-crocetin and recovery of 13-cis-crocetin from a mixture containing 13-cis-crocetin and all-trans-crocetin)
After crocetin is formed by hydrolysis of crocin, the crocetin-containing mixed solution after hydrolysis is acidified by mixing with acid to precipitate crocetin. When crocin is hydrolyzed with an acid, crocin dissolves in an acidic aqueous solution, but crocetin does not dissolve in an acidic aqueous solution, so crocetin precipitates as crocetin is formed by crocin hydrolysis. Therefore, a hydrolysis process can act as this precipitation process.

  As the acid, any known acid can be used, and preferably an acid that can be used for food or food production is used. Examples of acids that can be used include inorganic acids (eg, hydrochloric acid, sulfuric acid, phosphoric acid, etc.) and organic acids (eg, citric acid, acetic acid, etc.). The pH of the mixed solution during precipitation is preferably about 1.0 or more, more preferably about 2.0 or more, and most preferably about 2.8 or more. The pH of the mixture during precipitation is preferably about 5.0 or less, more preferably about 4.0 or less, and most preferably about 3.2 or less. The temperature and time for performing the precipitation can be arbitrarily set. For example, the temperature of the liquid mixture during precipitation is preferably about 0 ° C. or higher, more preferably about 4 ° C. or higher. The temperature of the mixed solution during precipitation is preferably about 30 ° C. or less, and most preferably about 20 ° C. or less. The time for performing the precipitation is preferably about 10 seconds or more, more preferably about 1 minute or more. The time for performing the precipitation is generally preferably about 1 hour or less, more preferably about 30 minutes or less, and most preferably about 5 minutes or less.

  The precipitated crocetin-containing solid is separated from the liquid by filtration with a filter paper, a filter or the like, or by centrifugation. When separated by filter paper or the like, a crocetin cake is formed on the filter paper or the like. Filter filtration may be performed by a filter press.

  When gardenia fruit is used as the crocin-containing raw material, the separated crocetin-containing solid (also referred to herein as “a mixture containing 13-cis-crocetin and all-trans-crocetin”) is all-trans-crocetin and Contains 13-cis-crocetin. The separated crocetin-containing solid often further contains an acid, a neutralized salt, impurities derived from the raw material, and the like. Therefore, the separated crocetin-containing solid can be washed with water. The separated crocetin-containing solid is preferably washed with water until substantially free of acid. The amount of water used at this time is preferably small so that 13-cis-crocetin does not elute excessively. For example, the amount of water used for washing is preferably no more than about 10 times the volume of the separated crocetin-containing solid, more preferably no more than about 5 times, and most preferably no more than about 3 times. preferable.

  Next, the separated crocetin-containing solid (which may or may not be washed with water) is dissolved under alkaline conditions to prepare a crocetin-containing mixed solution. For example, a crocetin-containing mixed solution (also referred to as a second crocetin-containing alkaline mixture) is obtained by dispersing a crocetin-containing solid in water and then mixing it with alkali to make it alkaline. Alternatively, a crocetin-containing mixed solution may be prepared by dispersing a crocetin-containing solid in an alkaline aqueous solution. The pH of the crocetin-containing mixed solution is preferably about 11.0 or more, and most preferably about 12.0 or more. The pH of the crocetin-containing mixed solution is preferably about 14.0 or less, and most preferably about 13.0 or less. When crocetin is sufficiently dissolved in the crocetin-containing mixed solution, the next step is performed. In this step, both all-trans-crocetin and 13-cis-crocetin are dissolved in the mixed solution containing crocetin.

  Next, all-trans-crocetin is precipitated by adjusting the pH of the crocetin-containing mixed solution to pH 7.0 to 8.0. For example, all-trans-crocetin is precipitated by mixing the crocetin-containing mixed solution with an acid to make the pH of the crocetin-containing mixed solution weakly alkaline. At this time, the pH is preferably adjusted to about 7.2 or higher, more preferably about 7.4 or higher, and particularly preferably about 7.5 or higher; pH is preferably about 7.9 or lower, Preferably, it is adjusted to about 7.7. The temperature and time for carrying out the precipitation of all-trans-crocetin can be arbitrarily set. For example, the temperature of the mixed solution during precipitation is preferably about 0 ° C. or higher, more preferably about 4 ° C. or higher, and particularly preferably about 10 ° C. or higher. The temperature of the mixed solution during precipitation is preferably about 30 ° C. or less, and most preferably about 20 ° C. or less. The lower the temperature of the mixed solution during the precipitation, the more the precipitation is promoted. Therefore, the temperature of the solvent is preferably as low as possible. The time for performing the precipitation is preferably about 10 seconds or more, more preferably about 1 minute or more. The time for performing the precipitation is generally preferably about 1 hour or less, more preferably about 30 minutes or less. Under this weakly alkaline condition, much of all-trans-crocetin precipitates, but much of 13-cis-crocetin remains dissolved. Therefore, a solution in which 13-cis-crocetin is concentrated with respect to all-trans-crocetin can be obtained by removing the precipitated solid.

  The precipitated all-trans-crocetin-containing solid is removed from the liquid by filtration with a filter paper, a filter, or the like, and centrifuged to obtain a 13-cis-crocetin-rich solution. When separated by filter paper or the like, an all-trans-crocetin-rich crocetin cake is formed on the filter paper or the like. At this time, the filter separation may be promoted by a filter press.

