CN102210786B - Method for extracting natural antioxidant from shells of camellia oleifera - Google Patents
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Abstract
The invention relates to a method for extracting a natural antioxidant from shells of camellia oleifera. With crushed shells of the camellia oleifera as raw materials and water as a solvent, the method comprises the following steps of: extracting by a microwave-assisting extract method to obtain an extracting solution, carrying out centrifugal separation on the extracting solution to obtain a supernatant, separating the supernatant by using a nonpolar or low-polarity macroporous adsorption resin, carrying out elution on the solution by using an aqueous solution of alcohol, and distilling the alcohol from an eluent to obtain a liquid natural antioxidant with procyanidins of the shells of the camellia oleifera as a main components; and carrying out freeze-drying on the obtained liquid natural antioxidant to obtain powdery natural antioxidant. The target product prepared by the method disclosed by the invention has stronger anti-oxidation biological activity.
Description
Technical field
The present invention relates to a kind of method, relate in particular to a kind of method of from shell of Camelliaoleifera Abel, extracting the natural anti-oxidation material from vegetable material extraction natural anti-oxidation active substance.
Background technology
Tea oil tree is the distinctive woody edible oil materials seeds of China, also is and one of Elaeis guineensis Jacq., Fructus Canarii albi, the Cortex cocois radicis world four big woody edible oil sources seeds equally celebrated for their achievements.In nearly 5,000 ten thousand mu of state-owned camellia oleifera lam, produce about 1,000,000 tons of Semen Camelliae per year, but the abundant edible camellia oil of production nutritive value.Shell of Camelliaoleifera Abel (also claiming shell of Camellia oleifera Abel, the peel of oil tea fruit) accounts for 60% of whole oil tea fruit weight, is the residue of oil tea secondary industry, and according to measuring and calculating, its annual growing amount has 1,600,000 tons approximately.Shell of Camelliaoleifera Abel is rich in plant polyphenol, and its main component is proanthocyanidin (procyanidins) class.It is active that proanthocyanidin has very strong antioxidation biology; Can remove harmful free radicals in the human body; Improve the immunity of human body; Can be used as the main effective ingredient of anti-cancer, mutation, control cardiovascular disease medicine and as the new type natural antioxidant of safety non-toxic, become important source material and the food of medicine, health product, the important additives of cosmetics.
Historical existing decades to the bioactive research of proanthocyanidin (mainly being procyanidin) abroad, particularly since the eighties, European countries such as Germany, France reach countries such as Japan, India, Korea S and have carried out number of research projects.China since the eighties development and use research of proanthocyanidin.
Nineteen fifty-one, France Jacques Masquelier found the antioxygenic property of procyanidin first, and successfully from the maritime pine bark, extracted procyanidin.U.S. Joslyn in 1967 etc. isolate the proanthocyanidin polyphenolic substance from Pericarpium Vitis viniferae and Semen Vitis viniferae.Bombardelli had invented the method for from Semen Vitis viniferae, extracting the high assay proto cyaniding mixture in 1976.Jacques Masquelier in 1979 further studies and from Semen Vitis viniferae, extracts procyanidin and realized industrialization.
Abroad mainly contain about the patent of extracting proanthocyanidin: French Jacques Masquellier in 1969 from the maritime pine bark with boiling water extraction, ethyl acetate extraction crude extract, chloroform precipitated product.Japan in 1989 have He Minming with
Aesculus hippocastanumBark is a raw material, uses water extraction, petroleum ether, resin absorption product.U.S. Tochiakl Ariga from Cortex Pini with methanol extraction, petroleum ether, ethyl acetate extraction, liquid-phase chromatographic column separate and to obtain product.Polish Oszmianski Jan in 1996 is a solvent with acetone, adopts ultrasound wave from Semen Vitis viniferae, to extract procyanidin, ethyl acetate extraction, chloroform precipitated product.U.S. Henkel company extracted procyanidin from Semen Vitis viniferae in 1997, with boiling water extraction, and ethyl acetate extraction crude extract, chloroform deposition product.
