Method for separating and purifying ceramide from hydration degumming byproduct

Abstract

The invention discloses a method for separating ceramide from hydrated degumming by-products, which comprises the following steps: (1) adding a certain amount of C1-C3 alcohol or C3-C5 ketone solvent into the hydrated degumming by-products containing ceramide to ensure that the concentration of alcohol or ketone in the system is more than or equal to 30%, centrifuging and collecting precipitate; and (2) adding 80%-100% by volume of C1-C3 alcohol or C3-C5 ketone solvent into the precipitate obtained in the step (1) for extraction, separation and concentration of a supernatant to obtain a ceramide product. The content of ceramide separated by the method can reach 9% or above, efficient conversion of byproducts is achieved, and the comprehensive utilization rate of resources is greatly increased.

Description

Technical Field

[0001] The invention relates to a method for separating and purifying ceramide from hydration degumming by-products. Background Art

[0002] Ceramide is a bioactive substance with important physiological functions and has great application value in the fields of skin science, medicine and food. With the improvement of living standards, people's demand for natural, safe and effective products continues to increase. Ceramide from natural sources is highly favored, which not only meets people's pursuit of health and beauty, but also conforms to the current concept of green and sustainable development.

[0003] At present, the relevant patent sources in the field of natural ceramide preparation are mainly konjac and there are many defects. For example, the patent with announcement number CN111499537B involves steps such as salting out, primary and secondary separation and crystallization of polymer magnetic microspheres, and the process is complicated. The patent application with publication number CN108947864A, when preparing ceramide, although ethanol is used to soak and remove the odorous substances of konjac powder, the solid is dried and used as feed, and the resource utilization is insufficient, and potassium hydroxide is needed to neutralize the acidic substances. The patent application with application number 201010254120.6 adopts organic reagents such as petroleum ether and ethyl acetate throughout the process, which is costly, toxic, flammable and explosive, and has prominent safety hazards. The patent application with application number 201110327965.8 adopts medium-pressure C18 column, which is only suitable for small-scale preparation and is difficult to meet large-scale production needs. The patent application with application number 2010105989816 needs to be extracted with ethanol for many times, and then repeatedly dissolved and extracted with ethyl acetate and petroleum ether, and the process is cumbersome.

[0004] In the process of oil processing, hydration degumming will produce by-products. In the past, these by-products were often regarded as waste and not fully utilized. With the advancement of technology and the emphasis on comprehensive resource utilization, it has become possible to separate and purify high-value ceramides from hydration degumming by-products. If ceramides can be effectively separated and purified from these by-products, it will not only achieve high-value utilization of by-products and improve the comprehensive utilization rate of resources, but also open up new ways for the production of ceramides. In view of this, it is particularly important to develop a method for separating and purifying ceramides from hydration degumming by-products. Summary of the invention

[0005] The present invention aims to provide a method for separating ceramide from hydration degumming by-products, wherein the purity of the separated ceramide can reach more than 9%, thereby achieving efficient conversion of the by-products and greatly improving the comprehensive utilization rate of resources.

[0006] To achieve the above purpose, the technical solution of the present invention is as follows:

[0007] A method for separating ceramide from hydration degumming by-products comprises the following steps:

[0008] (1) adding a C1-C3 alcohol or a C3-C5 ketone solvent to the hydrated degumming byproduct containing ceramide to make the concentration of the alcohol or ketone in the system ≥30%, centrifuging, and collecting the precipitate;

[0009] (2) adding 80% to 100% by volume of a C1-C3 alcohol or C3-C5 ketone solvent to the above precipitate for extraction, separation, and concentrating the supernatant to obtain a ceramide product.

[0010] Furthermore, the raw material sources of the hydration degumming by-products of the present invention include but are not limited to soybeans, rice bran, camellia seeds, and peppers.

[0011] Preferably, the concentration of alcohol or ketone in the system of step (1) of the present invention is further optimized to 30%-80%.

[0012] Furthermore, in step (2) of the present invention, the mass volume ratio of the precipitate to the extraction solvent (kg:L) is 1:1-10.

[0013] Furthermore, the extraction time in step (2) of the present invention is 0.5 to 1 h.

[0014] Furthermore, the concentration temperature in step (2) of the present invention is 60-70°C.

[0015] The beneficial technical effects of the technical solution of the present invention are:

[0016] The present invention provides a simple method for separating and purifying ceramide. The separation of ceramide can be achieved by controlling the alcohol / ketone concentration in the system and the volume fraction of the extraction solvent, and the content of the separated ceramide is greater than 9% and the yield is greater than 50%.

