Compound natural antioxidant, compound oil, antioxidant preparation and application thereof

Abstract

The application discloses a kind of compound natural antioxidant, compound grease, antioxidant preparation and application thereof.The compound natural antioxidant comprises the following components: oryzanol, vitamin E and phytosterol;Wherein, the mass ratio of oryzanol, vitamin E and phytosterol is (12.5-20):1:(10-25);Vitamin E includes tocotrienol;The mass percentage content of tocotrienol in vitamin E is not less than 30%;Phytosterol includes sitosterol;The mass percentage content of sitosterol in phytosterol is not less than 10%.The compound natural antioxidant of the application has excellent in vitro antioxidant performance, and can also have excellent in vivo antioxidant, anti-aging performance.

Description

Technical Field

[0001] This invention relates to a compound natural antioxidant, a compound oil, an antioxidant preparation, and their applications. Background Technology

[0002] Oryzanol is a mixture composed mainly of ferulic acid esters (primarily cycloartane alcohols) and sterol ferulic acid esters, primarily found in rice bran oil. Oryzanol can reduce endocrine imbalances, lower cholesterol and triglycerides, and has antioxidant effects. It can also regulate autonomic nervous system function and improve sleep. It has a significant therapeutic effect on autonomic nervous system disorders and also lowers blood lipids and cholesterol. As a drug, oryzanol can be used for sedation and sleep aid in conditions such as neurasthenia, premenstrual syndrome, and menopausal syndrome.

[0003] Both vitamin E and phytosterols can improve the body's immunity and enhance antioxidant capacity, but current evidence suggests that the combined use of antioxidants can have both synergistic and inhibitory effects.

[0004] Therefore, exploring the synergistic effects among various antioxidants is an important topic in the research and development of antioxidant products. Summary of the Invention

[0005] To address the shortcomings of existing technologies, this invention provides a compound natural antioxidant, a compound oil, an antioxidant preparation, and their applications. The compound natural antioxidant of this invention possesses excellent in vitro antioxidant properties as well as excellent in vivo anti-aging properties.

[0006] To achieve the above objectives, the technical method provided by this invention is as follows:

[0007] The first aspect of this invention provides a compound natural antioxidant comprising the following components: oryzanol, vitamin E, and phytosterols; wherein,

[0008] The mass ratio of oryzanol, vitamin E and phytosterol is (12.5-20):1:(10-25);

[0009] The vitamin E includes tocotrienols; the tocotrienols constitute not less than 30% of the vitamin E by mass.

[0010] The phytosterols include sitosterol; the sitosterol content in the phytosterols is not less than 10% by mass.

[0011] In this invention, the preferred mass ratio of oryzanol, vitamin E and phytosterol is (15-17.5):1:(15-25), for example, 17.5:1:15.

[0012] In this invention, the mass percentage of the tocotrienol in vitamin E is preferably 50%-100%, for example, 80%.

[0013] In this invention, the mass percentage of sitosterol in phytosterols is preferably 10%-50%, for example 20%, 30% or 40%.

[0014] In this invention, the vitamin E may further include tocopherol; the mass percentage of tocopherol in vitamin E may not exceed 70%, preferably 0-50%, for example 20%.

[0015] In this invention, the phytosterols may further include one or more of cholesterol, cholesterolanol, campesterol, brassosterol, Δ24-methylenecholesterol, brassosterol, brassosterol, stigmasterol, Δ7-brassenol, Δ5,23-stigmasterdienol, Δ5,24-stigmasterdienol, sterol, sitosterol, Δ5-amaresterol, Δ7-amaresterol, and Δ7-stigmasterenol; preferably, they include one or more of stigmasterol, campesterol, and brassosterol.

[0016] In this invention, the tocotrienol may include α-tocotrienol, β-tocotrienol and γ-tocotrienol; the mass ratio of α-tocotrienol, β-tocotrienol and γ-tocotrienol may be 1:1:1.

[0017] In some embodiments, the phytosterols may include 53% sitosterol, 7% stigmasterol, 26% campesterol and 13% brassosterol by weight percentage.

[0018] In some embodiments, the phytosterols may include, in addition to sitosterol, stigmasterol, campesterol and brassosterol in a mass ratio of 1:1:1.