Shell of Camelliaoleifera Abel is as the by-product of oil tea processing, in actual production, acts as a fuel usually to use or go out of use, and caused the pollution of environment and the waste of resource.In recent years, some researchs show that shell of Camelliaoleifera Abel has stronger antioxidant activity.The research that Lin Shen gives birth to (1997) shows that the acetone-water extract of shell of Camelliaoleifera Abel has very strong non-oxidizability, and its antioxidant content is mainly gallic acid and catechin.Chen Yue (2008) has set up the process of enriching of polyphenol in the shell of Camelliaoleifera Abel, adopts 60% acetone-water to extract, ethyl acetate extraction, and D101 type macroporous resin adsorption, behind 30% ethanol elution, the content of polyphenol has reached 45%.Shen Jianfu (2008) studies the ethanol extraction technology of total flavonoids substance in the shell of Camelliaoleifera Abel; Its research is illustrated in 30 times after 60% heavy soak with ethanol of appearance; 40 ℃ of following ultrasound wave auxiliary extraction 45 min; Extract 2 times continuously, total leaching rate of flavone can reach 97.34%, and the flavone stripping quantity in the shell of Camelliaoleifera Abel is 1.709%
The inventor's seminar has carried out the method research of extracting the proanthocyanidin antioxidant from plant parts such as hair Cortex Myricae Rubrae, Cortex Phyllanthis in recent years, has obtained relevant national inventing patent.But Shang Weijian extracts with the proanthocyanidin method of the natural anti-oxidation material that is the master from shell of Camelliaoleifera Abel.
Summary of the invention
, proanthocyanidin low for the utilization rate that solves the shell of Camelliaoleifera Abel that prior art exists can be used and extract the limited shortcoming of raw material type; The invention provides a kind of method of from shell of Camelliaoleifera Abel, extracting the natural anti-oxidation material; Can improve the added value of shell of Camelliaoleifera Abel, enlarge the raw material sources of proanthocyanidin.
Technical scheme of the present invention is: a kind of method of from shell of Camelliaoleifera Abel, extracting the natural anti-oxidation material; Shell of Camelliaoleifera Abel with after pulverizing is a raw material; With water is solvent, adopts the lixiviate of microwave-assisted extraction method to obtain extracting solution, and the supernatant that obtains after the extracting solution centrifugalize uses nonpolar or low polar macroporous adsorption resin to separate; Ethanol water carries out eluting, and eluent obtains the liquid natural anti-oxidation material that main component is the shell of Camelliaoleifera Abel proanthocyanidin after distilling out ethanol.
The granularity that described shell of Camelliaoleifera Abel is pulverized is 2～4mm.
Described microwave exposure temperature is 60～80 ℃, and exposure time is 15～35min, and the mass volume ratio of shell of Camelliaoleifera Abel and water is 1:15～25g/ml.
Described nonpolar or low polar macroporous adsorption resin is AB-8, D101, D4006, H103.
Macroporous adsorbent resin control upper prop liquid and eluent flow rate are 1.4～1.7 Bv/h.
Described ethanol water is that the volume ratio of second alcohol and water is 60%～70% solution.
Liquid natural anti-reflecting oxide plasmogamy obtains powdery natural anti-oxidation material through after the vacuum lyophilization after being made as aqueous solution again.
Described vacuum is≤0.01MPa.
Beneficial effect:
1. the current research relevant with proanthocyanidin both at home and abroad mainly concentrates on extraction separation natural anti-oxidation active substance from Semen Vitis viniferae, skin and Cortex Pini raw material, and the present invention extracts the natural anti-oxidation active substance from shell of Camelliaoleifera Abel.Oil tea has wide resource distribution in areas such as China Fujian, Guangxi, Jiangxi, Hunan, Zhejiang and Anhui.The annual shell of Camelliaoleifera Abel stock number that produces of China is huge, is rich in proanthocyanidin in the shell of Camelliaoleifera Abel, and therefore target product raw material sources of the present invention are abundant.At present shell of Camelliaoleifera Abel is as the forest products industrial wood waste, thereby is dropped contaminated environment usually.The present invention can further develop the new type natural antioxidant of high added value, realizes the efficient utilization of the forest reserves.
