CN106138130B - Mango seed flavone extract and preparation method thereof - Google Patents

Abstract

The invention discloses a mango seed flavone extract and a preparation method thereof, belonging to the field of food technology and biotechnology. The mango seed flavone extract is prepared by taking mango seeds as a raw material through alcohol extraction and a macroporous resin purification method. The mango seed flavone extract prepared by the invention has obvious bacteriostatic and antioxidant activity effects, and can be further researched and developed as an antioxidant health product or medicament.

Description

A kind of mango riboflavonoid extract and preparation method thereof

technical field

The invention relates to a mango riboflavonoid extract and a preparation method thereof, belonging to the fields of food technology and biotechnology.

Background technique

Mango (Latin name: Mangifera indica Linn), also known as citron fruit, stuffed fruit, honeydew, etc., belongs to the sumac family, mainly grows in tropical and subtropical coastal intertidal zones. The cultivation of mangoes is widely distributed. More than 70 countries in the world produce mangoes. The main provinces of my country are Guangxi, Guangdong, Taiwan, and Hainan, where mangoes are grown, and mangoes are rich in resources and varieties. In 2014, my country's mango planting area was 1.17 million hectares, with a total output of 880,000 tons. It is a large traditional tropical agricultural product in China. It has a huge potential for exporting foreign exchange and plays an important role in my country's foreign trade.

Mango is one of the famous tropical fruits, because of its delicate flesh (smooth and juicy), unique flavor (sweet taste), and rich in nutrients (rich in sugar, protein, crude fiber, vitamins, minerals and fats, etc.). Tropical fruit essence in one, known as the "king of tropical fruits". Mango has a very high medicinal value. Its peel can be used as medicine, which is a diuretic and dredging agent; nucleolus and mango leaves can also be used as medicine, which can detoxify, eliminate stagnation, and lower blood pressure. According to the analysis of traditional Chinese medicine, mango is a kind of fruit that is flat and sweet, quenches thirst and promotes fluid production, and has the effects of benefiting the stomach, relieving vomiting and preventing dizziness. Mangiferin contained in mango has obvious anti-peroxidation, protection of brain neurons and expectorant and cough.

As one of the few major provinces in my country that produces mangoes, Guangxi has the advantages of rich mango resources and a wide variety of varieties. However, at present, the market is mainly based on fresh sales from the origin, and a small amount is processed into juice, preserved fruit and pulp. , the mango deep processing industry is in urgent need of further development. Waste from mango production and processing used to be discarded or fed to livestock. In recent years, people have gradually carried out research on mango waste, including mango leaves, mango skin, mango stone, mango bark, etc., especially mango stone, which accounts for 20-60% of the total weight of mango, has attracted the attention of most researchers and entrepreneurs . According to reports, mango core is rich in amino acids, polypeptides, proteins, phenols, flavonoids, polysaccharides, organic acids, saponins, tannins, flavonoids, anthraquinones, alkaloids, coumarins and lactones, triterpenes and steroids, Alkaloids, volatile oils and oils and other active ingredients.

In recent years, more and more researches have been carried out in my country on active ingredients such as mango pit polyphenols, flavonoids, pyridone, terpenoids, and their antibacterial and antioxidant functions. However, there are still many problems. The separation effect is not obvious, the active ingredients obtained by identification are few, and the pharmacological effects of the active ingredients such as bacteriostatic and antioxidant effects are not clear.

The existing mango riboflavonoid extraction method also has problems such as low content of extracted total flavonoids and more organic solvent residues.

SUMMARY OF THE INVENTION

The purpose of the present invention is to make full use of natural resources, especially the characteristic fruit and vegetable resources of Guangxi, to provide a mango riboflavonoid extract and a preparation method thereof. At the same time, in order to solve the problems of low total flavonoid content and more organic solvent residues in the existing mango riboflavonoid extraction methods, the method of the present invention prepares the mango riboflavonoid extract through alcohol extraction and macroporous resin purification method.

