CN103571616A - Extracting composition of camellia and preparation method of extracting composition - Google Patents
Extracting composition of camellia and preparation method of extracting composition Download PDFInfo
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Abstract
The invention discloses an extracting composition of camellia. The extracting composition of the camellia comprises the following components in parts by mass: 15 to 25 parts of palmitic acid, 55 to 70 parts of oleic acid, 5 to 15 parts of a linoleic acid and linolenic acid mixture, 1.5 to 2.5 parts of squalene, 0.2 to 2.5 parts of beta-carotene and 0.5 to 1.5 parts of vitamin E, wherein the mass ratio of the linoleic acid to the linolenic acid is 6:1. The invention also discloses a preparation method of the extracting composition of the camellia. The preparation method comprises the following steps: (1) preparing camellia seed powder; (2) throwing the camellia seed powder into an extraction kettle, performing supercritical extraction and collecting extracts; (3) combining the extracts and performing liquid separation to obtain the extracting composition. The camellia extract provided by the invention has the advantages of clear natural components, high content of active components, zero solvent residue and stable quality; the method provided by the invention simplifies the flow, is high in yield and short in production period and is suitable for large-scale production; carbon dioxide is recycled, environmental pollution is avoided, and industrial 'three wastes' are avoided.
Description
Technical field
The present invention relates to camellia extractive technique field, the concrete extracts composition of a kind of camellia and preparation method thereof that refers to.
Background technology
Camellia (Camellia oleifera Abel) is the distinctive important woody oil tree species of China, and the tea oil that the camellia seed of usining is prepared as raw material is extensively eaten as a kind of edible oil of dietotherapy economic benefits and social benefits.Tea oil contains abundant unsaturated fatty acids, and its composition is similar to sweet oil with ratio, has the title of " east sweet oil ", " treasure in oil ".Tea seed in self-sow state is subject to agricultural chemicals, pollution by chemical fertilizer hardly., containing carcinogenic flavacin and harmful erucic acid and gossypol, not green health food.< < Compendium of Materia Medica > > records: " tea oil is cooler, has the effect of cool blood, hemostasis, heat-clearing, removing toxic substances, cures mainly liver blood loss, expelling parasite, beneficial stomach, improving eyesight ".According to < < Chinese Medicine voluminous dictionary > >, record, tea oil is not only nutritious, also there is important pharmaceutical use, can strengthen blood vessel elasticity and toughness, delay atherosclerosis, increase stomach absorptive function, promote internal secretion body of gland hormone secretion, control neural function declines, and improves the effects such as body immunity.It is multiple to human body beneficiating ingredient that modern study shows that tea oil contains vitamin-E, squalene and carotene etc., tea oil can strengthen blood vessel elasticity and toughness, promotion glandular secretion, improve body immunity, often ediblely can prevent arteriosclerosis, hypotensive, reducing blood-fat, cancer-resisting.Tea oil is a kind of edible oil very with DEVELOPMENT PROSPECT.
< < occupies diet spectrum > > thereupon and records, " tea oil cooking sumptuous courses at a meal, daily all suitable, cook food, there is light in pool, all oil only this most clearly, therefore all diseases are not avoided." visible tea oil is except edible, being used in China outward also has very long history.Tea oil has good skin care moisturizing and hair care effect.First, tea oil has the good absorptive function can be effectively sun-proof to ultraviolet shortwave, and secondly, it is identical with human body skin that the one-tenth of tea oil is grouped into 70%-80%, has good skin-friendly.In tea oil, abundant unsaturated fatty acids can make skin delicate moist.The 3rd, the vitamin-E containing in tea oil can increase skin elasticity and gloss, can also reduce the formation slowing down skin aging of lipofuscin.Tea oil has black effect of turning black widely to be applied as hair-dressing article by tea oil producing region women, through conventional tea oil, combs one's hair and can make hair pitch-black smooth glossy.Modern study show tea oil can also be anti-loss, treatment skin pruritus, chronic eczema.In addition, tea oil can also be bitten by mosquito control, application for the treatment of burn and scald among the people, elimination pouch, and prolonged application can also alleviate even eliminates striae of pregnancy.The good characteristic of tea oil starts the upsurge of a wild tea oil beauty treatment, beauty treatment in the world, and Development and Production has gone out take skin oil, hairdressing oil and the watch box wet goods tea oil cosmetic series product that tea oil is raw material.The makeup of natural senior beauty treatment various in style have been made by Japan with concise tea oil, make its benefit increase by tens times.Relevantly report that Japan reaches more than hundreds of tons for the tea oil of makeup every year.Therefore, tea-seed oil is applied to makeup for insider good.
The extraction of tea oil extensively adopts at present milling process, cold-press and solvent-extraction process.Three has a lot of drawbacks: (1) milling process high temperature steaming and frying technique is easily introduced trans fatty acid and carcinogenic substance benzopyrene, and first residual oil impurity is many, and post-treatment program is complicated.(2) cold-press has been avoided the impact of high temperature steaming and frying technique, but Oleum Camelliae extraction rate is low, and in the tea dregs of rice, resid amount is large, and post-treatment is complicated.(3) solvent method efficiency is low, and solvent loss is large, has dissolvent residual, has a strong impact on the quality of tea oil.(4) gained crude oil will be through coming unstuck, depickling, decolouring, deodorization, dewaxing, precipitation and winterization process, and the operational cycle is long, and productive rate is low, and the loss of nutritive substance is large, also produces a large amount of waste water and dregs.
Supercritical carbon dioxide extraction method is a kind of novel extractive technique, has that percentage extraction is high, a no solvent residue, simple to operate, environmental friendliness, and the advantage such as following process is simple, is used widely at Chinese medicine, chemical field at present.
