CN109481502A - A kind of preparation method of Chinese toon extract - Google Patents
A kind of preparation method of Chinese toon extract Download PDFInfo
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Abstract
The present invention discloses a kind of preparation method of Chinese toon extract, includes the following steps: ultrasonic wave assisted extraction Chinese toon in ethanol, obtains Chinese toon and extracts mixed liquor；Chinese toon extraction mixed liquor is evaporated into no ethanol flavor, is centrifuged, is taken precipitating, obtain Chinese toon extract crude product；Chinese toon extract crude product is prepared into Chinese toon crude extract dilution with water；Chinese toon crude extract dilution is crossed into macroporous resin column；Macroporous resin column is washed with distillation, until the liquid colorless of outflow；With ethanol elution macroporous resin column, until the liquid colorless of outflow, collects eluent；Eluent volatilization is concentrated into no alcohol taste, obtains Chinese toon extract sterling；Chinese toon extract sterling is lyophilized, Chinese toon extract sterling powder is obtained.The Chinese toon extract can be effectively reduced damage of the cis-platinum to body, increase internal glutathione content, reduce internal mda content.
Description
Technical field
The invention belongs to medical domains, are related to a kind of preparation method of Chinese toon extract, in particular to a kind of Chinese toona fruit mentions
Take the preparation method of object.
Background technique
Cis-platinum due to its anticancer spectrum is wide, effect is strong and can be with a variety of one same-actions of anti-tumor drug the characteristics of, be current
One of most common tumour medicine.But since cis-platinum does not have specificity to cell killing, plus cisplatin in treatment cancer is being used
While also normal cell can be caused to damage, generate stronger toxic side effect, which greatly limits the fortune of cis-platinum clinically
With.Micronucleus (micronucleus, MN) test has been obtained extensive as a kind of conventional genotoxicity detection method
Using.Its sample detected also expands to blood and tissue from marrow.In addition to conventional drug gene toxicity detection, change in gene
The diagnosis of degenerative disease, assessment and prevention of curative effect etc. play an important role[2]。
Chinese toona fruit can be described as Chinese toon, Chinese toon bell again, be the fruit of Chinese toon.Studies have shown that Chinese toona fruit contains polyphenol, volatilization
The a variety of active ingredients such as oil, alkaloid have the effects that stronger hypoglycemic.Curcumin (curcumin) is from Zingiber turmeric
A kind of Actions of Natural Polyphenol Pigments extracted in plant turmeric (Curcumin longa), has the characteristics that nontoxic[6].Research hair
Existing, curcumin is in addition to excellent coloring, also having a variety of nutritional activities and pharmacological effects, including anti-oxidant, drop blood
The effects of rouge and Immune-enhancing effect[5]。
Bibliography:
[1]Noha Ibrahim Said Salem,Magda Mohammad Noshy,Azza Ali
Said.Modulatory effect of curcumin against genotoxicity and oxidative stress
induced by cisplatin and methotrexate in male mice[J].Food and Chemical
Toxicology,2017,105.
[2]Agarwal,Rakhi.,Goel,Sudhir K.,Behari,Jai Raj..(2010)
.Detoxification and antioxidant effects of curcumin in rats experimentally
exposed to mercury.,JAppl Toxicol,30(5),457-68.
[3] Han Hedan, Wang Hai, Li Ailin, Han Zhaoyu, Koryo duckweed gadol extract cause nephrocyte toxicity to cis-platinum
Protective effect [J] canceration distortion mutation, 2018,30 (03): 204-208.
[4] Yu Mingze, Liu Yinong, Jiang Zhongren, Xie Huiping, Ge Yujie, Jing Mingwu, Guo Ming cyclophosphamide inducing mouse bone
Research [J] modern preventive medicine of the influence factor of myelocyte micronucleus, 2007 (05): 935-936+941.
[5] research and inquirement [J] the contemporary medical science of Li Lianying to curcumin antitumor action and mechanism, 2014,20 (32):
21-22.
