A process for the preparation of beta-carotene from paprika oleoresin and the product thereof

Abstract

The application provides a method for preparing beta-carotene from capsanthin and a product thereof, and specifically comprises the following steps: (1) performing enzymolysis on capsanthin paste; (2) extracting the capsanthin paste after enzymolysis to obtain capsanthin enzymolysis material solution; (3) performing precision filtration to obtain capsanthin enzymolysis filtrate; (4) performing adsorption by using a medium-pressure silica gel chromatographic column to perform chromatographic separation and collecting port effluent; (5) performing beta-carotene elution; (6) performing yellow pigment elution; (7) performing red pigment elution; and (8) performing orange pigment elution. The method combining enzymolysis and medium-pressure column chromatography is adopted for the first time, smaller particle size chromatographic silica gel is adsorbed, and four-stage elution treatment is performed, so that beta-carotene with a content of greater than 20% can be directly separated from the capsanthin paste after enzymolysis treatment, and capsanthin with a higher red color tone can be prepared. The prepared product can be widely applied in the fields of food and health food.

Description

Technical Field

[0001] This invention belongs to the field of food additive refining technology, specifically relating to a method for preparing β-carotene from capsicum pigment and its product. Background Technology

[0002] β-carotene (C 40 H 56 β-carotene is a type of carotenoid and an orange-yellow fat-soluble compound. It is one of the most common natural pigments in nature, and is found in a relatively large amount in green vegetables, sweet potatoes, carrots, peppers, Dunaliella salina, and palm trees.

[0003] Beta-carotene is mainly obtained through three methods: synthesis, microbial fermentation, and preparation from natural plants. Because beta-carotene has excellent health benefits, and the supply from natural sources cannot meet market demand, chemically synthesized beta-carotene has become extremely popular. However, because chemically synthesized beta-carotene cannot be completely absorbed by the human body and can produce certain toxic side effects, long-term use can cause irreversible damage. Therefore, it is not accepted by most food scientists and has been banned as a food additive in developed Western countries.

[0004] Naturally extracted beta-carotene is easily absorbed by the human body and possesses good antioxidant capabilities, leading to a surge in demand. Currently, palm oil and algae such as Dunaliella salina are excellent raw materials for beta-carotene. The key to obtaining natural beta-carotene from Dunaliella salina lies in its cultivation; however, Dunaliella salina cultivation is severely limited by external environmental conditions, making it difficult to scale up, and the production of natural beta-carotene falls far short of international market demand. Furthermore, the main production areas of palm oil are in Southeast Asia, which significantly restricts the development of natural beta-carotene in my country.

[0005] Currently, chili peppers are the most abundant and readily available source of β-carotene in China, with Xinjiang alone producing over 300,000 tons. The paprika red pigment it contains is primarily used in the food, beverage, cosmetics, and pharmaceutical industries, and is internationally recognized as the best Class A red pigment, with a color intensity 10 times that of other pigments. The main components of chili peppers are capsanthin, capsanthin, β-carotene, and zeaxanthin, with β-carotene accounting for over 13.9% of the total pigment. However, due to its predominantly food applications, its added value is too low, ultimately affecting the long-term development of the entire chili pepper industry.

[0006] In existing technologies, the complex pigment composition of capsanthin makes separation difficult, hindering its industrial-scale production. Particularly concerning is the separation of β-carotene from capsanthin. While some methods employ high-performance preparative liquid chromatography (HPLC) for β-carotene separation, these are limited to the preparation of reference standards. The expensive separation materials and complex elution systems are unsuitable for industrial production. Furthermore, the complex saponification process prior to separation transforms the main components of capsanthin, such as capsanthin, its ester derivatives, and lutein esters, leading to hydrolysis and transformation that compromises the natural characteristics of the remaining pigments after separation.

[0007] Furthermore, while the method for preparing β-carotene from chili peppers disclosed in "A Method for Preparing β-Carotene from Chili Peppers" involves first separating the chili pepper pigments into two shades, yellowish and reddish, using atmospheric pressure silica gel chromatography, and then refining the yellowish-red chili pepper red using saponification and other methods to finally obtain β-carotene, the column chromatography method used in this process cannot be used for continuous production. Each batch requires disassembling the column, activating the silica gel at high temperature, and then repacking the column for separation. Moreover, the elution cycle under low pressure is long, resulting in low separation efficiency, a complex process, and a long production cycle, becoming the main bottleneck for separating β-carotene from chili pepper red.

