Preparation method of liposome for encapsulating euphausia superba oil

Abstract

The invention discloses a preparation method of liposome for encapsulating euphausia superba oil, and belongs to the field of euphausia superba oil. Through material selection and hierarchical structure design, starch octenyl succinate, sodium alginate and pea protein are screened as composite wall materials, and the materials can achieve a synergistic effect on the molecule-interface-micro-nano scale to delay lipid oxidation, control the targeted release characteristic of active ingredients and improve the absorption efficiency of fat-soluble ingredients. Moreover, the liposome raw material of the euphausia superba oil is screened, and the liposome with low particle size and high stability is prepared.

Description

TECHNICAL FIELD

[0001] The application belongs to the field of Antarctic krill oil, and particularly relates to a preparation method of liposomes for encapsulating Antarctic krill oil. BACKGROUND

[0002] As a functional raw material rich in phospholipids (40%-80%) and omega-3 polyunsaturated fatty acids (EPA / DHA), Antarctic krill oil has physiological functions such as relieving inflammation and protecting the cardiovascular system, but its industrial application is limited by three major problems: 1) Poor stability: The double bond of omega-3 polyunsaturated fatty acids in krill oil is easily oxidized, and aldehydes and ketones harmful substances are generated under light and high temperature; at the same time, the active ingredients contained therein are sensitive to pH, and the gastric acid environment (pH 1-3) can cause phospholipid molecules to hydrolyze and break, resulting in a significant decrease in the retention rate of active ingredients in a short period of time.

[0003] 2) Low processing utilization rate: Under the traditional processing method, the compatibility of krill oil with the carrier is poor, and free oil is easily precipitated when prepared into liposomes, which not only causes waste of raw materials, but also affects the quality of the final product.

[0004] 3) Significant sensory defects: The inherent trimethylamine and oxidation products of krill oil result in strong fishy smell, which has extremely low consumer acceptance in health foods and functional foods, limiting its application scenarios.

[0005] Therefore, it is necessary to explore the embedding technology of Antarctic krill oil to find a high-stability Antarctic krill oil encapsulation method. SUMMARY

[0006] In order to solve the above technical problems, the present application selects octenyl succinate starch (OSA), sodium alginate (ALG) and pea protein (Pea) as the composite wall material through material selection and hierarchical structure design. The above-mentioned materials can realize synergistic effect on the molecular-interface-micro-nano scale to delay lipid oxidation, control the targeted release characteristics of active ingredients, and improve the absorption efficiency of lipid-soluble ingredients. Furthermore, the present application also selects the liposome raw material of Antarctic krill oil to prepare a liposome with low particle size and high stability.

[0007] The present application provides a preparation method of liposomes for encapsulating Antarctic krill oil, which comprises the following steps: 1. Mix the wall material with water to prepare a composite wall material mother liquor; 2. Mix the krill oil with phospholipids and steroids, then perform shearing, homogenization and micro-jet treatment to obtain the liposomes of Antarctic krill oil; 3. Mix the liposomes of Antarctic krill oil and the composite wall material mother liquor, then perform shearing and homogenization to obtain the liposomes for encapsulating Antarctic krill oil.

[0008] Further, the wall material in step 1 comprises one or more of sodium alginate, pea protein, hydroxypropyl methylcellulose, octenyl succinyl starch, soy protein, gelatin, whey protein, chitosan.

[0009] Preferably, the wall material in step 1 is sodium alginate, pea protein and octenyl succinyl starch.

[0010] Preferably, the complex wall material mother liquor in step 1 comprises 1-2wt% of octenyl succinyl starch, 0.5-1.5wt% of sodium alginate, 0.1-0.5wt% of pea protein, and the rest is water.

[0011] Specifically, the complex wall material mother liquor in step 1 comprises 1wt% of octenyl succinyl starch, 0.8wt% of sodium alginate, 0.2wt% of pea protein, and 98wt% of water.