[0019] In some embodiments, the compound natural antioxidants include oryzanol, vitamin E, and phytosterols; wherein the mass ratio of oryzanol, vitamin E, and phytosterols is (12.5-20):1:(10-25); the vitamin E includes tocotrienols and tocopherols; the mass percentage of tocotrienols in vitamin E is not less than 30%; the tocotrienols include α-tocotrienols, β-tocotrienols, and γ-tocotrienols; the phytosterols include sitosterol, stigmasterol, campesterol, and brassosterol; the mass percentage of sitosterols in phytosterols is not less than 10%.

[0020] A second aspect of the present invention provides a method for preparing the compound natural antioxidant as described above, comprising the following steps: uniformly mixing the components of the compound natural antioxidant to obtain the product.

[0021] A third aspect of the present invention provides a compound oil comprising a compound natural antioxidant and an oil as described above.

[0022] In this invention, the oil can be conventional in the art, and its main component is an ester formed by dehydration of higher fatty acids (such as synthetic and natural fatty acids) and glycerol; the oil is preferably one or more of rice bran oil, soybean oil, peanut oil, olive oil, rapeseed oil, sesame oil, castor oil and animal oil.

[0023] A fourth aspect of the present invention provides an antioxidant preparation comprising a compound of natural antioxidants and an organic solvent as described above.

[0024] In this invention, the organic solvent can be conventional in the art, and preferably one or more of ethanol, glycerol and acetic acid.

[0025] In this invention, the concentration of oryzanol in the compound oils and antioxidant preparations can be 10,000-30,000 ppm, preferably 15,000-25,000 ppm, such as 17,000 ppm, 19,000 ppm, 20,000 ppm, 21,000 ppm or 23,000 ppm.

[0026] In this invention, the concentration of vitamin E in the compound oils and antioxidant preparations can be 500-1800 ppm, preferably 800-1500 ppm, for example 971 ppm, 1086 ppm, 1142 ppm, 1200 ppm or 1314 ppm.

[0027] In this invention, the concentration of phytosterols in the compound oils and antioxidant preparations can be 8000-26000 ppm, preferably 12000-22000 ppm, for example 14565 ppm, 16290 ppm, 17310 ppm, 18000 ppm or 19710 ppm.

[0028] In some embodiments, the mass ratio of oryzanol, vitamin E, and phytosterol in the compound oil and antioxidant preparation is (12.5-17.5):1:(10-15); the concentration of oryzanol is 15000-25000 ppm; the concentration of vitamin E is 800-1500 ppm; and the concentration of phytosterol is 12000-22000 ppm.

[0029] In some embodiments, the mass ratio of oryzanol, vitamin E, and phytosterol in the compound oil and antioxidant preparation is 17.5:1:15; the concentration of oryzanol is 21,000 ppm; the concentration of vitamin E is 1,200 ppm; and the concentration of phytosterol is 18,000 ppm.

[0030] The fifth aspect of the present invention provides a method for preparing the compound oil or the antioxidant preparation as described above, comprising the following steps: mixing the compound natural antioxidant evenly in the oil or the organic solvent to obtain the product.

[0031] The sixth aspect of this invention provides the application of the compounded natural antioxidants as described above in food.

[0032] In this invention, the food can be general food, specialty food, or health product.

[0033] The seventh aspect of the present invention provides the application of the compounded natural antioxidants as described above in the preparation of anti-aging drugs.

[0034] In this invention, the dosage form of the drug may be an oral liquid preparation, tablet, capsule, granule or dry suspension.

[0035] In this invention, the daily dosage of the compound natural antioxidant can be 50-1000 mg, preferably 300-900 mg, for example 763 mg.

[0036] Based on common knowledge in the field, the above-mentioned preferred conditions can be combined arbitrarily to obtain various preferred embodiments of the present invention.

[0037] The reagents and raw materials used in this invention are all commercially available.

[0038] The positive and progressive effects of this invention are as follows:

[0039] This invention, based on the combination of three active ingredients—oryzanol, vitamin E, and phytosterols—investigates the effects of specific active ingredients in vitamin E (tocotrienol) and phytosterols (sitosterol) on the compounding effect. The results show that the compounded natural antioxidants of this invention possess excellent in vitro antioxidant properties and also exhibit excellent in vivo antioxidant and anti-aging properties. Furthermore, under specific ratios, the active ingredients show significant synergistic effects and have wide applications. Detailed Implementation

[0040] The invention will be further described through the following embodiments; however, the scope of the invention is not limited to the embodiments described below. Those skilled in the art will understand that various changes and modifications can be made to the invention without departing from its spirit and scope. The materials and methods used in the embodiments are described in a general and / or specific manner. Although many materials and methods of operation used to achieve the objectives of this invention are well known in the art, they are still described in as much detail as possible herein.