  The obtained 13-cis-crocetin-rich solution may be used as it is, or may be dried and dissolved in a solvent as necessary. The 13-cis-crocetin-rich solution can be dried using, for example, an air dryer, a vacuum dryer, a spray dryer, a freeze dryer, or the like. In this way, a 13-cis-crocetin-rich powder can be obtained.

  In the above, a method (also referred to as a pH adjustment method) in which all-trans-crocetin and 13-cis-crocetin are separated using a difference in solubility in water at a specific pH is described. However, the separation of all-trans-crocetin and 13-cis-crocetin may be performed by other methods known in the art. Examples of methods that can be used for this separation include solvent partitioning, recrystallization, adsorption chromatography, partition chromatography and the like.

  The 13-cis-crocetin-enriched powder may be further purified to make 13-cis-crocetin more pure or to remove impurities other than crocetin (eg, lipids, polyphenols, saccharides, etc.). Examples of purification methods include solvent partitioning, membrane fractionation (eg, membrane fractionation using ultrafiltration membranes), filtration, centrifugation, recrystallization and chromatography (eg, column chromatography). It is done. Examples of carriers that can be used for chromatography include gel filtration chromatography carriers, ligand exchange chromatography carriers, ion exchange chromatography carriers, adsorption chromatography carriers, and partition chromatography carriers.

  13-cis-crocetin may be purified to a higher purity. For example, it is preferable to purify until the purity is about 10% by weight or more, more preferably to be purified until the purity is about 25% by weight or more, and to be purified until the purity is about 30% by weight or more. It is particularly preferred. The purity of 13-cis-crocetin is, for example, about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, about 90% or more, or about 95%. You may refine | purify until it becomes more than%. The purity of 13-cis-crocetin can be determined, for example, by HPLC.

(3. Dye preparation of the present invention)
The dye preparation of the present invention is a dye preparation containing 13-cis-crocetin, which contains or does not contain all-trans-crocetin. When the peak area of lance-crocetin is 1, the peak area of 13-cis-crocetin is 1 or more.

  In this specification, “the peak area of 13-cis-crocetin when the peak area of all-trans-crocetin when analyzed by HPLC is 1” is the same conditions as in HPLC described in Example 1. The peak area of 13-cis-crocetin when the peak area of all-trans-crocetin is 1 as measured by 1. Preferably, Inertsil ODS-3 5 μm 4.6 × 250 mm (manufactured by GL Sciences) is used as a column, and liquid A (1% (v / v) acetic acid) and liquid B (acetonitrile) are used as a solvent, and the flow rate is 1 ml. At a column temperature of 40 ° C. for 0 minutes, the ratio of the B liquid at 20 minutes, the ratio of the B liquid at 20 minutes is 80%, the ratio of the B liquid at 30 minutes is 80%, When measured using a photodiode array (PDA) as a detector, time programmed so that the ratio of B liquid at 01 minutes is 20%, and the ratio of B liquid at 40 minutes is 20% The peak area of 13-cis-crocetin when the peak area of all-trans-crocetin is 1 in the chromatogram prepared at an absorbance of 417.0 nm. This area is determined by dividing the peak area of 13-cis-crocetin by the peak area of all-trans-crocetin. This value increases as the amount of all-trans-crocetin decreases.

  The peak area of 13-cis-crocetin when the peak area of all-trans-crocetin when analyzed by HPLC is 1, is preferably about 1 or more, more preferably about 1.5 or more, It is preferably about 1.8 or more, particularly preferably about 2 or more, and most preferably about 5 or more. The peak area of 13-cis-crocetin, when the peak area of all-trans-crocetin when analyzed by HPLC is 1, is not particularly limited because the higher the purity of 13-cis-crocetin, the larger the peak area. This value can be, for example, about 10,000 or less, about 5000 or less, about 3000 or less, about 2000 or less, about 1000 or less, about 500 or less, about 300 or less, about 100 or less, and the like. At this time, the molar extinction coefficient of 13-cis-crocetin is unknown. However, the molar extinction coefficient of cis-crocin: The molar extinction coefficient of trans-crocin is about 63350: about 89000 = about 0.7: 1 (J. Agric. Food Chem, 2008, 56, 1627-1637). In consideration, the molar extinction coefficient of 13-cis-crocetin is also considered to be considerably smaller than the molar extinction coefficient of all-trans-crocetin. Therefore, even if the peak area of all-trans-crocetin and the peak area of 13-cis-crocetin is 1: 1, the amount ratio is not 1: 1, but 13-cis-crocetin is considerably larger. It is estimated to be.

  Generally, the color value of the pigment preparation should be about 50 or more. The color value of the dye preparation of the present invention is preferably about 100 or more, more preferably about 200 or more. The color value of the dye preparation of the present invention may be, for example, about 500 or more, about 1000 or more, about 5000 or more. The color value of the dye preparation of the present invention is less than or equal to the color value of 13-cis-crocetin when the content of all-trans-crocetin is sufficiently low. Since all-trans-crocetin is considered to have a higher molar extinction coefficient than 13-cis-crocetin, the higher the all-trans-crocetin content, the more the dye compared to the case containing only 13-cis-crocetin. The color value of the formulation increases. The color value of the dye preparation of the present invention may be, for example, about 32000 or less, about 30000 or less, about 15000 or less, about 10,000 or less, about 5000 or less, about 1000 or less, and the like.

