Application of mandarin orange peel flavonoid substance in food for assisting blood pressure reduction and preparation method of mandarin orange peel flavonoid substance

Abstract

According to the application of the mandarin orange and pericarpium citri reticulatae flavonoid substance in auxiliary blood pressure lowering food, the effective concentration of neohesperidin of the flavonoid substance in mandarin orange and pericarpium citri reticulatae for achieving the blood pressure lowering effect in SHR bodies of spontaneously hypertensive rats is 25 mg / kg to 75 mg / kg, and the purity of the neohesperidin is 95.6%. The citrus reticulata blanco tangerine peel has the advantages that the citrus reticulata blanco tangerine peel is rich in neohesperidin components, can remarkably reduce the diastolic pressure and systolic pressure in SHR (spontaneous hypertensive rats), and can be used as functional food or special medical food for assisting in preventing and treating hypertension-related diseases.

Description

Technical Field

[0001] The invention relates to the technical field of blood pressure-assisting foods, in particular to the application of tangerine peel flavonoids in blood pressure-assisting foods, and also to a preparation method of tangerine peel flavonoids in blood pressure-assisting foods. Background Art

[0002] The pharmacopoeia records that tangerine peel has the functions of regulating qi, strengthening the spleen, drying dampness, and resolving phlegm. Tangerine peel is rich in flavonoids, essential oils, natural pigments, vitamins A, C, E, etc. Modern research shows that flavonoids have multiple effects such as lowering blood pressure, lowering blood lipids, and anti-oxidation. "Ou citrus" is a cultivated variety of citrus, native to southern Zhejiang, also known as bitter citrus. The fruit is bright golden, sweet and juicy, slightly bitter, and has high nutritional value. It has the effects of removing heat, promoting body fluid, resolving phlegm, relieving cough, cooling and detoxifying. There is a saying among the people that "Ou citrus during the Dragon Boat Festival is like an antelope", and it was a royal tribute in history.

[0003] In recent years, the cultivation area of ​​Ou oranges has been expanding, from Wenzhou to Fuding, Lishui and Taizhou, and Ou orange peel has become a new favorite in the market and a good project for farmers to increase their income. A clinical observation of 37 patients with primary hypertension treated with Ou orange fruit diet therapy was reported, and the effective rate reached 70% (Lin Xianrong, 2007), but there are few reports on the pharmacological effects of Ou orange peel, especially the research on the antihypertensive effect components of Ou orange peel.

[0004] In addition, it can be known from the existing literature that the content of neohesperidin is only 90.0% and the yield is 2.0% in Chinese patent application No. 2015108929773, "Method for Extracting Hesperidin and Neohesperidin from Citrus Peel and Fruit". Summary of the invention

[0005] In order to solve the above problems, the present invention provides the use of tangerine peel flavonoids in auxiliary blood pressure lowering food.

[0006] To achieve the above purpose, the present invention is solved by the following technical scheme: application of tangerine peel flavonoids in auxiliary blood pressure lowering food. The present invention purifies tangerine peel neohesperidin from crude flavonoids in tangerine peel with a purity of 95.6% and a yield of 6.5%, which is significantly better than the reported neohesperidin content of 90.0% and a yield of 2.0%.

[0007] Moreover, each concentration in the test (25 mg / kg to 75 mg / kg) had a significant (P<0.05) effect of lowering blood pressure in spontaneously hypertensive rats (SHR), with 50 mg / kg being the most economical and appropriate and reaching a very significant effect (P<0.01), and also having no effect on heart rate. This provides a basic basis for further research and development of Ougan tangerine peel as an auxiliary blood pressure-lowering food, and is novel, creative and practical.

[0008] The invention also provides a method for preparing the tangerine peel flavonoids in the blood pressure-reducing auxiliary food.

[0009] To achieve the above object, the present invention is solved by the following technical solutions:

[0010] Includes initial extraction;

[0011] The ultrasonic extraction of Ou orange peel water can obtain a crude flavonoid filtrate, which includes limonin, β-sitosterol, naringin and neohesperidin. The crude flavonoid filtrate is eluted with D101 macroporous resin, washed with water until colorless, and the effective components are eluted with 50% to 70% ethanol.

[0012] Furthermore, the initial extraction also includes an optimization process;

[0013] The 50% to 70% ethanol eluate is then separated by ultrafiltration using an ultrafiltration membrane system (molecular weight 3000-10000), the concentrated solution is allowed to crystallize, and the crystals are recrystallized with water to obtain neohesperidin. The yield of neohesperidin obtained by this method is 6.5%.

[0014] Furthermore, the purity of neohesperidin was 95.6% by HPLC.

[0015] Furthermore, 3 parts of Ou orange peel, 1 part of cicada flower and 1 part of wolfberry are compounded to prepare tea.

