CN106860498B - Method for simultaneously extracting polyphenol and flavone - Google Patents
Method for simultaneously extracting polyphenol and flavone Download PDFInfo
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- CN106860498B CN106860498B CN201710041502.2A CN201710041502A CN106860498B CN 106860498 B CN106860498 B CN 106860498B CN 201710041502 A CN201710041502 A CN 201710041502A CN 106860498 B CN106860498 B CN 106860498B
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- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
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- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
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Abstract
The invention belongs to the field of deep processing of agricultural products, and particularly relates to a method for simultaneously extracting polyphenol and flavone, which comprises the following steps: after the olive stones are shelled, the red skins and the nuts are separated, the separated red skins are dried, crushed and sieved, and are added into a mixed solution of ethanol and water, the mixed solution is heated, extracted and filtered, filtrate is collected, concentrated and dried to obtain flavone and polyphenol, and the extraction method has the advantages of high yield and purity, simple operation, convenience, rapidness, low cost and contribution to industrial production.
Description
Technical Field
The invention belongs to the field of deep processing of agricultural products, and particularly relates to a method for simultaneously extracting polyphenol and flavone.
Background
The polyphenol and flavonoid substances have strong physiological activities of resisting tumors, oxidation, bacteria and cancers, removing free radicals, preventing cardiovascular and cerebrovascular diseases and the like, and are natural antioxidants worthy of development.
The olive has more than 2000 years of planting history in China, and is mainly distributed in the provinces of Longnan, Fujian, Guangdong, Guangxi, Hainan and the like at present. A great deal of research shows that the olive contains a plurality of substances beneficial to human bodies, and the olive has great value to human health from the pulp to the kernel. The utilization of olives is mainly used for squeezing olive oil at present, but the deep development and utilization of the olive pit and the olive kernel are almost blank, such as the olive kernel red skin in the olive pit. Therefore, an economical and feasible processing technology of the olive red skin is developed and utilized in the olive industry chain, the added value of the product is improved, and the method has important economic and social significance for increasing the economic income of farmers.
Disclosure of Invention
The invention provides a method for simultaneously extracting polyphenol and flavone, which comprises the following steps:
(1) after the olive stones are shelled, the red skin is separated from the kernels, the separated red skin is dried, crushed and sieved,
wherein, after being crushed, the mixture is sieved by a 60-mesh sieve;
(2) adding the sieved red skins obtained in the step (1) into a mixed solution of ethanol and water, heating, extracting, filtering, collecting filtrate,
wherein, in the mixed solution of ethanol and water, the volume ratio of ethanol to water is 3: 1,
adding cortex Oroxyli into the mixed solution, extracting at 50 deg.C in a constant temperature water bath oscillator under stirring for 30 min;
(3) concentrating and drying the filtrate obtained in the step (2) to obtain flavone and polyphenol,
wherein the filtrate is desolventized and concentrated for 30min on a rotary evaporator, and the concentrated solution is subjected to vacuum freeze drying.
The invention has the beneficial effects that: the yield of polyphenol and flavone of the olive kernel red skin is high, and the purity is high; the extraction method has simple operation, convenience and rapidness, low cost and contribution to industrial production.
Drawings
FIG. 1 shows the specific results of the clearance test in example 1 of the present invention.
FIG. 2 is an absorption spectrum of olive kernel red skin extract in example 1 of the present invention.
Detailed Description
Example 1
(1) After the olive stones are shelled, separating the red skin from the kernels, naturally drying the separated red skin at room temperature (25 ℃), crushing, and sieving by a 60-mesh sieve;
(2) taking 50g of sieved red skin obtained in the step (1), placing the red skin in a beaker, and adding ethanol and water in a volume ratio of 3: 1, placing the beaker in a 50 ℃ constant temperature water bath oscillator, stirring and extracting the red skin for 30min, centrifugally filtering, collecting filtrate,
the detection means is as follows:
diluting the collected filtrate to a constant volume of 1000mL, putting 1mL of sample liquid into a clean 10mL volumetric flask, adding 4mL of ethanol aqueous solution with the mass fraction of 30%, then adding 4mL of aluminum chloride aqueous solution with the mass fraction of 1%, finally using 30% ethanol aqueous solution to a constant volume of 10mL, tightly covering, and shaking up by reversing. At the wavelength of 415nm, a blank reagent is used as a control, the absorbance is measured, and the following can be obtained through calculation: the yield of flavone is 1.5% (mass percentage of the obtained flavone relative to the raw material red skin);
or diluting the collected filtrate to 1000mL, taking 1mL sample solution, adding 5mL distilled water and 1mL forskolin phenol reagent (national drug group chemical reagent Co., Ltd.) into a clean 10mL volumetric flask, finally diluting with 7.5% sodium carbonate solution to 10mL, covering tightly, and shaking up by reversing. At the wavelength 765nm, a blank reagent is used as a control, the absorbance is measured, and the following can be obtained through calculation: the yield of polyphenol is 3% (mass percentage of the obtained polyphenol relative to the raw material red skin);
(3) and (3) desolventizing and concentrating the filtrate collected in the step (2) on a rotary evaporator for 30min, and then carrying out vacuum freeze drying on the concentrated solution to obtain polyphenol and flavone, wherein the total content of the polyphenol and the flavone in the dried product reaches 40% (the mass ratio of the flavone to the polyphenol to the dried product).