  In a preferred embodiment, the dye preparation of the present invention comprises 0.05% by weight or more of 13-cis-crocetin. The content of 13-cis-crocetin is preferably about 0.1% by weight or more, more preferably about 0.5% by weight or more, and further preferably about 1% by weight or more. The content of 13-cis-crocetin in the dye preparation of the present invention may be, for example, about 10% by weight or more, about 15% by weight or more in a specific embodiment. The content of 13-cis-crocetin in the pigment preparation of the present invention is, for example, about 90% by weight or less, about 85% by weight or less, about 80% by weight or less, about 75% by weight or less, about 70% by weight or less, about It may be 60 wt% or less, about 50 wt% or less, about 40 wt% or less, about 30 wt% or less.

  If the molar extinction coefficient of 13-cis-crocetin is known, the molar extinction coefficient of all-trans-crocetin, the peak of all-trans-crocetin on HPLC, It can be calculated from the peak area ratio to the area and the color value.

  In a preferred embodiment, the content of all-trans-crocetin in the dye preparation of the present invention is preferably about 1.0% by weight or less, more preferably about 0.7% by weight or less, and further preferably About 0.50% by weight or less, particularly preferably about 0.40% by weight or less, and most preferably about 0.35% by weight or less.

(3.1 Solid Dye Formulation of the Present Invention)
The dye formulations of the present invention are solid formulations in certain embodiments. The solid dye preparation of the present invention may contain, for example, an excipient. Examples of excipients include sugar, sugar alcohol, gum arabic and the like. Examples of sugar include dextrin and lactose.

  For example, a solid 13-cis-crocetin-containing dye preparation can be prepared by drying the 13-cis-crocetin-rich solution obtained by the production method of the present invention with or without an excipient. . The solid pigment preparation after drying can be crushed and sieved as necessary according to a method known in the art. By changing the mixing ratio of the excipient and the 13-cis-crocetin-rich solution, the content of 13-cis-crocetin in the obtained powder can be adjusted.

(3.2 Liquid dye preparation of the present invention)
The dye formulation of the present invention is a liquid formulation in certain embodiments. The solvent of the liquid dye preparation of the present invention preferably contains water. The solvent is preferably hydrous alcohol or water, more preferably hydrous ethanol or water.

  The proportion of the solvent in the liquid pigment preparation of the present invention can be, for example, about 85% by weight or more, about 90% by weight or more, about 95% by weight or more. The ratio of the solvent in the liquid pigment preparation of the present invention is, for example, about 99.5% by weight or less, about 99% by weight or less, about 98% by weight or less, about 97% by weight or less, about 96% by weight or less, about 95% by weight. % Or less.

  The liquid pigment preparation of the present invention may contain impurities derived from plant materials as long as the color and taste are not adversely affected. The 13-cis-crocetin-rich crocetin filtrate and its powder produced according to the method of the present invention may contain some impurities but have little adverse effect on color and taste. Therefore, the liquid pigment preparation of the present invention does not need to be costly for removing such impurities, and can be produced inexpensively and easily.

  The liquid pigment preparation of the present invention can be produced by obtaining 13-cis-crocetin-rich crocetin powder produced according to the method of the present invention and dissolving this powder in a solvent.

  In the present specification, the “liquid pigment preparation” refers to a liquid composition containing a pigment and used for coloring. In addition to all-trans-crocetin and a solvent, the liquid dye preparation of the present invention may contain other substances that can be used in liquid dye preparations usually in the art. Examples of such substances include preservatives, antioxidants, pH adjusters, sweeteners, food materials, and the like.

  The liquid pigment preparation of the present invention preferably contains an antioxidant. Examples of antioxidants include ascorbic acid, sodium ascorbate, enzyme-treated rutin, rosemary extract, green tea extract, chlorogenic acid and the like. It is preferable to use sodium ascorbate as an antioxidant. By including an antioxidant, the liquid pigment preparation of the present invention can improve stability and suppress a decrease in color value when stored for a long period of time. The liquid pigment preparation of the present invention preferably contains an antioxidant of about 0.05% by weight or more, more preferably about 0.1% by weight or more. The content of the antioxidant in the liquid pigment preparation of the present invention is preferably about 5% by weight or less, more preferably about 1% by weight or less. The liquid pigment preparation of the present invention may be a liquid pigment formulation that can be stored for a long period of time. The liquid pigment preparation of the present invention can be stored at room temperature, for example, for about 3 months or more or about 6 months or more.

(4. Use of 13-cis-crocetin-containing dye preparation of the present invention)
The 13-cis-crocetin-containing dye preparation and 13-cis-crocetin-containing dye composition of the present invention can be used for applications in which gardenia yellow is conventionally used. Furthermore, the 13-cis-crocetin-rich dye preparation and the 13-cis-crocetin-containing dye composition of the present invention are used for applications where a conventional crocetin dye preparation (ie, all-trans-crocetin) could not be used. be able to. Examples of uses for which crocetin pigment preparations could not be used conventionally include neutral foods and drinks.

  The foods and drinks to which the dye preparation and the dye composition of the present invention can be used can take any food form such as solid food, cream-like or jam-like semi-solid food, gel food, and beverage. Examples of food and drink include noodles (eg, Chinese noodles, cup noodles, chilled noodles, etc.), beverages (eg, neutral beverages (eg, neutral milk beverages), confectionery (eg, cookies, candy, gum, Western confectionery) , Sweet bread, chocolate, gummi, jelly confectionery), flower paste, cake sponge, bread dough, yogurt, ice cream, pudding, margarine, shortening, mayonnaise, dressing, etc. The pigment preparation and pigment composition of the present invention are used, for example, for foods mainly composed of wheat flour and starch and having a non-alkaline pH. Examples include pastes (creams sandwiched between breads), cake sponges, dough, etc. Examples of the stomach neutral foods and drinks, milk drinks, ice cream, such as sherbet and the like.

