A method for preparing capsanthin crystals and its application

Abstract

The present invention relates to the technical field of separation and purification of natural products, and specifically discloses a method and application for preparing capsanthin crystals. The method of the present invention includes: (1) preparing capsanthin ester using capsanthin as raw material; (2) saponification; (3) adsorbing the saponified solution with resin, and gradient desorption; (4) evaporation crystallization, specifically in the following manner: heating from room temperature to 35±0.5℃, with a heating rate of 1.5-2℃ / min; keeping warm at 35±0.5℃ for 15-20min; continuing to heat up to 40±0.5℃, with a heating rate of 0.25-0.5℃ / min; keeping warm at 40±0.5℃ for 20-30min; heating to a target crystallization temperature of 45-50℃, with a heating rate of 0.25-0.5℃ / min; continuing evaporation crystallization under the target crystallization temperature and stirring, and then cooling to room temperature at a rate of 0.3-0.6℃ / min, and controlling the solid content of the crystallization liquid to be 18-22%. The method of the invention can obtain high-purity capsanthin crystals with high trans-capsanthin content.

Description

Technical Field

[0001] The invention relates to the technical field of separation and purification of natural products, and in particular to a method for preparing capsanthin crystals and application thereof. Background Art

[0002] The red pigment esters in capsanthin are mainly esterified by short-chain, saturated fatty acids (such as lauric acid / myristic acid) in the form of monoesters or diesters; the yellow pigments are mainly esterified by long-chain, unsaturated fatty acids (such as linoleic acid) in the form of monoesters or diesters, and there are also unesterified red and yellow pigments and β-carotene without hydroxyl structure.

[0003] Due to the limitation of pepper varieties and the use of traditional organic solvent extraction and alcohol solvent despicating processes in industrial production, the obtained capsanthin contains a large amount of oil components, and the color-developing components account for a small proportion. It cannot meet the application needs of high-end customers for high-purity capsanthin. There are few studies on the purification of capsanthin, and there are very few studies on the separation and purification of capsanthin suitable for large-scale industrial production.

[0004] In Chinese patent CN104292091A, anhydrous ethanol is used to perform a preliminary extraction of red pepper residue, and the preliminary extract is dissolved in n-hexane and then capsanthin is extracted using two-dimensional liquid phase preparative chromatography. However, the amount of capsanthin extracted by preparative chromatography is small and is not suitable for large-scale production. In addition, preparative chromatography has high requirements for equipment and high maintenance costs.

[0005] Chinese patent CN104830096A uses column chromatography to separate capsanthin from capsanthin to prepare capsanthin. However, the separation effect of silica gel column needs to be improved.

[0006] Therefore, the above patents are all small-scale laboratory preparations. It is necessary to conduct further purification research on the separation of capsanthin crystals in capsanthin to meet the needs of large-scale preparation of high-purity capsanthin crystals. Summary of the invention

[0007] In view of the problems of the prior art, one of the purposes of the present invention is to provide a capsanthin crystal with high purity and high trans-capsanthin ratio and a preparation method and application thereof.

[0008] In order to achieve this object, the technical solution of the present invention is as follows:

[0009] A method for preparing capsanthin crystals, comprising:

[0010] (1) preparing capsanthin ester using capsanthin as raw material;

[0011] (2) saponifying the capsanthin ester to obtain a saponified solution;

[0012] (3) adsorbing the saponified solution with a resin, and obtaining a capsanthin desorption solution after gradient desorption;

[0013] (4) obtaining capsanthin crystals from the capsanthin desorption solution by evaporation crystallization;

[0014] The evaporation crystallization method is: heating from room temperature to 35±0.5°C at a heating rate of 1.5-2°C / min; keeping warm at 35±0.5°C for 15-20min; continuing to heat to 40±0.5°C at a heating rate of 0.25-0.5°C / min; keeping warm at 40±0.5°C for 20-30min; heating to a target crystallization temperature of 45-50°C at a heating rate of 0.25-0.5°C / min; continuing evaporation crystallization at the target crystallization temperature and under stirring, then cooling to room temperature at a rate of 0.3-0.6°C / min, and controlling the solid content of the crystallization liquid to be 18-22%.

[0015] The room temperature in the present invention refers to 20-25° C. During evaporation crystallization, the stirring rate is 60-80 r / min, and the evaporation crystallization time is 2-4 h.

[0016] The resin used in the present invention is a styrene-divinylbenzene skeleton adsorption separation resin.

[0017] The coloring components in capsanthin account for 8-10%, among which zeaxanthin and capsanthin have similar polarity and small molecular weight difference, so they are often difficult to separate. In addition, there is a lot of oil in capsanthin, and the efficiency is low when separated by traditional silica gel column chromatography. To this end, the present invention studies how to efficiently separate and obtain high-purity capsanthin crystals from capsanthin, and finds that capsanthin is first prepared as capsanthin ester, and then saponified, and resin can be used to efficiently separate zeaxanthin and capsanthin with similar polarity, and then cooperate with specific crystallization conditions, and control the formation process of crystals by specific gradient temperature change conditions, so as to achieve further purification of capsanthin, and obtain high-purity capsanthin crystals with a very high proportion of trans-capsanthin, which solves the problem of difficult separation of zeaxanthin and capsanthin.

