CN101731578A - Extract of carica papaya L. seeds produced in Hainan, preparation method and application thereof - Google Patents
Extract of carica papaya L. seeds produced in Hainan, preparation method and application thereof Download PDFInfo
- Publication number
- CN101731578A CN101731578A CN201010112814A CN201010112814A CN101731578A CN 101731578 A CN101731578 A CN 101731578A CN 201010112814 A CN201010112814 A CN 201010112814A CN 201010112814 A CN201010112814 A CN 201010112814A CN 101731578 A CN101731578 A CN 101731578A
- Authority
- CN
- China
- Prior art keywords
- extract
- papaya seed
- ethanol
- papaya
- preparation
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Landscapes
- Medicines Containing Plant Substances (AREA)
Abstract
The invention discloses an extract of carica papaya L. seeds, which is prepared by the following steps: dissolving the carica papaya L. seeds into ethanol and lixiviating the carica papaya L. seeds at the room temperature, rotationally steaming the filtered solution till the ethanol is evaporated to obtain a coarse extract of the carica papaya L. seeds, dissolving the coarse extract into water, then extracting the coarse extract by using an organic solvent, and concentrating the extract to obtain the extract of the carica papaya L. seeds. The invention also discloses a preparation method for the extract and application thereof. The extract is pure natural, has no toxic or side effect, has stability to illumination and metal ions (calcium, iron and zinc), and is suitable for acid conditions; the anti-oxidation activity of the extract is enhanced along with the rise of the temperature at the temperature of below 100 DEG C; the production method has the advantages of readily available raw materials, high waste utilization rate and low cost; and the prepared product can be widely used in the field of food, medicament, cosmetics, veterinary medicament, feed and the like.
Description
Technical field
The present invention relates to a kind of papaya seed extract, also relate to this preparation method of extract and, belong to natural plant product comprehensive development and utilization technical field as the purposes of food, medicine, cosmetics, feed or veterinary drug antioxidant.
Background technology
Papaya (Carica Papaya L.) is a kind of tropical evergreen fruit trees, originate in Mexico and Central America, mainly be distributed in provinces and regions such as Guangdong, Hainan, Guangxi, Yunnan, Taiwan, Fujian in China, in Xichang, Sichuan and Ganzhou a small amount of distribution is arranged also, wherein, Hainan is China papaya cultivation optimum district.After papaya is useful for consumption, often abandon a large amount of seeds, pericarp and plant residue, cause that the human settlement pollutes, its waste material for developing.In order to make full use of the papaya plant resources, avoid the human settlement to pollute, be necessary the papaya discarded object is carried out antioxidation activity research, and to papaya seed deep processing and utilization in addition.
Papaya is worth the high fruit except that being used to produce edible health, and also as medicinal, the trend of newtype drug also appears papaya fruit and discarded object are developed as in China recent years in African country, and papaya fruit also is commonly used for health products and cosmetics exploitation.Papaya is rich in protease, lipase, this be its be developed as medicine, as the material base of cosmetics and food additives etc., forefathers have mainly carried out comparatively deep theory and application research around papain and lipase.In fact, it also is that it suits as medicine, is used as one of major reason of cosmetics and food additives that melon tree extract has antioxidation activity, do not see research but have antioxidation activity, the papaya secondary metabolites is not appeared in the newspapers around the research that its antioxidation activity is developed yet about melon tree extract.
In addition, at present, the toxicity of food additives such as melamine and tonyred makes us food additives are turned pale at the mention of the tiger because of causing the fear among the crowd always, the safe storage of converted products is fresh-keeping also to be the focus that people pay close attention to, and therefore, developing nontoxic antioxidant is to be necessary very much.
Summary of the invention
The object of the present invention is to provide a kind of papaya seed extract, this extract is a raw material with the papaya seed after consuming, realize refuse reclamation, this extract also has antioxidation activity, stable to illumination, metal ion (calcium, iron and zinc), be suitable for acid condition, below 100 ℃ with temperature rising increased activity, and its safety non-toxic can solve the preservation and freshness of converted products and the social crowd terrified problem to additive.
The present invention also aims to provide the preparation method of above-mentioned papaya seed extract, this preparation method's technology is succinct, the extraction efficiency height.
In fact, the present invention also provides the purposes of above-mentioned papaya seed extract as food, medicine, cosmetics, feed or veterinary drug antioxidant.
Papaya seed extract provided by the invention is got the papaya seed and is dipped in the ethanol after the normal temperature lixiviate, filtrate revolving steamed to the ethanol evaporate to dryness, the crude extract of papaya seed, with this crude extract water dissolving, promptly get the papaya seed extract with concentrating behind the organic solvent extraction again.