  When the food or drink of the present invention is a neutral beverage, the neutral beverage can be of a chilled type.

  The food / beverage products in which the pigment preparation and the pigment composition of the present invention can be used are preferably neutral food / beverage products. As used herein, the term “neutral” refers to pH 6-8. The pH of the neutral food or drink is preferably about 6.5 or higher, more preferably about 6.7 or higher, and most preferably about 6.9 or higher. The pH of the neutral food or drink is preferably about 7.5 or less, more preferably about 7.3 or more, and most preferably about 7.1 or more.

  Colored food / beverage products can be manufactured by mixing the pigment preparation of this invention with food / beverage products or food-drinks raw materials, and coloring.

  In a specific embodiment, the food or drink for which the dye preparation and the dye composition of the present invention can be used is the above crocetin or a salt thereof as it is, or other food-and-drinkable additives, food materials, food ingredients, and further if necessary. Food additives and the like can be mixed as appropriate, and can be produced as a food or drink according to conventional methods. When the food or drink is a health food, the health food may be in a dosage form such as a liquid, powder, granule, tablet, microcapsule, soft capsule or hard capsule. A crocetin-containing composition such as an oil-and-fat composition mainly composed of 13-cis-crocetin or a salt thereof, an O / W type emulsion, a W / O type emulsion or a solubilized liquid is produced according to a conventional method. You may manufacture the food / beverage products of this invention by adding a cis-crocetin containing composition to food / beverage products.

  The amount of the dye preparation of the present invention added to the food or drink may be the same as the amount in which the dye preparation is usually used, or may be less than the conventional amount. For example, the added amount of the pigment preparation of the present invention is about 0.00001% by weight or more and about 0.00005% by weight or more with respect to the total weight of the food and drink in terms of 100-wt% 13-cis-crocetin. About 0.0001% or more, about 0.0005% or more, about 0.001% or more, about 0.005% or more, about 0.01% or more, about 0.05% or more, about It may be 0.1% by weight or more, about 0.5% by weight or more, about 1% by weight or more. The amount of the pigment preparation of the present invention is about 10% by weight or less, about 5% by weight or less, or about 1% by weight or less based on the total weight of the food or drink, in terms of 100% by weight of 13-cis-crocetin. About 0.5 wt% or less, about 0.1 wt% or less, about 0.05 wt% or less, about 0.01 wt% or less, about 0.0005 wt% or less, about 0.0001 wt% or less, about It may be 0.00005 wt% or less, about 0.00001 wt% or less.

When measuring the color value (E _{10%, 1 cm 2} ) in the following examples, etc., “Food Additives other than Chemically Synthetic Products Voluntary Standards (Second Edition)” edited by Japan Food Additives Association, The color value was measured with reference to:
(Color value measurement method)
The sample is precisely weighed so that the absorbance to be measured is in the range of 0.3 to 0.7, and dissolved in Kolthoff buffer solution (50 mM Na ₂ CO ₃ -50 mM Na ₂ B ₄ O ₇ , pH 10.0). Make exactly 500 ml. If it is difficult to dissolve, dissolve by sonication. Weigh 10 ml accurately and add Kolthoff buffer (50 mM Na ₂ CO ₃ -50 mM Na ₂ B ₄ O ₇ , pH 10.0) to make 50 ml, which is the test solution. Kolthoff said buffer _{(50mM Na 2 CO 3 -50mM Na} 2 B 4 O 7, pH10.0) as a control, by measuring the absorbance A at the maximum absorption of 420nm near the length 1cm of the liquid layer, by the following equation Find the color value.

  In the following examples, 13-cis-crocetin-rich crocetin powder was obtained using the production method described in Example 1.

(Example 1: Method for producing 13-cis-crocetin-rich crocetin powder)
25000 g of 50% (v / v) ethanol aqueous solution was added to 2500 g of dried gardenia fruits, and the mixture was extracted by stirring at 60 ° C. for 1 hour. An appropriate amount of 50% (v / v) ethanol aqueous solution was again added to the residue, followed by secondary extraction, and a crocin-containing extract was obtained. The obtained crocin-containing extract was concentrated under reduced pressure to obtain a crocin-containing concentrated extract having a color value of 500. 450 g of crocin-containing concentrated extract having a color value of 500 was diluted with 2050 g of water to obtain 2500 g of a crocin-containing solution having a color value of 90. A 10% aqueous potassium hydroxide solution was added to the crocin-containing solution to adjust the pH to 12.0. By stirring for 20 hours at 35 ° C., crocin was hydrolyzed and crocetinized. Thereafter, concentrated hydrochloric acid was added to adjust the pH to 3.0, followed by stirring for 60 minutes to precipitate crocetin. The precipitated crocetin was recovered as a crude crocetin cake by a filter press. The crude crocetin cake was washed with water until no acid was present. After washing with water, 50 g of crocetin cake was dispersed in 500 g of water, adjusted to pH 12.0 by adding 10% aqueous potassium hydroxide solution and stirred for 30 minutes to dissolve the crocetin cake to obtain a crocetin-containing alkaline mixture. . Concentrated hydrochloric acid is added to the crocetin-containing alkaline mixture to adjust the pH to 7.0 to 8.0 (the results of shaking the pH of “stage a” at this time are summarized in Table 3 below), thereby reducing the crocetin-containing weakness. An alkaline mixture was obtained. The adjusted crocetin-containing weak alkaline mixture was immediately filtered again with a filter press to obtain a 13-cis-crocetin-rich filtrate. The cake at this time was a cake containing a high amount of trans-crocetin. By drying and pulverizing this cake, a crocetin powder containing a high amount of trans-crocetin could be obtained. Hydrochloric acid was added to the 13-cis-crocetin-rich filtrate to adjust the pH to 3.0 for precipitation. The cake obtained by the filter press was dried to obtain 13-cis-crocetin-rich crocetin powder.