[0016] Furthermore, the orange peel extract is made into powder and mixed with other foods to prepare auxiliary blood pressure-lowering food.

[0017] The present invention is beneficial in that:

[0018] 1. The antihypertensive component neohesperidin is extracted from Ou orange tangerine peel, and the neohesperidin has a high yield and high purity, and the preparation process is relatively convenient. The Ou orange tangerine peel provided by the present invention has abundant sources, Ou oranges are cultivated in Zhejiang Province, and the preparation cost is low. It can be widely promoted and applied as a blood pressure auxiliary substance.

[0019] 2. Experiments have shown that orange peel is rich in neohesperidin, which can significantly reduce the diastolic and systolic blood pressure in spontaneously hypertensive SHR rats. It can be used as a functional food or a special medical food to assist in the prevention and treatment of hypertension-related diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 The graph is a graph showing the results of the inhibition of pyrogallol auto-oxidation by the crude flavonoid filtrate (OGCF) of the present invention. Note: OGN, Ou orange peel neohesperidin; # indicates a significant difference compared with the SHR control group P<0.05, ## indicates a very significant difference P<0.01. WKY, normotensive rats; SHR, spontaneously hypertensive rats.

[0021] Figure 2 The figure shows the changes of HR, SBP and DBP of SHR rats in each group after four weeks of drug administration of the present invention. DETAILED DESCRIPTION

[0022] The technical solutions in the embodiments of the present invention will be described clearly and completely below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all the embodiments.

[0023] Application of flavonoids from orange peel in auxiliary antihypertensive foods, about the extraction of crude flavonoids and neohesperidin from orange peel:

[0024] The ultrasonic extraction of Ougan tangerine peel water can obtain crude flavonoids, which contain limonin, β-sitosterol, naringin, neohesperidin and other hesperidin components. Four adsorption resins, AB-8, D101, NKA-9 and NKA-Ⅱ, were screened, and the most suitable macroporous resin for purifying flavonoids was D101.

[0025] Optimized process: Use water ultrasonic extraction to extract flavonoids from Ou orange peel raw materials, elute the filtrate with D101 macroporous resin, wash with water until colorless, elute the effective ingredients with 50% to 70% ethanol; then use an ultrafiltration membrane system (molecular weight 3000 to 10000) to ultrafilter and separate the 50% to 70% ethanol eluate, and place the concentrated solution for crystallization; recrystallize the crystals with water to obtain neohesperidin. The purity of the flavonoids was 95.6% as determined by HPLC. The maximum yield of neohesperidin obtained by this method is 6.5%.

[0026] Detection method for inhibiting pyrogallol auto-oxidation

[0027] Determination of the self-oxidation rate of pyrogallol: refer to the method reported by Li Yongli, Xu Shenhong, etc. and make some improvements. Add 9 ml Tris-HCl-EDTA buffer to each of the two test tubes. Add 40 μl of redistilled water to the control tube and mix well. Adjust the absorbance to zero at 420 nm with a 1 cm colorimetric cup. Add 40 μl of pyrogallol to the test tube and mix well. Immediately add a 1 cm colorimetric cup and read the absorbance A0 accurately at 1, 2, ... 10 minutes. Repeat 3 times and calculate the average value. Calculate the regression equation and correlation coefficient with time as the horizontal axis and absorbance as the vertical axis.

[0028] Experiment on inhibition of pyrogallol auto-oxidation by different doses of vitamin C and tangerine peel flavonoid solutions: add 9 ml of Tris-HCl-EDTA buffer and 0.1 (0.2, 0.3) ml of vitamin C (or flavonoid solution) to each of two test tubes; add 40 μl of redistilled water to the control tube and mix well; adjust the absorbance to zero at 420 nm; add 40 μl of pyrogallol to the measuring tube and mix well; immediately add a 1 cm colorimetric cup and read the absorbance accurately at 1, 2, ... 10 min. Repeat the measurement 3 times for each treatment group.

[0029] Data statistical processing:

[0030] The test results list the absorbance of each group X ± S to calculate the inhibition rate of pyrogallol auto-oxidation at 10 minutes X ± S. Inhibition rate (%) = (A0-AS) ÷ A0 × 100. The differences between the groups were analyzed by SSR multiple comparison analysis of variance.

[0031] The experiment found that the linear relationship between the self-oxidation rate and time of pyrogallol within 10 minutes was good and stable, so the Vc and tangerine peel flavonoids samples in this experiment were measured for absorbance changes within 10 minutes. Adding tangerine peel flavonoid solution had a significant inhibitory effect on the self-oxidation of pyrogallol (all P < 0.01), and it showed concentration dependence.