[0018] Crystallization is the process of solid matter precipitating from steam, solution or melt in a crystalline state. Since organic crystallization is a molecular assembly process in which similar molecules or ions are arranged in a regular pattern, it has a high degree of selectivity and can separate high-purity crystal products from a mixed solution containing more impurities. However, unlike other products, organic crystallization products have requirements for the crystal form, particle size distribution and crystal habit of organic crystalline substances in addition to purity and yield. The crystal form of organic matter reflects the internal microstructure of the crystal. The crystal habit of organic matter describes the external macroscopic morphology and can be considered as a morphological parameter of the crystallization process. Organic crystallization undergoes nucleation and growth, which is a complex multi-phase and multi-component heat and mass transfer process, and also involves surface reactions. Therefore, there are many factors that affect the crystallization process of organic matter. The choice of different crystallization methods and the change of crystallization temperature will lead to unexpected changes in the particle size distribution, crystal habit and crystal form of the crystal, thereby changing the quality of the product. At the same time, it is also related to the economy of subsequent processes (such as filtration, cleaning and drying). However, there is no general rule in the art for how to choose the crystallization method for different target products in order to effectively control the crystal configuration and ensure a good overall effect. This brings research and development obstacles to obtaining high-quality specific crystal products from different crystallization solution compositions. During the development of the present invention, attempts were made to proceed in other conventional ways, such as directly crystallizing the analytical solution using a conventional reduced pressure concentration method, but no matter how the concentration temperature, time, and vacuum degree were adjusted, the purity could not reach 98%, and high-purity capsanthin crystals with a very high proportion of trans-capsanthin could not be obtained. Finally, after changing the crystallization method and repeatedly exploring the crystallization temperature change curve, the high-purity product of the present invention was successfully obtained.

[0019] In step (2) of the present invention, saponification is achieved by reacting the capsanthin ester with an alkali in a diluent, wherein the mass ratio of the capsanthin ester to the alkali is 1:(0.2-0.4); the saponification temperature is 50-55° C., and the time is 6-8 hours;

[0020] The diluent is one or more of methanol, anhydrous ethanol, 95% ethanol, ethylene glycol or propylene glycol, and the mass volume ratio of the capsanthin ester to the diluent is 1: (5-6) g / ml.

[0021] During the saponification in step (2), the alkali is sodium hydroxide or potassium hydroxide.

[0022] The present invention further adjusts the composition state of fat and pigment and reduces free fatty acids by subjecting the capsanthin ester to a specific saponification reaction, thereby laying a foundation for subsequent efficient separation using resin. Preferably, after the saponification is completed, stirring is continued for 1.5-3 hours to allow the saponification reaction to proceed more thoroughly, and the saponified liquid can be fully mixed and cooled to a conventional temperature (room temperature) suitable for resin adsorption during stirring. In the present invention, resin adsorption can be directly performed after the saponified liquid is evenly mixed. The amount of resin used is 8-10 times the mass of the capsanthin ester before saponification.

[0023] In step (3) of the present invention, the resin adsorbs the saponified solution at a pH value of 4-4.5, and after adsorbing the saponified solution, the resin is first washed and then subjected to gradient desorption. The gradient desorption method is:

[0024] Firstly, a first desorption liquid is used for a first desorption, and then a second desorption liquid is used for a second desorption to obtain a capsanthin desorption liquid for subsequent operations;

[0025] The first desorption liquid is petroleum ether and ethyl acetate in a volume ratio of (20-20.5):1, and the second desorption liquid is petroleum ether and ethyl acetate in a volume ratio of (15-15.5):1;

[0026] The temperature of the first desorption liquid and the second desorption liquid is 35-40° C.;

[0027] During desorption, the flow rate of the first desorption liquid is 1-2 BV / h, and the flow rate of the second desorption liquid is 1-1.5 BV / h;

[0028] The volume mass ratio of the first desorption liquid to the saponification liquid is (2.5-4.5):1 L / kg, and the volume mass ratio of the second desorption liquid to the saponification liquid is (2.5-4.5):1 L / kg.

[0029] The present invention controls the desorption conditions to separate the target substance from the raw material in a gradient desorption manner. The selection and combination of the components of the gradient desorption solution of the present invention can effectively remove the yellow components (β-carotene, β-cryptoxanthin, epoxyzeaxanthin, etc.) in the impurities and pigments during the first desorption, and fully retain the target substance. During the second desorption, the components containing the target substance (including zeaxanthin and capsanthin) can be effectively eluted. The desorption solution obtained by the second desorption is combined with the subsequent crystallization step to further adjust the components, and finally the yield and purity of the capsanthin crystals are guaranteed.

[0030] During the research and development, the present invention tried to use a single system as the desorption liquid, but after using solvents such as petroleum ether, ethyl acetate, ethanol, methanol, and dichloromethane, the ideal desorption effect could not be achieved; then, the present invention also tried to use a variety of multi-systems as desorption liquids, such as dichloromethane and methanol, petroleum ether and ethanol systems, and finally found a more ideal combination of the present invention-petroleum ether and ethyl acetate system. However, different component ratios also significantly affect the desorption effect, so the present invention further tried repeatedly and finally determined that the composition and gradient of the desorption liquid of the present invention are preferred, which can better take into account the yield, purity and other effects.

[0031] Preferably, the present invention pre-treats the resin before resin adsorption, specifically in the following manner:

[0032] The resin is loaded into a resin chromatography column and compacted by nitrogen. The resin is activated by column treatment with 4% sodium hydroxide solution and 4% hydrochloric acid solution, respectively, at a flow rate of 1-2 BV / h and a treatment volume of 3-4 BV. After treatment, deionized water is added to wash until the pH of the outlet liquid is 4-4.5. Finally, the resin is soaked in 95-100% ethanol for 2-4 hours.

[0033] When the saponified liquid is adsorbed, the processing flow rate is 0.5-1 BV / h.

[0034] When the resin is washed after adsorbing the saponified liquid, deionized water is used, the treatment flow rate is 1-2 BV / h, and the washing time is 2-3 hours.

[0035] The specific operation method of step (1) of the present invention is:

[0036] ① Mix capsanthin, protective agent, composite alkali and alkali neutralization diluent to carry out alkali neutralization reaction; the temperature of the alkali neutralization reaction is 55-65° C. and the time is 0.5-2.5h;

[0037] The mass ratio of the capsanthin to the composite alkali is 1:(0.05-0.1); the composite alkali is composed of sodium hydroxide and calcium hydroxide in a mass ratio of 1:(0.5-2.5);

[0038] The protective agent is one or more of ethylenediaminetetraacetic acid, ethylenediaminetetraacetic acid disodium salt, ethylenediaminetetraacetic acid tetrasodium salt or vitamin C; the mass ratio of the capsanthin to the protective agent is 1: (0.002-0.005);

[0039] The alkaline neutralization diluent is one or more of methanol, anhydrous ethanol, 95% ethanol, acetone, isopropanol or 95% isopropanol; the mass volume ratio of the capsanthin to the alkaline neutralization diluent is 1: (3-6) g / ml;

[0040] ② The solution obtained after the alkali neutralization reaction is heated to 55-65°C at 4.0-6.0°C / min, maintained for 0.5-1.5h, then cooled to 15-20°C at 0.2-1°C / min, and crystallized at 15-20°C for 6-12h. After suction filtration and vacuum drying, capsanthin ester (powder) is obtained.