The preparation method of papaya seed extract provided by the invention comprises the steps:
(1) gets the papaya seed and clean to shine to eighty per cant and do, be dipped in the ethanol normal temperature lixiviate 10～20 days after the pulverizing, filter, filter residue is dipped in the ethanol extracts repeatedly again 1～3 time under the normal temperature, extracted 5～10 days at every turn, merging filtrate steams to the ethanol evaporate to dryness at 55 ℃ of backspins, gets papaya seed crude extract;
(2) with papaya seed crude extract water dissolving, use organic solvent extraction again 3～5 times, combining extraction liquid, revolve be concentrated into after the steaming remove behind the organic solvent the papaya seed extract.
The volumetric concentration of the described ethanol of step (1) is 20～90%.
Organic solvent described in the step (2) is one or more in benzinum, ethyl acetate and the n-butanol.
The temperature of revolving in the step (2) when steaming is 50～60 ℃.
Papaya seed extract provided by the invention is as the purposes of food, medicine, cosmetics, feed or veterinary drug antioxidant.
The present invention has following advantage:
(1) this papaya seed extract pure natural, have no side effect, and stable to illumination, metal ion (calcium, iron and zinc), be suitable for acid condition, its antioxidation activity below 100 ℃ with temperature rising increased activity;
(2) this papaya seed extract raw material is easy to get, and sharp useless rate height is with low cost, and preparation products obtained therefrom antioxidation activity height can be widely used in fields such as food, medicine, cosmetics, veterinary drug and feed.
Description of drawings
Fig. 1 is the ethanol crude extract of papaya seed and the antioxidation activity figure of different organic solvents extract;
Fig. 2 is that the ethanol crude extract and the different organic solvents extract of papaya seed removed and can be tried hard to OH.
The specific embodiment
Following examples only are used to set forth the present invention, and protection scope of the present invention is not only to be confined to following examples.The those of ordinary skill of described technical field all can be realized purpose of the present invention according to above content disclosed by the invention.
First's papaya seed extract and its production and application
The papaya seed extract that present embodiment provides, prepare by following method: getting the papaya seed, to be dipped in volumetric concentration be in 90% the ethanol after the normal temperature lixiviate, filtrate revolving steamed to the ethanol evaporate to dryness, get the crude extract of papaya seed, after this crude extract water dissolving, be the papaya seed extract with concentrating behind organic solvent benzinum, ethyl acetate and the extracting n-butyl alcohol respectively.
Embodiment 2
The papaya seed extract that present embodiment provides is with papaya seed running water wash clean; The seed of cleaning shines under the sun to eighty per cant dried, pulverizes with pulverizer; The employing volumetric concentration is 30% alcohol at normal temperature extraction extraction, lixiviate for the first time 20 days, be dipped in the ethanol each lixiviate for the 2nd and the 3rd time 10 days, in time filtering leaching liquor with filter cloth after each lixiviate reclaims, and filter residue turned back in the lixiviate container, the filtrate of reclaiming is merged the back in airtight container, preserve, under 55 ℃ of following constant temperature, revolve inspissation and contract, to the complete distilled-to-dryness of alcohol solvent, the ethanol crude extract then is accumulated in to revolve and steams in the bottle; Total crude extract with water-soluble, is taken up in order of priority with benzinum, ethyl acetate and extracting n-butyl alcohol, and continuous extraction is 5 times respectively, extract is revolved inspissation contract under 50 ℃, promptly gets the papaya seed extract.
The papaya seed extract that present embodiment provides is with papaya seed running water wash clean; The seed of cleaning shines under the sun to eighty per cant dried, pulverizes with pulverizer; The employing volumetric concentration is 50% alcohol at normal temperature extraction extraction, 2 weeks of the lixiviate first time, for the second time and be dipped in each 1 week of lixiviate in the ethanol for the third time, in time filtering leaching liquor with filter cloth after each lixiviate reclaims, and filter residue turned back in the lixiviate container, the filtrate that will reclaim merges the back and preserve in airtight container; Ethanol extract is revolved steaming be concentrated into the complete distilled-to-dryness of alcohol solvent under 55 ℃, the ethanol crude extract then is accumulated in to revolve and steams in the bottle; Total crude extract with water-soluble, is used extracting n-butyl alcohol, and continuous extraction is 3 times respectively, extract is revolved inspissation contract under 55 ℃, promptly gets the papaya seed extract.
