WO2009039716A1

WO2009039716A1 - Carotenoid formulations and use thereof, and feeds containing the formulation and processes for the preparation of the feeds

Info

Publication number: WO2009039716A1
Application number: PCT/CN2008/000545
Authority: WO
Inventors: Bin Shao; Xinde Xu; Di Zhou; Yanfeng Wang; Yimin Hong
Original assignee: Zhejiang Medicine Co., Ltd. Xinchang Pharmaceutical Factory
Priority date: 2007-09-27
Filing date: 2008-03-19
Publication date: 2009-04-02
Also published as: NO343960B1; CN101396068A; CL2008002231A1; CN101396068B; NO20083794L

Abstract

A carotenoid formulation in oil suspensions, comprises a carotenoid with an average particle size of 10μm or less, an antioxidant and an edible oil. A feed contains the said carotenoid formulation, a method for preparing the feed and use of the carotenoid formulation as an additive of animal feeds, foods, cosmetics and/or pharmaceuticals. The active ingredients in the feed granules are homogeneously distributed in the feed granules, majority of which are substantially distributed inside the granules, to reduce the loss caused by oxidation on the surface.

Description

CAROTENOID FORMULATIONS AND USE THEREOF₃ AND FEEDS

CONTAINING THE FORMULATION AND PROCESSES FOR THE

PREPARATION OF THE FEEDS

BACKGROUND OF THE INVENTION

Field of the Invention

This present invention relates to a novel carotenoid formulation, in particular, relates to a carotenoid formulation in oil suspensions, a feed containing the carotenoid formulation, and a method for preparing the feeds, and use of the carotenoid formulation as an additive of animal feeds, foods, cosmetics, and/or pharmaceuticals.

Description of the Related Art

According to whether oxygen atoms exist in the molecular structure or not, carotenoids can be divided into two major categories, i.e., carotenes and luteins. The former, such as β-carotene and lycopene, possesses no oxygen atom in the molecular structure, while the latter, such as canthaxanthin, lutein, zeaxanthin, and astaxanthin and the like, possesses hydroxyl groups or carbonyl groups or both of the oxygenic functional groups in the molecular structure. In the luteins, these with only hydroxyl groups in the molecular structure generally show a yellow color, these with carbonyl groups in the molecular structure generally show a red color.

Carotenoid is an important substance in the feed industry, it not only involves into the physiological metabolism of animal bodies, but also imparts various characteristic colors to animals bodies or feathers or hooves and gives rise to people an aesthetic feeling. Such carotenoids as lutein, zeaxanthin, and canthaxanthin are widely applied in the industry of laying hens, meat chicken, and laying ducks, which make skins and yolks of poultry show a favorable yellow and red respectively. While astaxanthin widely exists in some trout, salmon and other marine fishes, as well as in crabs and shrimps. However, animals are unable to synthesize carotenoids by themselves, they need to acquire these carotenoids through ingestion of wild plants or alga. Due to these reasons, a proper amount of carotenoid is generally added into feeds to supplement the levels of carotenoids inside the bodies of animals such as fishes and fowls in an industrialized fields, to achieve the purposes of nutrition and coloration.

Two problems frequently occur during the addition of carotenoids into feeds as follows: stabilities and low bioavailabilities of carotenoid. This is because carotenoid itself is a kind of very unstable substance, and the dissolubility of carotenoid in water even in oils is very low. Feeds, especially marine feeds go through expansion steps under the conditions of high-temperature and high-pressure in the course of processing. These extreme conditions are not conducive to the stability of carotenoid. It was reported by Berset (Ind. Aliment. Agric. (1987) 104: 529-533) that the loss of 18% of the active ingredients of canthaxanthin in the course of processing feeds. Hencken and Estermann (Aquaculture Ind. Develop. Report (1991) 91: 34-51) disclose extrusion and expansion result from a loss of 25%~48% of synthetic astaxanthin in the course of processing fish feed. Moreover, carotenoid formulations will greatly affect its bioavailability inside animal bodies. Therefore, there is a necessity to seek an appropriate carotenoid formulation, which is not only easy to add in the course of processing feeds but also stable, and more important, carotenoids added are required to be easily absorbed by the animal bodies with a typical high bioavailability.