[0032] refer to Figure 1 The results of the inhibition of pyrogallol auto-oxidation by Vc and Ougan tangerine peel crude flavonoids (OGCF) showed that the highest inhibition rate of pyrogallol auto-oxidation in vitro was 46.28±0.23 (P<0.01)%.

[0033] refer to Figure 2 , Antihypertensive effect of neohesperidin from orange peel on spontaneously hypertensive rats.

[0034] Experimental animals

[0035] Sixteen 9-week-old male spontaneously hypertensive rats (SHR) and four WKY (same strain but normal blood pressure) rats as controls, SPF grade, were purchased from Shanghai Weitong Lihua Biological Co., Ltd. Animal license number: SCXK (Zhe) 2017-149. The animals were kept in a standard animal room with a temperature of 24±2℃, a humidity of 50±5%, a dark-light cycle of 12h, free access to food and water, and adaptive feeding for one week.

[0036] Group dosing

[0037] 16 SHR rats were divided into 4 groups according to the initial blood pressure (there was no significant difference in blood pressure among rats in the groups, P>0.05), namely, low-concentration group (25mg / kg), medium-concentration group (50mg / kg), high-concentration group (75mg / kg) and hypertensive rat control group, which were fed orally for four weeks, and the control group was given an equal amount of normal saline. Blood pressure values ​​were measured on an empty stomach at 7-9 am every week at a fixed time, and heart rate (HR), systolic blood pressure (SBP) and diastolic blood pressure (DBP) were measured and recorded. Each rat was measured three times and the average value was taken. SPSS15.0 was used to analyze the data.

[0038] Results Analysis

[0039] Compared with the WKY normal blood pressure control group, there was no significant difference in heart rate (HR) among the groups; from the perspective of systolic blood pressure (SBP), compared with the hypertensive control group, there were significant differences in blood pressure between the OGD 25mg / kg and 75mg / kg groups (P<0.05), and there was a very significant difference in the 50mg / kg concentration group (P<0.01), indicating that OGD has a significant antihypertensive effect; from the perspective of diastolic blood pressure (DBP), compared with the hypertensive control group, the blood pressure values ​​of the blank control group and the drug-treated group were lower, among which the antihypertensive effect of the low, medium and high concentration groups of OGD changed in a gradient, but the effect was not significant.

[0040] It is obvious to those skilled in the art that the present invention is not limited to the details of the above-mentioned exemplary embodiments, and the present invention can be implemented in other specific forms without departing from the spirit or basic characteristics of the present invention. Therefore, no matter from which point of view, the embodiments should be regarded as exemplary and non-restrictive. The scope of the present invention is limited by the appended claims rather than the above description, and it is intended that all changes within the meaning and scope of the equivalent elements of the claims are included in the present invention.

Claims

1. The application of flavonoids from orange peel in auxiliary blood pressure lowering food is characterized in that: The effective concentration of neohesperidin, a flavonoid substance in orange peel, to achieve a blood pressure-lowering effect in spontaneously hypertensive rats (SHR) is 25 mg / kg to 75 mg / kg, and the purity of the neohesperidin is 95.6%.

2. The use of the flavonoids from Ou orange peel in auxiliary blood pressure lowering food according to claim 1, characterized in that: The optimal effective concentration of neohesperidin to achieve antihypertensive effect in spontaneously hypertensive rats (SHR) is 50 mg / kg.

3. A method for preparing flavonoids from Ougan orange peel in blood pressure-lowering food, characterized in that: Includes initial extraction; The ultrasonic extraction of Ou orange peel water can obtain a crude flavonoid filtrate, which includes limonin, β-sitosterol, naringin and neohesperidin. The crude flavonoid filtrate is eluted with D101 macroporous resin, washed with water until colorless, and the effective components are eluted with 50% to 70% ethanol.

4. The method for preparing the flavonoids from Ou orange peel in blood pressure-lowering food according to claim 3, characterized in that: After the initial extraction, it also includes optimization processing; The 50% to 70% ethanol eluate is then separated by ultrafiltration using an ultrafiltration membrane system (molecular weight 3000-10000), the concentrated solution is allowed to crystallize, and the crystals are recrystallized with water to obtain neohesperidin. The yield of neohesperidin obtained by this method is 6.5%.

5. The method for preparing the flavonoids from Ou orange peel in blood pressure-lowering food according to claim 4, characterized in that: The purity of the purified neohesperidin was determined to be 95.6% by HPLC.

6. The method for preparing the flavonoids from Ou orange peel in blood pressure-lowering food according to claim 5, characterized in that: Mix 3 parts of Ou orange peel, 1 part of cicada flower, and 1 part of wolfberry to make a tea.

7. The method for preparing the flavonoids from Ou orange peel in blood pressure-lowering food according to claim 5, characterized in that: The flavonoid extract from orange peel is made into powder and mixed with other foods to make an auxiliary blood pressure-lowering food.

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