The papaya seed extract that present embodiment provides is with papaya seed running water wash clean; The seed of cleaning shines under the sun to eighty per cant dried, pulverizes with pulverizer; The employing volumetric concentration is 70% alcohol at normal temperature extraction extraction, 10 days for the first time, 2nd, be dipped in the ethanol each lixiviate for 3 and 4 times 5 days, in time filtering leaching liquor with filter cloth after each lixiviate reclaims, and filter residue turned back in the lixiviate container, the filtrate that will reclaim merges the back and preserve in airtight container; Ethanol extract is revolved steaming be concentrated into the complete distilled-to-dryness of ethanol under 55 ℃, the ethanol crude extract then is accumulated in to revolve and steams in the bottle; Total crude extract with water-soluble, is used benzinum, ethyl acetate extraction successively, and continuous extraction is 3 times respectively, extract is revolved inspissation contract under 55 ℃, promptly gets the papaya seed extract.
Get the papaya seed extract that has prepared, the percentage by weight by 0.001%～0.1% joins in the food and goes, as antioxidant.
Embodiment 6
Get the above-mentioned papaya seed extract that has prepared, in the medicament preparation process, the percentage by weight by 0.01%～0.05% adds, as antioxidant.
Embodiment 7
Get the papaya seed extract that has prepared, when feed encapsulated, the percentage by weight by 0.001%～0.1% joined in the feed and goes, and stirred evenly, as antioxidant.
Get the papaya seed extract that has prepared, in the preparation process of cosmetics, the percentage by weight by 0.005%～0.1% joins in the cosmetics and goes, as antioxidant.
Embodiment 9
Get the papaya seed extract that has prepared, in the preparation process of veterinary drug, the percentage by weight by 0.01%～0.05% joins in the veterinary drug and goes, as antioxidant.
The antioxidation activity of second portion papaya seed extract
2.1 the antioxidation activity of melon tree extract test
Antioxidation activity for ease of each organic solvent of comparative analysis, in the preparation process of papaya seed extract, ethanol extract behind the above-mentioned evaporate to dryness is kept a minimum part as antioxidation activity, to environment and dielectric stability detection, to being left the total ethanol crude extract with water-soluble, use benzinum, ethyl acetate and extracting n-butyl alcohol successively, each solvent difference continuous extraction 3 times, each extract is revolved inspissation under 55 ℃ contract, together with the distillation of remaining aqueous solution, respectively above-mentioned 4 kinds of extracts.
When measuring antioxidation activity, the extract of ethanol crude extract, different organic solvents all dissolves with DMSO.
2.1.1 adopting the method for removing DPPH free radical ability measures the total antioxidant activity of ethanol crude extract and each extract.With absolute ethyl alcohol DPPH is mixed with 6.5 * 10
-4Mol.L
-1Solution, it is standby to place refrigerator and cooled to hide, and is diluted to 6.5 * 10 with absolute ethyl alcohol during use
-5Mol.L
-1Experiment is divided into 3 groups, and every group of cumulative volume is 3mL, and first group adds 2.5mL DPPH solution (6.5 * 10 in test tube
-5Mol.L
-1) and 0.5mL absolute ethyl alcohol (solvent of sample), survey absorbance at 517nm wavelength place behind the mixing, be designated as A
0Second group adds 2.5mL DPPH solution (6.5 * 10
-5Mol.L
-1) and the 0.5mL sample, shake up, survey absorbance behind the 50min of lucifuge reaction at room temperature, be designated as A
iThe 3rd group adds 2.5mL absolute ethyl alcohol (solvent of DPPH solution) and 0.5mL0.1mg/mL sample (extract of total ethanol crude extract, each organic solvent and positive control Vc), measures absorbance behind the mixing, is designated as A
jThe said sample collocation method is: the extract of total ethanol crude extract, different solvents and the DMSO solution of Vc are configured to 100mg.L with absolute ethyl alcohol
-1Mother liquor is diluted to 50,25,12.5,6.25 and 3.25mg.L with absolute ethyl alcohol more successively
-1, measure light absorption value A respectively
iBy formula calculate clearance rate: clearance rate %=[1-(A
i-A
j) A
o] * 100%.With sample quality concentration (C) is abscissa, and clearance rate (Y) is an ordinate, asks the workmanship to make curve, and obtains the mass concentration that clearance rate is 50% o'clock sample (EC50), and the active result of sample promptly represents with half clear quality concentration (EC50).
