Liquid chicken oil and duck oil, preparation method thereof and application of liquid chicken oil and duck oil in anti-liver cancer products

Abstract

The invention discloses liquid chicken oil and duck oil as well as a preparation method and application thereof in anti-liver cancer products, and relates to the technical field of oil fractionation, the liquid chicken oil or duck oil is heated to melt, cooled and crystallized, a surfactant and / or electrolyte aqueous solution is added when crystals appear, and the liquid chicken oil or the liquid duck oil is obtained through crystal growing, solid-liquid separation and oil-water separation; the surfactant and / or electrolyte are / is added in the oil fractionation process, so that the physicochemical properties of the chicken oil and the duck oil are improved, and the eating quality of the chicken oil and the duck oil is improved; pharmacological tests find that the prepared liquid chicken oil and liquid duck oil have certain inhibitory activity on liver cancer cells, and are expected to be applied to processing of anti-liver cancer foods or drugs, so that the economic values of the chicken oil and the duck oil are improved.

Description

Technical Field

[0001] The present invention relates to the technical field of oil fractionation, and particularly relates to a liquid chicken oil and duck oil, a preparation method thereof, and an application thereof in anti-hepatocarcinoma products. Background Art

[0002] Animal fats and oils are fats and oils obtained from fatty tissues in animals and are one of the main sources of edible fats and oils. They mainly include lard, beef tallow, mutton tallow, chicken oil, duck oil, fish oil, etc., and are usually in a solid or solidified state at room temperature. Most animal fats and oils contain a large amount of saturated fatty acids, unsaturated fatty acid oleic acid, cholesterol, vitamins and other components. At present, animal fats and oils have a wide range of applications in the food, daily chemical and industrial fields.

[0003] The processing of animal fats and oils can be divided into three levels: crude extraction processing, refining processing and deep processing. The main purpose of crude extraction processing is to separate the fats and oils from animal tissues. Refining processing is to carry out degumming, deacidification, decolorization and deodorization treatment on the crudely extracted animal fats and oils. Deep processing is to better serve the food industry production and improve the product grade. The deep processing of animal fats and oils adopts fractionation technology, blending technology, transesterification rearrangement technology, hydrogenation technology, etc. Among them, the main purpose of oil fractionation is to meet the requirements of food for fats and oils. At present, there are mainly the following three methods for oil fractionation: ① Dry fractionation. The melted fat is frozen, crystallized and separated by pressing. ② Wet fractionation. The fat is dissolved in a large amount of solvent (water or organic solvent), and then part of the high-melting triglycerides are crystallized at a suitable temperature, and then separated by decantation, filtration and centrifugation. The liquid oil yield of wet fractionation is higher than that of dry fractionation, but the production cost is higher. ③ Surfactant fractionation. An aqueous solution containing a surfactant and an electrolyte is added to the partially crystallized oil phase. The crystallized solid fat is preferentially wetted by the surfactant and enters the aqueous solution, and then the solid fat and liquid oil are separated by centrifugation. The separated solid fat is melted into liquid oil and centrifuged to obtain liquid and solid fats and oils, expanding the scope of use of animal fats and oils.

[0004] The acid value and peroxide value are key detection indicators reflecting the quality of animal fats and oils. The acid value refers to the percentage of free fatty acids in the total amount of fats and oils, and the peroxide value reflects the degree of oxidation during the processing, storage and transportation of fats and oils. In the national food safety standard "Edible Animal Fats and Oils" GB 10146-2015, it is stipulated that the acid value (KOH) of animal fats and oils ≤ 2.5 mg / g, and the peroxide value ≤ 0.2 g / 100 g. The present invention aims to prepare liquid chicken oil and liquid duck oil by adopting a suitable oil fractionation method, reduce their acid value and peroxide value, and improve their edible quality and medicinal value. Summary of the Invention

[0005] The technical problem to be solved by the present invention is to provide a preparation method of liquid chicken fat and duck fat, by adding surfactants and / or electrolytes during the fractionation process to prepare liquid chicken fat or liquid duck fat with low acid value and low peroxide value, and applying the obtained liquid chicken fat and liquid duck fat to the processing of anti-hepatocellular carcinoma foods or drugs to improve the economic value of chicken fat and duck fat.