[0012] Further, the phospholipid in step 2 comprises one or more of egg yolk phospholipid, rapeseed oil phospholipid, hydrogenated phospholipid, and soybean phospholipid.

[0013] Further, the steroid in step 2 comprises one or more of phytosterol and cholesterol.

[0014] Preferably, the phospholipid in step 2 is rapeseed oil phospholipid or soybean phospholipid.

[0015] Preferably, the steroid in step 2 is phytosterol.

[0016] Preferably, the liposome of Antarctic krill oil in step 2 comprises 1-5wt% of krill oil, 45-50wt% of phospholipid, 3-5wt% of phytosterol, and the rest is water.

[0017] Specifically, the liposome of Antarctic krill oil in step 2 comprises 1wt% of krill oil, 46wt% of phospholipid, 4wt% of phytosterol, and 49wt% of water.

[0018] Further, the rotation speed of shearing in steps 2 and 3 is 10000-20000rpm, and the time is 120-240s.

[0019] Further, the pressure of homogenization in steps 2 and 3 is 30-50MPa, and the temperature is 40-60℃, and the homogenization is repeated 2-4 times.

[0020] Further, the parameter of microfluidization treatment in step 2 is 20-30kpsi.

[0021] Further, the mass ratio of the liposome of Antarctic krill oil to the complex wall material mother liquor in step 3 is 3-10:97-90.

[0022] Specifically, the mass ratio of the liposome and the complex wall material mother liquor of the Euphausia superba oil in step 3 is 5:95.

[0023] The application provides the liposome encapsulating Euphausia superba oil prepared according to the method.

[0024] The application provides the application of the liposome encapsulating Euphausia superba oil in food preparation or health product preparation.

[0025] Beneficial effects The application screens wall materials and preparation raw materials of the liposome, and selects the optimal raw material combination from the aspects of thermal stability and particle size, so that the Euphausia superba oil can be stably embedded, the Euphausia superba oil has excellent stability and no fishy smell, and the industrialization problem of the Euphausia superba oil is effectively solved. BRIEF DESCRIPTION OF DRAWINGS

[0026] Figure 1 The particle size result of sodium alginate at pH 2.

[0027] Figure 2 The particle size result of sodium alginate at pH 7.

[0028] Figure 3 The particle size result of octenyl succinic acid starch at pH 2.

[0029] Figure 4 The particle size result of soybean protein at pH 7.

[0030] Figure 5 The potential test result of sodium alginate.

[0031] Figure 6 The potential test result of pea protein.

[0032] Figure 7 The potential test result of hydroxypropyl methyl cellulose 1500.

[0033] Figure 8 The potential test result of hydroxypropyl methyl cellulose 4000.

[0034] Figure 9 The potential test result of octenyl succinic acid starch.

[0035] Figure 10 The potential test result of soybean protein.

[0036] Figure 11 The potential test result of gelatin.

[0037] Figure 12 The potential test result of whey protein.

[0038] Figure 13Potentiometric test results for chitosan.

[0039] Figure 14 TSI stability analysis test results for Example 2.

[0040] Figure 15 TSI stability analysis test results for Example 2.

[0041] Figure 16 TSI stability analysis test results for Example 2.

[0042] Figure 17 Potentiometric test results for the gastric and oral environments in Example 2.

[0043] Figure 18 Potentiometric test results for the gastric and oral environments in Example 2.

[0044] Figures 19-22 TSI stability analysis test results for Example 3.

[0045] Figure 23 Particle size test results for the homogenized sample in Example 3.

[0046] Figure 24 Potentiometric test results for the homogenized sample in Example 3.

[0047] Figure 25 Particle size test results for the microfluidized sample in Example 3.