Patent (CN102641387A) is although relate to tea oil in supercritical carbon dioxide extraction tea seed, and carbon dioxide flow is bigger than normal its objective is that the tea oil of removing in camellia seed prepares for next step extracts tea-polyphenol., to the further separate study of tea oil, tea oil composition is not carried out to identification and analysis yet.Patent (CN101194713A) also relates to tea oil in supercritical carbon dioxide extraction tea seed, but steaming the easy carcinogenic substance of introducing of stir-fry operation, it also can cause that beneficiating ingredient runs off, post-treatment program complexity, and extraction temperature is higher, heat-sensitive substance may change, and extraction and parsing do not have the temperature difference.Patent (CN1557925A) also relates to tea oil in supercritical carbon dioxide extraction tea seed, but pulverizes the meticulous follow-up filtering and impurity removing of also wanting, and the tea oil of extraction only meets makeup, medical grade, tea oil is not formed further and is analyzed.Patent (CN103289816A) relates to supercritical carbon dioxide extraction tea oil, but just for simplifying the refining process of crude oil, and adopts cold-press to make crude oil cannot to solve the problem that oil yield is low.The document that relates to supercritical extraction tea-seed oil has Sun Jiping (supercritical CO
2extraction tea-seed oil preliminary study, grain and grease, the 5th phase in 2002), Wu Xuehui (technical study of supercritical carbon dioxide extraction tea oil, food science and technology, the 2nd phase in 2007), Fang Fang (supercritical CO
2the research of extraction tea-seed oil, grain and oil processing, the 10th phase in 2009) etc. but they only take oil yield and determine processing parameter as index, extraction temperature is higher, can not guarantee the quality of extract, also extract composition is not further studied.Zhou Xiao (the supercritical CO of Camellia oil
2extraction process is analyzed, popular science and technology, total the 95th phase in 2007) extract is not carried out to separation and purification and composition analysis.Li Li (supercritical carbon dioxide extraction camellia seed oil and GC/MS thereof analyze, Chemical Industry in Guangzhou, 39 11 phases of volume in 2011) has carried out GC-MS analysis to extract, but not to the further separation and purification of extract.Zhong Haiyan (supercritical CO
2the preliminary study of extraction tea oil, food & machinery).Zeng Hongyan (analyzes with GC-MS the tea oil lipid acid that different methods extracts, tropical and subtropical plant journal, 13 3 phases of volume in 2005) having compared lipid acid in Different Extraction Method tea oil forms, think that supercritical carbon dioxide extraction is the method for best extraction tea oil, but extraction process is not further optimized the content of activeconstituents in also undetermined oil.Their extraction process parameter is in Table 1:
The tea oil supercritical carbon dioxide extraction parameter of the existing patent of table 1 and document
Summary of the invention
The object of the invention is to provide extracts composition of a kind of camellia and preparation method thereof, utilize supercritical carbon dioxide fluid as extraction agent, camellia compositions useful in extraction camellia seed, make up the deficiency of Oleum Camelliae extraction technology in prior art, improve the quality of the extract of camellia, simplify refining procedure, utilize Camellia Sinensis Extract composition prepared by method of the present invention to form clear, active component content is high, zero dissolvent residual.
For achieving the above object, the extracts composition of camellia provided by the invention, it is characterized in that: by percentage to the quality, the extracts composition of described camellia comprises following component: the mixture of 15～25 palmitinic acids, 55～70 oleic acid, 5～15 linoleic acid plus linolenic acids, 1.5～2.5 squalenes, 0.2～2.5 β-carotene, 0.5～1.5 vitamin-E, and described linolic acid and linolenic mass ratio are 6:1; The combination extract of this camellia prepares by step:
(1) camellia seed is pulverized, and crosses 24 order medicine sieves, obtains camellia seed crushed material;
(2) camellia seed crushed material in step (1) is dropped in extraction kettle and carries out supercritical extraction, extracting pressure is 25～45MPa, and extraction temperature is 20～40 ℃, and extraction time is 90～240min, and separating still I pressure is 10～11MPa, and temperature is 45～50 ℃; Separating still II pressure is 7～8MPa, and temperature is 30～38 ℃; Carbon dioxide flow is 11～19kg/h, and isolated carbonic acid gas recycles, and collects extract;
(3) extract in separating still I and separating still II carry out separatory in combining step (2), the separated oyster white subnatant of removing, obtains the extracts composition of faint yellow transparent camellia.
As preferred version, by percentage to the quality, the extracts composition of described camellia comprises following component: the mixture of 22 palmitinic acids, 60 oleic acid, 12 linoleic acid plus linolenic acids, 2.5 squalenes, 2 β-carotenes, 1.5 vitamin-Es, and described linolic acid and linolenic mass ratio are 6:1; The combination extract of this camellia prepares by step:
(1) camellia seed is pulverized, and crosses 24 order medicine sieves, obtains camellia seed crushed material;
(2) camellia seed crushed material in step (1) is dropped in extraction kettle and carries out supercritical extraction, extracting pressure is 35MPa, and extraction temperature is 30 ℃, and extraction time is 120min, and separating still I pressure is 10MPa, and temperature is 48 ℃; Separating still II pressure is 8MPa, and temperature is 36 ℃; Carbon dioxide flow is 15kg/h, and isolated carbonic acid gas recycles, and collects extract;
(3) extract in separating still I and separating still II carry out separatory in combining step (2), the separated oyster white subnatant of removing, obtains the extracts composition of faint yellow transparent camellia.
Further, described camellia refers to the seed camellia seed of oil tea.
The present invention also provides extracts composition of a kind of camellia and preparation method thereof, and it comprises the following steps:
(1) camellia seed is pulverized, and crosses 24 order medicine sieves, obtains camellia seed crushed material;
(2) camellia seed crushed material in step (1) is dropped in extraction kettle and carries out supercritical extraction, extracting pressure is 25～45MPa, and extraction temperature is 20～40 ℃, and extraction time is 90～240min, and separating still I pressure is 10～11MPa, and temperature is 45～50 ℃; Separating still II pressure is 7～8MPa, and temperature is 30～38 ℃; Carbon dioxide flow is 11～19kg/h, and isolated carbonic acid gas recycles, and collects extract;
(3) extract in separating still I and separating still II carry out separatory in combining step (2), the separated oyster white subnatant of removing, obtains the extracts composition of faint yellow transparent camellia.
Further, it comprises the following steps:
(1) camellia seed is pulverized, and crosses 24 order medicine sieves, obtains camellia seed crushed material;
(2) camellia seed crushed material in step (1) is dropped in extraction kettle and carries out supercritical extraction, extracting pressure is 35MPa, and extraction temperature is 30 ℃, and extraction time is 120min, and separating still I pressure is 10MPa, and temperature is 48 ℃; Separating still II pressure is 8MPa, and temperature is 36 ℃; Carbon dioxide flow is 15kg/h, and isolated carbonic acid gas recycles, and collects extract;
(3) extract in separating still I and separating still II carry out separatory in combining step (2), the separated oyster white subnatant of removing, obtains the extracts composition of faint yellow transparent camellia.
Further, described camellia refers to the seed camellia seed of oil tea.
The invention has the advantages that:
One, this Camellia Sinensis Extract thing pure natural components is clear, and active component content is high, zero dissolvent residual, oxidizable composition, steady quality have been removed in separation.
Its two, service temperature is low, has avoided the loss to thermally labile component and oxidizable composition, has avoided the introducing of objectionable constituent.
Its three, simplified preparation flow, productive rate high, with short production cycle, be suitable for scale operation.
Its four, carbonic acid gas recycles, environmentally safe has been avoided the generation of Industrial " three Waste ".