  The color value E (10%, 1 cm) of the 13-cis-crocetin-rich crocetin powder produced under each of these conditions was measured with an absorptiometer U-3310 manufactured by HITACHI. The results are shown in Table 3 below. Further, these powders are analyzed by high performance liquid chromatography (HPLC) to determine the ratio of all-trans-crocetin to 13-cis-crocetin (herein referred to as “trans: cis ratio” or “trans : Cis)), the yield and purity of 13-cis-crocetin was determined. The HPLC measurement conditions were as follows:

  An HPLC chart for the crocetin powder when the pH at step a is 7.5 is shown in FIG. Peak 1 is all-trans-crocetin and peak 2 is 13-cis-crocetin. The pH of step a was most preferably 7.7, and the final 13-cis-crocetin-rich crocetin powder using the result was yield 5.0 g, color value 7675, trans: cis = 15: 85 ( Hereinafter, the powder when the pH of the stage a is 7.7 is also referred to as “crocetin powder of Example 1”).

  The 13-cis-crocetin high content crocetin powder obtained by this method passed all of the confirmation test and purity test for gardenia yellow pigment described in the Food Additives Official Document.

  The fact that the compound of peak 1 is all-trans-crocetin and that the compound of peak 2 is 13-cis-crocetin is obtained by fractionating these peak fractions and analyzing them by instrumental analysis such as NMRMS and UV spectrum. It was confirmed by analysis. The compound of peak 2 was fractionated and measured again under the same HPLC measurement conditions as above. As a result, trans: cis = 0.9: 99.1 and cis / trans was about 100 times. The results are shown in FIG.

The NMR conditions for the analysis of each peak compound were as follows: ¹ H spectrum measured at 600 MHz and ¹ H; ¹³ C spectrum measured at 150 MHz and ¹³ C; solvent: dimethyl sulfoxide-d6 ( DMSO-d6), the unit of spectrum is ppm, and the number of protons, multiplicity, and coupling constant (unit is Hertz) are shown in parentheses. The spectrum data of 13-cis-crocetin in ¹ H and ¹³ C NMR are shown in Table 4 below.

(Example 2. Production of liquid pigment preparation)
After mixing 0.8 g of 13-cis-crocetin-rich crocetin powder prepared in Example 1, 49.2 g of ethanol and 50 g of water to dissolve the crocetin powder, the pH was 6.8 with 10% aqueous sodium hydroxide solution. And filtered through a filter paper (ADVANTEC No. 5C) to obtain a liquid preparation containing 13-cis-crocetin in a high content. The color value E (10%, 1 cm) of this liquid preparation was measured with an absorptiometer. As a result, the color value E (10%, 1 cm) of this liquid preparation containing 13-cis-crocetin was 60. Since this liquid preparation uses the crocetin powder produced in Example 1, trans: cis = 15: 85. This liquid preparation has the advantages that it is easier to mix in the final product than the powder preparation, and that it is easy to handle without dusting at the production site.

(Example 3. Stability test of liquid pigment preparation)
The stability of the liquid pigment preparation produced in Example 2 was evaluated. Whether (1) no addition (Example 3-1) or (2) Na ascorbate is added to 0.1% by weight to the liquid pigment preparation produced in Example 2 (Example 3) -2), each preparation was prepared and stored refrigerated or at 55 ° C. for 7 days. Immediately after production, the color value of the liquid part was measured after 4 days and after 7 days of storage, and the presence or absence of precipitation was visually evaluated. The results are shown in Table 5 below.

  As a result, both in the case of no addition and in the case of addition of Na ascorbate, the liquid pigment preparation of the present invention is sufficiently stable as a liquid preparation with almost no decrease in color value after refrigerated storage for 7 days. all right. Further, even after the liquid preparation added with 0.1% by weight of sodium ascorbate was stored at 55 ° C. for 7 days, no significant decrease in color value was observed. The storage conditions for 7 days at 55 ° C. are generally considered as storage conditions equivalent to normal temperature half a year in this field. Therefore, it was found that the liquid dye preparation of the present invention can be stored for a long period of time by adding an antioxidant such as Na ascorbate. Also, no precipitation occurred in any sample. Therefore, it was found that the liquid pigment preparation of the present invention can be suitably used as a liquid formulation of crocetin pigment.

(Example 4.50% Solubility test in ethanol water)
All-trans-crocetin obtained by drying and grinding the 13-cis-crocetin-rich crocetin powder produced in Example 1 or the all-trans-crocetin-rich cake obtained during the production of the powder Any of the high content crocetin powders were used as samples. A sample (0.100 g for 13-cis-crocetin-rich crocetin powder; 0.0295 g for all-trans-crocetin-rich crocetin powder) is precisely measured and placed in a container where 50% (v / v) ethanol is added. 10 ml of aqueous solution was added. The container was heated to 60 ° C. with a water bath and held for about 10 minutes to dissolve the sample. Thereafter, the container was cooled with water and cooled to room temperature (about 20 ° C.). The liquid after cooling was centrifuged at 3000 rpm for 10 minutes to precipitate insoluble matter, and then the supernatant was passed through a 0.2 μm membrane filter to obtain a filtrate. The color value of this filtrate was measured. The results are shown in Table 6 below.