[0008] In summary, how to isolate β-carotene from capsicum pigment in high yield and high purity is a problem that urgently needs to be solved by those skilled in the art. Summary of the Invention

[0009] The purpose of this invention is to provide a method for preparing β-carotene from capsicum pigment, mainly utilizing enzymatic hydrolysis combined with medium-pressure column chromatography to prepare β-carotene from capsicum red pigment. This method is the first to combine enzymatic hydrolysis with medium-pressure column chromatography, using smaller particle size silica gel for adsorption and performing a four-stage elution process. This allows for the direct separation of β-carotene with a content ≥20% from the enzymatically hydrolyzed capsicum red oil extract, with a yield exceeding 92.5%, while simultaneously producing a higher quality reddish capsicum red.

[0010] To achieve the above objectives, the present invention provides a method for preparing β-carotene from capsaicin, specifically comprising the following steps:

[0011] (1) The chili oil paste was enzymatically hydrolyzed to obtain chili enzymatic hydrolysate;

[0012] (2) Extraction: Add an alkane solvent to the capsicum hydrolysate obtained in step (1) to dissolve it, let it stand to separate into layers, take the upper layer to obtain the capsicum hydrolysate solution;

[0013] (3) Fine filtration: The capsicum hydrolysate solution obtained in step (2) is finely filtered through a filter membrane to obtain the capsicum hydrolysate filtrate.

[0014] (4) Adsorption: The enzymatic hydrolysed capsanthin filtrate obtained in step (3) is separated by chromatography using a medium-pressure silica gel column, and the effluent from the end is collected;

[0015] (5) β-carotene elution: After the medium-pressure chromatography column loading is completed, the first eluent is used for elution, and the eluent is collected and concentrated to obtain β-carotene oil.

[0016] (6) Yellow pigment elution: After the β-carotene elution is completed, a second eluent is used for elution, and the eluent is concentrated to obtain yellow pigment ointment.

[0017] (7) Red pigment elution: After the yellow pigment elution is completed, the third eluent is used for elution, and the eluent is concentrated to obtain a reddish paprika oleoresin.

[0018] (8) Orange pigment elution: After the red pigment elution is completed, the silica gel column is eluted with the fourth eluent. The eluent is concentrated and then combined with yellow pigment oil to obtain a slightly orange-toned paprika red.

[0019] In a preferred embodiment, after step (8) is completed, step (9) further includes solvent replacement and equilibration:

[0020] After solvent replacement and equilibration treatment of the medium-pressure silica gel chromatography column using a displacement solvent, the next batch of separation production can be carried out.

[0021] In a preferred embodiment, step (1) specifically includes: adding membrane-filtered water and lipase to the chili oil paste and stirring the reaction.

[0022] In a preferred embodiment, in step (1), the mass ratio of the chili oil paste, membrane-filtered water, and lipase is 1:(0.4-0.6):(0.01-0.02);

[0023] More preferably, the mass ratio of the chili oil paste, membrane-filtered water, and lipase is 1:0.5:0.15;

[0024] Most preferably, the membrane conductivity of the water filtered by the membrane is <20 μs / cm.

[0025] In a preferred embodiment, in step (1), the stirring reaction temperature is 35-40°C and the stirring reaction time is 6-10h; more preferably, the stirring reaction temperature is 38°C and the stirring reaction time is 10h.

[0026] In this invention, by designing a suitable lipase to enzymatically hydrolyze the chili oil paste, the oils in the chili oil paste are enzymatically converted into fatty acids, thereby changing their polarity and enhancing the separation effect of β-carotene from oily impurities.

[0027] In a preferred embodiment, in step (2), the alkane solvent is an alkane containing 4 to 7 carbons;

[0028] More preferably, the alkane solvent containing 4 to 7 carbons includes one or more of n-hexane, petroleum ether, and vegetable oil extraction solvents.

[0029] In a preferred embodiment, in step (2), the amount of alkane solvent added is such that the color value of the capsicum enzymatic hydrolysate solution is adjusted to 60-120;

[0030] More preferably, the amount of alkane solvent added is used to adjust the color value of the solution to 80-100.

[0031] In this invention, alkane solvents are used as extraction solvents, which can directly extract capsanthin from the aqueous system, achieving the effect of oil-water separation.

[0032] In a preferred embodiment, in step (3), the function of fine filtration is to remove common impurities in the solution by physical methods. Therefore, conventional methods known to those skilled in the art can be used. Preferably, the fine filtration membrane is an organic filter membrane of 0.45 to 10 μm, and more preferably, the fine filtration membrane is an organic filter membrane of 1 μm.

[0033] In a preferred embodiment, in step (4), the stationary phase of the medium-pressure silica gel chromatography column is 300-500 mesh silica gel, and the mass ratio of silica gel to capsicum oil is 1:(0.25-0.4).

[0034] In a preferred embodiment, in step (4), the pressure of the medium-pressure silica gel chromatography column is 0.5-5 MPa, and the pigment adsorption capacity is 40-120 color value / g;

[0035] More preferably, the medium-pressure silica gel chromatography column has a pressure of 0.5–2 MPa and a pigment adsorption capacity of 60–100 color value / g.