[0041] Unless otherwise specified, all compounds used in the examples were purchased from Sigma.

[0042] Except for Example 3, the phytosterols in the other examples are mixtures of sitosterol, stigmasterol, campesterol and brassosterol, with each component having a mass percentage of 53% sitosterol, 7% stigmasterol, 26% campesterol and 13% brassosterol.

[0043] Except for Example 4, the vitamin E in the other examples was a mixture of tocopherol and tocotrienol, with each component having a mass percentage of 50% tocopherol and 50% tocotrienol.

[0044] The tocotrienols in the examples include α-tocotrienol, β-tocotrienol and γ-tocotrienol in a mass ratio of 1:1:1.

[0045] In the examples, PBS refers to phosphate buffer solution with a concentration of 0.1M and a pH of 7.2.

[0046] The room temperature in the embodiments is the conventional room temperature in the art, preferably 15-30°C.

[0047] The experimental results in the examples are expressed as mean ± standard error. After parametric or nonparametric variance tests, p < 0.05 is considered to be significant, and p < 0.01 is considered to be extremely significant.

[0048] Testing the ability of oils and fats to resist auto-oxidation

[0049] The ability of oils to resist auto-oxidation can be represented by the Oil Stability Index (OSI). This index reflects the sensitivity of oils to auto-oxidative deterioration and also indicates their storage stability. Oxidative stability was determined using a 743 Rancimat instrument (Metroën Instruments, Switzerland). Test samples (3g each) were prepared by adding compounded natural antioxidants to glycerol under different conditions. An airflow (20 L / h) was used to pass through the oil sample heated to 120°C. Volatile organic acids in the oil sample were collected in cold water to increase conductivity, and the conductivity was continuously recorded. The induction time [h] was automatically printed by the instrument software with an accuracy of 0.005. The induction time is related to the antioxidant effect of the oil sample; the longer the induction time, the stronger the antioxidant effect.

[0050] DPPH Oxygen Free Radical Scavenging Rate Determination

[0051] The test samples were prepared by adding compound natural antioxidants to glycerol under different conditions, and a 0.1 mmol / L DPPH solution was prepared using ethyl acetate. 0.1 g of the test sample was accurately weighed and dissolved completely in 4 mL of ethyl acetate. 2 mL of the prepared sample was mixed with 2 mL of DPPH solution and stored in the dark for 30 min. The absorbance at 517 nM was measured using a spectrophotometer. The DPPH oxygen free radical scavenging rate was calculated as [1 - (A...]. - A ) A0 ] 100%. A0, A and A The absorbance values ​​are ethyl acetate + DPPH, sample + DPPH, and sample + ethyl acetate, respectively.

[0052] Example 1: Effect of the mass ratio of oryzanol, vitamin E, and phytosterols

[0053] According to Table 1, compound natural antioxidants containing different mass ratios of oryzanol, vitamin E and phytosterols were added to glycerin to test the oil's ability to resist auto-oxidation and the DPPH oxygen free radical scavenging rate. The results are shown in Tables 1 and 2.

[0054] Table 1. Test results of compound natural antioxidants with different mass ratios 1

[0055]

[0056] Table 2. Test results of compound natural antioxidants with different mass ratios 2

[0057]

[0058] As can be seen from Table 1, when the mass ratio of oryzanol, vitamin E and phytosterol in the compound natural antioxidant of the present invention is in the range of (1-30):1:(0.1-25), even with low concentrations of the three substances, the OSI can still be increased from 3.8h to 7.7h and the DPPH free radical scavenging rate can still be increased from 23.6% to 72.3% as the mass ratio changes.

[0059] Furthermore, as shown in Table 2, when the mass ratio is (12.5-20):1:(10-25), the OSI is higher than 8h and the DPPH radical scavenging rate is higher than 65%; when the mass ratio is (15-17.5):1:(15-25), the OSI is not lower than 8.8h and the DPPH radical scavenging rate is not lower than 76%; the effect is most obvious when the mass ratio is 17.5:1:15, with the OSI reaching 9.1h and the DPPH radical scavenging rate reaching 79.6%.