  Thus, it was found that the 13-cis-crocetin-rich crocetin powder had about 16.3 times higher solubility in 50% ethanol aqueous solution than the all-trans-crocetin-rich powder. When the 13-cis-crocetin-rich crocetin powder was used, the color value was as high as 61.8, which was confirmed to be a level that can be sufficiently used as a pigment preparation. On the other hand, it was confirmed that the color value when the powder containing a high amount of all-trans-crocetin was used was as low as 3.8, which was a level that could not be used as a pigment preparation.

(Example 5. Coloring experiment 1: Flour coloring test)
13-cis-crocetin-rich crocetin powder (trans: cis = 20: 80) produced in the same manner as in Example 1 with the pH of step a being pH 7.6, or obtained during the production of the powder Trans-crocetin high content powder (trans: cis = 95: 5) obtained by drying and pulverizing the all-trans-crocetin high content cake, crocin pigment preparation (manufactured by Glico Nutrition Foods Co., Ltd .; trade name Kuchina Color 1600P) Either was used as a dye sample.

  The dye sample was added to water and mixed to obtain a 1% dye mixture liquid in terms of 450 (color value E (10%, 1 cm)). At this time, the sample was dissolved or dispersed in water. While dripping 25 g of the pigment mixture into a mixture of 50 g of wheat flour and 0.6 g of powdered brine, kneaded pear pigmented product of wheat flour (with powdered brine) was obtained. Similarly, a colored product was produced even in a system to which no powdered brine was added.

  The color of each gardenia pigment coloring matter (that is, a colored sample) was visually observed and L, a, and b values were measured with a color difference meter to confirm the color tone. The results are shown in Table 7 below.

  The photographs of these coloring tests are shown in FIG. From the above results, in the test using all-trans-crocetin high content powder, lemon yellow peculiar to crocetin was shown when powdered water was added, but the color turned orange when powdered water was not added. On the other hand, the 13-cis-crocetin-rich crocetin powder could be colored in lemon yellow specific to crocetin regardless of the presence of powdered water. The crocin pigment preparation does not change in color with or without powdered water, but it is a reddish yellow color that is different from the lemon yellow characteristic of crocetin.

  Thus, under powder irrigation conditions, the 13-cis-crocetin-rich crocetin powder exhibits a greenish yellow (lemon yellow) color tone, similar to the all-trans-crocetin-rich crocetin powder. Under the condition of no powder watering (pH around 6), the all-trans-crocetin-rich crocetin powder cannot give a greenish yellow color, but the 13-cis-crocetin-rich crocetin powder has a greenish color similar to the alkaline condition. Presents a certain yellow color. It is speculated that the all-trans-crocetin-rich crocetin powder turns orange because crocetin is not dissolved.

  That is, the 13-cis-crocetin-rich crocetin powder of the present invention can exhibit a preferable lemon yellow characteristic of crocetin even under weakly acidic conditions near the neutral range without powdered watering, which is clearly different from crocin. It is a color tone.

(Example 6. Coloring test 2: Milk coloring test)
A milk coloring test was conducted to compare the color tone in the neutral region when various yellow pigments were used. This milk coloring test assumes a chilled milk drink.

  Milk was colored yellow by adding 1 g of each yellow pigment sample to 100 g (pH 6.8) of commercially available milk in terms of color value E (10%, 1 cm) 50 and mixing. At this time, the color tone of the colored milk was confirmed visually and with a color difference meter as in Test 1.

  Further, 0.005 g of gardenia blue pigment (manufactured by Glico Nutrition Foods Co., Ltd .; trade name Kuchina Blue Color 1250PS; color value E (10%, 1 cm) 250) was added to 100 g of yellow colored milk, and colored green. Again, as in Test 1, the color of the milk colored green was confirmed visually and by a color difference meter. The yellow dye samples used are as follows and the results are shown in Table 8.

Yellow pigment sample used:
(A) Marigold dye preparation: Color value E (10%, 1 cm) 400; Prototype (b) Crocin dye preparation: Color value E (10%, 1 cm) 410; Product name Kuchina Color 1600P manufactured by Glico Nutrition Foods, Inc.
(C) 13-cis-crocetin-containing dye preparation: color value E (10%, 1 cm) 50; the liquid dye preparation prepared in Example 2 diluted 1.2 times with water (d) all-trans- Crocetin-containing pigment preparation: Color value E (10%, 1 cm) 50; obtained by drying and pulverizing the all-trans-crocetin-rich cake obtained in the production of 13-cis-crocetin prepared in Example 1 (E) safflower pigment preparation: color value E (10%, 1 cm) 195; trade name Carthamus Yellow Color manufactured by BIONATURE.

  The results of these coloring tests other than (12) are shown in FIG. (1) is the case where the marigold dye preparation is added only in yellow, (2) is the case where the crocin dye preparation is added only in yellow, and (3) and (10) are only in yellow. -When cis-crocetin pigment formulation is added, (9) is when yellow-only all-trans-crocetin pigment formulation is added, and (4) is when yellow-colored safflower pigment formulation is added (5) is the case where blue is added to the marigold dye preparation, (6) is the case where blue is added to the crocin dye preparation, and (7) is 13-cis. -When blue is added in addition to the crocetin pigment preparation, (8) is when blue is added in addition to the safflower pigment preparation. (11) is the case of milk only. (1) to (8) and (9) to (11) were separately tested.