[0036] In this step, the capsicum pigment components are adsorbed onto silica gel, and during the process, substances such as β-carotene are eluted. Because β-carotene has the least polarity in capsicum pigment, it is eluted first.

[0037] In a preferred embodiment, in step (5), the first eluent is an alkane containing 4 to 7 carbons, the elution volume is 1 to 5 BV, and the elution rate is 0.5 to 3 BV / h;

[0038] More preferably, the first eluent includes one or more of n-hexane, petroleum ether, and vegetable oil extraction solvent, with an elution volume of 1 to 3 BV and an elution rate of 0.5 to 2 BV / h.

[0039] In a preferred embodiment, in step (5), the concentration temperature is 40 to 100°C; the concentration vacuum degree is -0.03 to -0.09 MPa;

[0040] More preferably, the concentration temperature is 40 to 80°C; and the concentration vacuum degree is -0.05 to -0.09 MPa.

[0041] In this step, a low-polarity alkane eluent is used to achieve the elution and separation of β-carotene, thereby improving the product yield.

[0042] In a preferred embodiment, in step (6), the second eluent is obtained by mixing an alkane containing 4 to 7 carbons with acetone in a mass ratio of (8 to 50): 1;

[0043] More preferably, the alkane containing 4 to 7 carbons includes one or more of n-hexane, petroleum ether, and vegetable oil extraction solvents;

[0044] Most preferably, the alkane containing 4 to 7 carbons is mixed with acetone in a mass ratio of (8 to 20): 1.

[0045] In a preferred embodiment, in step (6), the elution volume is 1 to 5 BV and the elution rate is 0.5 to 3 BV / h;

[0046] More preferably, the elution volume is 1 to 3 BV and the elution rate is 0.5 to 2 BV / h.

[0047] In a preferred embodiment, in step (6), the concentration temperature is 40 to 100°C; the concentration vacuum degree is -0.03 to -0.09 MPa.

[0048] More preferably, the concentration temperature is 40 to 80°C; and the concentration vacuum degree is -0.05 to -0.09 MPa.

[0049] During the elution of β-carotene, the yellow pigment also moves, but the first eluent has low elution efficiency and the elution time is too long. It is necessary to increase the polarity of the eluent to the second eluent to achieve the elution and separation of the yellow pigment. Therefore, the aforementioned elution solvent and elution conditions were designed.

[0050] In a preferred embodiment, in step (7), the third eluent is obtained by mixing an alkane containing 4 to 7 carbons with acetone in a mass ratio of (8 to 50): 1;

[0051] More preferably, the alkane containing 4 to 7 carbons includes one or more of n-hexane, petroleum ether, and vegetable oil extraction solvents;

[0052] Most preferably, the alkane containing 4 to 7 carbons is mixed with acetone in a mass ratio of (8 to 20): 1.

[0053] In a preferred embodiment, in step (7), the elution volume is 1 to 5 BV and the elution rate is 0.5 to 3 BV / h;

[0054] More preferably, the elution volume is 1 to 3 BV and the elution rate is 0.5 to 2 BV / h.

[0055] In a preferred embodiment, in step (7), the concentration temperature is 40 to 100°C; the concentration vacuum degree is -0.03 to -0.09 MPa;

[0056] More preferably, the concentration temperature is 40 to 80°C; and the concentration vacuum degree is -0.05 to -0.09 MPa.

[0057] In the red pigment elution step, to separate the reddish pigment from the terminal orange pigment, it is not advisable to further increase the polarity of the eluent. The polarity of the second eluent is sufficient for eluting the reddish pigment. Furthermore, the third eluent and the second eluent produce different elution effects by adjusting the elution time (i.e., the amount of eluent used).

[0058] In a preferred embodiment, in step (8), the fourth eluent is acetone, the elution volume is 1-5 BV, and the elution rate is 0.5-3 BV / h;

[0059] More preferably, the elution volume is 1 to 3 BV and the elution rate is 0.5 to 2 BV / h.

[0060] In a preferred embodiment, in step (8), the concentration temperature is 40 to 100°C; the concentration vacuum degree is -0.03 to -0.09 MPa.

[0061] More preferably, the concentration temperature is 40 to 80°C; and the concentration vacuum degree is -0.05 to -0.09 MPa.

[0062] In this step, highly polar acetone is used to directly wash away all residual orange pigment and colorless impurities, thereby obtaining high-quality reddish paprika.

[0063] In a preferred embodiment, in step (9), the replacement solvent is an alkane containing 4 to 7 carbons, the solvent replacement and equilibrium volume is 1.5 to 5 BV, and the flow rate is 0.5 to 3 BV / h;

[0064] More preferably, the displacement solvent is an alkane containing 4 to 7 carbons, the solvent displacement and equilibrium volume is 2 to 4 BV, and the flow rate is 0.5 to 2 BV / h.