2. the present invention is solvent with water, adopts microwave-assisted to extract, and separates preparation shell of Camelliaoleifera Abel proanthocyanidin through macroporous resin adsorption, and easy and simple to handle, extraction time is short, only needs 15～35 min, and gained proanthocyanidin product oxidation resistance is strong.
Description of drawings
Fig. 1 is the HPLC chromatogram of shell of Camelliaoleifera Abel proanthocyanidin acid degradation product;
524 nm wavelength detect down, and 1, delphinidin (delphinidin), 2, anthocyanidin (cyanidin).
Fig. 2 is the positive ion mode ESI-MS collection of illustrative plates of shell of Camelliaoleifera Abel proanthocyanidin acid degradation product.
A, delphinidin (delphinidin), B, anthocyanidin (cyanidin).
Oil-tea camellia husks tannin component catabolite after the degraded of n-butyl alcohol-hydrochloric acid (95:5) system is analyzed through HPLC, mainly comprises two chromatograph absworption peak (see figure 1)s in the collection of illustrative plates demonstration catabolite.Catabolite shows its molecular ion peak [M] through positive ion mode ESI mass spectral analysis
+Be respectively
M/z303.1 [C
15H
11O
7]
+With 287.1 [C
15H
11O
6]
+(see figure 2) can identify that thus catabolite is respectively delphinidin (delphinidin) and anthocyanidin (cyanidin), can learn thus to have procyanidin and two kinds of structure types of former delphinidin in the shell of Camelliaoleifera Abel proanthocyanidin.
The specific embodiment
A kind of concrete grammar that from shell of Camelliaoleifera Abel, extracts the natural anti-oxidation material is following:
(1) pulverize: the shell of Camelliaoleifera Abel raw material after will purifying is pulverized with the plant pulverizer, sieve aperture Φ=2～4 mm;
(2) extract: with water is solvent, adopts microwave extraction, and the solid-liquid mass volume ratio is 1:15～25,60～80 ℃ of controlled microwave irradiation temperatures, and irradiation time 15～35 min get extracting solution;
(3) centrifugalize: extracting solution is isolated supernatant through centrifuge centrifugal 30 min under rotating speed 5000 rpm, and precipitate discards;
(4) resin absorption separates: with on the above-mentioned supernatant in advance through the nonpolar or low polar macroporous adsorption resin of purified treatment; The upper prop flow velocity is 1.4～1.7 Bv/h; The absorption back is with pure washing post; The reuse volume ratio is the flow velocity eluting of the ethanol water of 60～70% v/v with 1.4～1.7 Bv/h, collects eluent;
(5) desolventizing: at normal pressure (1.01 * 10
5Pa) from pure water elution liquid, steam ethanol under and get liquid natural anti-oxidation material, its main component is the shell of Camelliaoleifera Abel proanthocyanidin.
(6) vacuum lyophilization: use for ease and transport, can obtain powder through vacuum drying.Liquid natural anti-oxidation material is diluted with pure water, get product water solution; Product water solution carries out dehydrate under the vacuum low-pressure of≤0.01 Mpa after freezing, both got powdery natural anti-oxidation active substance.
Embodiment 1:
A kind of method of from shell of Camelliaoleifera Abel, extracting the natural anti-oxidation material:
(1) pulverize: the shell of Camelliaoleifera Abel raw material after will purifying is pulverized (sieve aperture Φ=2～4 mm) with the plant pulverizer.
(2) extract: with water is solvent, adopts microwave extraction, and the solid-liquid mass volume ratio is 1:15～25,60～80 ℃ of controlled microwave irradiation temperatures, and irradiation time 15～35 min get extracting solution; Wherein temperature can be: 60 ℃, 65 ℃, 70 ℃, 75 ℃, 80 ℃.
(3) centrifugalize: extracting solution is isolated supernatant through centrifuge centrifugal 30 min under rotating speed 5000 rpm, and precipitate discards.
(4) resin absorption separates: with on the above-mentioned supernatant in advance through the nonpolar or low polar macroporous adsorption resin of purified treatment; The upper prop flow velocity is 1.4～1.7 Bv/h; The absorption back is with pure washing post; The reuse volume ratio is the flow velocity eluting of the ethanol water of 60～70% v/v with 1.4～1.7 Bv/h, collects eluent.