  In the yellow-only test, the marigold pigment formulation and the crocin pigment formulation were visually classified as reddish yellow, and the 13-cis-crocetin-containing pigment formulation and safflower pigment formulation were classified as green pigments. . Furthermore, although the safflower pigment preparation was used at the same color value as the 13-cis-crocetin-containing pigment formulation in milk coloring, it felt that the coloring was lighter than the 13-cis-crocetin-containing pigment formulation. Therefore, although it is a problem of palatability, for example, it is preferable to use a 13-cis-crocetin-containing pigment preparation that exhibits lemon yellow and has better color development than safflower pigment in the image of vanilla or banana milk. The all-trans-crocetin pigment preparation was not dissolved, and orange fine particles were dispersed, and did not show the lemon yellow characteristic of dissolved crocetin.

  In the blue addition test, the marigold dye preparation had a dull yellow green color, and the crocin dye preparation had a strong yellowish and slightly dark yellow green color. Increasing the amount of blue pigment added can increase the greenness, but increasing the amount of the total pigment added increases darkness and increases costs. Both the 13-cis-crocetin-containing pigment preparation and safflower pigment preparation exhibited a preferred yellow-green color, but the safflower pigment preparation was white and melon milk-like and looked slightly thin. The 13-cis-crocetin-containing dye preparation had a yellowish green color with good color and a matcha milk style.

  Therefore, the 13-cis-crocetin-containing pigment preparation of the present invention is useful as a pigment capable of exhibiting lemon yellow peculiar to crocetin in the neutral range, which is also characteristic as compared with other yellow pigments.

(Example 7: Production of powder pigment preparation with high content of 13-cis-crocetin)
82 g of dextrin is dissolved in 400 g of hot water at 60 ° C., and is obtained in the process of Example 1 when the pH of step a is pH 7.7 while stirring at 10,000 rpm with a TK homomixer (Primix Plus Co., Ltd.), 13 -18g of cis-crocetin-rich crocetin cake (color value about 2500) was added to obtain a dispersion. This dispersion was pulverized by spray drying to obtain a 13-cis-crocetin-rich crocetin powder pigment preparation. The color value of this powder pigment preparation was measured and found to be 450.

(Comparative Example 7: Production of powder pigment preparation with high content of all-trans-crocetin)
94.3 g of dextrin is dissolved in 400 g of hot water at 60 ° C., and is obtained in the process of Example 1 when the pH of step a is pH 7.7 while stirring at 10000 rpm with a TK homomixer (Primix Plus Co., Ltd.). , 5.7 g of an all-trans-crocetin high content cake (color value about 8000) was added to obtain a dispersion. This dispersion was pulverized by spray drying to obtain a powder pigment preparation with a high all-trans-crocetin content. The color value of this powder pigment preparation was measured and found to be 450.

(Example 8. Coloring test 3: LL Chinese noodle coloring test)
Premixed 900 g of powerful powder and 100 g of starch (Glico Nutrition Foods Co., Ltd., trade name: Chemister 220) and Gluten 30 g (Glyco Nutrition Foods Co., Ltd., trade name: A-Glu WP) were put in a mixer. To this, 10 g of powdered Kansui, 10 g of sodium chloride, and 1 g of a cis-crocetin high content pigment preparation (color value 450) produced in Example 7 were uniformly added in 390 ml of water and mixed well.

  The resulting dough was banded with a composite roll machine, and then cut into a noodle band with a final noodle thickness of 1.4 mm with a rolling mill, with a cutting edge of 18th corner and a noodle length of 25 cm.

  The obtained noodle strings were divided into 200 g, boiled with hot water, rinsed with running water and cooled. This noodle was a preferred yellow noodle. After immersing this noodle in a 2% lactic acid solution for 1 minute, the lactic acid solution was thoroughly removed, 110 g was sealed in a pouch bag, and steam sterilized with a steamer at 90 ° C. for 30 minutes. This was cooled to make Long Life (LL) Chinese noodles. The obtained LL Chinese noodles were preferred yellow noodles. From this, it was confirmed that the cis-crocetin high content pigment preparation of the present invention can be suitably used for Chinese noodles and LL Chinese noodles.

(Example 9. Coloring test 4: Flower paste)
25.17 g of granulated sugar, 3.95 g of skim milk powder, 6.91 g of processed starch (trade name Chemister 420 manufactured by Glico Nutrition Foods Co., Ltd.), 0.44 g of citric acid, and 0.05 g of the following pigment preparation (1) or (2) Were mixed uniformly, 6.91 g of fats and oils that had been heated to 40 ° C. and 69.11 g of water were added and dissolved by heating at 60 ° C. with stirring. Then, after adjusting the amount of water to 100 g, the container was filled and cooled. The color tone after refrigeration for one day was measured with a color difference meter.

Dye preparation used:
(1) 13-cis-crocetin high-content pigment preparation prepared in Example 7 (color value 450)
(2) The all-trans-crocetin high content pigment preparation (color value 450) produced in Comparative Example 7.

  As a result, the flour paste using the 13-cis-crocetin high pigment preparation showed lemon yellow peculiar to crocetin, but the flour paste using the all-trans-crocetin high pigment formulation did not develop crocetin and was thin. Showed orange. The results are shown in Table 9 and FIG.