[0041] The vacuum drying temperature in step (1) is 45-60° C., the vacuum degree is -0.080-0.095 MPa, and the drying time is 8-12 h.

[0042] The present invention controls the composition of the alkali neutralization product by specifically selecting and matching the components participating in the alkali neutralization reaction, thereby facilitating the subsequent separation process and improving the separation effect.

[0043] The invention can help protect the color of the product by adding a protective agent in a specific proportion during the alkali neutralization reaction.

[0044] During the evaporation crystallization in step (4) of the present invention, it is preferred to first add seed crystals and / or an auxiliary crystallization agent to the capsanthin desorption solution, and then perform the evaporation crystallization operation; the auxiliary crystallization agent is an aliphatic polyoxyethylene alkylamine, and the amount of the auxiliary crystallization agent added is 0.1-0.3% of the mass of the capsanthin ester prepared in step (1); the amount of the seed crystals added is 0.1-0.5% of the mass of the capsanthin ester prepared in step (1).

[0045] More preferably, the auxiliary crystallization agent is polyoxyethylene octadecylamine and / or polyoxyethylene dodecylamine.

[0046] The seed crystals of the present invention can be commercially available capsanthin crystals, and preferably the capsanthin crystals prepared by the method of the present invention are used as the seed crystals. A product with higher purity can be obtained by adding seed crystals and / or auxiliary crystallization agents.

[0047] The method of the present invention further comprises the step of vacuum drying the filter cake after the evaporation and crystallization of the liquid obtained in the filtering step (4), wherein the vacuum drying temperature is 55-60°C, the vacuum drying time is 8-12h, and the vacuum degree is -0.080 to -0.095MPa.

[0048] In the present invention, the liquid after evaporation and crystallization can be filtered through a plate and frame filter.

[0049] The present invention further provides a capsanthin crystal, which is prepared by the above method. Preferably, in the capsanthin crystal, trans-capsanthin accounts for 97.5-98.0%, and trans-capsanthin, cis-capsanthin and capsanthin as a whole (red component) account for ≥98%.

[0050] The invention first prepares capsanthin into capsanthin ester, and then adopts saponification, resin separation method and evaporation crystallization method to separate the capsanthin in the capsanthin, so as to obtain capsanthin crystals with high purity and high trans-capsanthin content.

[0051] The present invention further provides capsanthin crystals prepared by the above method, or use of the capsanthin crystals in preparing a colorant.

[0052] The present invention also provides a colorant, which comprises the capsanthin crystals prepared by the above method, or the capsanthin crystals.

[0053] The colorant of the present invention has good coloring effect, and an ideal red coloring effect can be obtained by adding only a small amount.

[0054] The beneficial effects of the present invention are at least:

[0055] The preparation method of the present invention can obtain high-purity capsanthin crystals from capsanthin, with a purity of ≥98% after detection and extremely high trans-capsanthin content. No waste is generated in the preparation process, the operation is simple, the efficiency is high, and the preparation process is suitable for industrial large-scale production. DETAILED DESCRIPTION

[0056] The preferred embodiments of the present invention will be described in detail below in conjunction with examples. It should be understood that the following examples are provided only for the purpose of illustration and are not intended to limit the scope of the present invention. Those skilled in the art may make various modifications and substitutions to the present invention without departing from the purpose and spirit of the present invention.

[0057] The experimental methods used in the following examples are conventional methods unless otherwise specified. The materials, reagents, etc. used in the following examples are commercially available or prepared according to conventional methods in the art unless otherwise specified.

[0058] In the specific implementation method of the present invention, the method for detecting the content of the red component and the content of trans-capsanthin in capsanthin crystals is as follows:

[0059] Through high performance liquid chromatography, equipped with UV-visible detector, suitable for normal phase detection system, chromatographic column: silica gel column, 250×4.6mm, 5μm, column temperature: 30℃, injection volume: 20μL, running time: 40min, detection wavelength is 474nm. Specifically, the sample is weighed according to 22.5 / color value (unit is g), and the color value detection method is determined in accordance with "Food Additive Chili Red GB 1886.342015".

[0060] Weigh the sample into a 50mL volumetric flask. Make up to volume with acetone, shake well, and place it away from light for 10 minutes. Transfer 5mL of supernatant to a 100mL volumetric flask, shake gently, then add 30mL of n-hexane, make up to volume with 1mol / L sodium sulfate solution, and place it away from light for 1h. After taking it out, transfer 5mL of the upper clear liquid to a 50mL volumetric flask, make up to volume with n-hexane, and mix well. Take the treated upper clear liquid, pass it through a nylon filter membrane, inject it for detection, and calculate it according to the area normalization of the spectrum. Content = color component area / total area * 100%; when the red component content is determined, the color component is the component with a peak time of 19-45.0min, and when the trans-capsanthin content is determined, the color component is the component with a peak time of 22-28min.

[0061] Seed crystals were purchased from Shanghai Aladdin Biochemical Technology Co., Ltd. Resin was purchased from Xi'an Lanxiao Technology New Materials Co., Ltd.