Know-why of the present invention and technical study:
(1), supercritical carbon dioxide extraction parameter determines
1, single factor is investigated
The investigation of 1.1 extracting pressure
The camellia seed drying and crushing of shelling is crossed to 24 mesh sieves, take 450g and drop in extraction kettle, control that extracting pressure is respectively 25,30,35,40,45MPa, extraction temperature is 30 ℃, and carbon dioxide flow is 15kg/h, and extraction time is 120min, separating still I pressure 10～11MPa, 48 ℃ of temperature; Separating still II pressure 7～8MPa, 36 ℃ of temperature.Collect extract, separation, the calculating percentage extraction of weighing.(see figure 1)
As shown in Figure 1, in the situation that other parameters are definite, percentage extraction increases with the increase of extracting pressure, and after extracting pressure is 35MPa, the increase of percentage extraction is not obvious, therefore slective extraction pressure is 35MPa left and right.
1.2 the investigation of extraction temperature
The camellia seed drying and crushing of shelling is crossed to 24 mesh sieves, take 450g and drop in extraction kettle, control extraction temperature and be respectively 20,25,30,35,40 ℃, extracting pressure is 35MPa, and carbon dioxide flow is 15kg/h, and extraction time is 120min, separating still I pressure 10～11MPa, 48 ℃ of temperature; Separating still II pressure 7～8MPa, 36 ℃ of temperature.Collect extract, separation, the calculating percentage extraction of weighing.(see figure 2)
As shown in Figure 2, in the situation that other parameters are definite, percentage extraction increases with the rising of extraction temperature, and after temperature is 30 ℃, percentage extraction increases not obvious, therefore slective extraction temperature is 30 ℃ of left and right.
1.3 the investigation of carbon dioxide flow
The camellia seed drying and crushing of shelling is crossed to 24 mesh sieves, take 450g and drop in extraction kettle, control extract carbon dioxide flow and be 11,13,15,17,19kg/h, extracting pressure is 35MPa, and extraction temperature is 30 ℃, and extraction time is 120min, separating still I pressure 10～11MPa, 48 ℃ of temperature; Separating still II pressure 7～8MPa, 36 ℃ of temperature.Collect extract, separation, the calculating percentage extraction of weighing.(see figure 3)
As shown in Figure 3, in the situation that other parameters are definite, percentage extraction increases with the rising of carbon dioxide flow, and after carbon dioxide flow is 15kg/h, percentage extraction increases not obvious, therefore select carbon dioxide flow, is 15kg/h left and right.
1.4 the investigation of extraction time
The camellia seed drying and crushing of shelling is crossed to 24 mesh sieves, take 450g and drop in extraction kettle, control extracting pressure is 35MPa, and extraction temperature is 30 ℃, and carbon dioxide flow is 15kg/h,, separating still I pressure 10～11MPa, 48 ℃ of temperature; Separating still II pressure 7～8MPa, 36 ℃ of temperature.At extraction time, be 90,120,150,180, collect extract, separation, the calculating percentage extraction of weighing during 240min.(see figure 4)
As shown in Figure 4, in the situation that other parameters are definite, percentage extraction increases with the prolongation of extraction time, and after extraction time is 120min, percentage extraction increases not obvious, therefore the slective extraction time is 120min left and right.
2, orthogonal experiment
Camellia seed is shelled and is ground into meal, take 450g and drop in extraction kettle, according to orthogonal array, extracting pressure, extraction temperature and carbon dioxide flow are set, each parameter starts timing extraction after reaching parameters.
Percentage extraction %=(gained oil mass/charging capacity) * 100%
The orthogonal experiment level of factor table of table 2. camellia seed oil carbon dioxide upercritical fluid extraction
Table 3. camellia seed oil supercritical carbon dioxide extraction Orthogonal experiment results table
From range analysis result, four factors on the impact of percentage extraction are sequentially: extracting pressure ﹥ extraction temperature ﹥ extraction time ﹥ carbon dioxide flow.Percentage extraction increases with the rising of extracting pressure, with the rising of extraction temperature, increases, and with the increase of carbon dioxide flow, increases, and with the prolongation of extraction time, increases.
Table 4. variance analysis
From the results of analysis of variance, extracting pressure has remarkably influenced to percentage extraction, all the other there are no significant impact.The size that affects of each factor is: extracting pressure ﹥ extraction temperature ﹥ extraction time ﹥ carbon dioxide flow.
Consider actual production, increasing extracting pressure can improve equipment requirements, and the safety coefficient of operation reduces.The stability that rising extraction temperature can affect some composition declines quality product.Gained oil below 30 ℃ and 30 ℃ are golden yellow, and 35 ℃, 40 ℃ gained oil are slightly orange.Known based on the above results, best extraction parameters is: extracting pressure 35MPa, and 30 ℃ of extraction temperature, carbon dioxide flow 15L/h, extraction time is 2h.Separating still I pressure 10～11MPa, 48 ℃ of temperature; Separating still II pressure 7～8MPa, 36 ℃ of temperature.
(2) GC-MS analyzes camellia seed extract
1, the esterification of camellia seed extract
Take camellia seed extract 0.1g and be placed in 50ml Erlenmeyer flask, add 3ml1%KOH methanol solution, 2ml methanol solution, put 10min in 60 ℃ of water-baths, cooling, add 5ml normal hexane, jolt gently 1min, pour standing 15min in separating funnel into, minute get normal hexane layer, with a small amount of anhydrous sodium sulfate dehydration and get final product.
2, GC-MS conditional filtering
GC condition: capillary column (30m*0.25mm*0.25 μ m), carrier gas is helium, flow rate of carrier gas 1.0ml/min, temperature programming.
MS condition: ionization mode EI, sweep limit.Camellia seed extract carries out respectively GC-MS analysis before and after esterification, with Nist08 mass spectral database, carries out retrieval analysis moiety.Concrete screening parameter is in Table 5.
Table 5.GCMS conditional filtering
The best GCMS condition of selecting through testing sieve is: RTx-5MS capillary column (30m*0.25mm*0.25 μ m), and carrier gas is helium, and injector temperature is 250 ℃, and flow rate of carrier gas is 1.0ml/min, and splitting ratio is 60:1.Temperature programming: 100 ℃ of initial temperature, keep 2min, with 5 ℃/min, rise to 200 ℃, keep 1min.With 10 ℃/min, rise to 290 ℃, keep 10min.220 ℃ of MS interface temperature, 200 ℃ of ion source temperatures, ionization mode: EI, sweep limit 50-800m/z.The composition composition of camellia seed extract is in Table 6,7.
The extracts composition of table 6. camellia forms (before esterification)
In table 7. camellia seed, extracts composition forms (after esterification)
Two, efficacy study
Select 240 of the SPF level Kunming kind healthy male mices of Shanghai western pul-Bi Kai laboratory animal company limited breeding.Be divided into large group of V, 48 every group.Carry out respectively dinitrofluorobenzene inducing mouse DTH test, the clearance test of mouse carbon, Turnover of Mouse Peritoneal Macrophages is engulfed chicken red blood cell test, mouse antibodies founder cell and half hemolysis value (HC
50) mouse spleen lymphocyte of test and ConA induction transforms, NK cytoactive is tested.Every group is divided into 12 of 4 ，Mei groups of group more at random.