(5) desolventizing: at normal pressure (1.01 * 10
5Pa) from pure water elution liquid, steam ethanol under, obtaining main component is the liquid natural anti-oxidation material of shell of Camelliaoleifera Abel proanthocyanidin.
(6) vacuum lyophilization:, get product water solution with the pure water dilution.Product water solution carries out dehydrate under the vacuum low-pressure of 0～0.01 Mpa after freezing, both got powdery natural anti-oxidation active substance.
Embodiment 2:
(1) will through purify, air-dry and pulverize shell of Camelliaoleifera Abel raw material 50 g and pure water 750 mL of (using the plant pulverizer) through Φ 2 mm sieve plates, place 1000 mL there-necked flasks; Carry out microwave extraction; 80 ℃ of controlled microwave irradiation temperatures, irradiation time 35 min get extracting solution 620 mL.
(2) extracting solution is cooled off after the centrifuge centrifugalize is provided with centrifuge speed 5000 rpm, centrifugal 30 min isolate supernatant 550 mL.
(3) above-mentioned supernatant is added to the flow velocity of 1.4 Bv/h pass through pretreated AB-8 type macroporous resin (Bv=265 cm
3), then using the pure washing post of 2.4 Bv/h, ethanol-water solution 600 mL of reuse 70% merge eluent with the flow velocity eluting of 1.7 Bv/h.
(4) under normal pressure (1.01 * 10
5Pa) steam ethanol (reuse) in the alcohol-water eluent, obtaining main component is the liquid natural anti-oxidation material of shell of Camelliaoleifera Abel proanthocyanidin.
(5) with the pure water dilution, get product water solution.With the vacuum lyophilization at low temperatures of product water solution, obtain natural anti-oxidation active substance powder product 2.50 g, yield 5.47% (the shell raw material is by over dry).
Embodiment 3:
(1) will through purify, air-dry and pulverize shell of Camelliaoleifera Abel raw material 30 g and pure water 750 mL of (using the plant pulverizer) through Φ 2 mm sieve plates, place 1000 mL there-necked flasks; Carry out microwave extraction; 60 ℃ of controlled microwave irradiation temperatures, irradiation time 35 min get extracting solution 650 mL.
(2) extracting solution is cooled off after the centrifuge centrifugalize is provided with centrifuge speed 5000 rpm, centrifugal 30 min isolate supernatant 580 mL.
(3) above-mentioned supernatant is added to the flow velocity of 1.5 Bv/h pass through pretreated AB-8 type macroporous resin (Bv=265 cm
3), then using the pure washing post of 2.4 Bv/h, ethanol water 500 mL of reuse 70% merge eluent with the flow velocity eluting of 1.5 Bv/h.
(4) under normal pressure (1.01 * 10
5Pa) steam ethanol (reuse) in the ethanol water elution liquid, obtaining main component is the liquid natural anti-oxidation material of shell of Camelliaoleifera Abel proanthocyanidin.
(5) with the pure water dilution, get product water solution.With the vacuum lyophilization at low temperatures of product water solution, obtain natural anti-oxidation active substance powder product 1.60 g, yield 5.83% (the shell raw material is by over dry).
The product that above embodiment is made mixes the back as target product, measures oxidation resistance.
Attach 1: target product is measured the removing ability of free radical among the present invention
(DPPH, U.S. Sigman company) is reference material with diphenyl-picryl-diazanyl free radical, with the clearance rate of spectrophotometry target product of the present invention to the DPPH free radical.
Draw methanol solution (concentration is 0.025 mg/mL) 5 mL of DPPH, mix with methanol solution (concentration is 0.1 mg/mL) 1 mL of target product sample.
λ=517 nm places (is reference with methanol) tracking and measuring light absorption value.Absorbance continues to descend, and till maintenance was invariable, timing was also calculated.
Measure the result:
Absorbance drops to the time of steady state value: 12 min
DPPH clearance rate: 84.97%
DPPH clearance rate: 97 mg/gmin
Result data shows that target product has the removing ability to free radical (DPPH).