[0062] Example 1

[0063] This embodiment provides a high-purity capsanthin crystal and a preparation method thereof, which are specifically as follows:

[0064] (1) 50 kg of capsanthin (color value 222.5, purchased from Chenguang Biological Company), 50 g of vitamin C, 50 g of disodium ethylenediaminetetraacetic acid, 1.25 kg of sodium hydroxide and 1.25 kg of calcium hydroxide were added to an alkali neutralization reactor together with 300 L of anhydrous ethanol, dissolved at 20 ° C, and reacted at 55 ° C for 2.5 h. After the alkali neutralization was completed, the saponified liquid was poured into a crystallization kettle, heated to 65 ° C, the heating rate was 4 ° C / min, maintained for 1.5 h, then dropped to 15 ° C, the cooling rate was 0.2 ° C / min, crystallization was carried out for 12 h, suction filtered, vacuum dried at 55 ° C for 8 h, and the vacuum degree was -0.095 MPa to obtain 2.19 kg of capsanthin ester. Repeat the above steps to obtain 10 kg of capsanthin ester.

[0065] (2) Add 10 kg of the above-mentioned capsanthin ester, 2 kg of potassium hydroxide, and 60 L of anhydrous ethanol into a saponification reaction kettle, saponify at 50° C. for 6 h, continue stirring for 2 h, and set aside.

[0066] (3) 80 kg of LXC-03 resin was loaded into a chromatography column and compacted with nitrogen. The resin was activated by column treatment with 4% sodium hydroxide solution and 4% hydrochloric acid solution, respectively, at a flow rate of 1 BV / h and a treatment volume of 3 BV. After treatment, deionized water was added to wash until the outlet liquid pH = 4. Finally, the resin was soaked in ethanol for 2 hours, and the ethanol concentration was 95%.

[0067] The saponified liquid was added into the chromatography tower and directly adsorbed with the resin at a flow rate of 0.5 BV / h. After the adsorption was completed, it was washed with deionized water at a flow rate of 1 BV / h and a washing time of 2 h.

[0068] Gradient desorption, first pass 200L of petroleum ether: ethyl acetate = 20:1 desorption liquid (temperature is 40°C) into the tower, and the treatment flow rate is 2BV / h; then pass petroleum ether: ethyl acetate = 15:1 desorption liquid 200L (temperature is 40°C) into the tower again, and the treatment flow rate is 1BV / h. The capsanthin desorption liquid obtained by the second desorption is collected, passed into an evaporation crystallization tank, and 50g of seed crystals and 10g of auxiliary crystallization agent (polyoxyethylene octadecylamine AC-1805 produced by Jiangsu Hai'an Petrochemical) are added. The evaporation crystallization curve was set as follows: heating from room temperature 20℃ to 35℃, with a heating rate of 1.5℃ / min; keeping at 35℃ for 20min; continuing to heat to 40℃, with a heating rate of 0.25℃ / min; keeping at 40℃ for 30min; heating to the target crystallization temperature of 45℃, with a heating rate of 0.25℃ / min, evaporating and crystallizing for 4h at a stirring rate of 60r / min, then cooling to 20℃, with a cooling rate of 0.3℃ / min, and the final solid content of the crystallization liquid was 22.0%. The cooled crystallization liquid was passed through a plate and frame filter, and the filter cake was vacuum dried at 55℃ for 12h, with a vacuum degree of -0.095MPa. 2.2kg of capsanthin crystals were obtained, and the trans-capsanthin accounted for 98.02% and the total red component accounted for 98.53%.

[0069] Example 2

[0070] This embodiment provides a high-purity capsanthin crystal and a preparation method thereof, which are specifically as follows:

[0071] (1) 50 kg of capsanthin (color value 222.5), 250 g of tetrasodium ethylenediaminetetraacetic acid, 3.33 kg of sodium hydroxide and 1.67 kg of calcium hydroxide were added to an alkali neutralization reactor together with 150 L of 95% ethanol, dissolved at 20 ° C, and reacted at 65 ° C for 0.5 h. After the alkali neutralization, the saponified solution was poured into a crystallization reactor, heated to 55 ° C at a heating rate of 6 ° C / min, maintained for 0.5 h, then cooled to 20 ° C at a cooling rate of 1 ° C / min, crystallized for 6 h, filtered, and vacuum dried at 45 ° C for 8 h, with a vacuum degree of -0.080 MPa, to obtain 1.84 kg of capsanthin ester.

[0072] (2) Repeat the above step (1) to obtain 10 kg of capsanthin ester, add it, 4 kg of potassium hydroxide and 60 L of 95% ethanol into an alkali neutralization reaction kettle, saponify at 55°C for 8 h, continue stirring for 2 h, and set aside.

[0073] (3) 80 kg of LXC-03 resin was loaded into a chromatography column and compacted with nitrogen. The resin was activated by column treatment with 4% sodium hydroxide solution and 4% hydrochloric acid solution, respectively, at a flow rate of 2 BV / h and a treatment volume of 3 BV. After treatment, deionized water was added to wash until the outlet liquid pH = 4.5. Finally, the resin was soaked in ethanol for 4 hours, and the ethanol concentration was 95%.

[0074] The saponified liquid was added into the chromatography tower and directly adsorbed with resin at a flow rate of 1 BV / h. After the adsorption was completed, it was washed with deionized water at a flow rate of 2 BV / h and a washing time of 3 hours.

[0075] Gradient desorption, first pass 200L of desorption liquid of petroleum ether: ethyl acetate = 20:1 into the tower (temperature is 40°C), and the treatment flow rate is 1BV / h; then pass 200L of desorption liquid of petroleum ether: ethyl acetate = 15.5:1 into the tower (temperature is 40°C), and the treatment flow rate is 1.5BV / h. The capsanthin desorption liquid obtained by the second desorption is collected, passed into an evaporation crystallization tank, and 10g of seed crystals and 30g of auxiliary crystallization agent (polyoxyethylene octadecylamine AC-1805 produced by Jiangsu Hai'an Petrochemical) are added. The evaporation crystallization curve was set as follows: heating from room temperature 20℃ to 35.5℃, with a heating rate of 2℃ / min; keeping at 35.5℃ for 15min; continuing to heat to 40.5℃, with a heating rate of 0.5℃ / min; keeping at 40.5℃ for 20min; heating to the target crystallization temperature of 50℃, with a heating rate of 0.5℃ / min, evaporating and crystallizing for 2h at a stirring rate of 80r / min, then cooling to room temperature 20℃, with a cooling rate of 0.6℃ / min, and the final solid content of the crystallization liquid was 18.0%. The crystallization liquid was passed through a plate and frame filter, and the filter cake was vacuum dried at 55℃ for 12h, with a vacuum degree of -0.085Mpa. 1.88kg of capsanthin crystals were obtained, and the trans-capsanthin accounted for 97.64% and the total red component accounted for 98.01%.