Taking 7 cuvettes (numbered 1, 2, 3, 4, 5, 6 and 7) with 10mL, respectively adding 0.3mL of 7.5mmol/L ammonium ferrous sulfate solution, 0.3mL of 7.5mmol/L phenanthroline solution and 1mL of Tris-HCl buffer solution with pH of 7.4, respectively adding 0.2mL of 7.5mmol/L H into the cuvettes No. 2, 3, 4, 5, 6 and 72O2Adding 0.3mL of dispersion prepared from the dried mixture obtained in the step (3) into colorimetric tubes No. 3, 4, 5, 6 and 7 respectively to obtain a solution with the concentration of 1.0 mu g/mL, 5.0 mu g/mL, 10.0 mu g/mL, 20.0 mu g/mL and 40.0 mu g/mL in sequence, adding redistilled water to the volume of 10mL, reacting in a water bath at 37 ℃ for 1h, and reacting in a water bath at 52 DEG CThe absorbance was measured at a wavelength of 0nm, and the hydroxyl radical (. OH) clearance was calculated according to the following formula:
clearance ═ 100% [ (OD sample-OD loss)/(OD blank-OD loss) ]
(wherein "OD sample" is the absorbance detected by cuvettes 3, 4, 5, 6, 7 to which the extract was added; and "OD blank" is the absorbance detected by cuvettes to which no H was added2O2Absorbance detected by a colorimetric cylinder 1 of an aqueous solution; "OD loss" is the addition of H2O2Absorbance detected by cuvette 2 for aqueous solutions. )
The specific clearance calculation result is shown in the attached figure 1, wherein the abscissa represents the specific concentration of 0.3ml of the dispersion prepared from the dried substance obtained in the step (3) added to the relevant colorimetric tube; the ordinate represents the percentage of clearance. Therefore, the olive kernel red skin extract has good antioxidant activity.
Determination of the spectral properties of the product:
weighing 0.01g of the dried olive kernel red skin extract obtained in the step (3), preparing 0.10g/L solution by using 70% by volume of ethanol water solution, and scanning the solution within the range of 200-800 nm on an ultraviolet visible spectrophotometer by using the same solvent (70% by volume of ethanol water solution) as reference to obtain an absorption spectrogram of the olive kernel red skin extract.
As can be seen from the attached figure 2, two obvious absorption peaks are present near the wavelength of 300nm and 360nm in the ultraviolet region, which are typical absorption peaks of polyphenol and flavone.
Claims (5)
1. A method for simultaneously extracting polyphenol and flavone is characterized in that: the method comprises the following steps of,
(1) after the olive stones are shelled, separating the red skin from the kernels, drying, crushing and sieving the separated red skin;
(2) adding the sieved red skins obtained in the step (1) into a mixed solution of ethanol and water, heating, extracting, filtering and collecting filtrate;
wherein the volume ratio of ethanol to water is 3: 1;
the extraction is that the crushed red skin is added into the mixed solution and then extracted in a constant temperature water bath oscillator at 50 ℃;
(3) and (3) concentrating and drying the filtrate obtained in the step (2) to obtain flavone and polyphenol.
2. The method for simultaneously extracting polyphenols and flavonoids as claimed in claim 1, wherein: in the step (1), the mixture is crushed and then sieved by a 60-mesh sieve.
3. The method for simultaneously extracting polyphenols and flavonoids as claimed in claim 1, wherein: in the step (2), the extraction is carried out under a stirring state, and the extraction time is 30 min.
4. The method for simultaneously extracting polyphenols and flavonoids as claimed in claim 1, wherein: and (3) carrying out desolventizing concentration on the filtrate on a rotary evaporator for 30 min.
5. The method for simultaneously extracting polyphenols and flavonoids as claimed in claim 1, wherein: and (3) concentrating the filtrate, and then carrying out vacuum freeze drying.
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Non-Patent Citations (3)
| Title |
|---|
| 杏仁种皮中多酚类物质的提取研究;郭文娟;《食品工程》;20110930(第3期);第43-46页 * |
| 核桃仁种皮中的多酚类物质高压液相色谱分析;万政敏等;《食品工业科技》;20071231;第28卷(第7期);第212-213和224页 * |
| 红松子种皮提取物活性成分及抗氧化作用研究;苏晓雨等;《林产化学与工业》;20100831;第30卷(第4期);第99-102页 * |
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