Many studies on carotenoids are to achieve the above-mentioned two purposes. At the very beginning, it may obtain carotenoid microcapsules with soluble in warm water and good stability. Carotenoids are firstly mixed with other ingredients in the course of processing feeds, and then it proceeds with granulation process after curing treatment. Since the temperature is not required very high during granulating, the loss of active ingredients is still fairly low in the course of processing feeds, so long as carotenoid microcapsules have good stability and certain heat resistance. At present, certain amounts of carotenoid microcapsule and microgranule products are still added during mixture generally, when those feeds for poultry and other animals are supplemented with lutein, vitamin A, beta-carotene, and canthaxanthin in the course of mixing, curing, and granulating steps, but not need for approaches of high-temperature and high-pressure expansion. These products mainly include LaIi Red, LaIi Yellow, and Carophyll Red, etc. However, as for feeds such as shrimps and fishes, it is required to go through an expansion process under the conditions of high-temperatures and high-pressures in the course of processing feeds, it would be very easy to give rise to a quite bit loss of the active ingredients if carotenoid microcapsule products such as astaxanthin is added at a earlier stage. So, people have again developed the microcapsuled products of astaxanthin with soluble in cold water and a kind of a post spray-coating technique, namely, after expansion of feeds, astaxanthin microcapsules dissolved in cold water are homogeneously spray-coated into the expanded feeds, in order to avoid the destructive effect of high temperatures and high pressures on astaxanthin and to enhance the retention rate of astaxanthin in the course of processing feeds.

Now, there are several astaxanthin formulations suitable for this process, such as LaIi Pink from ZMC and Carophyll Pink from DSM, etc, and all of them are astaxanthin products with typical good stability and solubility in cold water. However, this process also gives rise to the following problems. Firstly, as well-known to all, a large quantity of oils, including a mixture of fish oils and plant oils, are always added into the fish feeds, in order to enhance the nutrition absorption of fishes and carotenoids. These oils are usually added through a post spray-coating, namely, after the fish feeds being extruded and expanded, a large number of oils with lipophilic is spray-coated on their surfaces, and afterwards astaxanthin dissolved in water to be added, thus it is very difficult for astaxanthin dissolved in water to be homogeneously spray-coated onto the feed granules that have been covered with a layer of oil on the surface. The presence of hydrophobic oils on the surface of granules has greatly increased the resistance for nanometer-grade astaxanthin dissolved in water to enter into the inner porous passages of feed granules. Moreover, it would assume that small granules might not only obstruct the interior passage of the granules but also hold back astaxanthin penetrating into the interior of granules and only is adsorbed on the surface of granules. Similar problems will also occurs if astaxanthin water solution is firstly spray-coated before oil being spray-coated, so that it would be very difficult for oils to be homogeneously distributed inside the feed granules.

For the above reasons, the added astaxanthin by a post spray-coating is generally adsorbed on the surface of feed granules but can not distributed inside itself, it will result in the loss of active ingredients due to oxidation. Moreover, it will result in the lack of reproducibility of the product quality due to the unhomogeneous distribution, and bring about certain trouble to feeding production.

In order to solve these problems, and the operation of spray-coating oils after extrusion and expansion in the process of producing feed granules of shrimps, the addition of astaxanthin can be combined into this step, that is, crystal astaxanthin is firstly made to a high-quality form of oil suspensions, and then the oil suspension is mixed and dispersed and dissolved in the oils, and the required amount of astaxanthin is added in the process of spray-coating.

US Patent No. 5,716,655 and 6,083,541 disclose a method for preparing granulated feeds, which relates to the post spray-coating of active ingredients therein. In particular, it relates to spray-coating the micropowder of carotenoid dispersed in oils into the expanding feed granules under normal pressure or vacuum environments. However, one of the distinct deficiencies of this method is that carotenoid is dispersed into oils in form of micropowder, with its diameter of particle as large as 20μm in maximum. As described above, such granules are so large enough to block the interior passages of the granulated feeds, to resist the oil suspension to further penetrate into the granules, and may lead to the fact that many carotenoid crystals will be directly adsorbed on the surface of feed granules. It not only easily results in unhomogeneous distribution of active ingredients but also greatly reduce the oil load of products. The products' quality lacks of reproducibility. And these carotenoids on the surface of feed granules are easily oxidated and degraded, and more importantly, from the beginning to the end, this will consequentially result in a quite low biological utilization inside animal bodies due to the presences of carotenoid in a form of micropowder crystals. Since the absorption and utilization of carotenoid in crystals are very lower for the organism, consequently, more carotenoid crystals have to be added to achieve the expected coloration effect. But it might cause a huge waste.