[0048] Figure 26 Potentiometric test results for the microfluidized sample in Example 3. DETAILED DESCRIPTION

[0049] Raw material sources Pea protein was purchased from Yuyang Plant Protein in Linyi, hydroxypropyl methylcellulose was purchased from Shandong Guangda Science and Technology Development, octenyl succinic anhydride starch was purchased from Hangzhou Prostar Starch, soy protein was purchased from Shandong Jiahua Biology, gelatin and chitosan were purchased from Nanjing Meiji Xin Biology Technology, whey protein was purchased from Hengtianr Natural, phytosterol was purchased from Xi'an Zebang Biology Technology, cholesterol was purchased from Shandong Juyong Biology, egg yolk lecithin and hydrogenated lecithin were purchased from Xi'an Xihai Biology Technology, rapeseed oil phospholipid and soybean lecithin were purchased from ADM. If not specified, other raw materials were ordinary commercially available raw materials.

[0050] Example 1 Screening of wall materials for krill oil was carried out. Different amounts of wall material raw materials (sodium alginate, pea protein, hydroxypropyl methyl cellulose (viscosity 1500 and viscosity 4000), octenyl succinic acid starch, soybean protein, gelatin, whey protein, chitosan) were added to water at 75 degrees Celsius, and then the pH was adjusted to 2-7 before the zeta potential test. The solution was transferred to a Malvern Zetasizer Nano sample pool for testing using a pipette.

[0051] The results are shown in Figures 1-13 . It was found that sodium alginate, pea protein, and octenyl succinic acid starch had large absolute values, so they had strong stability. Octenyl succinic acid starch had good particle size distribution uniformity under acidic conditions, pea protein had strong uniformity under neutral conditions, and sodium alginate had good uniformity under acidic and neutral conditions. Therefore, sodium alginate, pea protein, and octenyl succinic acid starch were compounded to prepare the embedding wall material for Antarctic krill oil.

[0052] Example 2 1. Sodium alginate + octenyl succinic acid starch, pea protein + octenyl succinic acid starch, and sodium alginate + pea protein + octenyl succinic acid starch (sodium alginate: pea protein = 1:4, 2:3, 1:1, 3:2, 4:1) were mixed with water at 75 degrees Celsius to obtain 7 different complex wall material mother liquor. The complex wall material (sodium alginate, soybean protein, and octenyl succinic acid starch) accounted for 2wt% in all complex wall material mother liquor, and the octenyl succinic acid starch accounted for 1wt%.

[0053] 2. The krill oil was mixed with the complex wall material mother liquor, and the sample was sheared using a high-speed shearing machine at a speed of 11000 rpm for 90s to obtain a primary emulsion. The krill oil accounted for 5wt% in the primary emulsion.

[0054] 3. The primary emulsion was passed into a high-pressure homogenizer, and the pressure was set to 40 MPa and the temperature was set to 45 degrees Celsius. The sample was homogenized for 3 cycles to improve the uniformity of the sample.

[0055] A control group was set as pure krill oil, which was not mixed with the complex wall material mother liquor and was directly sheared and homogenized. Then, the above samples were tested for TSI stability analysis, and the results are shown in Figure 14 , 15 It can be seen that the TSI stability of sodium alginate: pea protein = 1:4 was not good. The remaining samples had good high-temperature stability.

[0056] The above samples were subjected to zeta potential test under simulated gastric and oral environment (pH 2.4 and 6.8), and the results are shown in Figures 16-18As shown, the ratio of sodium alginate to pea protein is 4:1, which has a relatively large absolute value. Therefore, this material combination has better stability, making it the optimal choice for embedding wall material.

[0057] The particle size of the above samples was measured, and the results are shown in Table 1 below. It can be seen that as the amount of sodium alginate added increases, the particle size of the sample increases accordingly. At the same time, the PDI increases, indicating that the uniformity of the sample decreases.

[0058] Table 1

[0059] In summary, the composite wall material combination of krill oil + sodium alginate + octenyl succinate starch + pea protein (sodium alginate: pea protein = 4:1) simultaneously achieves high-temperature stability, acid resistance, and industrial applicability.