The preparation method of the mango riboflavonoid extract of the present invention comprises the following steps:

(1) dry and pulverize the mango pit;

(2) Extract the dried mango pit powder obtained in step (1) with an ethanol solution or methanol solution with an alcohol concentration of 20%-80% (v/v) as a solvent, and then concentrate the obtained extract under reduced pressure or vacuum Freeze-dried to extract;

(3) dissolving the extract obtained in the step (2) with water, an ethanol solution with an alcohol concentration of less than 50% (v/v) or a methanol solution with an alcohol concentration of less than 50% (v/v) to remove the insoluble matter to obtain the above sample solution;

(4) Take D101 macroporous adsorption resin, load the sample solution obtained in step (3), first wash off impurities with water, then elute with an eluent (ethanol solution or methanol solution), and collect the eluent; The weight ratio of the D101 macroporous adsorption resin and mango pit dry powder is 1:0.5-1:20;

(5) Concentrating the eluate collected in step (4) under reduced pressure or vacuum freeze-drying to obtain the mango riboflavonoid extract.

In an embodiment of the present invention, the extraction method in step (2) is to add 2-50L of ethanol solution or methanol solution with an alcohol concentration of 30%-80% per kilogram of mango core powder, and heat reflux or immersion or dipping extraction 2-5 times, 1-3 hours for each extraction during heat reflux extraction, 1-5 days for each extraction during immersion or dipping extraction, and the extracts are combined.

In an embodiment of the present invention, the extraction in the step (2) is to extract the mango pit powder with 40%-60% (v/v, the same below) ethanol.

In an embodiment of the present invention, the extraction in the step (2) is to extract the mango pit powder with ethanol for 1-4 times, most preferably 2-3 times.

In one embodiment of the present invention, in the extraction of the step (2), the ratio of solid to liquid (ratio of mango pit powder mass to ethanol solution, m/v) is 1:1-1:40, preferably 1:5 -1:30, more preferably 1:20-1:30.

In an embodiment of the present invention, in the extraction of the step (2), each extraction time is 0.5-4h, preferably, the extraction time is 1-3h.

In an embodiment of the present invention, the extraction of the step (2) is performed at 30-50°C.

In an embodiment of the present invention, the specific process conditions of the step (2) are: ethanol concentration of 50%, solid-liquid ratio of 1:30, extraction temperature of 40°C, and extraction time of 150 minutes.

In an embodiment of the present invention, the weight of the water, ethanol solution or methanol solution used for dissolving the extract in the step (3) is 3-50 times the weight of the extract.

In one embodiment of the present invention, the extraction method of step (3) is as follows: the extract obtained in step (2) is dissolved with 10 times the weight of the extract in water or 10% ethanol solution, and the insoluble matter is removed to obtain the sample solution.

In an embodiment of the present invention, the removal of insoluble matter in step (3) is performed by filtering or centrifuging.

In an embodiment of the present invention, in step (4), washing with water first to remove impurities refers to eluting with 2-5BV of water to remove impurities.

In an embodiment of the present invention, the elution flow rate of the step (4) is an eluent flow rate of 1BV/h-10BV/h, preferably 2BV/h-5Bv/h.

In an embodiment of the present invention, in the step (4), the concentration of the ethanol solution or methanol solution of the eluent is 5%-95%, preferably 20%-80%; the elution volume of the eluent is 1-10 column volumes, preferably 3-8 column volumes.

In an embodiment of the present invention, the step (4) is to use an ethanol solution with an alcohol concentration of 40-60% for elution.

In one embodiment of the present invention, the purification method of step (4) is as follows: take D101 macroporous resin whose weight ratio to mango stone powder is 1:5, and load the sample loading solution obtained in step (3) , 2-5BV of water is used to remove impurities, and then 3-10BV of 50% ethanol solution is used for elution, and the eluate obtained by elution with 50% ethanol solution is collected.

In an embodiment of the present invention, in the step (4), the loading amount of the mango riboflavonoid extract (mass of mango pip powder: mass of resin) is preferably 1:1-10:1.