[0017] Compared with other solutions in the prior art, the method of the present invention is simple to operate, short in steps, low in cost, and suitable for industrial large-scale production. In addition, the method of the present invention achieves efficient conversion of by-products, greatly improving the comprehensive utilization rate of resources. DETAILED DESCRIPTION

[0018] In order to make the purpose, technical solution and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with the embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0019] The content of ceramide in the present invention is detected by high performance liquid chromatography tandem time-of-flight mass spectrometry as follows: mobile phase: phase A is 1% formic acid-water solution, phase B is 1% formic acid-50% acetonitrile-50% isopropanol, and gradient elution is used for analysis (gradient elution conditions are shown in Table 1); control flow rate during elution: 0.2 mL / min; column temperature: 40°C; chromatographic column: BEH C18 column (2.1 mm×150 mm, particle size 1.7 μm); injection volume: 2.0 μL. The retention time of the ceramide component appears at 70-85.5 min.

[0020] Table 1 Gradient elution conditions during ceramide content detection

[0021] Time (min) Flow rate (mL / min) Phase A (%) Phase B (%) 0.00 0.2 40.0 60.0 2.00 0.2 40.0 60.0 50.00 0.2 16.0 84.0 120.00 0.2 16.0 84.0 121.00 0.2 0.0 100.0 122.00 0.2 40.0 60.0 124.00 0.2 40.0 60.0

[0022] The content of the ceramide in the present invention is measured using commercially available natural plant-derived ceramide (Shaanxi Yihaochuan Biotechnology Co., Ltd., ceramide content 5%) as a reference substance.

[0023] Example 1

[0024] 1) Take 10 L of hydrated degumming byproduct containing ceramide, the raw material of the hydrated degumming byproduct is derived from pepper, the mass concentration of solids in the hydrated degumming byproduct is 50 g / L, and the mass percentage of ceramide in the solid is 4.8%, add 40 L of methanol, stir evenly, centrifuge, and collect 0.58 kg of precipitate.

[0025] 2) Add 2.9 L of 90% methanol by volume to the above precipitate for extraction. After extraction for 1 hour, centrifuge and collect the supernatant. Concentrate the supernatant at 65° C. to obtain 0.146 kg of ceramide product.

[0026] After testing, the content of ceramide in the ceramide product obtained in this example is 10.7%. After calculation, it can be known that the yield of ceramide in this example is 65.2%.

[0027] Example 2

[0028] 1) Take 10 L of hydrated degumming byproduct containing ceramide, the raw material of the hydrated degumming byproduct is derived from pepper, the mass concentration of solids in the hydrated degumming byproduct is 50 g / L, and the mass percentage of ceramide in the solid is 4.8%, add 4.3 L of methanol, stir evenly, and collect 0.25 kg of precipitate by centrifugation.

[0029] 2) Add 1.25 L of 90% methanol by volume to the above precipitate for extraction. Centrifuge after extraction for 1 hour, collect the supernatant, and concentrate the supernatant at 65° C. to obtain 0.1 kg of ceramide product.

[0030] After testing, the content of ceramide in the ceramide product obtained in this example is 12.1%. After calculation, it can be known that the yield of ceramide in this example is 50.6%.

[0031] Example 3

[0032] 1) Take 10 L of hydrated degumming byproduct containing ceramide, the raw material of the hydrated degumming byproduct is derived from pepper, the mass concentration of solids in the hydrated degumming byproduct is 50 g / L, and the mass percentage of ceramide in the solid is 4.8%, add 10 L of methanol, stir evenly, and collect 0.4 kg of precipitate by centrifugation.

[0033] 2) Add 2 L of 90% methanol by volume to the above precipitate for extraction. After extraction for 1 hour, centrifuge and collect the supernatant. Concentrate the supernatant at 65° C. to obtain 0.12 kg of ceramide product.

[0034] According to the test, the content of ceramide in the ceramide product obtained in this example is 11.5%. According to the calculation, the yield of ceramide in this example is 57.7%.

[0035] Example 4

[0036] 1) 10 L of hydrated degumming byproduct containing ceramide is taken, wherein the raw material of the hydrated degumming byproduct is derived from soybean, the mass concentration of solids in the hydrated degumming byproduct is 85 g / L, and the mass percentage of ceramide in the solids is 3.4%, 40 L of methanol is added, and after stirring evenly, 0.58 kg of precipitate is collected by centrifugation.