The picked-up standard of 25 grams of edible oils for each person every day that Chinese Soclety of Nutrition is recommended.If 2.08g/kg*bw, 4.17g/kg*bw, tri-dosage groups of 8.33g/kg*bw (be equivalent to respectively human body (60kg meter) recommended intake 5 times, 10 times and 20 times) and commercially available edible oil group.With blank group.Given the test agent is mixed with basic, normal, high dose concentration with edible oil and is respectively 208mg/mL, 417mg/mL, 833mg/mL, and every day, gavage gave the given the test agent 5ml/kg*bw of respective concentration.Blank group gavage respective amount edible oil.Gavage is measured every strengthening immunity functional parameter after January continuously.
1 tests by the check of < < protective foods and the strengthening immunity functional check method of evaluation technique standard > > (version in 2003).
1.1 dinitrofluorobenzene inducing mouse Delayed onset transformation reactions (DTH)-ear swelling methods:
The continuous gavage of each dosage group mouse is after January, and every mouse shaves off belly wool with shaving a mao machine, the about 3cm * 3cm of scope, and with 10mg/mL dinitrofluorobenzene solution, 50 μ L evenly smear sensitization.Within 5th, with 10mg/mL dinitrofluorobenzene solution 10 μ L, be evenly applied in mouse right ear (two sides) afterwards and attack, after attacking, mouse is put to death in the dislocation of 24h cervical vertebra, cuts left and right two ears, with punch tool, takes off diameter 8mm auricle, weighs.
The degree that represents DTH by the difference of left and right ear weight.The weight difference of given the test agent group is significantly higher than the weight difference of control group, can judge this experimental result positive.
1.2 mouse carbon clearance tests:
The continuous gavage of each dosage group mouse is after January, and every caudal vein injects the india ink (0.1mL/10gbw) of 4 times of dilutions, treats that prepared Chinese ink injects timing immediately.After injection prepared Chinese ink, 2min and 10min get blood 20 μ L from intraocular corner of the eyes venous plexus respectively, and are added to 2mL0.1%Na
2cO
3in solution, with 722 spectrophotometers, at 600nm wavelength place's photometry density value, and get thymus gland, liver, spleen, utilize optical density value, liver is heavy and spleen re-computation phagocytic index a.Another thymus gland/body ratio, the spleen/body ratio of calculating.
With phagocytic index, represent the ability that mouse carbon is cleaned up.The phagocytic index of given the test agent group is significantly higher than the phagocytic index of control group, can judge this experimental result positive.
1.3 Turnover of Mouse Peritoneal Macrophages are engulfed chicken red blood cell test-half intracorporal method
The continuous gavage of each dosage group mouse is after January, preparation 20%(v/v) chicken erythrocyte suspension, every abdominal injection 1mL, interval 30min, mouse is put to death in cervical vertebra dislocation, being faced upward position is fixed on mouse plate, abdominal skin is cut off in center, through Intraperitoneal injection physiological saline 2mL, rotate mouse plate 1min, then sucking-off abdominal cavity washing lotion 1mL, average mark drips on 2 slide glasss, put into the enamel box that is lined with wet gauze, 37 ℃ of constant incubators of dislocation are hatched 30min, then rinsing in physiological saline, dry, with 1:1 acetone methanol solution, fix, 4%(v/v) Giemsa-phosphoric acid buffer dyeing 3min, with distilled water rinsing, dry again, mounting, light Microscopic observation.
The phagocytic percentage of given the test agent group or phagocytic index and control group comparison, difference all has significance, can judge this experimental result positive.
1.4 mouse antibodies founder cell detection-Jerne improvement slide methods
The continuous gavage of each dosage group mouse is after January, every mouse abdominal injection 2%(v/v) SRBC suspension 0.2mL carries out immunity, and after 4d, the dislocation of mouse cervical vertebra is put to death, take out spleen, be placed in the little plate that fills appropriate aseptic Hank ' s liquid, grind spleen, make cell suspension, through 200 eye mesh screens, filter, centrifugal (1000r/min) 10min, washes 2 times with Hank ' s liquid, finally by cell suspension in 5mLRPMI1640 nutrient solution, counting cells number, with RPMI1640 nutrient solution, adjusting cell concn is 5 * 10
6individual/mL.By after the substratum heating for dissolving of top layer, put 45 ℃ of water bath heat preservations, mix with Hank ' the s liquid of equivalent pH7.2-7.4 double strength, packing small test tube, every pipe 0.5mL, then add 10%SRBC(v/v in pipe, with the preparation of SA damping fluid) 50 μ L, 20 μ L splenocyte suspensions (5 * 10
6individual/mL), mix rapidly, be poured on the slide of brushing agarose thin layer, after agar solidifies, slide level is buckled and is placed on horse, put into 37 ℃, 5%CO
2in incubator, hatch 1.5h, then, by joining in slide frame groove after the complement 1:8 dilution preparing, continue to hatch after 1.5h, counting hemolysis plaque number.
With plaque number/10
6splenocyte represents, the plaque number of given the test agent group is significantly higher than the plaque number of control group, can judge this experimental result positive.
1.5 serum hemolysin mensuration-half hemolysis value (HC
50)
The continuous gavage of each dosage group mouse, after January, is prepared 2%(v/v) SRBC suspension, every mouse abdominal injection 0.2mL carries out immunity, plucks eyeball and get blood in centrifuge tube after 4d, places 1h, the centrifugal 10min of 2000r/min, the separated serum of also collecting.After 200 times of dilutions of serum, the optical density value during by method of inspection working sample pipe and SRBC HD50.The amount of hemolysin is with half hemolysis value (HC
50) represent.
The HC of given the test agent group
50be significantly higher than the HC of control group
50, can judge this experimental result positive.
Mouse spleen lymphocyte transformation experiment-the mtt assay of 1.6ConA induction
The continuous gavage of each dosage group mouse is after January, cervical vertebra dislocation method is put to death animal, and the aseptic spleen of getting, is placed in the little plate that fills appropriate aseptic Hank ' s liquid, grind spleen, make cell suspension, through 200 eye mesh screens, filter centrifugal (1000r/min) 10min, with Hank ' s liquid, wash 2 times, then by cell suspension in 1mLRPMI1640 complete culture solution, the blue dyeing counting viable count (all more than 95%) of platform phenol, is 3 * 10 with RPMI1640 nutrient solution adjustment cell concn
6individual/mL.Divide two holes to add in 24 well culture plates cell suspension, 1mL，Yi hole, every hole adds 75 μ LConA liquid, and another hole in contrast, is placed in 37 ℃, 5%CO
2in incubator, cultivate 72h.Cultivation finishes front 4h, and every hole sucks supernatant liquor 0.7mL gently, adds 0.7mL not containing the RPMI1640 nutrient solution of calf serum, adds MTT(5mg/mL simultaneously) 50 μ L/ holes, continue to cultivate 4h.After cultivation finishes, every hole adds 1mL acid isopropyl alcohol, and piping and druming mixes, and purple crystal is dissolved completely.Solution is moved in 96 well culture plates, by microplate reader, under wavelength 570nm, measure each sample hose optical density value.