[0006] The technical problem to be solved by the present invention is achieved by the following technical solutions:

[0007] The first object of the present invention is to provide a preparation method of liquid chicken fat and duck fat, heating and melting chicken fat or duck fat, cooling, crystallizing, adding an aqueous solution of surfactant and / or electrolyte when crystals appear, crystal growing, solid-liquid separation, and oil-water separation to obtain liquid chicken fat or liquid duck fat.

[0008] The second object of the present invention is to provide liquid chicken fat or liquid duck fat obtained by the aforementioned preparation method.

[0009] The third object of the present invention is to provide the application of the liquid chicken fat or liquid duck fat in the processing of anti-hepatocellular carcinoma products.

[0010] The beneficial effects of the present invention are as follows: By adding surfactants and / or electrolytes during the oil fractionation process, the present invention improves the physical and chemical properties of chicken fat and duck fat, thereby improving the edible quality of chicken fat and duck fat; and through pharmacological experiments, it is found that the liquid chicken fat and liquid duck fat prepared by the present invention have certain inhibitory activities against hepatocellular carcinoma cells and are expected to be applied to the processing of anti-hepatocellular carcinoma foods or drugs, thereby improving the economic value of chicken fat and duck fat. Description of the Drawings

[0011] Figure 1 Shows the effects of the liquid chicken fat and liquid duck fat prepared in the examples of the present invention on the cell viability of different cancer cells (A) and the cytotoxicity to different hepatocellular carcinoma cells and normal hepatocytes (B). Detailed Embodiments

[0012] In order to make the technical means, creative features, achieved purposes and effects of the present invention easy to understand, the present invention will be further described below in conjunction with specific examples and diagrams.

[0013] The present invention provides a preparation method of liquid chicken fat and duck fat, heating and melting chicken fat or duck fat, cooling, crystallizing, adding an aqueous solution of surfactant and / or electrolyte when crystals appear, crystal growing, solid-liquid separation, and oil-water separation to obtain liquid chicken fat or liquid duck fat.

[0014] Further, the temperature for heating and melting is 55 - 65°C. When setting the heating temperature, it is necessary to not only ensure that the solid fats originally contained in chicken fat and duck fat are completely destroyed, but also avoid the oxidation of chicken fat and duck fat. Because the oxidation of chicken fat and duck fat will destroy the nutritional components and even produce harmful substances, and also for the consideration of energy consumption.

[0015] Further, the cooling and crystallization is carried out by cooling at a rate of 1 - 10°C / h to 20 - 25°C for crystallization. Preferably, the cooling and crystallization is carried out by cooling at a rate of 2 - 5°C / h to 20 - 25°C for crystallization. The cooling rate and the cooling temperature are decisive conditions affecting the crystallization process, which not only affect the fractionation effect of liquid oils and fats, but also affect the stability and application effect of liquid oils and fats.

[0016] Further, the surfactant is one or more of Tween and sucrose esters. The main functions of surfactants in the fractionation of oils and fats are emulsification and wetting. On the one hand, surfactants can reduce the surface tension at the oil - water interface, making the oil form uniform fine droplets in water to ensure the uniform distribution of the oil during the fractionation process; on the other hand, surfactants can reduce the surface tension of the liquid on the solid surface, and through the wetting effect, the liquid oil is separated from the solid surface and emulsified and dispersed in water.

[0017] Further, the electrolyte is one or more of sodium chloride, magnesium sulfate, and sodium sulfate. The main functions of electrolytes in the fractionation of oils and fats are to inhibit excessive emulsification and promote crystal growth.

[0018] Further, the concentration of the surfactant in the aqueous solution is 0.5 - 1.5 wt%. The surfactant or the electrolyte can be added alone, or the surfactant and the electrolyte can be added simultaneously, and the surfactant and the electrolyte are added in the form of an aqueous solution.

[0019] Further, the concentration of the electrolyte in the aqueous solution is 1 - 3 wt%.