[0060] Example 3 After selecting suitable wall materials, the next step is to prepare krill oil liposomes.

[0061] First, phospholipids, krill oil, and phytosterols from different sources were mixed with water at 80 degrees Celsius and stirred evenly. Specifically, the following mixtures were prepared: 0.25g krill oil + 1g phytosterol (low krill oil addition), 11.5g krill oil + 1g phytosterol (high krill oil addition), 0.25g krill oil + 11.5g egg yolk lecithin + 1g phytosterol, 0.25g krill oil + 11.5g rapeseed oil phospholipids + 1g phytosterol, 0.25g krill oil + 11.5g rapeseed oil phospholipids + 1g cholesterol, 0.25g krill oil + 11.5g hydrogenated lecithin + 1g phytosterol, and 0.25g krill oil + 11.5g soybean lecithin + 1g phytosterol. These were named groups 1 to 7. The amount of water added must meet the requirement that the krill oil content in the high krill oil addition group (group 2) is 5 wt%, and the krill oil content in the other groups is 1 wt%.

[0062] The sample was sheared using a high-speed shearing machine at 15,000 rpm for 180 seconds to obtain a primary emulsion. The primary emulsion was then passed through a high-pressure homogenizer at 40 MPa and 45°C for three cycles to improve sample homogeneity, resulting in liposomes of Antarctic krill oil, phospholipids, and a mixture of phospholipids from different sources.

[0063] Then, the above samples were subjected to TSI stability analysis tests, and the results are as follows: Figures 19-22 As shown, it can be seen that the TSI stability of the mixture of 1% krill oil and phytosterol in group 1 is not good, while the TSI stability of the mixture of 1% krill oil and hydrogenated phospholipids and phytosterol is slightly better than that of the control group containing the mixture of 1% krill oil and phytosterol.

[0064] Figures 23-24 Comparing the emulsion particle size of the above samples before and after microfluidization (20 kpsi) with Table 2, it is proved that microfluidization can effectively reduce the emulsion particle size of the samples. However, the particle size of the low phosphorus krill oil and phytosterol group and the hydrogenated phospholipid and phytosterol group samples increased significantly.

[0065] Figures 25-26 As shown in Table 3, microfluidization can reduce the absolute value of the sample potential, but the rapeseed phospholipid and phytosterol group and the soybean phospholipid and phytosterol group can increase the absolute value of the potential after microfluidization, and at the same time, improve the stability of the sample. Although the particle size and absolute value of the potential of the soybean phospholipid and phytosterol group are the most prominent, the TSI stability of the soybean phospholipid and phytosterol group significantly decreases after microfluidization. At the same time, the rapeseed phospholipid effectively reduces the emulsion particle size and increases the sample stability (absolute value of particle size increases) through microfluidization, and the rapeseed phospholipid has better high-temperature stability. In products that need long-time high-temperature processing, rapeseed phospholipid can be used as a more stable liposome material. If krill oil does not need long-time high-temperature processing, soybean phospholipid can be used as a liposome material.

[0066] Table 2

[0067] Table 3

[0068] Example 4 Finally, after screening suitable wall materials and liposome raw materials, the embedding of krill oil liposomes is carried out.

[0069] 1. Mix sodium alginate, pea protein, and octenyl succinic acid starch with water at 75 degrees Celsius to obtain a composite wall material mother liquor; the octenyl succinic acid starch in the composite wall material mother liquor accounts for 1 wt%, the sodium alginate accounts for 0.8 wt%, the pea protein accounts for 0.2 wt%, and the water accounts for 98 wt%.