By the optical density value that adds ConA hole, deduct the optical density value that does not add ConA hole and represent lymphocytic multiplication capacity, the optical density(OD) difference of given the test agent group is significantly higher than the optical density(OD) difference of control group, can judge this experimental result positive.
1.7NK cytoactive detection-determination of lactate dehydrogenase method
The continuous gavage of each dosage group mouse is after January, cervical vertebra dislocation method is put to death animal, the aseptic spleen of getting, be placed in the little plate that fills appropriate aseptic Hank ' s liquid, grind spleen, make cell suspension, through 200 eye mesh screens, filter, with Hank ' s liquid, wash 2 times, centrifugal (1000r/min) 10min at every turn, abandoning supernatant upsprings cytoplasm, add 0.5mL aqua sterilisa within 20 seconds, after splitting erythrocyte, to add again 0.5mL2 times of Hank ' s liquid and 8mLHank ' s liquid, centrifugal 10min(1000r/min), resuspended containing 10% calf serum RPMI1640 complete culture solution with 1.0mL, with counting after 1% glacial acetic acid dilution, the blue dyeing counting viable count (all more than 95%) of platform phenol, with RPMI1640 nutrient solution, adjusting cell concn is 2 * 10
7individual/mL.
Before experiment, 24h, by target cell (YAC-1 cell) cultivations of going down to posterity, washes 3 times with Hank ' s liquid before application, with RPMI1640 complete culture solution adjustment cell concn, is 4 * 10
5individual/mL. gets YAC-1 cell and each 100 μ L(of splenocyte is imitated target than 50:1) add in U-shaped 96 well culture plates, YAC-1 cell Spontaneous release hole adds YAC-1 cell and each 100 μ L of nutrient solution, the maximum release aperture of YAC-1 cell adds YAC-1 cell and each 100 μ L of 1%NP40, above-mentioned every three parallel holes of all establishing, in 37 ℃, 5%CO
2in incubator, cultivate 4h, then by 96 well culture plates with the centrifugal 15min of 1500r/min, draw at the bottom of supernatant liquor 100 μ L horizontalizations in 96 well culture plates in every hole, add LDH matrix liquid 100 μ L simultaneously, reaction 10min, every hole adds the HCl30 μ L of 1mol/L, measures optical density value at microplate reader 490nm place.
The NK cytoactive of given the test agent group is significantly higher than the NK cytoactive of control group, can judge this experimental result positive.
1.8 data analysis
With spss10.0 software, each experiment raw data is carried out to homogeneity test of variance, meet the data information that homoscedasticity requires, with the comparative approach between two of mean between a control group of a plurality of experimental group in one-way analysis of variance method, carry out statistical treatment; The data information of skewed distribution or heterogeneity of variance is carried out to suitable variable conversion, after meeting the neat requirement of normal state or variance, by the data of conversion gained, carry out statistical treatment.
1.9 results are judged
Aspect four of cellular immune functions, humoral immune function, monocytes/macrophages function, NK cytoactive, any two aspect results are positive, can judge that this given the test agent has strengthening immunity function.
1, result
In experimentation, drinking water for animals is ingested normally, and outward appearance is without extremely.
1.1 impacts of camellia seed extract on Mouse Weight
From table 8, relatively, there are no significant for difference for initial body weight 2.08g/kg*bw, the 4.17g/kg*bw of mouse, 8.33g/kg*bw and blank group.The initial body weight that shows mouse is comparatively balanced between each group.
Gavage gives the Camellia Sinensis Extract thing of mouse various dose after January, each dosage group weight gain value is through normal distribution, homogeneity test of variance, meet normal distribution, homoscedasticity requirement, with the comparative approach between two of mean between a plurality of experimental group in one-way analysis of variance method and a control group, carry out statistical treatment.Relatively, there are no significant for difference for 2.08g/kg*bw, 4.17g/kg*bw, 8.33g/kg*bw and blank group.
The impact of table 8 camellia seed extract on Mouse Weight
Continued 8
Continued 8
Continued 8
Continued 8
The impact of 2.2 camellia seed extracts on mouse thymus, spleen organ
Gavage gives the Camellia Sinensis Extract thing of mouse various dose after January, each dosage group thymus gland/body weight, spleen/body weight are through homogeneity test of variance, meet homoscedasticity requirement, with the comparative approach between two of mean between a plurality of experimental group in one-way analysis of variance method and a control group, carry out statistical treatment.Relatively, there are no significant for difference for 2.08g/kg*bw, 4.17g/kg*bw, 8.33g/kg*bw and blank group.In Table 9:
The impact of table 9 camellia seed extract on mouse thymus, spleen organ
The impact that 2.3 camellia seed extracts transform the mouse spleen lymphocyte of ConA induction
Gavage gives the Camellia Sinensis Extract thing of mouse various dose after January, the difference that adds ConA hole and do not add ConA hole absorbancy is through homogeneity test of variance, meet homoscedasticity requirement, with the comparative approach between two of mean between a plurality of experimental group in one-way analysis of variance method and a control group, carry out statistical treatment.8.33g/kg*bw group compares with blank group, and difference has significance.In Table 10:
The impact that table 10 camellia seed extract transforms the mouse spleen lymphocyte of ConA induction
The impact on DNFB inducing mouse DTH on mouse of 2.4 camellia seed extracts
Gavage gives the Camellia Sinensis Extract thing of mouse various dose after January, and auricular concha increases weight through homogeneity test of variance, meets homoscedasticity requirement, with the comparative approach between two of mean between a plurality of experimental group in one-way analysis of variance method and a control group, carries out statistical treatment.8.33g/kg*bw group compares with blank group, and difference has significance.