The extract antioxidation activity of papaya seed total ethanol runic thing and variant solvent as shown in Figure 1.The extract antioxidation activity of total ethanol crude extract and different solvents and the linear regression relation of its concentration are all extremely remarkable or remarkable, say that the extract of bright total crude extract of ethanol and different solvents all has strong and weak different antioxidation activities.From linear regression coeffficient, ethyl acetate, n-butyl alcohol extract and positive control bigger, total ethanol crude extract placed in the middle, benzinum and water extract then minimum.Illustrate that the former three antioxidation activity significantly is subjected to its concentration affects, has stronger antioxidation activity; Total ethanol crude extract antioxidation activity is placed in the middle; Benzinum and water extract antioxidation activity are faint.Anti-oxidant EC50 value according to regression equation calculation acetic acid ethyl ester extract, n-butyl alcohol extract, total ethanol crude extract, petroleum ether extract, water extract and positive control is respectively: 64.61,109.30,248.63,1009.5,1628.33 and 66.96mg.mL
-1As seen, the acetic acid ethyl ester extract antioxidation activity is a little more than positive control, n-butyl alcohol extract is about 61.26% of positive control antioxidation activity, total crude extract is about 26.93% of positive control antioxidation activity, benzinum and water extract then antioxidation activity are extremely faint with respect to positive control, are respectively 6.63% and 4.11% of positive control.
2.1.2 adopt TEAC method mensuration ethanol crude extract and each extract to ABTS
+The removing activity.With 7mmol.L
-1ABTS
+5mL and 140mmol.L
-1K
2S
2O
888 μ L mixed liquors spend the night 3 ℃ of following lucifuges, preparation ABTS
+Stock solution.When this stock solution is used in experiment, use 20mmol.L
-1Sodium-acetate buffer slowly dilutes, preparation ABTS
+Working solution is when being diluted to λ=734nm, till absorbance A=0.70 ± 0.02.Get the extract 0.1mg.ml of total crude extract, different solvents extraction preparation respectively
-1 Sample liquid 30 μ L are with ABTS
+Working solution 3mL mixes, and shakes up reaction 10s, the preparation test sample.With the sodium-acetate buffer is reference, test sample is left standstill 6min under 3 ℃, measures absorbance A under λ=734nm.With 0,0.25,0.315,0.417,0.625,1.25 and 2.50mmol.L
-1Trolox replace above-mentioned test sample, measure the absorbance A under each concentration, the drawing curve is asked the equation of linear regression of absorbance A about the test sample concentration C.Working curve converses with mmol.L according to this
-1Trox is that each test sample of unit is removed ABTS
+Active.
The total antioxidant activity of table 1TEAC method and FRAP method and the active performance of anti-2 kinds of active oxygens
Annotate: the different capitalization English letters of digital heel are represented difference extremely significantly (p＜0.01), represent difference not remarkable (p＜0.01) with identical capitalization.
The result is as shown in table 1.As seen the n-butyl alcohol extract antioxidation activity utmost point is significantly higher than other extract and total ethanol crude extract; Acetic acid ethyl ester extract and total crude extract antioxidation activity do not have significant difference, take second place; The petroleum ether extract antioxidation activity utmost point is significantly higher than water extract, extremely significantly is lower than other each extract and total crude extract; The water extract antioxidation activity is minimum.
Measure the total antioxidant activity of ethanol crude extract and each extract 2.1.3 adopt the FRAP method.With 20mmol.L
-1FeCl
36H
2O2.5ml and 10mmol.L
-1TPTZ2.5ml mix, incorporate 300mmol.L again
-1, among the pH3.6 acetate buffer 25ml, the preparation mixed liquor.Get above-mentioned mixed liquor 1.8ml, under 37 ℃ of water-baths, add distilled water 180 μ L, 0.1mg.ml successively
-1Total ethanol crude extract and each extract sample liquid 60 μ L, mixing afterreaction 30min.With methyl alcohol is reference, when λ=593nm, reads absorbance A.Configuration 0,100,300,500,700,900 and 1000 μ mol.L
-1FeSO
4Gradient solution is an abscissa with concentration, and absorbance A is an ordinate, asks the workmanship to make curve and calculates each crude extract antioxidation activity mutually.Antioxidation activity FRAP value representation, promptly every gram fruit dry powder is equivalent to FeSO
4Amount of substance (μ mol.L
-1).Test sample repeats for 3 times.
Testing result is as shown in table 1.As seen the acetic acid ethyl ester extract antioxidation activity utmost point is significantly higher than other each extracts and total crude extract; And total crude extract, n-butanol and the mutual difference of water extract are not remarkable; The petroleum ether extract antioxidation activity is extremely significantly minimum.