[0076] Example 3

[0077] This embodiment provides a high-purity capsanthin crystal and a preparation method thereof, which are specifically as follows:

[0078] (1) 50 kg of capsanthin (color value 222.5), 100 g of vitamin C, 100 g of ethylenediaminetetraacetic acid, 1.14 kg of sodium hydroxide and 2.86 kg of calcium hydroxide were added to an alkali neutralization reactor together with 200 L of acetone, dissolved at 20 ° C, and reacted at 60 ° C for 2 h. After the alkali neutralization, the saponified liquid was poured into a separation kettle, heated to 60 ° C at a heating rate of 4.5 ° C / min, maintained for 1 h, then cooled to 18 ° C at a cooling rate of 0.5 ° C / min, crystallized for 8 h, filtered, and vacuum dried at 60 ° C for 8 h, with a vacuum degree of -0.090 MPa, to obtain 2.07 kg of capsanthin ester.

[0079] (2) Repeat step (1) to prepare 10 kg of capsanthin ester, add it, 3 kg of potassium hydroxide and 55 L of methanol into an alkali neutralization reactor, saponify at 52° C. for 6 h, continue stirring for 2 h, and set aside.

[0080] (3) 80 kg of LXC-03 resin was loaded into a chromatography column and compacted with nitrogen. The resin was activated by column treatment with 4% sodium hydroxide solution and 4% hydrochloric acid solution, respectively, at a flow rate of 2 BV / h and a treatment volume of 3 BV. After treatment, deionized water was added to wash until the outlet liquid pH = 4.2. Finally, the resin was soaked in ethanol for 4 hours, and the ethanol concentration was 95%.

[0081] The saponified liquid was added into the chromatography tower and directly adsorbed with resin at a flow rate of 1 BV / h. After the adsorption was completed, it was washed with deionized water at a flow rate of 2 BV / h and a washing time of 3 hours.

[0082] Gradient desorption, first pass 205L of petroleum ether: ethyl acetate = 20:1 desorption liquid (temperature is 35°C) into the tower, and the treatment flow rate is 2BV / h; then pass petroleum ether: ethyl acetate = 15:1 desorption liquid 250L (temperature is 35°C) into the tower, and the treatment flow rate is 1BV / h. The capsanthin desorption liquid obtained by the second desorption is collected, passed into an evaporation crystallization tank, and 20g of seed crystals and 40g of auxiliary crystallization agent (polyoxyethylene dodecylamine AC-1215 produced by Jiangsu Haian Petrochemical) are added. The evaporation crystallization curve was set as follows: heating from room temperature 20℃ to 35℃, with a heating rate of 1.6℃ / min; keeping at 35℃ for 20min; continuing to heat to 39.5℃, with a heating rate of 0.45℃ / min; keeping at 39.5℃ for 25min; heating to the target crystallization temperature of 48℃, with a heating rate of 0.35℃ / min, evaporating and crystallizing for 3.5h at a stirring rate of 60r / min, then cooling to room temperature 20℃, with a cooling rate of 0.5℃ / min, and the final solid content of the crystallization liquid was 21.1%. The cooled liquid was passed through a plate and frame filter, and the filter cake was vacuum dried at 55℃ for 12h, with a vacuum degree of -0.087MPa. 1.97kg of high-purity capsanthin crystals were obtained, and the trans-capsanthin accounted for 97.91% and the total red component accounted for 98.15% after testing.

[0083] Example 4

[0084] This embodiment provides a high-purity capsanthin crystal and a preparation method thereof, which are specifically as follows:

[0085] (1) 50 kg of capsanthin (color value 222.5), 100 g of ethylenediaminetetraacetic acid, 50 g of disodium ethylenediaminetetraacetic acid, 1 kg of sodium hydroxide and 2 kg of calcium hydroxide were added to an alkali neutralization reactor together with 250 L of isopropanol, dissolved at 20 ° C, and reacted at 55 ° C for 1.5 h. After the alkali neutralization was completed, the saponified liquid was poured into a separation kettle, heated to 55 ° C, the heating rate was 5 ° C / min, maintained for 1.2 h, then cooled to 17 ° C, the cooling rate was 0.5 ° C / min, crystallized for 10 h, filtered, and vacuum dried at 55 ° C for 8 h, the vacuum degree was -0.095 MPa, and 1.96 kg of capsanthin ester was obtained.

[0086] (2) Repeat step (1) to prepare 10 kg of capsanthin ester, add it, 2 kg of sodium hydroxide and 50 L of ethylene glycol into an alkali neutralization reactor, saponify at 50° C. for 6 h, continue stirring for 2 h, and set aside.

[0087] (3) 80 kg of LXC-03 resin was loaded into a chromatography column and compacted with nitrogen. The resin was activated by column treatment with 4% sodium hydroxide solution and 4% hydrochloric acid solution, respectively, at a flow rate of 1 BV / h and a treatment volume of 3 BV. After treatment, deionized water was added to wash until the outlet liquid pH = 4.3. Finally, the resin was soaked in ethanol for 2 hours, and the ethanol concentration was 95%.

[0088] The saponified liquid was added into the chromatography tower and directly adsorbed with the resin at a flow rate of 0.5 BV / h. After the adsorption was completed, it was washed with deionized water at a flow rate of 1 BV / h and a washing time of 2 h.