To overcome these deficiencies in techniques, the present invention herein provides a novel carotenoid formulation in oil suspensions, wherein the diameter of particle of carotenoid micropowder in the oil suspension is small enough to be easily and fully dissolved in spray-coating oils at a lower temperature during the follow-up operation, and homogeneously distributed into the interior porous passages of the feed granules, consequently, it might obtain a product with a high degree of oil load and stable mass. Furthermore, the addition of certain natural antioxidant in the oil suspension is beneficial not only to the stability of carotenoid in the feed granules but also to its absorption and utilization by animal bodies.

SUMMARY OF THE INVENTION

The present invention herein discloses a novel oil suspension of carotenoid formulation used in feed granules, and the carotenoid can be added in combination with the addition of oils in the process of preparation of feeds. As the diameter of particles of the carotenoid in oil suspensions is very small, it can be fully dissolved by a slight rise of temperature before being added into the feed granules. It is beneficial to penetration of carotenoid into the interior passages of feed granules, to achieve a homogeneous distribution, a higher oil load, and stable quality of products, and to increase the bioavailability of carotenoid for organism, moreover, adding certain amount of antioxidant in the oil suspension is to reduce the loss of carotenoid's oxidation and to increase its bioavailability in animal bodies.

One aspect of the present invention is to provide a carotenoid formulation in oil suspensions, the formulation comprises carotenoid with an average particle diameter of less than or equal to lOμm, an antioxidant, and an edible oil. Wherein, the carotenoid comprises astaxanthin, lutein, zeaxanthin, β-carotene, α-carotene, cryptoxanthin, lycopene , canthaxanthin and tartrazine. The average particle diameter of the carotenoid granules in the oil suspension is 4~6μm.

Wherein the edible oil comprises plant oils and animal oils. The plant oils comprise soybean oils, sunflower oils, rapeseed oils, corn oil, peanut oils, olive oils; while animal oils comprise fish oils.

Wherein the formulation also comprises an antioxidant. The antioxidant comprises one or more than one ingredient selected from the group consisting of natural Vitamin E, synthetic Vitamin E, 2.6-butylated hydroxytoluene (BHT), and butylated hydroxyanisole (BHA). The amount of addition of the antioxidant is l~10wt% by weight of oil suspensions, preferably, the amount of addition of the antioxidant is 2~5wt% by weight of oil suspensions.

Wherein the amount of the carotenoid is 10~30wt% by weight of oil suspensions.

Another aspect of the present invention is to provide a feed containing the carotenoid formulation, the amount of carotenoids in the final granules of the feeds is about 15 ~100mg/kg dry matter. The average diameter of the final granules of the feeds is about 9mm, with a length of 0.9-1.2 mm, or other relative shape feed granules.

A further aspect of the present invention is to provide a method for preparing the feeds, and the method comprises the following steps of:

(1) Mixing carotenoids with the edible oil, grinding or ball milling the mixture, and getting the oil suspension.

(2) Stirring and raising the temperature to make carotenoids dissolved in the oil suspensions, getting a transparent oil solution with carotenoid's particle diameter of Oμm.

(3) Spray-coating the oil solution onto the surface of feed granules under normal pressure or in vacuum atmosphere, getting the final feed granules.

Wherein, the carotenoid is firstly premixed with some plant oils and ground, to reach the required fineness, and then fully mixed with residual plant oils, or the carotenoid is mixed with all the plant oils and then ground .

The temperature of dissolution is 30-70⁰C, preferably, 40-60⁰C; and the time of heat preservation is 0.5~5hr under the temperature, preferably, 0.5-2hr.

Another aspect of this invention is to provide the use of the carotenoid formulation as an additive for animal feeds, foods, cosmetics, and/or pharmaceuticals.