[6] anti-oxidant and antitumor activity [J] the modern food science and technology of Chen Jianping, Li Lin, Su Jianyu curcumin,
2014,30(12):11-15+6.
[7] Qin Mo, Wei Meiyan, Zhou Qin, Fang Ling, Wei Lijia, Zheng's cyperus Yao's .6 kind gelatine capsule medicine shell is to bone marrow cells in mice
The Guangdong influence [J] the medicine of micronuclear rates, 2015,36 (13): 1982-1986.
Summary of the invention
Chinese toona fruit [Toona sinensis (A.Juss.) Roem seed] is the fruit of Meliaceae plant Chinese toon, is studied herein
Chinese toon seed extract (Extractfrom toona sinensis seeds, ETSS) extract is to cis-platinum (Cis-
Diamminedichloroplatinum, CDDP) inducing mouse bone marrow cell toxicity protective action.This experiment is fragrant by extracting
Chinese toon seed extract pre-processes mouse, is to intervene object, while using curcumin with Chinese toon seed extract by experiment in vivo
(Curcumin, CMN) is used as positive control, after giving continuous three days intraperitoneal injection Chinese toon seed extracts of mouse, then by suitable
Platinum induction processing, puts to death mouse afterwards for 24 hours, takes that bone marrow cells in mice compares each group micronuclear rates and renal tissue GSH, MDA biochemistry refers to
Scale value, protection effect of the research Chinese toon seed extract to cisplatin induction mouse cytotoxic.The result shows that with cis-platinum model group phase
Than Chinese toon seed extract group, curcumin group and Chinese toon seed extract and curcumin joint group micronuclear rates significantly reduce, and GSH contains
Amount is significant to be increased, and MDA content significantly reduces (p ﹤ 0.05 or p ﹤ 0.01).Conclusion: Chinese toon seed extract is to cisplatin induction Mouse Bone
Myelocyte and nephrocyte toxicity have protective action, can reduce intracorporal oxidative stress degree, improve intrinsic antioxidative ability,
And joint curcumin effect is more significant.
This application provides a kind of preparation method of Chinese toon extract, the preparation method includes the following steps:
(a) ultrasonic wave assisted extraction Chinese toon in ethanol obtains Chinese toon and extracts mixed liquor；
(b) Chinese toon extraction mixed liquor is evaporated into no ethanol flavor, is centrifuged, takes precipitating, obtain Chinese toon extract crude product.
In some embodiments, the preparation method further includes following steps:
(c) the Chinese toon extract crude product is prepared into Chinese toon crude extract dilution with water；
(d) the Chinese toon crude extract dilution is crossed into macroporous resin column；
(e) macroporous resin column is washed with distillation, until the liquid colorless of outflow；
(f) macroporous resin column described in ethanol elution, until the liquid colorless of outflow, collects eluent；
(g) eluent volatilization is concentrated into no alcohol taste, obtains Chinese toon extract sterling.
In some embodiments, the preparation method further includes following steps:
(h) the Chinese toon extract sterling is lyophilized, obtains Chinese toon extract sterling powder.
In some embodiments, in step (a), concentration of alcohol used be 60-80%, preferably 70%；And/or
In step (a), the temperature of the ultrasonic wave assisted extraction is 65-75 DEG C, preferably 70 DEG C；And/or
In step (a), the power of the ultrasonic wave assisted extraction is 200-400W, preferably 320W；And/or
In step (a), the time of the ultrasonic wave assisted extraction is 30-120min, preferably 60min, 45min；And/or
In step (a), the extraction solid-liquid ratio of the ultrasonic wave assisted extraction is 1:10-50, preferably 1:30 or 1:25；
And/or
In step (a), in ethanol ultrasonic wave assisted extraction Chinese toon two or more times, the Chinese toon being used in mixed way
Extract mixed liquor；And/or
In step (b), the speed of the centrifugation is 3500-4500r/min, preferably 4000r/min；And/or
In step (b), the time of the centrifugation is 10-30min, preferably 20min.