  As described above, the dye preparation of the present invention has a remarkable effect that it is very easy to use when added to foods and has good color development.

(Example 10: Production of powder pigment preparation with high content of 13-cis-crocetin)
13-cis prepared in the same manner as in Example 1 with the pH of step a being pH 7.0 instead of the 13-cis-crocetin-rich crocetin cake when pH of step a of Example 1 is pH 7.7 -Using crocetin-rich crocetin powder, a 13-cis-crocetin-rich crocetin powder pigment preparation was obtained in the same manner as in Example 7. This 13-cis-crocetin high content crocetin powder was trans: cis = 40: 60.

  That is, 82 g of dextrin is dissolved in 400 g of hot water at 60 ° C., and is obtained in the process of Example 1 when the pH of step a is pH 7.7 while stirring at 10000 rpm with a TK homomixer (Primix Plus Co., Ltd.). Then, 18 g of 13-cis-crocetin high content crocetin cake (color value about 2500) was added to obtain a dispersion. This dispersion was pulverized by spray drying to obtain a 13-cis-crocetin-rich crocetin powder pigment preparation. The color value of this powder pigment preparation was measured and found to be 450.

(Example 11. Coloring test 4: flour paste)
A flour paste was prepared in the same manner as in Example 9 using the 13-cis-crocetin high-content pigment preparation prepared in Example 10 instead of the 13-cis-crocetin high-content pigment preparation prepared in Example 7. . As a result, the color tone of the flower paste after refrigeration for one day showed lemon yellow peculiar to crocetin as in Example 9. From this, even when trans: cis = 50: 50, unlike conventional all-trans-crocetin high pigment preparation, it is possible to develop lemon yellow peculiar to crocetin without coloring orange. all right.

  As mentioned above, although this invention has been illustrated using preferable embodiment of this invention, this invention should not be limited and limited to this embodiment. It is understood that the scope of the present invention should be construed only by the claims. It is understood that those skilled in the art can implement an equivalent range based on the description of the present invention and the common general technical knowledge from the description of specific preferred embodiments of the present invention. Patents, patent applications, and documents cited herein should be incorporated by reference in their entirety, as if the contents themselves were specifically described herein. Understood.

  The pigment preparation of the present invention has a remarkable effect that it can develop a lemon yellow characteristic of crocetin not only in the alkaline region but also in the neutral region. The pigment preparation of the present invention can be used well not only for alkaline foods but also for neutral foods.

  The liquid pigment preparation of the present invention is easier to mix in the final product than the powder pigment formulation, and is easy to handle without dust at the production site.

Claims (17)

  In the case of a pigment preparation containing 13-cis-crocetin with or without all-trans-crocetin and all-trans-crocetin, the peak area of all-trans-crocetin as analyzed by HPLC is calculated. A pigment preparation having a peak area of 13-cis-crocetin of 1 or more when 1.   The pigment preparation according to claim 1, wherein the peak area of 13-cis-crocetin when the peak area of all-trans-crocetin is 1 when analyzed by HPLC is 1 or more and 10,000 or less.   The pigment preparation according to claim 1 or 2, wherein the color value is 100 or more and 32000 or less.   The pigment preparation according to any one of claims 1 to 3, wherein a content of the 13-cis-crocetin is 0.05% by weight or more.   The pigment preparation according to any one of claims 1 to 4, wherein a content of the 13-cis-crocetin is 0.1 wt% or more and 75 wt% or less.   The pigment preparation according to any one of claims 1 to 5, which is a powder preparation.   The pigment preparation according to any one of claims 1 to 5, which is a liquid preparation.   The pigment preparation according to claim 7, comprising water or alcohol as a solvent. A method for producing a dye composition with a high content of 13-cis-crocetin,
Dissolving a mixture containing 13-cis-crocetin and all-trans-crocetin under alkaline conditions to obtain a mixed solution containing crocetin;
A step of precipitating all-trans-crocetin by adjusting the pH of the crocetin-containing mixed solution to pH 7.0-8.0; and removing the precipitated all-trans-crocetin to contain a high content of 13-cis-crocetin A method comprising the step of obtaining a dye composition. The mixture containing 13-cis-crocetin and all-trans-crocetin is
Extracting a crocin-containing raw material with a solvent to obtain a crocin extract; and hydrolyzing crocin in the crocin extract or a concentrate thereof to obtain a mixture containing 13-cis-crocetin and all-trans-crocetin 10. A method according to claim 9, obtained by a process.   The method according to claim 9 or 10, wherein the pH of the precipitation step is 7.2 to 7.8.   The manufacturing method of the colored food-drinks including the process of mixing and coloring the pigment preparation of any one of Claims 1-9 with food-drinks or food-drinks raw materials.   The manufacturing method of Claim 12 whose said food / beverage products are neutral food / beverage products.   The manufacturing method of Claim 13 whose said neutral food / beverage products are milk drinks or flour paste.   In the case of food and drink containing 13-cis-crocetin, with or without all-trans-crocetin and all-trans-crocetin, the peak area of all-trans-crocetin when analyzed by HPLC is The food / beverage products whose peak area of 13-cis-crocetin when set to 1 is 1 or more.   The food / beverage products of Claim 15 whose said food / beverage products are neutral food / beverage products.   The food / beverage products of Claim 16 whose said neutral food / beverage products are milk drinks or a flower paste.