As described above, in the course of processing conventional feeds, especially marine feeds, a substantial loss will be caused by additions of carotenoid before expansion due to a high temperature and high pressure process. After a water-soluble carotenoid in dry powder form is dispersed in water, the loss of active ingredients may be reduced by spray-coating the carotenoids dispersed in water onto the surface of feed granules through a post spray-coating technique due to a high temperature and high pressure process. However, the course of processing the feeds is a process of additions of oils through a post spray-coating technique itself. Thus the process is inconvenient due to two steps of post spray-coating process; on the other hand, due to the completely opposite hydrophilicity and hydrophobicity of post spray-coating substances in two steps, the water-soluble ingredients firstly spray-coated on the surface of feed granules will surely prevent the oil spray-coated in second step from transferring inside the granules, or the oils spray-coated on the surface of the granules will further prevent the carotenoid subsequently spray-coated and dissolved in water from distributing homogeneously on the surface of feed granules and interior thereof due to the hydrophobic action, to affect the stability of product quality. In the subsequent process, the above-mentioned drawbacks can be avoided by dispersing immediately the carotenoid such as astaxanthin crystals into the oils by a post spray-coating, and in combination with the addition of carotenoids in the post spray process. Thereby the process is simple and reduces the loss of carotenoids in the course of processing carotenoid. However, since carotenoids in form of crystals of the above-mentioned method are adsorbed on the surface of feed granules, these crystals may possibly block the interior passage of the feed granules, to result in unhomogeneous distribution of active ingredients inside feeds, and more importantly, the carotenoids ingested by animal bodies exist in the form of crystals, it may greatly reduce the biological utilization of the carotenoids and give rise to a rather poor coloring effect.

Therefore, in view of the above-mentioned drawbacks in each process, the present invention discloses a method of effectively adding carotenoids to feed granules, to make the carotenoids kept in a dissolved state before a post spray-coating. Thus, it ensures both a simple operation and an increase in the stability of product quality, and more importantly to achieve a very good coloring effect in animal breeding.

The detailed description of the present invention is as follows: In the present invention, the carotenoid is firstly ultra-fine pulverized in a certain way to a particle diameter of less than lOμm, preferably 4-6μm. Then the pulverized powder is substantially dispersed in plant oils, wherein the plant oils comprise soybean oils, sunflower oils, rapeseed oils and other oils purchased in a commercial articles. The carotenoid crystals can be premixed with some plant oils and ground to reach a required fineness, and then are substantially mixed with the residual plant oils; or the carotenoid crystals are substantially mixed with all plant oils, and then are ground. The mode of grinding may be directly grinding or ball grinding and others. The final product in oil suspensions shows an excellent fluidity, wherein the particle diameter of carotenoids is generally about 4~6μm, less than lOμm in maximum. The distribution of such particle is beneficial to good dissolution of granules in subsequent process. Distribution of particle diameter of the typical carotenoid in oil suspensions is shown in Figure 1. The concentration of the carotenoids is about 10~30wt%. The amount of active ingredients may be higher, under the premise of the good fluidity of liquid.

According to the present invention, a certain amount of antioxidants is added into the carotenoids in oil suspensions described above, to reduce the loss of carotenoids in the process of preparation and preservation. Said antioxidants comprise natural vitamin E, synthetic vitamin E, BHT, BHA, or other antioxidants legally available, preferably vitamin E, so that it is helpful to the absorption and utilization of carotenoid by animal bodies. The added amount of the antioxidant is l~10wt% for the total amount of the oil suspensions, preferably 2-5 wt%. Prior to spray-coating carotenoids in oil suspensions into the feed granules after the extrusion and expansion process, some or all of the oil suspensions is homogeneously mixed with the required oils (plant oils or fish oils or mixtures thereof) for spray-coating, and dissolved under stirring at an temperature to obtain a transparent solution. The temperature is about 30-70 "C, preferably 40~60°C. It is held at the temperature for 0.5~5hr, preferably, 0.5~2hr. The amounts of the carotenoid and its isomers have no any change in the course of processing.

The carotenoid in the above-mentioned oils may directly be spray-coated onto the surface of feed granules together with oils under normal pressure or vacuum conditions. Prior to spray-coating according to the formulations of the feeds, additional oils may be supplemented or not, these oils comprise plant oils or fish oils or mixtures thereof, and then the temperature may be reduced to a normal temperature, or the temperature may be kept at initial temperature at which the carotenoids in oil suspensions may be kept in a dissolved state in oils. In brief, the carotenoid is in a dissolved state before spray-coating. Distribution of a typical particle diameter of the carotenoids in oils is shown in Figure 2.

In the present invention, the feed components can be appropriately regulated depending on requirements of practical applications. The purpose is to meet nutritional requirements of different animals. The parameters of feed granules such as appearance, shape, size, weight, porosity and others are also determined depending on different feeding subjects as well as feeding characters.