In one embodiment of the present invention, the method is as follows: extracting mango riboflavonoids by heating and refluxing an ethanol solution, recovering ethanol, concentrating, adsorbing the mango riboflavonoids extract on a macroporous resin, eluting, and collecting the eluate , concentrated and dried to obtain purified mango riboflavonoids.

In one embodiment of the present invention, the method is specifically: (1) dry and pulverize the mango pit; (2) weigh 1 kg of mango pit powder, add 20L to 25L of ethanol solution with a concentration of 50%, and extract three times under heat reflux , heat reflux extraction for 1 to 3 hours each time, combine the extracts, and concentrate the extracts under reduced pressure; (3) dissolve the extracts with water 5 to 10 times the weight of the extracts, and filter out the insolubles to obtain a sample loading solution; (4) Load the sample solution on the column, control the flow rate of the effluent to be 0.5BV/h, and the volume ratio of the sample load to the resin AB-8 to be 2:1, until all the extraction solution enters the resin bed; use 2-5BV of water to carry out Elution to remove impurities, then rinse the resin bed with a 40-60% ethanol solution of 3-10BV, control the flow rate of the effluent to 2BV/h, and collect the eluent of the ethanol solution; (5) Depressurize the collected eluent Concentrated to a certain volume and spray-dried to obtain the purified product.

The present invention also claims the application of the mango riboflavonoid extract obtained according to the preparation method or purification method in the preparation of food, health care product or medicine.

The health care product or drug is a health care product or drug with antibacterial and antioxidant activity.

Beneficial effects of the present invention:

1. The present invention effectively improves the extraction efficiency of flavonoids by improving the extraction steps of mango ribose flavonoids, and the content of flavonoids in the obtained extract can reach 9.5-12.8 mg/g (that is, 9.5 mg/g can be extracted from the dry powder of 1kg mango pips). -12.8g flavonoids). Moreover, the method of the present invention uses water or ethanol as a solvent, which is cheap and non-toxic.

2. The present invention adopts the method of macroporous resin purification to further purify the extract. The purification method is easy and fast to operate, and the selected macroporous resin has the characteristics of large adsorption capacity and high desorption rate, and effectively removes impurities in the extract. The problem of organic solvent residue is prevented.

3. The mango riboflavonoid extract obtained according to the method of the present invention has a yield of 9.5-12.8 mg/g (dry mango pit), and the obtained flavonoid extract has obvious scavenging effects on hydroxyl and superoxide radical anions.

Description of drawings

Figure 1 is a graph of the standard curve of rutin.

Detailed ways

The technical content of the present invention will be further described below in conjunction with the embodiments. The following embodiments are illustrative, not restrictive, and the protection scope of the present invention cannot be limited by the following embodiments.

The equipment, chemical reagents and detection methods used in each embodiment are as follows:

Instruments and equipment: FW100 high-speed universal pulverizer, Tianjin Test Instrument Co., Ltd.; UPC-Ⅱ-20T Youpu Series Ultrapure Water Device, Sichuan Youpu Ultrapure Technology Co., Ltd.; SQP Electronic Balance, Sartorius Scientific Instruments (Beijing) Co., Ltd.; ZWYR-D2403 Oscillating Incubator, Shanghai Zhicheng Analytical Instrument Manufacturing Co., Ltd.; SHZ-DⅢ Yuhua Brand Circulating Water Vacuum Pump, Gongyi Yuhua Instrument Co., Ltd.; HHS-6S Electronic Constant Temperature Stainless Steel Water Bath Pot, Shanghai Yichang Instrument Yarn Sieve Factory; UV1901PC UV-Vis Spectrophotometer, Shanghai Aoyan Scientific Instrument Co., Ltd.;

Chemical reagents: anhydrous sodium carbonate, methanol, ethanol, acetone, ethyl acetate, Folin phenol reagent, gallic acid standard, various types of macroporous resins, etc. are all provided by Liuzhou Suli Co., Ltd.