[0089] Gradient desorption, first pass 200L of desorption liquid of petroleum ether: ethyl acetate = 20:1 (temperature is 38°C) into the tower, and the treatment flow rate is 2BV / h; then pass 200L of desorption liquid of petroleum ether: ethyl acetate = 15.1:1 (temperature is 38°C) into the tower again, and the treatment flow rate is 1BV / h. The capsanthin desorption liquid obtained by the second desorption is collected, passed into an evaporation crystallization tank, and 30g of seed crystals and 20g of auxiliary crystallization agent (polyoxyethylene octadecylamine AC-1805 produced by Jiangsu Hai'an Petrochemical) are added. The evaporation crystallization curve was set as follows: heating from room temperature 20℃ to 34.5℃, with a heating rate of 1.5℃ / min; keeping at 34.5℃ for 15min; continuing to heat to 40℃, with a heating rate of 0.3℃ / min; keeping at 40℃ for 20min; heating to the target crystallization temperature of 47℃, with a heating rate of 0.5℃ / min, evaporating and crystallizing for 2.5h at a stirring rate of 60r / min, then cooling to room temperature 20℃, with a cooling rate of 0.5℃ / min, and the final solid content of the crystallization liquid was 18.7%. The crystallization liquid was passed through a plate and frame filter, and the filter cake was vacuum dried at 60℃ for 8h, with a vacuum degree of -0.085MPa. 2.08kg of high-purity capsanthin crystals were obtained, and the trans-capsanthin accounted for 97.85% and the total red component accounted for 98.27%.

[0090] Example 5

[0091] This embodiment provides a high-purity capsanthin crystal and a preparation method thereof, which are specifically as follows:

[0092] Repeat step (1) of Example 1 to prepare 10 kg of capsanthin ester, add it, 2 kg of potassium hydroxide and 60 L of anhydrous ethanol into an alkali neutralization reactor, saponify at 53° C. for 7.5 h, continue stirring for 3 h, and set aside.

[0093] 80kg of LXC-03 resin was loaded into the chromatography column and compacted with nitrogen. The resin was activated by column treatment with 4% sodium hydroxide solution and 4% hydrochloric acid solution, respectively, at a flow rate of 1BV / h and a treatment volume of 3BV. After treatment, deionized water was added to wash until the outlet liquid pH = 4.5. Finally, the resin was soaked in ethanol for 2 hours, and the ethanol concentration was 95%.

[0094] The saponified liquid was added into the chromatography tower and directly adsorbed with the resin at a flow rate of 0.5 BV / h. After the adsorption was completed, it was washed with deionized water at a flow rate of 1 BV / h and a washing time of 2 h.

[0095] Gradient desorption, first pass 200L of petroleum ether: ethyl acetate = 20:1 desorption liquid (temperature is 40°C) into the tower, with a treatment flow rate of 1.8BV / h; then pass 200L of petroleum ether: ethyl acetate = 15:1 desorption liquid (temperature is 40°C) into the tower again, with a treatment flow rate of 1BV / h, collect the capsanthin desorption liquid obtained by the second desorption, pass it into an evaporation crystallization tank, and add 10g of seed crystals and 30g of auxiliary crystallization agent (polyoxyethylene octadecylamine AC-1805 produced by Jiangsu Hai'an Petrochemical). The evaporation crystallization curve was set as follows: heating from room temperature 20℃ to 35℃, with a heating rate of 1.8℃ / min; keeping at 35℃ for 18min; continuing to heat to 40.2℃, with a heating rate of 0.25℃ / min; keeping at 40.2℃ for 20min; heating to the target crystallization temperature of 48℃, with a heating rate of 0.3℃ / min, evaporating and crystallizing for 3h at a stirring rate of 80r / min, then cooling to room temperature 20℃, with a cooling rate of 0.3℃ / min, and the final solid content of the crystallization liquid was 19.9%. The crystallization liquid was passed through a plate and frame filter, and the filter cake was vacuum dried at 55℃ for 12h, with a vacuum degree of -0.085MPa. 2.01kg of capsanthin crystals were obtained, and the trans-capsanthin accounted for 97.76% after testing, and the total proportion of red components was 98.38% pure.

[0096] Example 6

[0097] This embodiment provides a high-purity capsanthin crystal and a preparation method thereof, which is substantially the same as that of Example 1, except that during evaporation and crystallization, no seed crystal and auxiliary crystallization agent are added, and the remaining steps are the same as those of Example 1. Finally, 1.8 kg of capsanthin crystals were obtained, and after testing, the proportion of trans-capsanthin was 94.51%, and the total proportion of red components was 95.57%.

[0098] Example 7

[0099] This embodiment provides a high-purity capsanthin crystal and a preparation method thereof, which is basically the same as Example 1, except that during evaporation crystallization, polyoxyethylene (20) sorbitan monostearate is used instead of polyoxyethylene octadecylamine as an auxiliary crystallization agent, and the remaining steps are the same as Example 1.

[0100] Finally, 1.75 kg of capsanthin crystals were obtained. After testing, the proportion of trans-capsanthin was 97.33%, and the total proportion of red components was 97.65%.

[0101] Example 8

[0102] This embodiment provides a capsanthin crystal and a preparation method thereof, which is substantially the same as that of embodiment 1, except that the method for preparing capsanthin into capsanthin ester in step (1) is a solvent crystallization method, which is as follows:

[0103] Dissolve capsanthin in a solvent, the mass volume ratio of capsanthin to solvent is 1:10, the solvent is a mixed solvent of petroleum ether and propanol, the volume ratio of petroleum ether to propanol is 1:10. Add capsanthin first, then add the dissolving solvent, stir for 30 minutes to fully dissolve, then cool to 5 degrees Celsius, crystallize for 12 hours, and filter to obtain capsanthin ester.

[0104] The remaining steps remain unchanged. Finally, 0.85 kg of capsanthin crystals were obtained, and the trans-capsanthin accounted for 97.42% and the total red component accounted for 97.94%.