According to requirements, the added carotenoid may comprise astaxanthin, canthaxanthin, lutein, zeaxanthin as well as other fatty-soluble vitamin monomers or a mixture of one or more thereof. The amount of the carotenoid in the final feed granules is about 15mg/kg~ 100mg/kg dry matter. The total amount of added oils depends on the requirements of animal growth.

The active ingredients of feed granules according to the present invention are homogeneously distributed in it, and most of which distributed in the interior of feed granules, thereby to reduce the loss of surface oxidations. The carotenoid exists in a dissolved state to be beneficial to absorption and utilization of animals. Furthermore, the results of animal testing also indicate the coloring effect on fishes is very well after feeding the feeds. The features of the present invention will be more clearly understood by reference to the following embodiments, which are not to be construed as limiting the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

Fig. 1 shows the distribution of particle diameter of the carotenoid in oil suspensions.

Fig. 2 shows the distribution of particle diameter of the carotenoid prior to spray-coating.

DETAILED DESCRIPTION OF THE PRESENT INVENTION AND PREFERRED EMBODIMENTS THEREOF

Example 1

Preparation of 20% Astaxanthin in Oil Suspensions

2.2kg of pure astaxanthin (Content: 97.2%), 0.3kg of natural vitamin E and 5.0kg of sunflower oils were weighted & mixed homogeneously and ground in a ball-miller for 1.5hr, to obtain a dense fluid. And then 2.5kg of sunflower oils is supplemented to the dense fluid and homegeneously mixed to obtain astaxanthin in oil suspensions with good fluidity.

The amount of astaxanthin in the oil suspensions is 21.2wt% through measurement with UV spectrophotometric method. The average particle diameter of the feed granules in the oil suspension is about 4.68μm through measurement with laser particle size distribution instrument.

Dissolution of Astaxanthin Prior to Spray-coating

8.0kg of the above mentioned astaxanthin in oil suspensions is mixed with the required part (4800kg) of oils (a mixture of fish and soybean salad oils with a proportion of 2:1). The temperature is raised to 60 °C and kept for 1.5hr under stirring, to make the suspended astaxanthin particle dissolved substantially to get a transparent solution. And then 4800kg of the residual oils are added into the above mentioned ^

transparent solution, and stirred homogeneously to obtain a homogeneous solution and the temperature is dropped to 40⁰C. The particle diameter of astaxanthin in the solution is Oμm determined by laser particle size distribution instrument. The result indicates the astaxanthin is completely dissolved in oils.

Spray-coating of Trout Feeds

1200kg of the above mentioned oil solution of astaxanthin (0.2kg of astaxanthin) is spray-coated onto 2800kg of the semi-products of feed granules under vacuum condition (The feeds have been grained and expanded, but not spray-coated with oils). The feed granules have a diameter of 9mm, a length of 0.9-1.2 mm and a weight of about 350mg/granule. The process of spray-coating needs about l.Ohr. The color on surface of each granule of the obtained feeds is homogeneous, and the color in the interior and exterior of feed granules is homogeneous. The result shows that astaxanthin completely penetrate into the interior of the feed granules together with oils.

The amount of the astaxanthin in the final feed after treatment is 50mg/kg dry matter through measurement with HPLC.

Test of Feeding Trout 5000 Atlantic salmon with 6 months old are fed with the above-mentioned feed granules of adding astaxanthin and oils by a post spray-coating technique. 10 salmon of them are randomly selected at regular intervals. And then their back muscles are respectively treated and determined by HPLC to obtain the amount of astaxanthin thereof. The average value is applicable herein. The amounts of astaxanthin in Atlantic salmon muscle are respectively 8.5,

10.4 and 14.7mg/kg determined by the method above, after 50, 100 and 150 days of feeding. Besides, the color is fresh and bright, it shows significant coloring effects. Comparative Example 2

Feed granules containing 50mg/kg dry matter of astaxanthin is prepared according to the method of former US. Patent. There are no any antioxidants such as natural Vitamin E and others in the oil suspension. No process of heating and dissolving substantially presents prior to spray-coating oils. Due to the blockage of undissolved granules in the internal porous passages of feed granules, it makes the internal and the apparent surface colors of feed granules unhomogeneous.

Animal testing is performed with feed granules of this method according to the Example 1. After 50, 100 and 150 days of feeding, the amounts of astaxanthin in Atlantic salmon muscle are respectively 5.9, 8.8 and 11.5mg/kg. Thus, the rate of utilization of astaxanthin for salmon is respectively lower by 30.6%, 15.4% and 21.8% in different stages, in comparison with those of Example 1.