[0020] Further, the mass ratio of the chicken fat or duck fat to the aqueous solution is 1:(0.5 - 1). The amount of water used affects the separation effect of liquid oils and fats on the one hand and determines the amount of waste liquid generated on the other hand. Therefore, it is necessary to control the amount of the aqueous solution and the concentrations of the surfactant and the electrolyte in the aqueous solution.

[0021] Further, the time for crystal ripening is 1 - 24 h. Preferably, the time for crystal ripening is 4 - 8 h.

[0022] In the present invention, the solid - liquid separation method is preferably centrifugal separation, which has the advantages of high separation efficiency and good separation effect; first, the solid and liquid are separated by solid - liquid separation, and then the liquid oil is separated by oil - water separation.

[0023] The present invention provides a liquid chicken fat or liquid duck fat obtained by the foregoing preparation method.

[0024] The present invention provides the application of the liquid chicken fat or liquid duck fat in the processing of anti-hepatocellular carcinoma products.

[0025] Furthermore, the anti-hepatocellular carcinoma products include anti-hepatocellular carcinoma foods and anti-hepatocellular carcinoma drugs, that is, the liquid chicken fat or liquid duck fat can be used for the processing of foods or drugs with anti-hepatocellular carcinoma efficacy, and it has the characteristics of wide sources and high safety.

[0026] In the following examples, the raw chicken fat and duck fat are the chicken fat and duck fat provided by Anhui Tianxiang Grain and Oil Food Co., Ltd., which are prepared from the same batch of chicken and duck fat by the same crude extraction and refining process.

[0027] Example 1

[0028] Heat 100 g of chicken fat to 60 °C until it melts completely, cool it to 25 °C at a cooling rate of 5 °C / h for crystallization. When crystals appear, add 50 g of a Tween 20 aqueous solution (concentration 0.5 wt%), carry out crystal growth for 8 h, separate solid from liquid, and separate oil from water to obtain liquid chicken fat.

[0029] Example 2

[0030] Heat 100 g of chicken fat to 55 °C until it melts completely, cool it to 23 °C at a cooling rate of 2 °C / h for crystallization. When crystals appear, add 100 g of a sucrose ester SE15 aqueous solution (concentration 1 wt%), carry out crystal growth for 4 h, separate solid from liquid, and separate oil from water to obtain liquid chicken fat.

[0031] Example 3

[0032] Heat 100 g of chicken fat to 65 °C until it melts completely, cool it to 20 °C at a cooling rate of 3 °C / h for crystallization. When crystals appear, add 100 g of a sodium chloride aqueous solution (concentration 1 wt%), carry out crystal growth for 6 h, separate solid from liquid, and separate oil from water to obtain liquid chicken fat.

[0033] Example 4

[0034] Heat 100 g of duck fat to 60 °C until it melts completely, cool it to 24 °C at a cooling rate of 4 °C / h for crystallization. When crystals appear, add 100 g of a magnesium sulfate aqueous solution (concentration 2 wt%), carry out crystal growth for 5 h, separate solid from liquid, and separate oil from water to obtain liquid duck fat.

[0035] Example 5

[0036] Heat 100 g of duck fat to 60 °C until it melts completely, cool it to 25 °C at a cooling rate of 5 °C / h for crystallization. When crystals appear, add 100 g of a sodium sulfate aqueous solution (concentration 3 wt%), carry out crystal growth for 8 h, separate solid from liquid, and separate oil from water to obtain liquid duck fat.

[0037] Example 6

[0038] Heat 100 g of chicken fat to 60 °C until it melts completely, cool it to 22 °C at a cooling rate of 4 °C / h for crystallization. When crystals appear, add 100 g of an aqueous solution prepared from Tween 20 (concentration 1 wt%) and sodium chloride (concentration 2 wt%), carry out crystal growth for 7 h, separate solid from liquid, and separate oil from water to obtain liquid chicken fat.

[0039] Example 7

[0040] Heat 100 g of duck fat to 60 °C until it melts completely, cool it to 23 °C at a cooling rate of 5 °C / h for crystallization. When crystals appear, add 100 g of an aqueous solution prepared from sucrose ester SE15 (concentration 1 wt%) and sodium sulfate (concentration 3 wt%), carry out crystal growth for 6 h, separate solid from liquid, and separate oil from water to obtain liquid duck fat.