[0065] Another objective of this invention is to provide β-carotene prepared by any of the above methods. This invention employs medium-pressure column chromatography to directly separate β-carotene from enzymatically hydrolyzed capsaicin oil, simultaneously preparing a higher quality reddish capsaicin. Furthermore, it is the first time that continuous medium-pressure silica gel column chromatography has been applied to the industrial-scale separation and production of capsaicin, achieving industrial-scale production of over 20% β-carotene directly separated from capsaicin oil. The overall process is simple and efficient. The yield of the obtained β-carotene is over 92.5%, and the purity is over 20%.

[0066] Compared with the prior art, the technical solution of the present invention has the following advantages:

[0067] This invention is the first to combine enzymatic hydrolysis technology with medium-pressure column chromatography using silica gel as the stationary phase to directly separate β-carotene from capsicum oil. The capsicum oil is enzymatically hydrolyzed to obtain capsicum enzymatic hydrolysate. The hydrolysate is dissolved in a solvent, finely filtered, and then subjected to continuous wet chromatography under medium pressure to obtain two eluents. The eluents are concentrated to yield β-carotene and capsicum red pigments with yellow, orange, and reddish hues, respectively.

[0068] In this invention, silica gel column chromatography employs normal-phase separation. The elution order of substances depends on their polarity. The polarity of the elution solvent increases from lowest to highest, and the substances are eluted in order of decreasing polarity. Therefore, β-carotene has the lowest polarity, followed by other yellow pigments, then reddish pigments, and finally orange pigments with the highest polarity. This method optimizes the separation of different hues of paprika oleoresin, yielding yellowish, orangeish, and reddish paprika oleoresin, respectively, and increasing the absorbance ratio of the red pigment from 1.02 to over 1.08. Furthermore, this invention simplifies the direct separation of β-carotene from paprika oleoresin extract, enabling large-scale industrial production of β-carotene.

[0069] In summary, this invention is the first to employ a separation technique combining enzymatic hydrolysis and medium-pressure column chromatography to directly prepare a 20% β-carotene product. Simultaneously, it significantly improves the quality of capsicum red, resulting in a substantial increase in its overall quality and added value, making it widely applicable in food and health food products. The technical effects of this invention not only significantly enhance the product scale and market advantage of β-carotene products derived from chili peppers but also lay the foundation for expanding new development opportunities in the chili pepper industry. Detailed Implementation

[0070] To enable those skilled in the art to better understand the present invention, the present invention will be further described in detail below with reference to specific embodiments, but it should be understood that the scope of protection of the present invention is not limited to the specific embodiments.

[0071] This invention provides a method for preparing β-carotene from capsicum pigment and the resulting product, thus solving the technical problem that the complex pigment composition and difficult separation of capsicum red pigment in existing technologies have prevented the industrial-scale production of capsicum red pigment.

[0072] The technical solution of this application will be described in detail below through specific embodiments:

[0073] Unless otherwise specified, the technical means used in this invention are conventional means well known to those skilled in the art. All raw materials, reagents, instruments, and equipment used in this invention can be purchased from the market or prepared using existing methods. Unless otherwise specified, all reagents used in this invention are of analytical grade. The capsicum oil paste used in this invention was purchased from Henan Zhongda Hengyuan Biotechnology Co., Ltd., and its preparation method is as follows: mature chilies are dried at 30-50°C, pulverized, and granulated to obtain chili granules; the chili granules are conveyed into a continuous extractor, and during the conveying process, an extraction solvent is sprayed through a spray pipe to ensure uniform mixing between the extraction solvent and the chili granules. The capsicum is extracted using the extraction solvent to obtain a pigment extract; the pigment extract is concentrated under reduced pressure until the solvent content is <0.5%, yielding the capsicum oil paste.

[0074] Example 1

[0075] a) Take 100kg of chili oil paste {color value 273.34, absorbance (A)} 470nm / A 454nm The concentration of β-carotene was 0.979, and the content of β-carotene was 1.07%. 0.5 times the amount of membrane filter water was added, and 1.5% of the weight of lipase was added. The mixture was stirred and reacted at 38°C for 10 hours to obtain the enzymatic hydrolysate of capsicum.

[0076] b) After the enzymatic hydrolysis reaction is complete, add 190 kg of vegetable oil extraction solvent to the enzymatic hydrolysate, adjust the color value to 98, stir and dissolve at room temperature for 30 min, let stand for 60 min, and slowly remove the lower aqueous phase; the upper pigment phase is finely filtered through a 1 μm organic filter membrane to obtain the enzymatic hydrolysate of capsicum red.