Establishment of rutin standard curve: Accurately take 20 mg of rutin standard powder, dissolve it in a 50 mL beaker with 60% ethanol, and place it in a 100 mL volumetric flask, then dilute it to the mark with 60% ethanol, and shake it up; Rutin 0.2mg/mL solution ^[9] ; then get rutin solution 0.0mL, 0.25mL, 0.50mL, 1.0mL, 1.5mL, 2.0mL, 2.5mL respectively and place in 25mL volumetric flask, then add 60% Ethanol solution to 10mL, then add 1mL 5% sodium nitrite solution in turn, shake well, and let stand for 6min; then add 1mL 10% aluminum nitrate solution, shake well, let stand for 6min; add 4% sodium hydroxide solution 1mL; add 60% The volume of ethanol was adjusted to the scale, shaken well, and placed for 15 minutes, and the absorbance value was measured with a spectrophotometer at a wavelength of 500 nm; a standard curve of rutin was prepared, with the concentration of rutin (mg/mL) as the abscissa, and the absorbance value was vertical. Coordinates, draw a standard curve (Figure 1), and perform regression analysis. The results showed that there was a good linear relationship between rutin and its absorbance value in the concentration range of 0-0.07mg/ml, and the linear regression equation in this range was: y=13.403x-0.01063, R ² =0.9991.

Determination of total flavonoid content: Weigh the mango pit powder sample and add ethanol extractant, calculate the total flavonoid concentration in the sample solution according to the rutin standard curve, and then calculate the total flavonoid content in the mango pit powder according to the formula. Calculated as follows:

Total flavonoid content (mg/g)=C×V/W

In the formula, C——the sample concentration value (mg/mL) obtained according to the standard curve;

V——total volume of sample solution (mL);

W - weight of raw material (g).

Determination of Antioxidant Activity:

Determination of the ability of scavenging hydroxyl radicals: Take the sample solutions of different mass concentrations in 5 test tubes, add ferrous sulfate and hydrogen peroxide to each, let stand, add salicylic acid after 10 minutes, and measure the absorbance A at 510 nm after standing _1. Use distilled water instead of salicylic acid to measure the absorbance value A ₂ according to the above method, use distilled water to replace the liquid to be tested and measure the absorbance value A ₀ according to the above method, V _C is used as a control, and calculate the scavenging rate of flavonoids to hydroxyl radicals according to the following formula : hydroxyl radical (%)=[A ₀ -(A ₁ -A ₂ )]/(A ₀ )*100 (%).

Determination of superoxide anion free radical scavenging ability: take 4.5ml Tris-HCl buffer solution, preheat in 20℃ water bath for 20min, add 1ml sample solution of different concentrations and 0.4ml 25mmol/L pyrogallol solution respectively, mix well Then, react in a 25°C water bath for 5 min, then add 1 ml of 8 mol/L HCl solution to terminate the reaction, and then measure the absorbance A ₁ at 325 nm, replace the sample solution with distilled water for the blank, and measure the absorbance A ₀ . The scavenging rate of samples to superoxide anion radicals: scavenging rate (%)=(1-A ₁ /A ₀ )*100 (%).

Determination of DPPH scavenging ability: Take 2.0 mL of different mass concentrations (1.0, 5.0, 10.0, 15.0, mango pit active ingredient sample solution respectively in 5 test tubes, add 2 mL of DPPH (0.04 mg/ml) to each, and let stand for 20 min and detect at 517 nm Absorbance A ₁ ; replace DPPH with absolute ethanol, measure absorbance A ₂ according to the above-mentioned method; do blank group measure absorbance A ₀ by the above-mentioned method, take _VC as a control, the scavenging rate of sample to DPPH free radical: DPPH scavenging rate (% )=[1-(A ₁ -A ₂ )/A ₀ ]*100(%).