2.1.4 detect each crude extract to O
2 -The removing ability.With pH7.8,65mmol.L
-1Phosphate buffer 1.0mL, 7.5mmol.L
-1Xanthine 0.1mL, 10mmol.L
-1Chlorination oxyammonia 0.1mL, 0.1mg.mL
-1Extract sample liquid 0.1mL, redistilled water 0.4mL and the 20 μ g.mL of total crude extract and each organic solvent
-1The xanthine oxidase 0.3mL of albumen mixes successively, reacts 20min behind the mixing in 25 ℃ of water-baths.Get above-mentioned reactant liquor 0.5mL, add 19mmol.L
-1Aminobenzenesulfonic acid 0.5mL and 1.0% alpha-naphthylamine 0.5mL are placed on after fully mixing under the room temperature behind the reaction 20min, measure absorbance A when λ=530nm
1Replace sample liquid to carry out above-mentioned reaction with deionized water, measure the blank absorbance A
0Alpha-tocopherol with the variable concentrations gradient replaces sample liquid, asks and makes calibration curve, and the alpha-tocopherol concentration gradient is 0,10,100,200,500,800,1000 μ mol.L
-1O
2 -Removing ability amount (the μ mol.L of alpha-tocopherol
-1) expression.Test sample repeats for 3 times.
Testing result is as shown in table 1.As seen total ethanol crude extract O
2 -The removing ability utmost point is significantly higher than other each extract; The O of ethyl acetate and water extract
2 -It is not remarkable to remove capacity variance, takes second place; The O of n-butyl alcohol extract
2 -The removing ability extremely significantly is lower than total extract, ethyl acetate and water extract, and only the utmost point is significantly higher than petroleum ether extract; The O of petroleum ether extract
2 -The removing ability is extremely significantly minimum.
2.1.5 detect total ethanol crude extract and each extract to H
2O
2The removing ability.With 20%TiCl
4Concentrated hydrochloric acid solution 0.135mL, 0.1mg.mL
-1The 0.17mol.L of total crude extract and each extract sample liquid 0.1mL, pH7.4
-1Phosphate buffer 0.185mL and 17.0mol.L
-1NH
3H
2O0.2mL mixes, and at room temperature reacts 5min, produce white floccule after, use 3mol.L
-1H
2SO
4The 3mL dissolving; When λ=410nm, measure absorbance A
1Replace sample liquid with deionized water, repeat above-mentioned reaction, measure blank absorbency A
00,20,40,60,80,100mg.mL replace sample liquid to make calibration curve with Vc, the Vc concentration gradient is:
-1, dissolve with DMSO.Test sample repeats 3 times.
Testing result is as shown in table 1.As seen the H of total ethanol crude extract and acetic acid ethyl ester extract
2O
2It is not remarkable to remove capacity variance; The H of total crude extract
2O
2The removing ability utmost point is significantly higher than n-butanol, water and petroleum ether extract; The H of acetic acid ethyl ester extract
2O
2The removing ability is not simultaneously remarkable with the difference of n-butanol and water extract yet, and the utmost point is significantly higher than petroleum ether extract; The H of petroleum ether extract
2O
2The removing ability is then minimum.
2.1.6 detect total ethanol crude extract and each extract OH is removed ability.With 10mmol.L
-1FeSO
4-EDTA 0.2mL, 10mmol.L
-1Total crude extract of D-deoxyribose 0.5ml, variable concentrations gradient and the sample liquid 0.1mL of each extract mix, be settled to 1.8mL with phosphate buffer; In above-mentioned mixed liquor, add 10mmol.L
-1H
2O
20.2mL, under 37 ℃ of water-baths, react 1h; Continue to add 2.8%TCA1.0mL, 1.0%TBA10mL, boiling water bath 15min behind the mixing; After the cooling, under λ=532nm, colorimetric estimation A
sApplication of sample liquid not in above-mentioned steps, all the other are all identical, record the blank absorbance A
CIn above-mentioned steps, application of sample liquid and not reacting in 37 ℃ of water-baths does not record the blank background absorbance A
0Replace sample liquid with Sodium Benzoate solution, make positive control, measure its A according to above-mentioned steps
sCalculate the OH clearance rate under the variable concentrations gradient, computing formula is: clearance rate %=(A
c-A
s) * 100/ (A
c-A
0).Set up sample liquid, positive control concentration gradient and clearance rate equation of linear regression respectively, and obtain the EC50 value, according to this total ethanol crude extract, each extract and positive control are compared OH and remove ability.The sample liquid concentration gradient is: 0.1,0.2,0.4,0.6,0.8 and 1.0mg.mL
-1Positive control sodium benzoate concentration gradient is: 0.01,0.05,0.10,0.15,0.20 and 0.25mg.mL
-1
Testing result as shown in Figure 2.As seen, remove the petroleum ether extraction beyond the region of objective existence, the OH of all the other each extracts, total crude extract and positive control removes ability and all extremely remarkable or remarkable positive correlation of its concentration.Show and remove the petroleum ether extraction beyond the region of objective existence that the OH that all the other each extracts and total crude extract all have in various degree removes ability.According to equation of linear regression, the OH of acetic acid ethyl ester extract, n-butyl alcohol extract, water extract, total ethanol crude extract and positive control removes ability EC50 value and is respectively: 0.09,0.21,0.33,0.76 and 0.10mg.mL
-1, the OH of visible ethyl acetate extract removes slightly that ability is better than positive control, and n-butanol, water extract OH removing ability are about 47.62% and 30.30% of positive control, and total ethanol crude extract OH removing ability is about 13.16% of positive control.