[0077] c) The enzymatically hydrolyzed capsanthin filtrate was subjected to silica gel column chromatography at a flow rate of 1 Bv / h using a continuous wet loading method, with a chromatographic pressure of 0.7–0.8 MPa. Elution was performed using 1500 L of vegetable oil extractant at a flow rate of 1 Bv / h to obtain β-carotene eluent. The β-carotene eluent was concentrated under reduced pressure to obtain 4.64 kg of orange-yellow oil paste with a color value of 510.15 and an absorbance ratio (A). 470nm / A 454nm The content of β-carotene was 0.850, with a yield of 8.68% based on capsicum red; the content of β-carotene was 21.36%, with a yield of 92.63% based on β-carotene content.

[0078] d) After β-carotene elution is complete, elute 750 L of a mixture of vegetable oil extraction solvent and acetone (mass ratio 10:1) at a flow rate of 1 Bv / h to obtain a yellow pigment eluent. Concentrate the yellow pigment eluent under reduced pressure to obtain 30.14 kg of yellow pigment oil paste with a color value of 107.42 and an absorbance ratio (A). 470nm / A 454nm The content of β-carotene was 0.835, and the yield was 11.86%; the content of β-carotene was 0.05%, and the yield based on the β-carotene content was 1.52%.

[0079] e) After the yellow pigment elution is complete, continue eluting with a mixture of vegetable oil extraction solvent and acetone (mass ratio 10:1) at a flow rate of 1 Bv / h for 1500 L to obtain the red pigment eluent. Concentrate the red pigment eluent under reduced pressure to obtain 35.97 kg of a reddish-brown paprika oil paste, with a color value of 411.89 and an absorbance ratio (A). 470nm / A 454nm The yield was 1.080, and the efficiency was 54.27%.

[0080] f) After the red pigment elution is complete, elute 900 L of acetone at a flow rate of 1 Bv / h to obtain the orange pigment eluent. Concentrate the orange pigment eluent under reduced pressure to obtain 18.63 kg of a slightly orange-toned paprika oil paste, with a color value of 324.44 and an absorbance ratio (A). 470nm / A 454nm The yield was 0.951, and the efficiency was 22.14%.

[0081] g) After acetone elution is complete, the medium-pressure chromatography column is subjected to solvent replacement and equilibration treatment using vegetable oil extraction solvent at a rate of 1 Bv / h. The solvent volume is 1500 L (2 Bv). After solvent replacement and equilibration treatment, the next batch of separation production can be carried out.

[0082] Example 2

[0083] a) Take 100kg of chili oil paste {color value 304.17, absorbance (A)} 470nm / A454nm The concentration of β-carotene was 0.987, and the content of β-carotene was 1.09%. 0.5 times the amount of membrane filter water was added, and 1.5% of the weight of lipase was added. The mixture was stirred and reacted at 38°C for 10 hours to obtain the enzymatic hydrolysate of capsicum.

[0084] b) After the enzymatic hydrolysis reaction is complete, add 210 kg of petroleum ether to the enzymatic hydrolysate, adjust the color value to 100, stir and dissolve at room temperature for 30 min, let stand for 60 min, and slowly remove the lower aqueous phase; the upper pigment phase is filtered through a 0.8 μm organic filter membrane to obtain the enzymatic hydrolysate of capsicum red.

[0085] c) The enzymatically hydrolyzed capsanthin filtrate was subjected to silica gel column chromatography at a flow rate of 1.5 Bv / h using a continuous wet loading method, with a chromatographic pressure of 1.0–1.2 MPa. Elution was performed with 1500 L of petroleum ether at a flow rate of 1.0 Bv / h to obtain a β-carotene eluent. The β-carotene eluent was concentrated under reduced pressure to obtain 4.50 kg of an orange-yellow oil paste with a color value of 540.76 and an absorbance ratio (A). 470nm / A 454nm The content of β-carotene was 0.855, with a yield of 8.01% based on capsicum red; the content of β-carotene was 23.08%, with a yield of 95.34% based on β-carotene content.

[0086] d) After β-carotene elution is complete, elute 1000 L of a mixed solvent of petroleum ether and acetone (mass ratio 12:1) at a flow rate of 1 Bv / h to obtain a yellow pigment eluent. Concentrate the yellow pigment eluent under reduced pressure to obtain 31.62 kg of yellow pigment oil, with a color value of 89.22 and an absorbance ratio (A). 470nm / A 454nm The content of β-carotene was 0.841, with a yield of 9.28% based on capsicum red; the content of β-carotene was 0.10%, with a yield of 2.86% based on β-carotene content.

[0087] e) After the yellow pigment elution is complete, continue eluting with a mixed solvent of petroleum ether and acetone (mass ratio 12:1) at a flow rate of 1 Bv / h for 2000 L to obtain the red pigment eluent. Concentrate the red pigment eluent under reduced pressure to obtain 36.48 kg of a reddish-brown paprika oil paste, with a color value of 466.92 and an absorbance ratio (A). 470nm / A 454nm The yield was 1.083, and the efficiency was 56.03%.