Embodiment 1: the preparation method of the mango riboflavonoid extract of the present invention

(1) Extraction of mango riboflavonoids: dry and pulverize mango pits, weigh 1 kg of mango pit powder, add 22 L of ethanol with a concentration of 50%, extract three times under heat reflux, extract under heat reflux for 1.5 hours each time, combine the extracts, and concentrate the extracts under reduced pressure Juice extract, the extract is dissolved in water 10 times the weight of the extract, and the insolubles are filtered out to obtain a sample loading solution with a total flavonoid content of about 11.5g/kg mango pit powder.

(2) Upper column: the mango riboflavonoid extract was loaded onto the column, the flow rate of the effluent was controlled to be 0.5BV/h, and the volume ratio of the loading volume to the resin DM101 was 2:1, until all the extract entered the resin bed.

(3) Elution: use 2-5BV of water to elute to remove impurities, then use 3-10BV of 5%-80% ethanol to rinse the resin bed, control the flow rate of the effluent to 2BV/h, and collect the eluent, Respectively concentrated under reduced pressure to a certain volume, and spray-dried to obtain purified products of different components, wherein the total flavonoid content of all components is about 11.5g/kg, and in the purified product, the flavonoid content in the water eluent is about 1.1g /kg, the content of flavonoids in 5% ethanol eluent is about 1.9g/kg, the content of flavonoids in 10% ethanol eluent is about 2.4g/kg, and the content of flavonoids in 20% ethanol eluent is about 3.8g/kg , the flavonoid content in 30% ethanol eluent is about 4.8g/kg, the flavonoid content in 40% ethanol eluent is about 7.6g/kg, the flavonoid content in 50% ethanol eluent is about 8.1g/kg, the 60 The content of flavonoids in % ethanol eluent is about 7.4g/kg, the content of flavonoids in 70% ethanol eluent is about 5.6g/kg, and the content of flavonoids in 80% ethanol eluent is about 4.6g/kg.

The above data shows that the ethanol solution with an alcohol concentration of 40-60% is used for elution, and the obtained component has the highest flavonoid content.

Example 2: Select different conditions for extraction

Using the method provided by the present invention, the extraction experiment of mango riboflavonoids (sample is 1 g dried and pulverized of mango pits) is selected without conditions. On the basis of single factor experiment, four main factors, ethanol concentration, material-to-liquid ratio, extraction time and extraction temperature, are selected. Do the L ₉ (3 ⁴ ) orthogonal test (Table 1), and the orthogonal result analysis is shown in Table 2.

Table 1 Design of the header of the orthogonal test of the extraction process of mango riboflavonoids

Table 2 The results of orthogonal experiment of L ₉ (3 ⁴ ) extraction process of mango riboflavonoids

Table 3 Mango Riboflavonoid Extraction Verification Test

It can be seen from Table 2 that the order of the influence of various factors on the extraction of total flavonoids from mango pips is: D>A>C>B, that is, solid-liquid ratio>extraction temperature>extraction time>ethanol concentration. The effect was the most significant, and the effect of ethanol concentration on the extraction of mango riboflavonoids was the least significant. From the orthogonal test results, it can be concluded that the best test combination is A ₂ B ₃ C ₂ D ₃ , that is, the extraction temperature is 40°C, the extraction time is 2.5h, the ethanol concentration is 50%, and the solid-liquid ratio is 1:30. Since the optimal process combination is not in the combination of the orthogonal table, it is necessary to do a verification test for analysis. As a result, the flavonoid content obtained by the verification test is 1.91 mg/g, so the optimal process condition for the extraction of mango riboflavonoids is 50% ethanol concentration. The ratio of solid to liquid was 1:30, the extraction temperature was 40 °C, and the extraction time was 150 min.

Example 3: Macroporous resin purification process

The inventors compared the processing effects of different resin types, sample concentration, adsorption rate, sample loading, ethanol elution concentration, elution rate, etc. on the mango riboflavonoid extract.