[0070] 2. Mix 0.25 g of krill oil, 11.5 g of rapeseed oil phospholipid, 1 g of phytosterol, and water, and then use a high-speed shearing machine to shear the sample, set the speed to 15000 rpm, and shear for 180 s to obtain a primary emulsion 1; the mass of krill oil in the primary emulsion 1 accounts for 1 wt%, the mass of rapeseed oil phospholipid accounts for 46 wt%, the mass of phytosterol accounts for 4 wt%, and the mass of water accounts for 49 wt%.

[0071] 3. Pass the primary emulsion into a high-pressure homogenizer, set the pressure to 40 MPa and the temperature to 45 degrees Celsius, and circulate homogenization 3 times to improve the uniformity of the sample, and then perform microfluidization treatment with a treatment parameter of 20 kpsi to obtain krill oil liposomes.

[0072] 4、Mix the liposome of Antarctic krill oil with the complex wall material mother liquor, use a high-speed shearing machine to shear the sample, set the rotation speed to 11000 rpm, shear for 90 s, to obtain a primary emulsion 2; the liposome of Antarctic krill oil accounts for 5wt% in the primary emulsion 2.

[0073] 5、Pass the primary emulsion 2 into a high-pressure homogenizer, set the pressure to 40 MPa, the temperature to 45℃, circulate homogenization 3 times, improve the sample uniformity, finally obtain the liposome encapsulating Antarctic krill oil.

[0074] The above provided examples are not used to limit the scope covered by the present application, and the described steps are not used to limit the execution order. The person skilled in the art makes obvious improvements to the present application in combination with the existing common knowledge, which also falls within the protection scope defined by the claims of the present application.

Claims

1. A method for preparing liposomes encapsulating oil of Euphausia superba, characterized in that, The preparation method comprises the following steps: (1) mixing octenyl succinate starch, sodium alginate, pea protein and water to prepare a composite wall material mother liquor; the octenyl succinate starch accounts for 1-2 wt%, the sodium alginate accounts for 0.5-1.5 wt%, the pea protein accounts for 0.1-0.5 wt%, and the rest is water; (2) mixing krill oil, phospholipid and phytosterol, and then performing shearing, homogenization and microfluidization treatment to obtain a krill oil liposome; the phospholipid is rapeseed oil phospholipid or soybean phospholipid; the krill oil liposome contains 1-5 wt% krill oil, 45-50 wt% phospholipid and 3-5 wt% phytosterol, and the rest is water; (3) mixing the krill oil liposome and the composite wall material mother liquor, and then performing shearing and homogenization to obtain a krill oil encapsulated liposome.

2. The preparation method according to claim 1, characterized in that, In step (1), the octenyl succinate starch accounts for 1 wt%, the sodium alginate accounts for 0.8 wt%, the pea protein accounts for 0.2 wt%, and the water accounts for 98 wt%.

3. The preparation method according to claim 1, characterized in that, In step (2), the krill oil liposome contains 1 wt% krill oil, 46 wt% rapeseed oil phospholipid, 4 wt% phytosterol and 49 wt% water.

4. The preparation method according to claim 1, characterized in that, In steps (2) and (3), the shearing speed is 10,000-20,000 rpm, and the time is 120-240 s.

5. The preparation method according to claim 1, characterized in that, In steps (2) and (3), the homogenization pressure is 30-50 MPa, the temperature is 40-60°C, and the homogenization cycle is 2-4 times.

6. The preparation method according to claim 1, characterized in that, In step (2), the microfluidization parameter is 20-30 kpsi.

7. The preparation method according to claim 1, characterized in that, In step (3), the mass ratio of the krill oil liposome to the composite wall material mother liquor is 3-10:97-90.

8. The method of claim 1, wherein the method further comprises the step of: In step (3), the mass ratio of the krill oil liposome to the composite wall material mother liquor is 5:

95.

9. Liposomes encapsulating oil of Euphausia superba, characterized in that, The krill oil encapsulated liposome is prepared according to the preparation method of any one of claims 1-8.

10. The krill oil encapsulated liposome of claim 9 is used in food preparation or health product preparation.

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