Attach 2: the target product TAC is measured among the present invention
(U.S. Sigman company) is tester with ascorbic acid, measures the TAC of target product of the present invention with iron ion reduction/resistance to oxidation (FRAP) algoscopy.
Get 0.1 mL target product sample methanol solution (concentration is 0.1 mg/mL), with 3.0 mL TPTZ working solutions (by 0.3 mol/L acetate buffer, 25 mL, 10 mmol/L TPTZ solution, 2.5 mL, 20 mmol/L FeCl
3Solution 2.5 mL form) reaction 5 min in 25 ℃ of waters bath with thermostatic control, 593 nm places record absorbance, are reference with the deionized water.Experimental result is represented with the amount (mmol AAE/g) of the ascorbic acid that reaches identical oxidation resistance.
Measure the result:
TAC: 6.46 mmol AAE/g
Result data shows that target product has stronger oxidation resistance.
Therefore: target product has antioxidant activity among the present invention; Remove free radical ability and iron ion reduction/resistance to oxidation.
Claims (3)
1. method of from shell of Camelliaoleifera Abel, extracting the natural anti-oxidation material; It is characterized in that: the shell of Camelliaoleifera Abel with after pulverizing is a raw material; With water is solvent, adopts the lixiviate of microwave-assisted extraction method to obtain extracting solution, and the supernatant that obtains after the extracting solution centrifugalize uses nonpolar or low polar macroporous adsorption resin to separate; Ethanol water carries out eluting, and eluent obtains the liquid natural anti-oxidation material that main component is the shell of Camelliaoleifera Abel proanthocyanidin after distilling out ethanol; Described microwave exposure temperature is 60～80 ℃, and exposure time is 15～35min, and the mass volume ratio of shell of Camelliaoleifera Abel and water is 1: 15～25g/ml; Described nonpolar or low polar macroporous adsorption resin is AB-8, D101, D4006, H103; The granularity that described shell of Camelliaoleifera Abel is pulverized is 2～4mm; Macroporous adsorbent resin control upper prop liquid and eluent flow rate are 1.4～1.7Bv/h; Described ethanol water is that the volume ratio of second alcohol and water is 60%～70% solution.
2. the method for from shell of Camelliaoleifera Abel, extracting the natural anti-oxidation material as claimed in claim 1 is characterized in that: liquid natural anti-reflecting oxide plasmogamy obtains powdery natural anti-oxidation material through after the vacuum lyophilization after being made as aqueous solution again.
3. the method for from shell of Camelliaoleifera Abel, extracting the natural anti-oxidation material as claimed in claim 2 is characterized in that: described vacuum is≤0.01MPa.
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Families Citing this family (8)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103030710B (en) * | 2013-01-05 | 2015-03-18 | 广西大学 | Resource utilization method of camellia oleifera shells |
| CN104072468A (en) * | 2013-03-26 | 2014-10-01 | 宜春元博山茶油科技农业开发有限公司 | Proanthocyanidin extraction process from camellia seed hull |
| CN104789354A (en) * | 2015-03-30 | 2015-07-22 | 浙江大学 | Application of myrica rubra leaf proanthocyanidin to lipid antioxidation |
| CN107198668B (en) * | 2017-05-25 | 2020-10-23 | 中南大学 | Extract of antioxidant components in camellia seed hulls as well as extraction method and application thereof |
| CN110408404B (en) * | 2019-08-26 | 2020-06-02 | 江西农业大学 | Application of camellia oleifera shell leaching liquor in reducing soil greenhouse gas emission |
| CN112618430B (en) * | 2020-12-16 | 2023-12-08 | 湖南林之神林韵油茶科技发展有限公司 | Camellia procyanidine mask and preparation method thereof |
| CN112522341A (en) * | 2020-12-30 | 2021-03-19 | 江西神州通油茶科技有限公司 | Co-production method for extracting xylo-oligosaccharide and procyanidine from camellia oleifera shells |
| CN114874173B (en) * | 2022-05-24 | 2024-07-19 | 湖南金昌生物技术有限公司 | Method for extracting procyanidine from fresh camellia oleifera shells |
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