The mensuration of natural drug and food antioxidation activity in vitro does not still have standard method at present, and current methods respectively has limitation, adopts distinct methods to estimate the different result of the normal performance of natural products antioxidation activity.Therefore, all adopt several different methods to detect the natural products antioxidation activity in present many reports, the antioxidation activity of material is done comprehensive the evaluation.The application adopts multiple assay method testing result to show, papaya seed acetic acid ethyl ester extract has very strong antioxidation activity, and n-butyl alcohol extract has stronger antioxidation activity, and the total ethanol crude extract then has very strong removing O
2 -And H
2O
2Ability, this explanation papaya seed oxidation-resistant active ingredient mainly concentrates on ethyl acetate and n-butyl alcohol extract, and to the wherein worth separation and purification of anti-oxidant monomeric compound, the total ethanol crude extract shows the strongest removing O
2 -And H
2O
2Ability illustrates and removes O
2 -And H
2O
2The time, complex interactions may take place in the composition in the different extracts, causes each extract after the rough segmentation to remove O
2 -And H
2O
2Ability drop.This has the monomeric compound of the antioxidation activity phenomenon more weak than the crude extract antioxidation activity that contains this monomeric compound with forefathers and comes down to consistent.
In a word, ethyl acetate and n-butyl alcohol extract have very strong and stronger antioxidation activity, and ethanol extract has the anti-O of very strong removing
2 -And H
2O
2Ability.
2.2 the security performance of papaya seed extract
2.2.1 the studies on acute toxicity of papaya seed extract is got Kunming mouse, irritates stomach by 10g/kg and gives papaya seed extract in the foregoing description, feeds continuously 4 times in 1 day, each 6h at interval, give normal diet then, each organ is carried out the pathologic inspection, do not find organ disease.
2.2.2 Kunming mouse is got in the research of the long term toxicity of papaya seed extract, in pressing, heavy dose of group (4g/kg, 8g/kg) irritate stomach and give papaya seed extract in the foregoing description, every day 1 time, the continuous irrigation stomach was handled after 14 days, measure indexs such as mouse body weight, heart function, liver function, renal function, electrocardiogram, the administration group compares with control group respectively, shows no obvious abnormalities.Internal organs such as the pathological examination heart, liver,kidney,spleen, lung, stomach, duodenum, large intestine, small intestine, adrenal gland and genitals, the crude extract group of feeding compare with control group respectively, and all not having obviously poisons sexually revises.
Edible papaya seed ethyl acetate of above results suggest Kunming mouse and n-butyl alcohol extract safety non-toxic can be used as the antioxidant that food, medicine, cosmetics, feed and veterinary drug are processed.
2.3 the papaya seed extract is to the stability study of environment and common medium
As mentioned above, above-mentioned papaya seed extract has very strong and stronger DPPH antioxidation activity, therefore, behind this extract enforcement light, heat, soda acid and metal ion processing certain hour, the stability of observation DPPH antioxidation activity (EC50).The DPPH method detects identical with preceding method.The same papaya seed ethanol extract of handling, O is removed in observation
2-The stability of ability (every milligram of crude extract is equivalent to the amount of tocopherol).DPPH method and removing O
2-The ability detection method is identical with preceding method.
2.3.1 the influence of illumination is dissolved above-mentioned papaya seed ethyl acetate, n-butyl alcohol extract and ethanol extract with DMSO, shine under daylight, by light application time 12h every day, light application time as shown in table 2 is respectively handled.The result shows in different time, total ethanol crude extract and each extract antioxidation activity all do not have significant difference, the antioxidation activity that ethanol crude extract, acetic acid ethyl ester extract and n-butyl alcohol extract are described is stable to illumination, need not consider the influence of illumination to extract in processed and applied.