[0088] f) After the red pigment elution is complete, elute with acetone at a flow rate of 0.8 Bv / h for 750 L to obtain the orange pigment eluent. Concentrate the orange pigment eluent under reduced pressure to obtain 18.31 kg of a slightly orange-toned paprika oil paste, with a color value of 361.77 and an absorbance ratio (A). 470nm / A454nm The yield was 0.949, and the efficiency was 21.79%.

[0089] g) After acetone elution is complete, the medium-pressure chromatography column is subjected to solvent replacement and equilibration using petroleum ether at a rate of 1.5 Bv / h, with a solvent volume of 2250 L (3 Bv). After solvent replacement and equilibration, the next batch of separation production can proceed.

[0090] Example 3

[0091] a) Take 100kg of chili oil paste {color value 267.63, absorbance (A)} 470nm / A 454nm The concentration of β-carotene was 0.979, and the content of β-carotene was 1.03%. 0.5 times the amount of membrane filter water was added, and 1.5% of the weight of lipase was added. The mixture was stirred and reacted at 38°C for 10 hours to obtain the enzymatic hydrolysate of capsicum.

[0092] b) After the enzymatic hydrolysis reaction is complete, add 180 kg of n-hexane to the enzymatic hydrolysate, adjust the color value to 100, stir and dissolve at room temperature for 30 min, let stand for 60 min, and slowly remove the lower aqueous phase; the upper pigment phase is finely filtered through a 0.45 μm organic filter membrane to obtain the enzymatic hydrolysate of capsicum red.

[0093] c) The enzymatically hydrolyzed capsanthin filtrate was subjected to silica gel column chromatography at a flow rate of 2.0 Bv / h using a continuous wet loading method, with a chromatographic pressure of 1.5–1.7 MPa. Elution was performed with 2250 L of n-hexane at a flow rate of 1.5 Bv / h to obtain a β-carotene eluent. The β-carotene eluent was concentrated under reduced pressure to obtain 4.74 kg of an orange-yellow oil paste with a color value of 489.23 and an absorbance ratio (A). 470nm / A 454nm The content of β-carotene was 0.848, with a yield of 9.03% based on capsicum red; the content of β-carotene was 20.38%, with a yield of 93.79% based on β-carotene content.

[0094] d) After β-carotene elution is complete, elute 600 L of a mixed solvent of n-hexane and acetone (mass ratio 8:1) at a flow rate of 1 Bv / h to obtain a yellow pigment eluent. Concentrate the yellow pigment eluent under reduced pressure to obtain 30.36 kg of yellow pigment paste with a color value of 88.77 and an absorbance ratio (A). 470nm / A 454nm The content of β-carotene was 0.841, and the yield was 10.07%; the content of β-carotene was 0.03%, and the yield based on the β-carotene content was 1.02%.

[0095] e) After the yellow pigment elution is complete, continue eluting with a mixed solvent of n-hexane and acetone (mass ratio 8:1) at a flow rate of 1 Bv / h for 1300 L to obtain the red pigment eluent. Concentrate the red pigment eluent under reduced pressure to obtain 34.34 kg of a reddish-brown paprika oleoresin oil, with a color value of 416.02 and an absorbance ratio (A). 470nm / A 454nm The yield was 1.081, and the efficiency was 53.38%.

[0096] f) After the red pigment elution is complete, elute 1000 L of acetone at a flow rate of 1.2 Bv / h to obtain the orange pigment eluent. Concentrate the orange pigment eluent under reduced pressure to obtain 18.49 kg of a slightly orange-toned paprika oil paste, with a color value of 338.94 and an absorbance ratio (A). 470nm / A 454nm The yield was 0.957, and the efficiency was 23.42%.

[0097] g) After acetone elution is complete, the medium-pressure chromatography column is subjected to solvent replacement and equilibration with n-hexane at a rate of 1.2 Bv / h, using 1875 L (2.5 Bv) of solvent. After solvent replacement and equilibration, the next batch of separation production can proceed.

[0098] Comparative Example 1

[0099] a) Take 100kg of chili oil paste {color value 267.63, absorbance (A)} 470nm / A 454nm The concentration of the red pigment was 0.979, and the β-carotene content was 1.03%. 180 kg of n-hexane was added and stirred to dissolve the red pigment. The color value was adjusted to 96, and then filtered through a 300-mesh filter cloth to obtain the paprika red solution.