In some embodiments, in step (c), the weight ratio of Chinese toon raw material used in water used and step (a) is 1:
2-10, preferably 1:5；And/or
In step (d), the macroreticular resin is AB-8 macroreticular resin；And/or
Before executing step (d), the small fashionable dress column of 12-36 is impregnated with 90-98% ethyl alcohol, is then washed with 90-98% ethyl alcohol
It washs, is finally washed with water to no alcohol taste, it is preferable that impregnate 24 small fashionable dress columns with 95% ethyl alcohol and finally used then with 95% ethanol washing
It is washed to no alcohol taste；And/or
In step (d), the Chinese toona fruit crude extract dilution is crossed into institute with the speed of the preferred 8mL/min of 5-10mL/min
State macroporous resin column.
In some embodiments, in step (f), concentration of alcohol used be 60-80%, preferably 70%.
In some embodiments, before step (h), the Chinese toon extract sterling is freezed in -80 DEG C of refrigerators
5-10 hours, preferably 8 hours.
In some embodiments, step (h) is small for the Chinese toon extract sterling is placed in 12-36 in freeze drier
Shi Youxuan obtains the Chinese toon extract powder after 24 hours.
In some embodiments, the Chinese toon is selected from Chinese toona fruit.
In some embodiments, the Chinese toon uses powder.
The Chinese toon extract can be effectively reduced damage of the cis-platinum to body, increase internal glutathione content, reduce
Internal mda content.
Detailed description of the invention
Fig. 1 is blank control bone marrow cell micronucleus stained photographs；
Fig. 2 is Chinese toona fruit extract-treated bone marrow cell micronucleus stained photographs；
Fig. 3 is that curcumin handles bone marrow cell micronucleus stained photographs；
Fig. 4 is that Chinese toon seed extract+curcumin handles bone marrow cell micronucleus stained photographs；
Fig. 5 is Chinese toon seed extract+cis-platinum extract-treated bone marrow cell micronucleus stained photographs；
Fig. 6 is curcumin+cisplatin treated bone marrow cell micronucleus stained photographs；
Fig. 7 is Chinese toon seed extract+curcumin+cisplatin treated bone marrow cell micronucleus stained photographs；
Fig. 8 is cisplatin treated bone marrow cell micronucleus stained photographs.
Specific embodiment
Technical solution in order to preferably explain the present invention, is discussed in detail the embodiment of the present invention with reference to the accompanying drawing.With
Lower embodiment should not be construed as further illustrating the present invention to fixation or limitation of the invention.Unless otherwise specified, real
Applying technical characteristic used in example and could alternatively be has equivalent or identity function or effect under the premise of without departing substantially from inventive concept
Other techniques known in the art features.
1 materials and methods
1.1 material
Chinese toona fruit is purchased from Hui nationality's medicinal material market；SPF grades of Kunming kind adult male mices 80,6~8 week old, weight
25~30g is freely eaten and drinks water, provided by Anhui Province's anti-aging Chinese herbal medicine Industry Technology Center.
1.2 instruments and reagent
Instrument: Motic BA400 type biomicroscope (band digital photographing and image analysis system), medicinal herb grinder (Zhejiang
The Yongkang City Jiang Sheng small stream bank hardware medical instrument factory), electronic balance (Shanghai Ao Haosi Instrument Ltd.), desk centrifuge (Town in Shanghai
Pavilion scientific instrument factory), thermostat water bath (Beijing Orient Jing Rui development in science and technology Co., Ltd), microplate reader (Thermo), freezing it is dry
Dry machine (SIM international Group), drying box (the upper macro experimental facilities Co., Ltd of Nereid), -80 DEG C of refrigerators
(Thermo), experimental water is the ultrapure water of Millipore ultrapure water machine preparation, Ultrasound Instrument (the limited public affairs of city of Kunshan's ultrasonic instrument
Department).
Reagent: Giemsa pigment, cis-platinum (CDDP, freeze-dried type) be originated from Shandong pharmacy (Hainan) Co., Ltd (lot number:
2WA2A1307047B, 20mg/ bottles), curcumin (green skies Biotechnology Co., Ltd, product number SC0299-25mg), GSH
Kit and MDA kit are purchased from Nanjing and build up Bioengineering Research Institute.