It may show from Example 1 and Comparative Example 2 that the present invention has the following advantages. Firstly, the size distribution of the specific particles in oil suspensions is beneficial to completely dissolve at lower temperatures before spray-coating. Secondly, the complete dissolution of astaxanthin before spray-coating makes it difficult to block the interior porous passages of feed granules, thereby it makes the active ingredients homogeneously distributed on the surface and interior of the feed granules, and is beneficial to the stability of the product quality. Moreover, the absorption of astaxanthin in the dissolved state for fish is largely enhanced. Finally, the presence of antioxidants such as natural vitamin E in oil suspensions is also beneficial to the fish growth as well as absorption and utilization of astaxanthin. Furthermore, the animal testing shows that coloring effect is very well after feeding the feed granules according to the present invention.

Examples 3-6

The process for preparations of oil suspensions in Examples 3~6 are listed in Table 1:

Although the present invention has been described in connection with the above embodiments, it should be understood that the present invention is not limited to such preferred embodiments and procedures set forth above. The embodiments and procedures were chosen and described in order to best explain the principles of the invention and its practical application, to thereby enable others skilled in the art to best utilize the invention. It will be apparent to those skilled in the art that various substitution, modifications and changes may be thereto without departing from the scope and spirit of the invention. Therefore, the intention is intended to cover all alternative constructions and equivalents falling within the spirit and scope of the invention as defined only by the appended claims and equivalents thereto.

Claims

1. A carotenoid formulation in oil suspensions, comprises a carotenoid with an average particle size of lOμm or less, an antioxidant, and an edible oil.

2. A carotenoid formulation according to the claim 1 , wherein the carotenoid comprises astaxanthin, lutein, zeaxanthin, β-carotene, α-carotene, cryptoxanthin, lycopene, canthaxanthin and citranaxanthin.

3. A carotenoid formulation according to the claim 1, wherein the edible oil comprises plant oils and/or animal oils; wherein the plant oils comprise soybean oils, sunflower oils, rapeseed oil, corn oils, peanut oils and olive oils, and the animal oils comprise fish oils.

4. A carotenoid formulation according to the claim 2, wherein the carotenoid has an average particle size of 4~6μm.

5. A carotenoid formulation according to the claim 1, further comprises an antioxidant.

6. A carotenoid formulation according to the claim 5, wherein the antioxidant comprises one or more ingredients selected from the group consisting of Natural Vitamin E, Synthetic Vitamin E, 2, 6-di-tert-butyl-p-cresol (BHT), and butylated hydroxyanisole(BHA).

7. A carotenoid formulation according to the claim 6, wherein the amount of the antioxidant added is l~10wt% by weight of the oil suspensions.

8. A carotenoid formulation according to the claim 6, wherein the amount of the antioxidant added is 2~5wt % by weight of the oil suspensions.

9. A carotenoid formulation according to any one of the claims 1~8, wherein the amount of the carotenoid is 10-30 wt % by weight of the oil suspensions.

10. A feed comprising the carotenoid formulation according to any one of the claims 1-9, wherein the amount of carotenoids in the final granules is about ^

15mg/kg~100mg/kg dry matter.

11. A method for preparing the feed according to the claim 10, comprises the following steps of:

(2) Stirring and raising the temperature to make carotenoids dissolved in the oil suspension, getting a transparent oil solution with carotenoid's particle diameter of Oμm.

(3) Spray-coating the oil solution onto the surface of feed granules under normal pressure or in vacuum atmosphere, getting the final feed granule.

12. The method according to the claim 11, wherein the carotenoid is firstly premixed with some plant oils and ground, to reach the required fineness, and then fully mixed with residual plant oils, or the carotenoid is mixed with all the plant oils and then ground.

13. The method according to the claim 11, wherein the dissolution temperature is about 30~70^°C.

14. The method according to the claim 13, wherein the dissolution temperature is about 40-60 °C .

15. The method according to the claim 13 or 14, wherein the time of heat preservation is about 0.5~5hr under the temperature.

16. The method according to claims 13 or 14, wherein the time of heat preservation is 0.5~2hr under the temperature.

17. A use of the carotenoid formulations according to any one of claims 1-9 as an additive for animal feeds, foods, cosmetics, and/or pharmaceuticals. FIGURES MISSING UPON FILING OF RECORD COPY