[0037] 2) Add 2.9 L of 90% methanol by volume to the above precipitate for extraction. Centrifuge after extraction for 1 hour, collect the supernatant, and concentrate the supernatant at 65° C. to obtain 0.207 kg of ceramide product.

[0038] According to the test, the content of ceramide in the ceramide product obtained in this example is 9.5%. According to the calculation, the yield of ceramide in this example is 68.2%.

[0039] Example 5

[0040] 1) 10 L of hydrated degumming byproduct containing ceramide is taken, wherein the raw material of the hydrated degumming byproduct is derived from camellia seeds, the mass concentration of solids in the hydrated degumming byproduct is 63 g / L, the mass percentage of ceramide in the solids is 3.7%, and the solid content in the byproduct is 5%, 40 L of methanol is added, stirred evenly, and 0.58 kg of precipitate is collected by centrifugation.

[0041] 2) Add 2.9 L of 100% methanol by volume to the above precipitate for extraction. Centrifuge after 0.5 h of extraction, collect the supernatant, and concentrate the supernatant at 65° C. to obtain 0.126 kg of ceramide product.

[0042] According to the test, the content of ceramide in the ceramide product obtained in this example is 11.4%. According to the calculation, the yield of ceramide in this example is 61.5%.

[0043] Example 6

[0044] 1) 10 L of hydrated degumming byproduct containing ceramide is taken, wherein the raw material of the hydrated degumming byproduct is derived from camellia seeds, the mass concentration of solids in the hydrated degumming byproduct is 63 g / L, and the mass percentage of ceramide in the solids is 3.7%, 40 L of ethanol is added, stirred evenly, and 0.55 kg of precipitate is collected by centrifugation.

[0045] 2) Add 2.75 L of 90% ethanol by volume to the above precipitate for extraction. Centrifuge after extraction for 1 hour, collect the supernatant, and concentrate the supernatant at 65° C. to obtain the ceramide product.

[0046] According to the test, the content of ceramide in the ceramide product obtained in this example is 10.9%. According to the calculation, the yield of ceramide in this example is 63.5%.

[0047] Example 7

[0048] 1) 10 L of hydrated degumming byproduct containing ceramide is taken, wherein the raw material of the hydrated degumming byproduct is derived from rice bran, the mass concentration of solids in the hydrated degumming byproduct is 45 g / L, and the mass percentage of ceramide in the solids is 6.3%, 40 L of propanol is added, the mixture is stirred evenly, and 0.55 kg of precipitate is collected by centrifugation.

[0049] 2) Add 2.75 L of 90% propanol by volume to the above precipitate for extraction. After extraction for 1 hour, centrifuge and collect the supernatant. Concentrate the supernatant at 65° C. to obtain 0.158 kg of ceramide product.

[0050] According to the test, the content of ceramide in the ceramide product obtained in this example is 11.2%. According to the calculation, the yield of ceramide in this example is 62.4%.

[0051] Example 8

[0052] 1) 10 L of hydrated degumming byproduct containing ceramide is taken, wherein the raw material of the hydrated degumming byproduct is derived from camellia seeds, the mass concentration of the solid in the hydrated degumming byproduct is 63 g / L, and the mass percentage of ceramide in the solid is 3.7%, 40 L of acetone is added, stirred evenly, and 0.53 kg of precipitate is collected by centrifugation.

[0053] 2) Add 2.65 L of 90% by volume acetone to the above precipitate for extraction. After extraction for 1 hour, centrifuge and collect the supernatant. Concentrate the supernatant at 65° C. to obtain 0.122 kg of ceramide product.

[0054] According to the test, the content of ceramide in the ceramide product obtained in this example is 11.6%. According to the calculation, the yield of ceramide in this example is 60.7%.

[0055] Example 9

[0056] 1) 10 L of hydrated degumming byproduct containing ceramide is taken, wherein the raw material of the hydrated degumming byproduct is derived from pepper, the mass concentration of the solid in the hydrated degumming byproduct is 50 g / L, and the mass percentage of ceramide in the solid is 4.8%, 40 L of butanone is added, stirred evenly, and 0.53 kg of precipitate is collected by centrifugation.

[0057] 2) Add 2.65 L of 90% by volume butanone to the above precipitate for extraction. After extraction for 1 hour, centrifuge and collect the supernatant. Concentrate the supernatant at 65° C. to obtain 0.113 kg of ceramide product.