1.3 method
1.3.1 the extraction of Chinese toon seed extract
Chinese toona fruit powder 20g is weighed in beaker, with 70% EtOH Sonicate ripple assisted extraction (70 DEG C, 320W), repeats two
Secondary, extracting solid-liquid ratio is 1:30,1:25 respectively, and extraction time is 60min, 45min respectively, and merging filtrate volatilizees and is concentrated into nothing
Ethanol flavor, 4000r/min are centrifuged 20min, take its precipitating, i.e. Chinese toon extract crude product, and addition 100mL distilled water dilution is spare,
As Chinese toona fruit crude extract dilution.
It takes 100g AB-8 macroreticular resin, impregnates 24 small fashionable dress columns with 95% ethyl alcohol first, then with 95% ethanol washing,
It is finally washed with water to no alcohol taste, Chinese toona fruit crude extract dilution is crossed into AB-8 large pore resin absorption column with the speed of 8mL/min, inhale
After attached, first with distillation washing column, extra pigment and impurity washed away, until efflux is colourless, is then washed with 70% ethyl alcohol
Take off to the liquid colorless of outflow, eluent volatilization be finally concentrated into no alcohol taste, -80 DEG C refrigerator freezing 8 hours, then be placed in cold
Chinese toona fruit extract powder, i.e. Chinese toon seed extract sterling are obtained in lyophilizer after 24 hours, is stored for future use in 4 DEG C of refrigerators.
Chinese toon seed extract described in following tests uses the Chinese toon seed extract sterling.
1.3.2 the toxicity preliminary experiment of mouse peritoneal injection Chinese toon seed extract, curcumin
Male mice 32 are taken, is divided into 4 groups at random, is grouped situation are as follows: (1) blank control group；(2) Chinese toon seed extract
Group；(3) curcumin group；(4) Chinese toon seed extract group, curcumin joint group.After adapting to feeding 1 week, each group is handled as follows:
(1) group gives intraperitoneal injection of saline 0.1ml/kg bw for three days on end；(2) group is given for three days on end is injected intraperitoneally
Chinese toon seed extract 150mg/kg bw；(3) group gives intraperitoneal injection curcumin 150mg/kg bw for three days on end；(4) continuous 3 are organized
It gives intraperitoneal injection Chinese toon seed extract and curcumin 150mg/kg bw (the ratio between Chinese toon seed extract and curcumin are 1:1).
Operation is primary daily for above-mentioned each group intraperitoneal injection.
Every mouse last time administration is put to death afterwards for 24 hours, is taken out two sides femur with the PBS solution of pH6.8 and is rinsed marrow, takes
Bone marrow cell is respectively used to following micronucleus test, takes nephridial tissue to be prepared into 10% homogenate spare, measures malonaldehyde in renal tissue
(MDA) and glutathione (GSH) content.
1.3.3 the influence of mouse peritoneal injection Chinese toon seed extract, curcumin combination with cisplatin to mouse
Male mice 32, it is divided into 4 groups at random, are as follows: (1) Chinese toon extract and cis-platinum combined processing group；(2) curcumin
With cis-platinum combined processing group；(3) Chinese toon extract, curcumin joint and cisplatin treated group；(4) cis-platinum model group.
Parallel to adapt to after feeding 1 week, each group is handled as follows:
(1) group is given for three days on end is injected intraperitoneally after Chinese toon seed extract 150mg/kg bw single intraperitoneal injection cis-platinum again
6.5mg/kg bw；(2) group is given for three days on end is injected intraperitoneally after curcumin 150mg/kg bw single intraperitoneal injection cis-platinum again
6.5mg/kg bw；(3) group gives intraperitoneal injection Chinese toon seed extract and the total 150mg/kg bw (Chinese toona fruit of curcumin for three days on end
The ratio between extract and curcumin are 1:1) after single intraperitoneal injection cis-platinum 6.5mg/kg bw again；(4) group gives abdominal cavity for three days on end
Injecting normal saline (0.9% sodium chloride solution) single intraperitoneal injection cis-platinum 6.5mg/kg bw afterwards.Above-mentioned each group intraperitoneal injection is equal
Operation is primary daily.