Preparation of sample solution: Weigh 20 g of mango pit powder, according to ethanol concentration of 50%, solid-liquid ratio of 1:30, extraction temperature of 40 °C, each extraction for 1 hour, and extraction for 3 times. After extraction, the filtrates were combined and concentrated to dryness under reduced pressure. Vacuum dry to constant weight, accurately weigh an appropriate amount of dry paste, dissolve it in a 10ml volumetric flask with methanol or ethanol solution, add methanol or ethanol solution to dilute to the mark, and shake well for later use. Pipette 1ml of filtrate into a 10ml volumetric flask, add methanol or ethanol solution to dilute to the mark, and shake well. The content of flavonoids was determined according to the above method.

Static adsorption: Take 10 g of each of 10 pretreated macroporous adsorption resins, add 10 ml of mango riboflavonoid solution to each, shake once every 10 minutes, and take the resin-adsorbed solution after 2 hours to measure the flavonoid content, and calculate the amount of flavonoids in each resin. Adsorption rate of mango riboflavonoids.

Static desorption:

Filter and drain the statically adsorbed resin, add 30 ml of 50% ethanol for desorption, shake every 10 min, and after 2 h, take the resin-adsorbed solution to measure the flavonoid content, and calculate the desorption rate of mango riboflavonoids by each resin. See Table 4. It can be seen from the table that the adsorption rates of the 10 resins are all large (greater than 90%), and the desorption rates are all greater than 40%. Among them, the desorption rates of AB-8, DM21 and D101 are all greater than 60%. DM21, D101 do dynamic adsorption experiments.

Table 4 Adsorption and desorption rates of mango riboflavonoids by macroporous resin

Dynamic adsorption:

Take 15mL of each of the three preferred resins AB-8, DM21, and D101 for static adsorption after treatment in the column, add the mango riboflavonoid extract on the top of the column, carry out dynamic adsorption at a flow rate of 0.5BV/h, and collect the outflow according to the volume of the resin. The flavonoid content was measured, and the adsorption rate of mango riboflavonoids by each resin was calculated. The results are shown in Table 5, indicating that when AB-8 and D101 reach saturation adsorption, the amount of total flavonoids adsorbed is larger than that of DM21, which was selected for desorption experiments.

Table 5 Dynamic adsorption results of mango riboflavonoids by macroporous resin

Dynamic desorption: take 20ml of each of the treated AB-8 and D101 resins in the column, add mango riboflavonoid extracts to the top of the column, adsorb at a flow rate of 0.5BV/h, and use 50% ethanol at a flow rate of 2BV/h. The flow rate was eluted, and the eluate was collected according to the volume of the resin, the flavonoid content was determined, and the adsorption rate of each resin to mango riboflavonoids was calculated. The results are shown in Table 6. It can be seen that D101 has better desorption effect than AB-8, and it is selected as the best resin.

Table 6 Dynamic desorption results of mango riboflavonoids by macroporous resin

Ethanol concentration: take 10 parts of 20ml of the treated DM101 resin in the column, add mango riboflavonoid extract on the top of the column, carry out adsorption at a flow rate of 0.5BV/h, and then use 5BV of water, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% ethanol was eluted separately at a flow rate of 2BV/h, and the eluate was collected according to the volume of the resin, and the flavonoid content was determined. The results are shown in Table 7. It can be seen that when the ethanol elution concentration is 40%-60%, the eluted total flavonoids have the highest yield.

Table 7 Investigation of ethanol elution concentration

Example 4: Preparation method of mango riboflavonoid extract

Dry and pulverize the mango pits, weigh 1 kg of mango pit powder, add 20 L of ethanol with a concentration of 50%, extract three times under heat reflux, and extract under heat reflux for 1 h each time, combine the extracts, concentrate the extracts under reduced pressure, and extract the extract. 10 times the weight of the paste was dissolved in water, and the insoluble matter was filtered out to obtain a sample loading solution with a total flavonoid content of 12.8 g/kg mango pit powder.