[0041] Measure the acid value and peroxide value of the liquid chicken fat and liquid duck fat prepared in the above Examples 1 - 7, and the results are shown in Table 1.

[0042] Determine the acid value according to the second method of Standard GB 5009.229 - 2016;

[0043] Determine the peroxide value according to the second method of Standard GB 5009.227 - 2023.

[0044] Table 1

[0045] Acid value (mg / g) Peroxide value (g / 100g) Example 1 0.93 0.08 Example 2 0.97 0.09 Example 3 1.05 0.12 Example 4 0.12 0.10 Example 5 0.10 0.08 Example 6 0.84 0.06 Example 7 0.08 0.05

[0046] As can be seen from Table 1, the acid values and peroxide values of the liquid chicken fat and liquid duck fat prepared in Examples 1 - 7 all meet the national food safety standard for animal fats GB 10146 - 2015.

[0047] In order to further optimize the fractionation effect of liquid chicken fat and liquid duck fat, the inventors abandoned the commonly used surfactants in the art and screened new surfactants for the fractionation of liquid chicken fat and liquid duck fat.

[0048] Preferably, the surfactant is Boc-L-lysine.

[0049] Preferably, the surfactant is tert-butyl 2-(tert-butoxycarbonylamino)-6-hydroxyhexanoate.

[0050] The inventors found through research that Boc-L-lysine and tert-butyl 2-(tert-butoxycarbonylamino)-6-hydroxyhexanoate can replace Tween and sucrose ester as surfactants for the fractionation of liquid chicken fat and liquid duck fat to obtain higher-quality liquid chicken fat and liquid duck fat.

[0051] Example 8

[0052] Example 8 only replaces Tween 20 in Example 6 with Boc-L-lysine.

[0053] Heat 100 g of chicken fat to 60 °C until it melts completely, cool it to 22 °C at a cooling rate of 4 °C / h for crystallization. When crystals appear, add 100 g of an aqueous solution prepared from Boc-L-lysine (concentration 1 wt%) and sodium chloride (concentration 2 wt%), carry out crystal growth for 7 h, perform solid-liquid separation and oil-water separation to obtain liquid chicken fat.

[0054] According to the above standards, the acid value of the liquid chicken fat is 0.75 mg / g and the peroxide value is 0.04 g / 100 g.

[0055] Example 9

[0056] Example 9 only replaces sucrose ester SE15 in Example 7 with tert-butyl 2-(tert-butoxycarbonylamino)-6-hydroxyhexanoate.

[0057] Heat 100 g of duck fat to 60 °C until it melts completely, cool it to 23 °C at a cooling rate of 5 °C / h for crystallization. When crystals appear, add 100 g of an aqueous solution prepared from tert-butyl 2-(tert-butoxycarbonylamino)-6-hydroxyhexanoate (concentration 1 wt%) and sodium sulfate (concentration 3 wt%), carry out crystal growth for 6 h, perform solid-liquid separation and oil-water separation to obtain liquid duck fat.

[0058] According to the above standards, the acid value of the liquid duck fat is 0.05 mg / g and the peroxide value is 0.03 g / 100 g.

[0059] Perform MTT experiments on the liquid chicken fat prepared in Example 6 and the liquid duck fat prepared in Example 7. The specific steps are as follows:

[0060] (1) Prepare stock solutions of liquid chicken fat and liquid duck fat at a concentration of 500 mg / mL with dimethyl sulfoxide (DMSO) respectively, pass them through a microporous membrane with a pore size of 0.22 μm, and then dilute the stock solutions with complete medium to 1, 3, 5, and 7 mg / mL.

[0061] (2) When cancer cells or normal cells grow to the logarithmic growth phase, treat them with trypsin-EDTA solution, add cell suspension to a 96-well plate, 6000 - 11000 cells / well. After culturing for 24 h, discard the old medium, then add serum-free culture medium and continue culturing for 12 h. Discard the serum-free culture medium, and then add 100 μL of chicken fat or duck fat with concentrations of 1, 3, 5, and 7 mg / mL respectively, and continue culturing for 48 h. At the same time, set a zero-adjustment group with only culture medium, a blank control group with only cells and culture medium, and a solvent DMSO control group, with 4 parallels in each group.