Gavage gives the Camellia Sinensis Extract thing of mouse various dose after January, and auricular concha increases weight through homogeneity test of variance, meets homoscedasticity requirement, with the comparative approach between two of mean between a plurality of experimental group in one-way analysis of variance method and a control group, carries out statistical treatment.8.33g/kg*bw compares with blank group, and difference has significance.In Table 11:
The impact on DNFB inducing mouse DTH on mouse of table 11 camellia seed extract
The impact of 2.5 camellia seed extracts on mouse antibodies founder cell (hemolysis plaque number)
Gavage gives the Camellia Sinensis Extract thing of mouse various dose after January, hemolysis plaque number is through normal distribution, homogeneity test of variance, meet normal distribution, homoscedasticity requirement, with the comparative approach between two of mean between a plurality of experimental group in one-way analysis of variance method and a control group, carry out statistical treatment.8.33g/kg*bw compares with blank group, and difference has significance.In Table 12:
The impact of table 12 camellia seed extract on mouse antibodies founder cell (hemolysis plaque number)
2.6 camellia seed extracts are to mice serum half hemolysis value (HC
50) impact
Gavage gives the Camellia Sinensis Extract thing of mouse various dose after January, HC
50through normal distribution, homogeneity test of variance, meet normal distribution, homoscedasticity requirement, with the comparative approach between two of mean between a plurality of experimental group in one-way analysis of variance method and a control group, carry out statistical treatment.8.33g/kg*bw compares with blank group, and difference has significance.In Table 13:
Table 13 camellia seed extract is to mice serum half hemolysis value (HC
50) impact
2.7 camellia seed extracts are cleaned up the impact of ability on mouse carbon
Gavage gives the Camellia Sinensis Extract thing of mouse various dose after January, and phagocytic index a, through homogeneity test of variance, meets homoscedasticity requirement, with the comparative approach between two of mean between a plurality of experimental group in one-way analysis of variance method and a control group, carries out statistical treatment.Relatively, there are no significant for difference for 2.08g/kg*bw, 4.17g/kg*bw, 8.33g/kg*bw and blank group.In Table 14:
Table 14 camellia seed extract is cleaned up the impact of ability on mouse carbon
2.8 camellia seed extracts are engulfed the phagocytic percentage of chicken red blood cell and the impact of phagocytic index to Turnover of Mouse Peritoneal Macrophages
Gavage gives the Camellia Sinensis Extract thing of mouse various dose after January, and phagocytic index, phagocytic percentage are through sin
-1p
1/2(P is phagocytic percentage, represents decimally) carries out homogeneity test of variance after transforming, and meets homoscedasticity requirement, with the comparative approach between two of mean between a plurality of experimental group in one-way analysis of variance method and a control group, carries out statistical treatment.8.33g/kg*bw group compares with blank group, and difference has significance.In Table 15:
Table 15 camellia seed extract is engulfed the phagocytic percentage of chicken red blood cell and the impact of phagocytic index to Turnover of Mouse Peritoneal Macrophages
2.9 impacts of camellia seed extract on NK cells in mice activity
Gavage gives the Camellia Sinensis Extract thing of mouse various dose after January, and NK cytoactive is through sin
-1p
1/2(P is NK cytoactive, represents decimally) carries out homogeneity test of variance after transforming, and meets homoscedasticity requirement, with the comparative approach between two of mean between a plurality of experimental group in one-way analysis of variance method and a control group, carries out statistical treatment.Relatively, there are no significant for difference for 2.08g/kg*bw, 4.17g/kg*bw, 8.33g/kg*bw and blank group.In Table 16:
The impact of table 16 camellia seed extract on NK cells in mice activity
3, conclusion
Camellia seed extract is with 2.08g/kg*bw, 4.17g/kg*bw, 8.33g/kg*bw successive administration January, and result shows:
(1) in the mouse spleen lymphocyte transformation experiment of cellular immune function: ConA induction, 8.33g/kg*bw group adds ConA hole and is significantly higher than blank group with the difference that does not add ConA hole absorbancy; In the experiment of dinitrofluorobenzene inducing mouse Delayed onset transformation reactions, the weightening finish of 8.33g/kg*bw group auricular concha is significantly higher than blank group.
(2) humoral immune function: in antibody-producting cell test experience, 8.33g/kg*bw group hemolysis plaque number is significantly higher than blank group; 8.33g/kg*bw group half hemolysis value (HC in the determination experiment of serum hemolysin
50) be significantly higher than blank group.
(3) monocytes/macrophages function: Turnover of Mouse Peritoneal Macrophages is engulfed 8.33g/kg*bw group phagocytic percentage and phagocytic index in chicken red blood cell experiment and is significantly higher than blank group.
Therefore think that camellia seed extract has strengthening immunity function.
Three, comprehensively analyze
(1) optimal processing parameter of the extract of co_2 supercritical fluid extraction camellia is: extracting pressure 35MPa, and 30 ℃ of extraction temperature, carbon dioxide flow 15L/h, extraction time is 2h.Separating still I pressure 10-11MPa, 48 ℃ of temperature; Separating still II pressure 7-8MPa, 36 ℃ of temperature.
(2) GC-MS analytical results shows that the extract of camellia contains the unsaturated fatty acidss such as abundant oleic acid, linoleic acid plus linolenic acid, and ratio is appropriate,, containing erucic acid, is not a kind of desirable edible oil.
(3) in camellia seed extract, also contain the material of the multiple beneficial HUMAN HEALTH such as squalene, β-sitosterol, β-carotene, vitamin-E, in addition, the main component tea oil of extract also has good skin-friendly, stronger uv-absorbing function and shinny function of crow.Therefore the extract of camellia has good DEVELOPMENT PROSPECT aspect food, medicine and cosmetology.
(4) before extraction, need not strictly control water content, moisture and some water-soluble impurities can be removed in extract in separation.For simplifying technological operation, provide foundation.
(5) extract lower floor is a small amount of milky white liquid, and upper strata is faint yellow oil.Subnatant oxidation stain in air after separated, affects the quality of the extract of camellia, therefore remove.The extract of the camellia obtaining after separation is faint yellow transparent stable homogeneous.
(6) camellia seed extract has the function of strengthening immunity.
Accompanying drawing explanation
Fig. 1 is that percentage extraction of the present invention is with the variation diagram of extracting pressure.
Fig. 2 is that percentage extraction of the present invention is with the variation diagram of extraction temperature.
Fig. 3 is that percentage extraction of the present invention is with the variation diagram of carbon dioxide flow.
Fig. 4 is that percentage extraction of the present invention is with the variation diagram of extraction time.
Embodiment
Below in conjunction with embodiment, the present invention is described in further detail.
Embodiment mono-:
Get the camellia seed 400g separated with shell, pulverize, cross 24 mesh sieves, obtain camellia seed crushed material 386g.Camellia seed crushed material is dropped in 1L extraction kettle, and setting extracting pressure is 25MPa, and extraction temperature is 20 ℃, and separating still I pressure is 11MPa, and temperature is 45 ℃; Separating still II pressure is 7MPa, and temperature is 38 ℃; Carbon dioxide flow 11kg/h, isolated carbonic acid gas recycles, extraction 90min after equipment is stable, and the extract of collecting in separating still I, separating still II merges, further separation obtains flaxen transparent oily Camellia Sinensis Extract thing 133g again, and yield is 34.46%.Analyze the extracts composition of this camellia, its component is: the mixture of 15% palmitinic acid, 70% oleic acid, 8.5% linoleic acid plus linolenic acid, 2.5% squalene, 2.5% β-carotene, 1.5% vitamin-E.(yield=extract content/charging capacity)
Embodiment bis-:
Get the camellia seed 500g separated with shell, pulverize, cross 24 mesh sieves, obtain camellia seed crushed material 453g.Camellia seed crushed material is dropped in 1L extraction kettle, and setting extracting pressure is 35MPa, and extraction temperature is 30 ℃, and separating still I pressure is 10MPa, and temperature is 48 ℃; Separating still II pressure is 8MPa, and temperature is 36 ℃; Carbon dioxide flow 15kg/h, isolated carbonic acid gas recycles, extraction 120min after equipment is stable, and the extract of collecting in separating still I, separating still II merges, further separation obtains flaxen transparent oily Camellia Sinensis Extract thing 140.4g again, and yield is 30.99%.Analyze the extracts composition of this camellia, its component is: the mixture of 25% palmitinic acid, 55% oleic acid, 14.5% linoleic acid plus linolenic acid, 1.5% squalene, 2.5% β-carotene, 1.5% vitamin-E.