The table 2 different light time is to the influence of papaya seed extract antioxidation activity
2.3.2 Temperature Influence is dissolved above-mentioned papaya seed ethyl acetate, n-butyl alcohol extract and total ethanol CE with DMSO, water bath with thermostatic control 4h under different temperatures is cooled to room temperature (30 ℃) back and detects antioxidation activity.Treatment of different temperature is as shown in table 3 respectively.There is utmost point significant difference in the crude extract antioxidation activity, and generally below 100 ℃, it is stronger to show as each extract antioxidation activity under the high temperature, illustrates that melon tree extract is as the heating condition in the antioxidant ability process.
Table 3 different temperatures is to the influence of the antioxidation activity of melon tree extract
2.3.3pH influence will state papaya seed ethyl acetate, n-butyl alcohol extract and ethanol extract and dissolve with DMSO, under different pH, measure the crude extract antioxidation activity, analyze that antioxidation activity changes characteristics under the different pH, pH handles as shown in table 4.The result shows that each extract antioxidation activity under acid condition is stable, and the next antioxidation activity of alkali condition extremely significantly weakens.Illustrate that each extract is suitable for doing the antioxidant of acid processed goods, and to the alkali condition sensitivity.
The different pH of table 4 are to the influence of papaya seed extract antioxidation activity
2.3.4 the influence of metal ion is dissolved above-mentioned papaya seed ethyl acetate, n-butyl alcohol extract and ethanol extract with DMSO, detect antioxidation activity under the different metal ionic medium, illustrates that common metal ion influences the crude extract antioxidation activity.At present, in health food, medicine and the processing of refining feed, the normal metal cation components such as calcium, iron and zinc that add, with the metallic element of growing by these adjustings of dietary supplementation and assurance body growth, therefore, not add metal ion is contrast, has inquired into the influence of this three metal ion species to the crude extract antioxidation activity.Result in the table 5 shows that the different metal ion processing does not all make significant difference to each extract antioxidation activity, the antioxidation activity that the said extracted thing is described is stable to this 3 metal ion species, can directly use these extracts as antioxidant in present health products processed and applied.
The influence of table 5 different metal ion pair papaya seed extract antioxidation activity
Claims (6)
1. a papaya seed extract is characterized in that, gets the papaya seed and is dipped in the ethanol after the normal temperature lixiviate, filtrate revolving steamed to the ethanol evaporate to dryness, get the crude extract of papaya seed,, promptly get the papaya seed extract with concentrating behind the organic solvent extraction again this crude extract water dissolving.
2. the preparation method of the described papaya seed extract of claim 1 is characterized in that, comprises the steps:
(1) gets the papaya seed and clean to shine to eighty per cant and do, be dipped in the ethanol normal temperature lixiviate 10～20 days after the pulverizing, filter, filter residue is dipped in the ethanol extracts repeatedly again 1～3 time under the normal temperature, extracted 5～10 days at every turn, merging filtrate steams to the ethanol evaporate to dryness at 55 ℃ of backspins, gets papaya seed crude extract;
(2) with papaya seed crude extract water dissolving, use organic solvent extraction again 3～5 times, combining extraction liquid, revolve be concentrated into after the steaming remove behind the organic solvent the papaya seed extract.
3. the preparation method of papaya seed extract according to claim 2 is characterized in that, the volumetric concentration of the ethanol described in the step (1) is 20～90%.
4. the preparation method of papaya seed extract according to claim 2 is characterized in that, the organic solvent described in the step (2) is one or more in benzinum, ethyl acetate and the n-butanol.
5. the preparation method of papaya seed extract according to claim 2 is characterized in that, the temperature of revolving in the step (2) when steaming is 50～60 ℃.