Every mouse last time administration is put to death afterwards for 24 hours, is taken out two sides femur with the PBS solution of pH6.8 and is rinsed marrow, takes
Bone marrow cell is respectively used to following micronucleus test, takes nephridial tissue to be prepared into 10% homogenate spare, measures malonaldehyde in renal tissue
(MDA) and glutathione (GSH) content.
1.4 micronucleus test
It takes appropriate marrow in shrinkage pool porcelain plate, is mixed with the fetal calf serum of inactivation, is added drop-wise to push jack on glass slide, it is naturally dry
It is dry.Each preparation of specimen 2.Push jack is put into methanol and fixes 10min, be put into after taking-up in Giemsa dye liquor dyeing 15~
30min, pure water rinsing are dried.Selection cell is complete, smear is uniform, dyeing region appropriate, in oily microscopic observation.First low
It is observed under times mirror, selection is evenly distributed, and dyes preferable region, then count under the microscope oily.Polychromatic erythrocyte
(Polychromatic erythrocytes, PCE) is in dusty blue, and one may occur in which one or more micronucleus into the cell[7].Out
The PCE of existing multiple micronucleus counts the PCE number (MnPCEs/ containing micronucleus in 1000PCE by a calculating, for every mouse
1000PCEs) and the percentage of the PCE containing micronucleus (%MNPCEs).
1.5 statistical method
It is analyzed using 12.0 statistical software of SPSS.Enumeration data Chi-square Test, continuous data variance analysis into
Row compares.It is that difference has significant meaning with P < 0.05, is that difference has extremely significant meaning with P < 0.01.
2 experimental results
The influence of 2.1 Chinese toon seed extracts and curcumin to Micronucleus
Normal mouse PCE micronuclear rates are 5 ‰ hereinafter, it is the positive that micronuclear rates, which are greater than 5 ‰,[4].About drug toxicity, according to
The pertinent regulations of Chinese herbal and crude drugs preparations, every kind of Chinese medicine are recorded nontoxic in two or more ancient books, are just considered as free of toxic effects, the experiment
The single medicinal material zingiberaceous plant of selection, Chinese toon are being showed no toxicity record in document " book on Chinese herbal medicine ", Compendium of Material Medica, " other mesh ",
The drug of description selection is safe and reliable[3].As shown in Table 1, compared with blank control group, simple injection curcumin or Chinese toona fruit are mentioned
It takes object not and will lead to the increase of micronuclear rates, there was no significant difference, and each cell mean is 5 ‰ hereinafter, for feminine gender.Preliminary proof
Chinese toon seed extract and curcumin will not generate toxic action to mouse.Fig. 1 to Fig. 4 is each group part bone marrow cell micronucleus dye respectively
Pornographic condition, Rate of Appearance Micronuclei is extremely low or even occurs without micronucleus, and arrow show micronucleus, and Fig. 1-4 is the photo of each group PCE micronucleus
(Giemsa, × 1000 oil mirrors).
1 Chinese toon seed extract of table, influence of the curcumin to Micronucleus
2.2 Chinese toon seed extracts, influence of the curcumin to cisplatin induction Micronucleus
As shown in Table 2, compared with blank control group, cis-platinum model group micronuclear rates significantly increase, and micronuclear rates are greater than 5 ‰ and are
The positive, and difference is extremely significant (P < 0.01).The different micronuclear rates positive of degree is also presented in remaining administration group, illustrates a certain amount of suitable
Platinum can inducing mouse generation cytotoxicity.But compared with cis-platinum model group, mouse is given at Chinese toon seed extract and curcumin
Its micronuclear rates is substantially reduced after reason, individual Chinese toon seed extract and curcumin processing group significant difference (P < 0.05), Chinese toona fruit
Extract and curcumin Combined Treatment difference are extremely significant (P < 0.01).Illustrate processed by Chinese toon seed extract and curcumin
Mouse has certain protective action to the cytotoxicity of cisplatin induction, and Chinese toon seed extract and curcumin Combined Treatment are than single
Only high treating effect.Fig. 5 to Fig. 8 is the photo of each group part bone marrow cell micronucleus staining conditions respectively.