Embodiment tri-:
Get the camellia seed 600g separated with shell, pulverize, cross 24 mesh sieves, obtain camellia seed crushed material 568g.Camellia seed crushed material 450g is dropped in 1L extraction kettle, and setting extracting pressure is 45MPa, and extraction temperature is 40 ℃, and separating still I pressure is 10MPa, and temperature is 50 ℃; Separating still II pressure is 8MPa, and temperature is 35 ℃; Carbon dioxide flow 19kg/h, isolated carbonic acid gas recycles, extraction 120min after equipment is stable, and the extract of collecting in separating still I, separating still II merges, further separation obtains flaxen transparent oily Camellia Sinensis Extract thing 149.3g again, and yield is 33.18%.Analyze the extracts composition of this camellia, its component is: the mixture of 25% palmitinic acid, 67.3% oleic acid, 5% linoleic acid plus linolenic acid, 2% squalene, 0.2% β-carotene, 0.5% vitamin-E.
Embodiment tetra-:
Get the camellia seed 400g separated with shell, pulverize, cross 24 mesh sieves, obtain camellia seed crushed material 386g.Camellia seed crushed material is dropped in 1L extraction kettle, and setting extracting pressure is 35MPa, and extraction temperature is 35 ℃, and separating still I pressure is 11MPa, and temperature is 45 ℃; Separating still II pressure is 7MPa, and temperature is 30 ℃; Carbon dioxide flow 15kg/h, isolated carbonic acid gas recycles, extraction 120min after equipment is stable, and the extract of collecting in separating still I, separating still II merges, further separation obtains flaxen transparent oily Camellia Sinensis Extract thing 130.5g again, and yield is 33.81%.Analyze the extracts composition of this camellia, its component is: the mixture of 22.5% palmitinic acid, 56% oleic acid, 15% linoleic acid plus linolenic acid, 2.5% squalene, 2.5% β-carotene, 1.5% vitamin-E.
Embodiment five:
Get the camellia seed 500g separated with shell, pulverize, cross 24 mesh sieves, obtain camellia seed crushed material 453g.Camellia seed crushed material is dropped in 1L extraction kettle, and setting extracting pressure is 40MPa, and extraction temperature is 36 ℃, and separating still I pressure is 10MPa, and temperature is 49 ℃; Separating still II pressure is 7MPa, and temperature is 37 ℃; Carbon dioxide flow 16kg/h, isolated carbonic acid gas recycles, extraction 120min after equipment is stable, and the extract of collecting in separating still I, separating still II merges, further separation obtains flaxen transparent oily Camellia Sinensis Extract thing 149.4g again, and yield is 32.98%.Analyze the extracts composition of this camellia, its component is: the mixture of 20% palmitinic acid, 60% oleic acid, 14% linoleic acid plus linolenic acid, 2.5% squalene, 2.3% β-carotene, 1.2% vitamin-E.
Embodiment six:
Get the camellia seed 600g separated with shell, pulverize, cross 24 mesh sieves, obtain camellia seed crushed material 568g.Camellia seed crushed material 450g is dropped in 1L extraction kettle, and setting extracting pressure is 30MPa, and extraction temperature is 30 ℃, and separating still I pressure is 10MPa, and temperature is 48 ℃; Separating still II pressure is 8MPa, and temperature is 36 ℃; Carbon dioxide flow 15kg/h, isolated carbonic acid gas recycles, extraction 120min after equipment is stable, and the extract of collecting in separating still I, separating still II merges, further separation obtains flaxen transparent oily Camellia Sinensis Extract thing 135.8g again, and yield is 30.18%.Analyze the extracts composition of this camellia, its component is: the mixture of 22% palmitinic acid, 60% oleic acid, 12% linoleic acid plus linolenic acid, 2.5% squalene, 2% β-carotene, 1.5% vitamin-E.
Embodiment seven:
Get the camellia seed 400g separated with shell, pulverize, cross 24 mesh sieves, obtain camellia seed crushed material 386g.Camellia seed crushed material is dropped in 1L extraction kettle, and setting extracting pressure is 35MPa, and extraction temperature is 30 ℃, and separating still I pressure is 10MPa, and temperature is 48 ℃; Separating still II pressure is 8MPa, and temperature is 36 ℃; Carbon dioxide flow 15kg/h, isolated carbonic acid gas recycles, extraction 120min after equipment is stable, and the extract of collecting in separating still I, separating still II merges, further separation obtains flaxen transparent oily Camellia Sinensis Extract thing 122.7g again, and yield is 31.79%.Analyze the extracts composition of this camellia, its component is: the mixture of 23% palmitinic acid, 67% oleic acid, 5% linoleic acid plus linolenic acid, 1.5% squalene, 2% β-carotene, 1.5% vitamin-E.
Claims (6)
1. the extracts composition of a camellia, it is characterized in that: by percentage to the quality, the extracts composition of described camellia comprises following component: the mixture of 15～25 palmitinic acids, 55～70 oleic acid, 5～15 linoleic acid plus linolenic acids, 1.5～2.5 squalenes, 0.2～2.5 β-carotene, 0.5～1.5 vitamin-E, and described linolic acid and linolenic mass ratio are 6:1; The combination extract of this camellia prepares by step:
(1) camellia seed is pulverized, and crosses 24 order medicine sieves, obtains camellia seed crushed material;
(2) camellia seed crushed material in step (1) is dropped in extraction kettle and carries out supercritical extraction, extracting pressure is 25～45MPa, and extraction temperature is 20～40, and ℃ extraction time is 90～240min, and separating still I pressure is 10～11MPa, and temperature is 45～50 ℃; Separating still II pressure is 7～8MPa, and temperature is 30～38 ℃; Carbon dioxide flow is 11～19kg/h, and isolated carbonic acid gas recycles, and collects extract;
(3) extract in separating still I and separating still II carry out separatory in combining step (2), the separated oyster white subnatant of removing, obtains the extracts composition of faint yellow transparent camellia.