[0100] b) The capsanthin solution was subjected to atmospheric pressure silica gel column chromatography at a flow rate of 1 Bv / h using a continuous wet loading method. The stationary phase of the column was 60-100 mesh silica gel. Elution was performed with 1500 L of n-hexane at a flow rate of 1 Bv / h to obtain a yellow pigment eluent. The yellow pigment eluent was concentrated under reduced pressure to obtain 43.8 kg of a deep yellow paste with a color value of 138.52 and an absorbance ratio (A). 470nm / A 454nm The content of β-carotene was 0.887, with a yield of 22.68% based on capsicum red, and the content of β-carotene was 2.26%, with a yield of 96.31% based on β-carotene content.

[0101] c) After the yellow pigment elution is complete, elute 1500 L of acetone solvent at a flow rate of 1 Bv / h to obtain the red pigment eluent. Concentrate the red pigment eluent under reduced pressure to obtain 54.4 kg of red pigment paste with a color value of 363.35 and an absorbance ratio (A). 470nm / A 454nmThe yield was 1.021, and the efficiency was 73.83%.

[0102] g) After acetone elution is complete, use nitrogen to force the solvent out of the chromatography column, remove the silica gel from the chromatography column, dry it under reduced pressure at 60℃ and -0.06Mpa until there is no obvious solvent, then activate it at 105℃ for 2 hours, and then pack it into the column for the next batch of separation production.

[0103] Comparative Example 2

[0104] a) Take 100kg of chili oil paste {color value 294.38, absorbance (A)} 470nm / A 454nm The concentration of β-carotene was 0.983, and the content of β-carotene was 1.05%. 0.5 times the amount of membrane filter water was added, and 1.5% of the weight of lipase was added. The mixture was stirred and reacted at 38°C for 10 hours to obtain the enzymatic hydrolysate of capsicum.

[0105] b) After the enzymatic hydrolysis reaction is complete, add 200 kg of vegetable oil extraction solvent to the enzymatic hydrolysate, adjust the color value to 100, stir and dissolve at room temperature for 30 min, let stand for 60 min, and slowly discharge the lower aqueous phase; the upper pigment phase is then filtered through a 300-mesh filter cloth to obtain the enzymatic hydrolysate of paprika oleoresin.

[0106] b) The enzymatically hydrolyzed capsanthin filtrate was subjected to atmospheric pressure silica gel column chromatography at a flow rate of 1.0 Bv / h using a continuous wet loading method. The stationary phase of the column was 200-300 mesh silica gel. Elution was performed with 750 L of vegetable oil extractant at a flow rate of 1.0 Bv / h to obtain yellow pigment eluent I. The yellow pigment eluent was concentrated under reduced pressure to obtain 11.1 kg of orange-yellow oil paste with a color value of 307.76 and an absorbance (A) ratio of [missing value]. 470nm / A 454nm The content of β-carotene was 0.849, with a yield of 11.63% based on capsicum red, and the content of β-carotene was 5.32%, with a yield of 79.48% based on β-carotene content.

[0107] c) Elution was continued using 1500 L of vegetable oil extraction solvent at a flow rate of 1.0 Bv / h to obtain yellow pigment eluent II. The yellow pigment eluent II was concentrated under reduced pressure to obtain 24.51 kg of deep yellow oil paste with a color value of 147.25 and an absorbance ratio (A). 470nm / A 454nm The content of β-carotene was 0.891, with a yield of 12.26% based on capsicum red; the content of β-carotene was 0.70%, with a yield of 16.28% based on β-carotene content.

[0108] d) After the yellow pigment elution is complete, elute 1500 L of acetone solvent at a flow rate of 1 Bv / h to obtain the red pigment eluent. Concentrate the red pigment eluent under reduced pressure to obtain 53.28 kg of red pigment paste, with a color value of 393.83 and an absorbance ratio (A). 470nm / A 454nm The yield was 1.024, and the efficiency was 71.28%.

[0109] e) After acetone elution is complete, nitrogen gas is used to force the solvent out of the chromatography column, the silica gel in the chromatography column is removed, and it is first dried under reduced pressure at 60℃ and -0.06Mpa until there is no obvious solvent. Then it is activated at 105℃ for 2 hours before being packed into the column for the next batch of separation production.

[0110] Table 1 compares the analytical and detection results of the samples prepared in Examples 1-3 and Comparative Examples 1-2.

[0111]

[0112]

[0113] Based on the results in Table 1, we can conclude that:

[0114] (1) The paprika oleoresin in the embodiment can be separated into yellowish (A) 470nm / A 454nm ≤0.855), orange-toned (A) 470nm / A 454nm (0.949–0.957) and reddish (A) 470nm / A 454nm The three hues (≥1.080) can be separated into three shades, while the contrast ratio can only separate them into a yellowish hue (A). 470nm / A 454nm ≤0.890) and reddish (A) 470nm / A 454nm Hue (≥1.020);

[0115] (2) Example A: Reddish paprika 470nm / A 454nm The ratio can be increased to over 1.080, while the comparative A 470nm / A 454nm The reason why it can only reach above 1.020 is because the orange pigment part failed to separate from the red pigment;

[0116] (3) The examples can directly separate β-carotene with a content of more than 20% from the enzymatically hydrolyzed chili oil. The β-carotene content separated in Comparative Example 1 was only 2.31%, while Comparative Example 2, after enzymatic hydrolysis, showed a significant improvement, but the β-carotene content was only 5.32%. The average β-carotene content of the examples was more than 9 times that of Comparative Example 1 and more than 4 times that of Comparative Example 2.