[0058] After testing, the content of ceramide in the ceramide product obtained in this example is 12.2%. After calculation, it can be known that the yield of ceramide in this example is 57.3%.

[0059] Example 10

[0060] 1) 10 L of hydrated degumming byproduct containing ceramide is taken, wherein the raw material of the hydrated degumming byproduct is derived from camellia seeds, the mass concentration of the solid in the hydrated degumming byproduct is 63 g / L, and the mass percentage of ceramide in the solid is 3.7%, 40 L of pentanone is added, the mixture is stirred evenly, and 0.53 kg of precipitate is collected by centrifugation.

[0061] 2) Add 2.65 L of 90% by volume pentanone to the above precipitate for extraction. After extraction for 1 hour, centrifuge and collect the supernatant. Concentrate the supernatant at 65° C. to obtain 0.102 kg of ceramide product.

[0062] According to the test, the content of ceramide in the ceramide product obtained in this example is 12.5%. According to the calculation, the yield of ceramide in this example is 54.9%.

[0063] Embodiment 11

[0064] 1) 10 L of hydrated degumming byproduct containing ceramide is taken, wherein the raw material of the hydrated degumming byproduct is derived from rice bran, the mass concentration of solids in the hydrated degumming byproduct is 45 g / L, and the mass percentage of ceramide in the solids is 6.3%, 40 L of methanol is added, and after stirring evenly, 0.58 kg of precipitate is collected by centrifugation.

[0065] 2) Add 0.58 L of 90% methanol by volume to the above precipitate for extraction. Centrifuge after extraction for 1 hour, collect the supernatant, and concentrate the supernatant at 65° C. to obtain 0.139 kg of ceramide product.

[0066] According to the test, the content of ceramide in the ceramide product obtained in this example is 11%. According to the calculation, the yield of ceramide in this example is 53.8%.

[0067] Example 12

[0068] 1) 10 L of hydrated degumming byproduct containing ceramide is taken, wherein the raw material of the hydrated degumming byproduct is derived from rice bran, the mass concentration of solids in the hydrated degumming byproduct is 45 g / L, and the mass percentage of ceramide in the solids is 6.3%, 40 L of methanol is added, and after stirring evenly, 0.58 kg of precipitate is collected by centrifugation.

[0069] 2) Add 5.8 L of 80% methanol by volume to the above precipitate for extraction. Centrifuge after extraction for 1 hour, collect the supernatant, and concentrate the supernatant at 65° C. to obtain 0.237 kg of ceramide product.

[0070] According to the test, the content of ceramide in the ceramide product obtained in this example is 9%. According to the calculation, the yield of ceramide in this example is 75.3%.

[0071] Example 13

[0072] 1) Take 10 L of hydrated degumming byproduct containing ceramide, the raw material of the hydrated degumming byproduct is derived from pepper, the mass concentration of solids in the hydrated degumming byproduct is 50 g / L, and the mass percentage of ceramide in the solid is 4.8%, add 40 L of methanol, stir evenly, and collect 0.58 kg of precipitate by centrifugation.

[0073] 2) Add 2.65 L of 90% by volume acetone to the above precipitate for extraction. After extraction for 1 hour, centrifuge and collect the supernatant. Concentrate the supernatant at 65° C. to obtain 0.143 kg of ceramide product.

[0074] According to the test, the content of ceramide in the ceramide product obtained in this example is 10.7%. According to the calculation, the yield of ceramide in this example is 63.7%.

[0075] Comparative Example 1

[0076] 1) Take 10 L of hydrated degumming byproduct containing ceramide, the raw material of the hydrated degumming byproduct is derived from pepper, the mass concentration of solids in the hydrated degumming byproduct is 50 g / L, and the mass percentage of ceramide in the solids is 4.8%, add 0.526 L of methanol, stir evenly, and collect 0.05 kg of precipitate by centrifugation.

[0077] 2) Add 0.25 L of 90% methanol by volume to the above precipitate for extraction. Centrifuge after extraction for 1 hour, collect the supernatant, and concentrate the supernatant at 65° C. to obtain 0.011 kg of ceramide product.

[0078] According to the test, the content of ceramide in the ceramide product obtained in this comparative example is 9.4%. According to the calculation, the yield of ceramide in this comparative example is 4.25%.