6. the described papaya seed extract of claim 1 is as the purposes of food, medicine, cosmetics, feed or veterinary drug antioxidant.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201010112814A CN101731578A (en) | 2010-02-09 | 2010-02-09 | Extract of carica papaya L. seeds produced in Hainan, preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201010112814A CN101731578A (en) | 2010-02-09 | 2010-02-09 | Extract of carica papaya L. seeds produced in Hainan, preparation method and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101731578A true CN101731578A (en) | 2010-06-16 |
Family
ID=42455991
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201010112814A Pending CN101731578A (en) | 2010-02-09 | 2010-02-09 | Extract of carica papaya L. seeds produced in Hainan, preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101731578A (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101935295A (en) * | 2010-08-31 | 2011-01-05 | 中国热带农业科学院热带生物技术研究所 | Method for extracting active ingredients from papaya seeds |
| CN102846763A (en) * | 2012-09-25 | 2013-01-02 | 长春理工大学 | Pawpaw seed extraction method, and liver-protecting drug containing pawpaw seed extract |
| CN103951598A (en) * | 2014-05-06 | 2014-07-30 | 中国热带农业科学院椰子研究所 | Method for extracting benzyl isothiocyanate from papaya seeds |
| CN104928013A (en) * | 2015-06-15 | 2015-09-23 | 贵州师范大学 | Method for extracting pawpaw seed oil |
| CN107156657A (en) * | 2017-05-05 | 2017-09-15 | 安徽秋果食品有限公司 | A kind of dilated food rich in papaya extractives and preparation method thereof |
-
2010
- 2010-02-09 CN CN201010112814A patent/CN101731578A/en active Pending
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101935295A (en) * | 2010-08-31 | 2011-01-05 | 中国热带农业科学院热带生物技术研究所 | Method for extracting active ingredients from papaya seeds |
| CN101935295B (en) * | 2010-08-31 | 2013-01-16 | 中国热带农业科学院热带生物技术研究所 | Method for extracting active ingredients from papaya seeds |
| CN102846763A (en) * | 2012-09-25 | 2013-01-02 | 长春理工大学 | Pawpaw seed extraction method, and liver-protecting drug containing pawpaw seed extract |
| CN103951598A (en) * | 2014-05-06 | 2014-07-30 | 中国热带农业科学院椰子研究所 | Method for extracting benzyl isothiocyanate from papaya seeds |
| CN103951598B (en) * | 2014-05-06 | 2016-02-17 | 中国热带农业科学院椰子研究所 | A kind of method extracting benzyl isothiocyanide from Semen Fructus Chaenomelis |
| CN104928013A (en) * | 2015-06-15 | 2015-09-23 | 贵州师范大学 | Method for extracting pawpaw seed oil |
| CN107156657A (en) * | 2017-05-05 | 2017-09-15 | 安徽秋果食品有限公司 | A kind of dilated food rich in papaya extractives and preparation method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Lama-Muñoz et al. | Content of phenolic compounds and mannitol in olive leaves extracts from six Spanish cultivars: Extraction with the Soxhlet method and pressurized liquids | |
| Offor et al. | Proximate nutritional analysis and heavy metal composition of dried Moringa oleifera leaves from Oshiri Onicha LGA, Ebonyi State, Nigeria | |
| CN103070400B (en) | Use of lotus procyanidin as advanced glycosylation end product formation inhibitor | |
| Weidner et al. | Extracts of phenolic compounds from seeds of three wild grapevines—Comparison of their antioxidant activities and the content of phenolic compounds | |
| CN101731578A (en) | Extract of carica papaya L. seeds produced in Hainan, preparation method and application thereof | |
| CN102613281B (en) | Preparation method for south China pine needle extractives and application of south China pine needle extractive in freshness of eggs | |
| CN104116132B (en) | The application in cigarette of a kind of agate card enzymatic hydrolyzed extract | |
| Arenas et al. | Fate of polyphenols in pili (Canarium ovatum Engl.) pomace after in vitro simulated digestion | |
| CN102940597A (en) | Natural compound botanical antioxidant, as well as preparation method and application thereof in cosmetics | |
| CN102715620A (en) | Method for producing fast-food fruit and vegetable frozen-dried powder | |
| Weidner et al. | Phenolic compounds and the antioxidant properties in seeds of green-and yellow-podded bean (Phaseolus vulgaris L.) varieties | |
| CN110201043A (en) | A kind of olibanum composite essential oil and application | |
| Boukhalfa et al. | Antioxidant activity and Hypolipidemic effect of Ficus carica leaf and twig extracts in Triton WR-1339-induced hyperlipidemic mice | |
| CN111840156A (en) | Extraction method of active substance of cinnamomum camphora flower and determination method of oxidation resistance | |
| Meagher et al. | Insecticide Residues, Colorimetric Determination of Guthion Residues in Crops | |
| CN104106627A (en) | Vegetable fresh-keeping composition containing fructus forsythiae leaf extract and preparation method thereof | |
| US20190216718A1 (en) | Cosmetic composition containing complex extract of sargassum horneri and enteromorpha prolifera | |
| CN108186400A (en) | A kind of moisturizer and preparation method thereof | |
| Lekar et al. | Subcritical water extraction of chlorogenic acid from green coffee beans | |
| CN111000838A (en) | Use of protocatechuic acid and composition containing protocatechuic acid | |
| Maduka et al. | Protective role of Moringa oleifera Lam aqueous leaf extract on some excretory products and hematological parameters in acetaminophen induced albino rats. IOSR J | |
| CN101791113A (en) | Peel extractive of papaya produced in Hainan as well as preparation method and application thereof | |
| CN106667833B (en) | Camellia oleifera seed coat extract and preparation method and application thereof | |
| CN102119993B (en) | Preparation method of onion extract, | |
| KR20100039534A (en) | The processing method of laminaria extracts with chlorophyll |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20100616 |