[0062] (3) After culturing for 48 h, add 20 μL of MTT solution with a concentration of 5 mg / mL to each well, continue culturing for 4 h, discard the culture medium, add 150 μL of DMSO, place it on a shaker and shake at a low speed for 10 min, measure the absorbance value at a wavelength of 490 nm with a multifunctional microplate reader, and calculate the IC 50 .

[0063] Figure 1 A shows the effects of liquid chicken fat and liquid duck fat on the cell viability of different cancer cells. From Figure 1 A, it can be seen that liquid chicken fat and duck fat can inhibit the cell viability of hepatocarcinoma cell line H22, lung cancer cell lines PC-9 and PC-9 / GR. Among them, the IC 50 values of chicken fat and duck fat for hepatocarcinoma cell line H22 are 4.53 and 4.48 mg / mL respectively, and the IC 50 values for lung cancer cell line PC-9 are 4.93 and 4.45 mg / mL respectively, and they have a lower IC 50 for the cytotoxicity to hepatocarcinoma cell line H22.

[0064] Figure 1 B shows the cytotoxicity of liquid chicken fat and liquid duck fat to different hepatocarcinoma cells and normal hepatocytes. From Figure 1 B, it can be seen that liquid chicken fat and liquid duck fat can inhibit the cell viability of hepatocarcinoma cell lines SK-Hep-1, HepG2 and HuH-7, while having no significant effect on the cell viability of normal hepatocyte line HL-7702.

[0065] The above shows and describes the basic principles, main features and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited by the above embodiments. What is described in the above embodiments and the specification only illustrates the principle of the present invention. Without departing from the spirit and scope of the present invention, the present invention will have various changes and improvements, and these changes and improvements all fall within the scope of the present invention claimed. The scope of the present invention claimed is defined by the appended claims and their equivalents.

Claims

1. A method for preparing liquid chicken fat and duck fat, characterized in that: The chicken fat or duck fat is heated to melt, cooled, and crystallized. When crystals appear, a surfactant and / or an aqueous solution of an electrolyte is added, and the crystals are grown, and the solid-liquid separation and the oil-water separation are performed to obtain liquid chicken fat or liquid duck fat.

2. The method for preparing liquid chicken fat and duck fat as claimed in claim 1, characterized in that: The heating and melting temperature is 55-65°C.

3. The method for preparing liquid chicken fat and duck fat as claimed in claim 1, characterized in that: The cooling crystallization is performed by cooling the temperature to 20-25°C at a cooling rate of 1-10°C / h; preferably, the cooling crystallization is performed by cooling the temperature to 20-25°C at a cooling rate of 2-5°C / h.

4. The method for preparing liquid chicken fat and duck fat as claimed in claim 1, characterized in that: The surfactant is one or more of Tween and sucrose fatty acid ester; preferably, the concentration of the surfactant in the aqueous solution is 0.5-1.5 wt %.

5. The method for preparing liquid chicken fat and duck fat as claimed in claim 1, characterized in that: The electrolyte is one or more of sodium chloride, magnesium sulfate, and sodium sulfate; preferably, the concentration of the electrolyte in the aqueous solution is 1-3 wt %.

6. The method for preparing liquid chicken fat and duck fat as claimed in claim 1, characterized in that: The mass ratio of the chicken fat or duck fat to the aqueous solution is 1:(0.5-1).

7. The method for preparing liquid chicken fat and duck fat as claimed in claim 1, characterized in that: The crystal growing time is 1 to 24 hours; preferably, the crystal growing time is 4 to 8 hours.

8. Liquid chicken fat or liquid duck fat obtained by the method for preparing liquid chicken fat or duck fat according to any one of claims 1 to 7.

9. Use of the liquid chicken fat or liquid duck fat according to claim 8 in the processing of anti-liver cancer products.

10. The use according to claim 9, characterized in that: The anti-liver cancer products include anti-liver cancer foods and anti-liver cancer medicines.

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