[0105] Example 9

[0106] This embodiment provides a high-purity capsanthin crystal and a preparation method thereof, which is substantially the same as that of Example 1, except that, during the gradient desorption in step (2), the temperature of the desorption liquid used for the two desorptions is the room temperature of 25°C. The other conditions and steps remain unchanged. Finally, 1.83 kg of capsanthin crystals were obtained, and the trans-capsanthin accounted for 97.28% and the total red component accounted for 97.89%.

[0107] Example 10

[0108] The present embodiment provides a high-purity capsanthin crystal and a preparation method thereof, which is substantially the same as Example 1, except that, during the gradient desorption in step (2), petroleum ether: ethyl acetate = 22: 1 desorption liquid 200L (temperature is 40°C) is first introduced into the tower, and the treatment flow rate is 2BV / h; and then petroleum ether: ethyl acetate = 16.5: 1 desorption liquid 200L (temperature is 40°C) is introduced into the tower again, and the treatment flow rate is 1BV / h, and the capsanthin desorption liquid obtained by the second desorption is collected, and subsequent evaporation and crystallization are continued. The remaining conditions and steps remain unchanged. Finally, 2.05kg of capsanthin crystals are obtained, and the trans-capsanthin accounts for 96.70% after testing, and the total proportion of red components is 97.73%.

[0109] Embodiment 11

[0110] The present embodiment provides a high-purity capsanthin crystal and a preparation method thereof, which are as follows: Repeat step (1) of Example 1 to prepare 10 kg of the above-mentioned capsanthin ester, add it and 3.5 kg of sodium hydroxide and 58 L of ethylene glycol into a saponification reaction kettle, saponify at 52° C. for 7 hours, continue stirring for 2 hours, and set aside.

[0111] 80kg of LXC-03 resin was loaded into a chromatography column and compacted with nitrogen. The resin was activated by column treatment with 4% sodium hydroxide solution and 4% hydrochloric acid solution, respectively, at a flow rate of 1BV / h and a treatment volume of 3BV. After treatment, deionized water was added to wash until the outlet liquid pH = 4. Finally, the resin was soaked in ethanol for 2h, and the ethanol concentration was 95%.

[0112] The saponified liquid was added into the chromatography tower and directly adsorbed with the resin at a flow rate of 0.5 BV / h. After the adsorption was completed, it was washed with deionized water at a flow rate of 1 BV / h and a washing time of 2 h.

[0113] Gradient desorption, first pass 200L of petroleum ether: ethyl acetate = 20:1 desorption liquid (temperature is 40°C) into the tower, and the treatment flow rate is 2BV / h; then pass petroleum ether: ethyl acetate = 15:1 desorption liquid 200L (temperature is 40°C) into the tower again, and the treatment flow rate is 1BV / h. The capsanthin desorption liquid obtained by the second desorption is collected, passed into an evaporation crystallization tank, and 50g of seed crystals and 10g of auxiliary crystallization agent (polyoxyethylene octadecylamine AC-1805 produced by Jiangsu Hai'an Petrochemical) are added. The evaporation crystallization curve was set as follows: heating from room temperature 20℃ to 35℃, with a heating rate of 1.5℃ / min; keeping at 35℃ for 20min; continuing to heat to 39.5℃, with a heating rate of 0.25℃ / min; keeping at 39.5℃ for 30min; heating to the target crystallization temperature of 48℃, with a heating rate of 0.25℃ / min, evaporating and crystallizing for 2.8h at a stirring rate of 60r / min, then cooling to 20℃, with a cooling rate of 0.4℃ / min, and the final solid content of the crystallization liquid was 19.8%. The cooled crystallization liquid was passed through a plate and frame filter, and the filter cake was vacuum dried at 55℃ for 12h, with a vacuum degree of -0.095MPa. 2.6kg of capsanthin crystals were obtained, and the trans-capsanthin accounted for 97.81% and the total red component accounted for 98.44%.

[0114] Comparative Example 1

[0115] This comparative example provides a capsanthin crystal and a preparation method thereof, which is substantially the same as Example 1, except that the operation of step (1) and the saponification operation in step (2) are not performed, and the capsanthin (color value 222.5) in Example 1 is directly used as a raw material for resin separation. The specific separation conditions are substantially the same as those described in Example 1, and the desorption liquid flow rate is reduced during desorption to better separate and prolong the desorption time. Specifically, the treatment flow rate when desorbing the desorbent using petroleum ether: ethyl acetate = 20:1 is 0.5 BV / h, and the treatment flow rate when desorbing the desorbent using petroleum ether: ethyl acetate = 15:1 is 0.5 BV / h. Other steps such as evaporation and crystallization are the same as those in Example 1.

[0116] Finally, 1.2 kg of high-purity capsanthin crystals were obtained. After testing, the trans-capsanthin accounted for 89.53% and the total red component accounted for 90.96%.

[0117] Comparative Example 2

[0118] This comparative example provides a capsanthin crystal and a preparation method thereof, which is basically the same as Example 1, except that the evaporation crystallization curve is: directly heating from 20°C to the target crystallization temperature of 45°C, with a heating rate of 1°C / min. At a stirring rate of 60r / min, the evaporation crystallization is carried out for 4h, and the temperature is cooled to 20°C, with a cooling rate of 0.3°C / min. The final solid content of the crystallization liquid is 22.5%. The cooled crystallization liquid is passed through a plate and frame filter, and the filter cake is vacuum dried at 55°C for 12h. 1.6kg of capsanthin crystals are obtained, and after testing, the trans-capsanthin accounts for 89.07%, and the total red component accounts for 92.58%.

[0119] Comparative Example 3

[0120] This comparative example provides a capsanthin crystal and a preparation method thereof, which is substantially the same as Example 1, except that the evaporation crystallization curve is:

[0121] The temperature was raised from room temperature 20°C to 35°C at a rate of 3°C / min; the temperature was kept at 35°C for 20 minutes; the temperature was further raised to 40°C at a rate of 0.5°C / min; the temperature was kept at 40°C for 30 minutes; the temperature was raised to the target crystallization temperature of 40.5°C at a rate of 0.5°C / min, and the crystallization was evaporated for 4 hours at a stirring rate of 60r / min, and then the temperature was lowered to 20°C at a rate of 0.3°C / min, and the solid content of the final crystallization liquid was 21.1%. The cooled crystallization liquid was passed through a plate and frame filter, and the filter cake was vacuum dried at 55°C (vacuum degree was -0.080MPa) for 12 hours. 2.1kg of capsanthin crystals were obtained, and the trans-capsanthin accounted for 96.94% and the total red component accounted for 97.37%.