The mango riboflavonoid extract was loaded onto the column, the flow rate of the effluent was controlled to be 0.5BV/h, and the volume ratio of the loading volume to the resin DM101 was 2:1, until all the extract entered the resin bed. Use 3BV of water to remove impurities, and then use 8BV of 50% ethanol to rinse the resin bed, control the flow rate of the effluent to 2BV/h, collect the eluate, and concentrate to a certain volume under reduced pressure, respectively, and spray-drying to obtain the purified product - mango core Flavonoid extract, the flavonoid content is 10.9g/kg mango pit powder.

Example 5: Preparation method of mango riboflavonoids extract

Dry and pulverize the mango pits, weigh 1kg of mango pit powder, add 25L of 50% ethanol, extract three times under heat reflux, extract under heat reflux for 1 hour each time, combine the extracts, concentrate the extracts under reduced pressure, and extract the extract. 10 times the weight of the paste was dissolved in water, and the insolubles were filtered out to obtain a sample loading solution with a total flavonoid content of 12.1 g/kg.

The mango riboflavonoid extract was loaded onto the column, the flow rate of the effluent was controlled to be 0.5BV/h, and the volume ratio of the loading volume to the resin DM101 was 2:1, until all the extract entered the resin bed. Use 2BV of water to remove impurities, then use 10BV of 40% ethanol to rinse the resin bed, control the flow rate of the effluent to 2BV/h, collect the eluate, concentrate under reduced pressure to a certain volume, and spray dry to obtain the purified product - mango core Flavonoid extract, the flavonoid content is 9.8g/kg mango pit powder.

Example 6: Preparation method of mango riboflavonoid extract

Dry and pulverize the mango pits, weigh 1 kg of mango pit powder, add 20 L of 50% ethanol, extract under heat reflux for three times, and extract under heat reflux for 1.5 hours each time, combine the extracts, and concentrate the extracts under reduced pressure. 8 times the weight of the extract was dissolved in water, and the insolubles were filtered out to obtain a sample loading solution with a total flavonoid content of 11.2 g/kg.

The mango riboflavonoid extract was loaded onto the column, the flow rate of the effluent was controlled to be 0.5BV/h, and the volume ratio of the loading volume to the resin DM101 was 2:1, until all the extract entered the resin bed. Use 5BV of water to remove impurities, then use 8BV of 60% ethanol to rinse the resin bed, control the flow rate of the effluent to 2BV/h, collect the eluate, concentrate under reduced pressure to a certain volume, and spray dry to obtain the purified product - mango core Flavonoid extract, the flavonoid content is 9.1g/kg mango pit powder.

Example 7: Antibacterial and antioxidant properties of the mango riboflavonoid extract obtained by the method of the present invention

Determination of antibacterial effect:

的培养皿中，每皿15-20mL培养基，待平板冷却后，每皿中加入0.5mL菌悬液，用三角玻璃涂棒均匀涂成薄板备用。将无菌滤纸片浸入配好的药液其中12h，沥干。每一含菌平板上，呈正三角形排布3片带药无菌纸片，细菌于37℃条件下培养24h，啤酒酵母和黑曲霉于30℃条件下培养5d，测其抑菌圈直径。根据抑菌圈大小来确定其抑菌效果。以不加芒果核黄酮溶液只加菌悬液为对照。结果显示，本发明得到的芒果核黄酮提取物具有强烈的抑菌效果。The test strain was activated on the slant medium, and 1-2 rings were picked with an inoculation ring and shaken in a 50mL sterile saline flask for 10min (with several glass beads in it), and the bacterial suspension was required to be about 10 ⁸ CFU/mL Spare left and right. Weigh a certain amount of the mango riboflavonoid extract samples obtained in Examples 4-6 by vacuum freeze-drying, dissolve the samples with sterile 40% ethanol aqueous solution on the ultra-clean workbench to the required concentration, and set aside for later use. The freshly prepared medium is sterilized at 121°C. When the medium is cooled to 50-60°C, pour the medium into the sterilized medium on the ultra-clean workbench.