[0060] Example 2: Effects of oryzanol, vitamin E, and phytosterol concentrations

[0061] Based on the results in Example 1, a fixed ratio of oryzanol, vitamin E, and phytosterols of 17.5:1:15 was selected and added to glycerol at different concentrations as shown in Tables 3 and 4. The oil's ability to resist auto-oxidation and the DPPH oxygen free radical scavenging rate were tested, and the results are shown in Tables 3 and 4.

[0062] Table 3. Test results of different concentrations of compound natural antioxidants 1

[0063]

[0064] Table 4. Test results of different concentrations of compound natural antioxidants 2

[0065]

[0066] As can be seen from Tables 3 and 4, when the mass ratio of oryzanol, vitamin E and phytosterol is constant, the antioxidant capacity of the compound natural antioxidant of the present invention gradually increases with the increase of the concentration of the three active ingredients.

[0067] According to Table 3, when the concentration of oryzanol was 10,000-30,000 ppm, the OSI was higher than 9h, and the DPPH free radical scavenging rate was higher than 80%. As the concentration of oryzanol exceeded 25,000 ppm, the antioxidant capacity decreased slightly, proving that the antioxidant effect of compound natural antioxidants has a capping effect.

[0068] Furthermore, according to Table 4, when the content of oryzanol was between 19,000 and 25,000 ppm, the OSI was higher than 10 h, and the DPPH free radical scavenging rate was higher than 85%, showing stronger antioxidant capacity.

[0069] Example 3: Effects of sterol types and contents in plant sterols

[0070] Sitosterol, a type of phytosterol, is an important antioxidant. To determine the effect of sitosterol on the compound natural antioxidant of this invention, based on the results of Examples 1 and 2, a compound natural antioxidant with a oryzanol concentration of 21,000 ppm, a vitamin E concentration of 1,200 ppm, and a phytosterol concentration of 18,000 ppm was prepared and added to glycerol, with a mass ratio of oryzanol, vitamin E, and phytosterol of 17.5:1:15 as a fixed ratio. The oil's ability to resist auto-oxidation and the DPPH oxygen free radical scavenging rate were then tested.

[0071] The content of sitosterol among the phytosterols is shown in Table 5. Among the remaining phytosterols, stigmasterol, campesterol, and brassosterol each account for 33.3%.

[0072] Table 5. Effects of different sitosterol contents on plant sterols

[0073]

[0074] The results showed that the antioxidant effect of the compound natural antioxidants gradually increased with the increase of sitosterol content. When the sitosterol content in phytosterols exceeded 10%, the OSI was higher than 8.5h, and the DPPH free radical scavenging rate was higher than 80%.

[0075] Based on Group 6 in Table 5, this invention further investigated the types of phytosterols. As shown in Table 6, ergosterol and cholesterol were used to replace the sitosterol of this invention. Among the remaining phytosterols, stigmasterol, campesterol and brassicasterol each accounted for 33.3%. The test results showed that the scavenging rates of OSI and DPPH oxygen free radicals were significantly reduced.

[0076] Table 6. Effects of different types of phytosterols

[0077]

[0078] Example 4: Effect of tocotrienol content in vitamin E

[0079] Vitamin E mainly contains two important antioxidants: tocopherols and tocotrienols. To determine the effect of tocotrienols on the compound natural antioxidant of this invention, based on the results of Examples 1 and 2, a compound natural antioxidant with a concentration of 21,000 ppm oryzanol, 1,200 ppm vitamin E, and 18,000 ppm phytosterols was prepared and added to glycerol using a fixed ratio of 17.5:1:15 of oryzanol, vitamin E, and phytosterols. The oil's ability to resist auto-oxidation and the DPPH oxygen free radical scavenging rate were then tested.

[0080] The content of tocotrienols in vitamin E is shown in Table 7, with α-tocotrienol, β-tocotrienol, and γ-tocotrienol each accounting for 33.3%.

[0081] Table 7. Effect of different tocotrienol contents in vitamin E

[0082]

[0083] The results showed that the antioxidant effect of the compound natural antioxidants gradually increased with the increase of tocotrienol content. When the content of tocotrienol in vitamin E exceeded 30%, the OSI was higher than 8.5h, and the DPPH free radical scavenging rate was higher than 85%.