[0117] Based on the above analysis, it is evident that the process of the embodiment has obvious advantages in terms of separation effect and product quality. The 20% β-carotene product produced by this invention can be widely used in food and health food.

[0118] The foregoing description of specific exemplary embodiments of the invention is for illustrative and explanatory purposes. These descriptions are not intended to limit the invention to the precise forms disclosed, and it will be apparent that many changes and variations can be made in accordance with the foregoing teachings. The exemplary embodiments were chosen and described in order to explain the specific principles of the invention and its practical application, thereby enabling those skilled in the art to implement and utilize various different exemplary embodiments of the invention, as well as various different choices and variations. The scope of the invention is intended to be defined by the claims and their equivalents.

Claims

1. A method for preparing β-carotene from capsanthin, characterized by, The method comprises the following steps: (1) enzymatic hydrolysis of the paprika oleoresin to obtain paprika enzymatic hydrolysate; (2) extraction: adding an enzyme hydrolysate extraction solvent to the paprika enzymatic hydrolysate obtained in step (1) to dissolve, standing and separating, taking the upper layer to obtain a paprika enzymatic hydrolysate solution; (3) fine filtration: the paprika enzymatic hydrolysate solution obtained in step (2) is subjected to fine filtration through a filter membrane to obtain an enzymatic paprika filtrate; (4) adsorption: the enzymatic paprika filtrate obtained in step (3) is subjected to chromatographic separation by a medium-pressure silica gel chromatographic column, and the effluent at the port is collected; (5) beta-carotene elution: after the sample loading on the medium-pressure chromatographic column is completed, the first eluent is used for elution, and the eluate is concentrated to obtain a beta-carotene oleoresin; (6) yellow pigment elution: after the beta-carotene elution is completed, the second eluent is used for elution, and the eluate is concentrated to obtain a yellow pigment oleoresin; (7) red pigment elution: after the yellow pigment elution is completed, the third eluent is used for elution, and the eluate is concentrated to obtain a reddish paprika red; (8) orange pigment elution: after the red pigment elution is completed, the fourth eluent is used for elution of the silica gel column, and the eluate is concentrated and combined with the yellow pigment oleoresin to obtain a reddish paprika red; In step (1), the enzymatic hydrolysis specifically comprises: adding membrane filtration water and lipase to the paprika oleoresin, and stirring and reacting, wherein the mass ratio of the paprika oleoresin, the membrane filtration water and the lipase is 1:(0.4-0.6):(0.01-0.02); In step (2), the enzyme hydrolysate extraction solvent comprises one or more of n-hexane, petroleum ether and plant oil extraction solvent; the enzyme hydrolysate extraction solvent is added in an amount to adjust the color value of the paprika enzymatic hydrolysate solution to 60-120; In step (4), the stationary phase of the medium-pressure silica gel chromatographic column is 300-500 mesh chromatographic silica gel, and the mass ratio of the amount of silica gel to the paprika oleoresin is 1:(0.25-0.4); In step (4), the pressure of the medium-pressure silica gel chromatographic column is 0.5-5 MPa; In step (5), the first eluent comprises one or more of n-hexane, petroleum ether and plant oil extraction solvent, the elution volume is 1-5 BV, and the elution speed is 0.5-3 BV / h; In step (6), the second eluent is obtained by mixing an alkane containing 4-7 carbons and acetone at a mass ratio of (8-50):1; In step (7), the third eluent is obtained by mixing an alkane containing 4-7 carbons and acetone at a mass ratio of (8-50):1; In step (8), the fourth eluent is acetone, the elution volume is 1-5 BV, and the elution speed is 0.5-3 BV / h.

2. The process for preparing β-carotene from capsanthin as claimed in claim 1, wherein, After step (8) is completed, step (9) of solvent replacement and balance is further included: After the medium-pressure silica gel chromatographic column is subjected to solvent replacement and balance treatment with a replacement solvent, the next batch of separation production can be carried out.

3. The method for preparing β-carotene from capsanthin according to claim 2, wherein the capsanthin is capsanthin extracted from red pepper. In step (9), the replacement solvent is an alkane containing 4-7 carbons, the solvent replacement and balance volume is 1.5-5 BV, and the flow rate is 0.5-3 BV / h.

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