2 Chinese toon seed extract of table, influence of the curcumin to cisplatin induction Micronucleus
Note: compared with the control group, * P < 0.05, P < 0.01 * *；Compared with model group, #P < 0.05, ##P < 0.01.Arrow
It show micronucleus, Fig. 5-8 each group PCE micronucleus (Giemsa, × 1000 oil mirrors)
In conjunction with the result of 2.1 sections and 2.2 sections it is found that the combination of Chinese toon seed extract and curcumin or both is to common mouse
Micronucleus in myeloid tissue, which accounts for PCE ratio, to be influenced unobvious, illustrate on the physiological function of healthy mice influence very little.And in cis-platinum
In the bone marrow of mice of processing, micronucleus content can be significantly reduced.It follows that Chinese toon seed extract and turmeric or both
Combination at least can be used in preventing, treating or slowing down the damage of the mouse body as caused by cis-platinum.
The influence of 2.3 Chinese toon seed extracts and curcumin to mouse kidney tissue GSH, MDA content
As shown in Table 3, compared with blank control group, it is simple inject curcumin or Chinese toon seed extract will not cause GSH,
MDA content is abnormal, and there was no significant difference compared with the control group.Chinese toon seed extract and curcumin are further demonstrated to mouse
Toxic action will not be generated.
3 Chinese toon seed extract of table, influence of the curcumin to mouse kidney tissue GSH, MDA content
The influence of 2.4 Chinese toon seed extracts, curcumin to cisplatin induction mouse kidney tissue GSH, MDA value
As shown in Table 4, compared with blank control group, cis-platinum model group GSH content is significantly reduced, and MDA content significantly increases,
And difference is extremely significant (P < 0.01).Compared with cis-platinum model group, mouse gives its GSH after Chinese toon seed extract and curcumin processing
Content is significantly raised, individual Chinese toon seed extract and curcumin processing group significant difference (P < 0.05), Chinese toon seed extract with
Curcumin Combined Treatment difference is extremely significant (P < 0.01)；Mouse gives its MDA content after Chinese toon seed extract and curcumin processing
It is substantially reduced, individual Chinese toon seed extract and curcumin processing group significant difference (P < 0.05), Chinese toon seed extract and turmeric
Plain Combined Treatment difference is extremely significant (P < 0.01).Illustrate to lure cis-platinum by Chinese toon seed extract and the processed mouse of curcumin
The cytotoxicity and renal toxicity and injury of kidney led have certain protective action, and Chinese toon seed extract and curcumin Combined Treatment
Than independent high treating effect, illustrate that Chinese toon extract and curcumin can generate synergistic effect.
4 Chinese toon seed extract of table, curcumin are to the content of cisplatin induction mouse kidney tissue GSH, MDA
Note: compared with the control group, * P < 0.05, P < 0.01 * *；Compared with model group, #P < 0.05, ##P < 0.01.
In conjunction with the result of 2.3 sections and 2.4 sections it is found that the combination of Chinese toon seed extract and curcumin or both is to common mouse
The content influence of GSH, MDA in nephridial tissue are unobvious, illustrate to influence very little to the physiological function of healthy mice.And at cis-platinum
In the kidney of mouse of reason, GSH content is dramatically increased, and MDA content substantially reduces.
GSH content dramatically increases and can activate or improve many removing toxic substances for relying on GSH, anti-oxidant etc. and play protection to body
The biochemical reaction of effect.MDA content, which substantially reduces, can reduce MDA to the crosslinking denaturation of the large biological molecules such as the promotion albumen of body
Harm.