[0084] Example 5: In vivo antioxidant effects of different doses of compound natural antioxidants

[0085] A compound natural antioxidant with a concentration of 21,000 ppm oryzanol, 1,200 ppm vitamin E, and 18,000 ppm phytosterol was prepared by using a fixed mass ratio of 17.5:1:15 for oryzanol, vitamin E, and phytosterols. This compound was added to glycerol and administered to aging mice via gavage at different doses for prophylaxis. Mice were also injected subcutaneously into the back of the mice with 120 mg / kg D-galactose once daily for 6 weeks to establish an aging model. The in vivo antioxidant effect of the compound natural antioxidant was observed by detecting the level of superoxide dismutase (SOD) in serum. The results are shown in Table 7.

[0086] Table 8 shows that a total dose of 0.35-3.5 mg / day of the compound natural antioxidants exhibited good antioxidant effects in vivo, with an optimal dose range of 1.05-3.15 mg / day and an optimal dose of 2.8 mg / day. Based on the dosage conversion ratio between mice and humans, the human body showed good antioxidant capacity at 50-1000 mg / day, with an optimal dose range of 300-900 mg / day and an optimal dose of 763 mg / day.

[0087] Table 8. In vivo antioxidant results of different doses of compound natural antioxidants

[0088]

Claims

1. A compound natural antioxidant, characterized in that, It contains the following components: oryzanol, vitamin E, and phytosterols; among which, The mass ratio of oryzanol, vitamin E, and phytosterol is (15-17.5):1:(15-25). The vitamin E includes tocotrienols; the tocotrienols constitute not less than 30% of the vitamin E by mass. The phytosterols include sitosterol; the sitosterol content in the phytosterols is not less than 10% by mass.

2. The compound natural antioxidant as described in claim 1, characterized in that, The tocotrienols contained in vitamin E comprised 50%-100% by mass. And / or, the sitosterol has a mass percentage of 10%-50% in phytosterols.

3. The compound natural antioxidant as described in claim 1, characterized in that, The vitamin E also includes tocopherol; the tocopherol content in vitamin E does not exceed 70% by mass.

4. The compound natural antioxidant as described in claim 3, characterized in that, The tocopherol content in vitamin E does not exceed 50% by mass.

5. The compound natural antioxidant as described in claim 1, characterized in that, The phytosterols also include one or more of the following: cholesterol, cholesterolanol, campesterol, rapeseed sterol, Δ24-methylenecholesterol, brassosterol, brassosterol, stigmasterol, Δ7-brassenol, Δ5,23-stigmasterdienol, Δ5,24-stigmasterdienol, sterol, sitosterol, Δ5-amaresterol, Δ7-amaresterol, and Δ7-stigmasterol.

6. The compound natural antioxidant as described in claim 5, characterized in that, The phytosterols include one or more of stigmasterol, campesterol, and brassosterol.

7. A compound oil, characterized in that, It comprises a compound of natural antioxidants and oils as described in any one of claims 1-6.

8. The compound oil as described in claim 7, characterized in that, The oil is one or more of the following: rice bran oil, soybean oil, peanut oil, olive oil, rapeseed oil, sesame oil, castor oil, and animal oil.

9. An antioxidant preparation, characterized in that, It comprises a compound of natural antioxidants and organic solvents as described in any one of claims 1-6.

10. The antioxidant preparation according to claim 9, characterized in that, The organic solvent is one or more of ethanol, glycerol, and acetic acid.

11. The compound oil as described in claim 7 or 8, or the antioxidant preparation as described in claim 9 or 10, characterized in that, The concentration of the oryzanol is 10,000-30,000 ppm; And / or, the concentration of said vitamin E is 500-1800 ppm; And / or, the concentration of the phytosterol is 8000-26000 ppm.

12. The compound oil as described in claim 7 or 8, or the antioxidant preparation as described in claim 9 or 10, characterized in that, The concentration of the oryzanol is 15,000-25,000 ppm; And / or, the concentration of said vitamin E is 800-1500 ppm; And / or, the concentration of the phytosterol is 12,000-22,000 ppm.

13. The application of a compound natural antioxidant as described in any one of claims 1-6 in the preparation of food.

14. The use of a compound natural antioxidant as described in any one of claims 1-6 in the preparation of an anti-aging drug.

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