Add 0.5 mL of bacterial suspension to each dish after the plate is cooled, and evenly coat it into a thin plate with a triangular glass coating rod for later use. Immerse the sterile filter paper in the prepared medicinal solution for 12h and drain. On each bacteria-containing plate, three sterile paper sheets with medicine were arranged in a equilateral triangle. The bacteria were incubated at 37°C for 24 hours, and the yeast and Aspergillus niger were incubated at 30°C for 5 days, and the diameter of the inhibition zone was measured. The bacteriostatic effect was determined according to the size of the inhibition zone. To control without adding mango riboflavonoid solution and only adding bacterial suspension. The results show that the mango riboflavonoid extract obtained by the present invention has a strong antibacterial effect.

Research on antioxidant activity of mango riboflavonoids:

Hydroxyl is the most active active oxidative free radical, which can induce oxidative damage in the body, and its scavenging rate is often an important indicator of the antioxidant effect of the reaction drug. Similarly, superoxide anion is also the main reactive oxygen radical in the organism, and the lipid peroxidation caused by it is an important cause of aging, cardiovascular disease and tumorigenesis. The determination of the hydroxyl radical scavenging ability of mango riboflavonoids found that its scavenging ability increased with the increase of the concentration of flavonoids. When the concentration of mango riboflavonoids reached 0.38-0.45mg/ml, the scavenging rate could reach about 90%, indicating that mango riboflavonoids have a relatively high scavenging ability. Strong scavenging of hydroxyl radicals. The determination of the superoxide anion radical scavenging ability of mango riboflavonoids showed that the higher the concentration of mango riboflavonoids, the stronger the superoxide anion radical scavenging ability. When the concentration of mango riboflavonoids reached 0.32-0.43mg/mL, the scavenging ability tended to Stable, the clearance rate reached 83%, but the clearance capacity was weaker than that of Vc. The DPPH free radical scavenging ability of mango riboflavonoid extract was measured in experiments and found that its scavenging ability increased with the increase of flavonoid concentration. Mango riboflavonoids have a strong scavenging effect on DPPH free radicals. The above data show that mango riboflavonoids have good scavenging ability to both hydroxyl radicals and superoxide anion radicals. The research results will provide a reference for the comprehensive utilization of mango seed resources and the development of corresponding functional foods.

The content of the present invention has been described in detail above through the embodiments, but the content is only a preferred embodiment of the present invention and cannot be considered to limit the scope of implementation of the present invention. Anyone who is familiar with this technology can make various changes and modifications without departing from the spirit and scope of the present invention. Therefore, the protection scope of the present invention should be defined by the claims.

Claims (2)

1. A preparation method of a mango seed flavone extract is characterized by comprising the following steps:

(1) drying and crushing mango kernels;

(2) extracting the mango seed dry powder obtained in the step (1) by using an ethanol solution with the alcohol concentration of 50% v/v as a solvent, and then concentrating the obtained extracting solution under reduced pressure or freeze-drying the extracting solution in vacuum to obtain an extract;

(3) dissolving the extract obtained in the step (2) with water, an ethanol solution with the alcohol concentration of less than 50% v/v or a methanol solution with the alcohol concentration of less than 50% v/v, and removing insoluble substances to obtain a loading solution;

(4) taking D101 macroporous resin with the weight ratio of 1:5 to mango seed powder, sampling the sampling solution obtained in the step (3), washing with 2-5BV of water to remove impurities, eluting with 3-10BV of 50% ethanol solution, and collecting the eluent obtained by eluting with 50% ethanol solution;

(5) concentrating the eluate collected in the step (4) under reduced pressure or freeze-drying in vacuum to obtain a mango seed flavone extract;

the specific process conditions of the step (2) are as follows: the ethanol concentration is 50%, the ratio of the materials to the liquid is 1:30, the extraction temperature is 40 ℃, and the extraction time is 150 min.

2. The mango kernel flavone extract obtained by the method of claim 1.

Images