[0079] Comparative Example 2

[0080] 1) Take 10 L of hydrated degumming byproducts, wherein the raw material of the hydrated degumming byproducts is derived from peppers, the mass concentration of solids in the hydrated degumming byproducts is 50 g / L, and the mass percentage of ceramide in the solids is 4.8%, add 40 L of methanol, stir evenly, and collect 0.58 kg of precipitate by centrifugation.

[0081] 2) Add 2.9 L of water to the above precipitate for extraction. After extraction for 1 hour, centrifuge and collect the supernatant. Concentrate the supernatant at 65° C. to obtain 0.323 kg of ceramide product.

[0082] According to the test, the content of ceramide in the ceramide product obtained in this comparative example is 5.3%. According to the calculation, the yield of ceramide in this comparative example is 71.3%.

[0083] Comparative Example 3

[0084] 1) Take 10 L of hydrated degumming byproducts, wherein the raw material of the hydrated degumming byproducts is derived from peppers, the mass concentration of solids in the hydrated degumming byproducts is 50 g / L, and the mass percentage of ceramide in the solids is 4.8%, add 40 L of methanol, stir evenly, and collect 0.58 kg of precipitate by centrifugation.

[0085] 2) 17.4 L of 90% by volume methanol was added to the above precipitate for extraction. After extraction for 1 hour, the mixture was centrifuged, the supernatant was collected, and the supernatant was concentrated at 65° C. to obtain 0.285 kg of ceramide product.

[0086] According to the test, the content of ceramide in the ceramide product obtained in this comparative example is 6.2%. According to the calculation, the yield of ceramide in this comparative example is 73.5%.

[0087] Comparative Example 4

[0088] 1) Take 10 L of hydrated degumming byproducts, wherein the raw material of the hydrated degumming byproducts is derived from peppers, the mass concentration of solids in the hydrated degumming byproducts is 50 g / L, and the mass percentage of ceramide in the solids is 7.66%, add 40 L of methanol, stir evenly, and collect 0.58 kg of precipitate by centrifugation.

[0089] According to the test, the content of ceramide in the ceramide product obtained in this comparative example is 5.5%. According to the calculation, the yield of ceramide in this comparative example is 83.3%.

[0090] It can be seen from the above embodiments and comparative examples that the method of the present invention can well separate the ceramide component in the by-product of hydration degumming. If the method of the present invention is not adopted, for example, in comparative example 1, the alcohol concentration in the first extraction system is controlled to 5%, the yield of ceramide is significantly reduced; in comparative example 2, the extraction solvent in step (2) is changed to water, in comparative example 3, the volume ratio of the precipitate to the extraction solvent is controlled to 1:30, and in comparative example 4, only one extraction is performed, and the purity of ceramide is significantly reduced. It shows that the technical solution of the present invention has outstanding technical effects.

[0091] The above is only a specific embodiment of the present invention, but the protection scope of the present invention is not limited thereto. Any changes or substitutions that can be easily thought of by a person skilled in the art within the technical scope disclosed by the present invention should be included in the protection scope of the present invention. Therefore, the protection scope of the present invention should be based on the protection scope of the claims.

Claims

1. A method for separating ceramide from hydration degumming by-products, characterized in that: The following steps are involved: (1) adding a certain amount of C1-C3 alcohol or C3-C5 ketone solvent to the hydrated degumming byproduct containing ceramide so that the concentration of alcohol or ketone in the system is ≥30%, centrifuging and collecting the precipitate; (2) adding 80% to 100% by volume of a C1-C3 alcohol or C3-C5 ketone solvent to the precipitate of step (1) for extraction, separation, and concentrating the supernatant to obtain a ceramide product.

2. A method for separating ceramide from hydration degumming by-products according to claim 1, characterized in that: The raw material sources of the hydration degumming by-products include but are not limited to soybeans, rice bran, camellia seeds, and peppers.

3. The method for separating ceramide from hydration degumming byproducts according to claim 1, characterized in that: The concentration of alcohol or ketone in the system of step (1) is 30%-80%.

4. The method for separating ceramide from hydration degumming byproducts according to claim 1, characterized in that: The mass volume ratio of the precipitate to the extraction solvent in step (2) is kg:L in a range of 1:1 to 10.

5. The method for separating ceramide from hydration degumming byproducts according to claim 1, characterized in that: The extraction time in step (2) is 0.5 to 1 h.

6. The method for separating ceramide from hydration degumming byproducts according to claim 1, characterized in that: The concentration temperature in step (2) is 60-70°C.