2. the extracts composition of camellia according to claim 1, it is characterized in that: by percentage to the quality, the extracts composition of described camellia comprises following component: the mixture of 22 palmitinic acids, 60 oleic acid, 12 linoleic acid plus linolenic acids, 2.5 squalenes, 2 β-carotenes, 1.5 vitamin-Es, and described linolic acid and linolenic mass ratio are 6:1; The combination extract of this camellia prepares by step:
(1) camellia seed is pulverized, and crosses 24 order medicine sieves, obtains camellia seed crushed material;
(2) camellia seed crushed material in step (1) is dropped in extraction kettle and carries out supercritical extraction, extracting pressure is 35MPa, and extraction temperature is 30, and ℃ extraction time is 120min, and separating still I pressure is 10MPa, and temperature is 48; ℃ separating still II pressure is 8MPa, and temperature is 36; ℃ carbon dioxide flow is 15kg/h, and isolated carbonic acid gas recycles, and collects extract;
(3) extract in separating still I and separating still II carry out separatory in combining step (2), the separated oyster white subnatant of removing, obtains the extracts composition of faint yellow transparent camellia.
3. the extracts composition of camellia according to claim 1 and 2, is characterized in that: described camellia refers to the seed camellia seed of oil tea.
4. extracts composition of camellia and preparation method thereof according to claim 1, it comprises the following steps:
(1) camellia seed is pulverized, and crosses 24 order medicine sieves, obtains camellia seed crushed material;
(2) camellia seed crushed material in step (1) is dropped in extraction kettle and carries out supercritical extraction, extracting pressure is 25～45MPa, and extraction temperature is 20～40, and ℃ extraction time is 90～240min, and separating still I pressure is 10～11MPa, and temperature is 45～50 ℃; Separating still II pressure is 7～8MPa, and temperature is 30～38 ℃; Carbon dioxide flow is 11～19kg/h, and isolated carbonic acid gas recycles, and collects extract;
(3) extract in separating still I and separating still II carry out separatory in combining step (2), the separated oyster white subnatant of removing, obtains the extracts composition of faint yellow transparent camellia.
5. extracts composition of camellia according to claim 2 and preparation method thereof, is characterized in that: it comprises the following steps:
(1) camellia seed is pulverized, and crosses 24 order medicine sieves, obtains camellia seed crushed material;
(2) camellia seed crushed material in step (1) is dropped in extraction kettle and carries out supercritical extraction, extracting pressure is 35MPa, and extraction temperature is 30, and ℃ extraction time is 120min, and separating still I pressure is 10MPa, and temperature is 48; ℃ separating still II pressure is 8MPa, and temperature is 36; ℃ carbon dioxide flow is 15kg/h, and isolated carbonic acid gas recycles, and collects extract;
(3) extract in separating still I and separating still II carry out separatory in combining step (2), the separated oyster white subnatant of removing, obtains the extracts composition of faint yellow transparent camellia.
6. according to extracts composition of the camellia described in claim 4 or 5 and preparation method thereof, it is characterized in that: described camellia refers to the seed camellia seed of oil tea.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104673485A (en) * | 2015-01-29 | 2015-06-03 | 衢州市质量技术监督检测中心 | Method for removing polyaromatic hydrocarbon pollutants from camellia oil |
| CN104758331A (en) * | 2015-04-30 | 2015-07-08 | 溧阳市天目湖保健品有限公司 | Chinese medicine composition capable of enhancing immunity and preparation method thereof |
| CN107114491A (en) * | 2017-05-08 | 2017-09-01 | 湖北鑫尔绿色食品有限公司 | The elderly's tea oil and its production method |
| CN109157454A (en) * | 2018-10-25 | 2019-01-08 | 上海清轩生物科技有限公司 | A kind of Camellia combined extracts and its preparation and the application in cosmetics |
| CN112316484A (en) * | 2020-10-31 | 2021-02-05 | 洞口县亿丰农林牧科技有限公司 | Method for extracting camellia oleifera polyphenol from tea oil |
| CN113068750A (en) * | 2021-04-10 | 2021-07-06 | 贵州长基供应链管理有限公司 | Camellia blend oil for improving immunity and processing method thereof |
| CN117778095A (en) * | 2023-08-07 | 2024-03-29 | 湖南金昌生物技术有限公司 | Preparation method of camellia seed extract |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103289816A (en) * | 2013-05-14 | 2013-09-11 | 广州绿和缘生物科技有限公司 | Tea oil processing method and device special for tea oil processing method |
-
2013
- 2013-11-22 CN CN201310594946.0A patent/CN103571616A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103289816A (en) * | 2013-05-14 | 2013-09-11 | 广州绿和缘生物科技有限公司 | Tea oil processing method and device special for tea oil processing method |
Non-Patent Citations (5)
| Title |
|---|
| 曾虹燕等: "用GC-MS分析不同方法提取的茶油脂肪酸", 《热带亚热带植物学报》, vol. 13, no. 3, 31 December 2005 (2005-12-31), pages 271 - 274 * |
| 李莉等: "超临界二氧化碳流体萃取油茶籽油及其GC/MS分析", 《广州化工》, vol. 39, no. 11, 31 December 2011 (2011-12-31), pages 49 - 52 * |
| 邢立民: "CO2 超临界萃取技术在油脂生产中的应用", 《内蒙古科技与经济》, no. 23, 31 December 2009 (2009-12-31) * |
| 黄翠莉，吴苏喜: "响应面法对超临界ＣＯ2萃取茶籽仁油的工艺优化", 《生物加工过程》, vol. 8, no. 3, 31 May 2010 (2010-05-31), pages 13 - 17 * |
| 黄翠莉: "油茶籽油的超临界CO_2萃取及其功能评价", 《中国优秀硕士学位论文全文数据库 工程科技Ⅰ辑》, no. 6, 15 June 2012 (2012-06-15), pages 024 - 235 * |
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| CN104673485A (en) * | 2015-01-29 | 2015-06-03 | 衢州市质量技术监督检测中心 | Method for removing polyaromatic hydrocarbon pollutants from camellia oil |
| CN104758331A (en) * | 2015-04-30 | 2015-07-08 | 溧阳市天目湖保健品有限公司 | Chinese medicine composition capable of enhancing immunity and preparation method thereof |
| CN107114491A (en) * | 2017-05-08 | 2017-09-01 | 湖北鑫尔绿色食品有限公司 | The elderly's tea oil and its production method |
| CN109157454A (en) * | 2018-10-25 | 2019-01-08 | 上海清轩生物科技有限公司 | A kind of Camellia combined extracts and its preparation and the application in cosmetics |
| CN112316484A (en) * | 2020-10-31 | 2021-02-05 | 洞口县亿丰农林牧科技有限公司 | Method for extracting camellia oleifera polyphenol from tea oil |
| CN113068750A (en) * | 2021-04-10 | 2021-07-06 | 贵州长基供应链管理有限公司 | Camellia blend oil for improving immunity and processing method thereof |
| CN117778095A (en) * | 2023-08-07 | 2024-03-29 | 湖南金昌生物技术有限公司 | Preparation method of camellia seed extract |
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