[0122] Although the present invention has been described in detail above with general descriptions and specific embodiments, it is obvious to those skilled in the art that some modifications or improvements may be made thereto based on the present invention. Therefore, these modifications or improvements made without departing from the spirit of the present invention all fall within the scope of protection claimed by the present invention.

Claims

1. A method for preparing capsanthin crystals, characterized in that: include: (1) Preparation of capsanthin ester using capsanthin as raw material; (2) saponifying the capsanthin ester to obtain a saponified solution; In step (2), saponification is achieved by reacting the capsanthin ester with an alkali in a diluent, wherein the mass ratio of the capsanthin ester to the alkali is 1:(0.2-0.4); the saponification temperature is 50-55°C, and the time is 6-8h; The diluent is one or more of methanol, anhydrous ethanol, 95% ethanol, ethylene glycol or propylene glycol, and the mass volume ratio of the capsanthin ester to the diluent is 1: (5-6) g / ml; (3) adsorbing the saponified solution with a resin, and obtaining a capsanthin desorption solution after gradient desorption; In step (3), the resin is a styrene-divinylbenzene skeleton adsorption separation resin, and the resin adsorbs the saponified liquid at a pH value of 4-4.

5. After adsorbing the saponified liquid, the resin is first washed and then gradient desorption is performed. The gradient desorption method is: Firstly, a first desorption liquid is used for a first desorption, and then a second desorption liquid is used for a second desorption to obtain a capsanthin desorption liquid for subsequent operations; The first desorption liquid is petroleum ether and ethyl acetate in a volume ratio of (20-20.5):1, and the second desorption liquid is petroleum ether and ethyl acetate in a volume ratio of (15-15.5):1; (4) obtaining capsanthin crystals from the capsanthin desorption solution by evaporation crystallization; The evaporation crystallization method is: heating from room temperature to 35±0.5°C at a heating rate of 1.5-2°C / min; keeping at 35±0.5°C for 15-20min; Continue to heat up to 40±0.5°C at a heating rate of 0.25-0.5°C / min; keep warm at 40±0.5°C for 20-30min; heat up to the target crystallization temperature of 45-50°C at a heating rate of 0.25-0.5°C / min; continue evaporation and crystallization at the target crystallization temperature and under stirring, then cool to room temperature at a rate of 0.3-0.6°C / min, and control the solid content of the crystallization liquid to be 18-22%.

2. The method according to claim 1, characterized in that In step (3), the temperature of the first desorption liquid and the second desorption liquid is 35-40°C; During desorption, the flow rate of the first desorption liquid is 1-2 BV / h, and the flow rate of the second desorption liquid is 1-1.5 BV / h; And / or, the volume mass ratio of the first desorption liquid to the saponification liquid is (2.5-4.5): 1 L / kg, and the volume mass ratio of the second desorption liquid to the saponification liquid is (2.5-4.5): 1 L / kg.

3. The method according to claim 1, characterized in that The specific operation method of step (1) is as follows: ① Mix capsanthin, protective agent, composite alkali and alkali neutralization diluent to carry out alkali neutralization reaction; the temperature of the alkali neutralization reaction is 55-65° C. and the time is 0.5-2.5h; The mass ratio of the capsanthin to the composite alkali is 1: (0.05-0.1); the composite alkali is composed of sodium hydroxide and calcium hydroxide in a mass ratio of 1: (0.5-2.5); The protective agent is one or more of ethylenediaminetetraacetic acid, disodium ethylenediaminetetraacetic acid, tetrasodium ethylenediaminetetraacetic acid or vitamin C; the mass ratio of the capsanthin to the protective agent is 1: (0.002-0.005); The alkaline neutralization diluent is one or more of methanol, anhydrous ethanol, 95% ethanol, acetone, isopropanol or 95% isopropanol; the mass volume ratio of the capsanthin to the alkaline neutralization diluent is 1: (3-6) g / ml; ② The solution obtained after the alkali neutralization reaction is heated to 55-65°C at 4.0-6.0°C / min, maintained for 0.5-1.5h, then cooled to 15-20°C at 0.2-1°C / min, and crystallized at 15-20°C for 6-12h. After suction filtration and vacuum drying, capsanthin ester is obtained.

4. The method according to any one of claims 1 to 3, characterized in that: During the evaporation crystallization in step (4), seed crystals and / or an auxiliary crystallization agent are first added to the capsanthin desorption solution, and then the evaporation crystallization operation is performed; the auxiliary crystallization agent is an aliphatic polyoxyethylene alkylamine; And / or, the amount of the auxiliary crystallization agent added is 0.1-0.3% of the mass of the capsanthin ester prepared in step (1); the amount of the seed crystal added is 0.1-0.5% of the mass of the capsanthin ester prepared in step (1).

5. The method according to claim 4, characterized in that The auxiliary crystallization agent is polyoxyethylene octadecylamine and / or polyoxyethylene dodecylamine.

6. The method according to any one of claims 1 to 3 and 5, characterized in that: The method further comprises the step of vacuum drying the filter cake after the slurry obtained by evaporating the crystals in the filtering step (4), wherein the vacuum drying temperature is 55-60° C., the vacuum drying time is 8-12 hours, and the vacuum degree is -0.080 to -0.095 MPa.

7. The method according to claim 4, characterized in that The method further comprises the step of vacuum drying the filter cake after the slurry obtained by evaporating the crystals in the filtering step (4), wherein the vacuum drying temperature is 55-60° C., the vacuum drying time is 8-12 hours, and the vacuum degree is -0.080 to -0.095 MPa.