It follows that the combination of Chinese toon seed extract and turmeric or both at least can be used in preventing, treating or slow down by
The damage of mouse body caused by cis-platinum.
3 discuss
Research shows that oxidative stress is one of main mechanism of toxicity of cisplatin, the oxidative stress that reactive oxygen species (ROS) mediate
It is played an important role in toxicity of cisplatin.For cis-platinum in vivo mainly through kidney excretion, metal platinum ion can be accumulated in kidney, therefore
Its renal toxicity is most obvious.Cis-platinum is toxic to the different parts of kidney, glomerulus, renal tubular function and morphology can be caused to change
Become.Cis-platinum can interact with antioxidant system, influence antioxidant system and Antioxidant Enzymes, different tissues is caused to aoxidize
The weight of damage, response to oxidative stress degree and institutional framework loss is positively correlated in apparent.
Mouse marrow cell micro nuclear test is considered as the prefered method for carrying out internal genetic toxicity test, for detecting dye
The damaging action of colour solid clastogen and aneuploid inducer to chromosome[1].The microkernel incidence of measurement marrow PCE is detection
One highly useful index of cytogenetics damage, it is all chromosome to be made to be broken, or it is coupled chromosome and spindle
The genetic damage substance destroyed can be detected with micronucleus test.
Studies have shown that containing phenolic hydroxyl group in the structure of curcumin, when peroxidatic reaction of lipid occurs for cell membrane, phenol
Oxidation reaction can occur for hydroxyl, can effectively terminate radical reaction[1], thus it shows many physiological activity, such as antioxygen
Change, is antitumor, many effects such as prevent aging[6], become the research hotspot in natural drug in recent years.Chinese toon seed extract pair
The cytotoxicity caused protective action of cis-platinum may be related with the anti-oxidant and free-radical scavenging activity of Chinese toon antioxidant content.
This experiment establishes CDDP inducing mouse model of oxidative using bone marrow cells in mice as research object, it was demonstrated that
Chinese toon seed extract and curcumin further demonstrate Chinese toona fruit according to experimental result and extract to the protective action of cytotoxicity
Object and curcumin have synergy, this research will provide new thinking and money to develop new natural harmless antioxidant
Source.
The result of study of this test shows two kinds of natural plants phenolic active components --- Chinese toon seed extract and curcumin
Administering drug combinations can generate synergistic effect, can be effectively reduced the heredity damage of the cytotoxicity and Mouse Somatic Cells by cisplatin induced
Wound.Preliminary judgement Chinese toon seed extract can antagonism cis-platinum induced mice chromosome mutation, and Chinese toon seed extract and curcumin
With synergistic effect.In conclusion certain density Chinese toon seed extract can significantly improve oxidation resistance in Mice Body, right
Genotoxicity caused by the such anticancer drug of cis-platinum has good adjustment effect.
Above each embodiment is only intended to further illustrate the present invention, is not for limiting protection model of the invention
It encloses, it is all obviously to change based on equivalents made by design of the invention and to each technical solution of the invention
Into each falling within protection scope of the present invention.
Claims (10)
1. a kind of preparation method of Chinese toon extract, the preparation method include the following steps:
(a) ultrasonic wave assisted extraction Chinese toon in ethanol obtains Chinese toon and extracts mixed liquor；
(b) Chinese toon extraction mixed liquor is evaporated into no ethanol flavor, is centrifuged, takes precipitating, obtain Chinese toon extract crude product.
2. the preparation method of Chinese toon extract as described in claim 1, which is characterized in that the preparation method further includes walking as follows
It is rapid:
(c) the Chinese toon extract crude product is prepared into Chinese toon crude extract dilution with water；
(d) the Chinese toon crude extract dilution is crossed into macroporous resin column；
(e) macroporous resin column is washed with distillation, until the liquid colorless of outflow；
(f) macroporous resin column described in ethanol elution, until the liquid colorless of outflow, collects eluent；
(g) eluent volatilization is concentrated into no alcohol taste, obtains Chinese toon extract sterling.
3. the preparation method of Chinese toon extract as claimed in claim 2, which is characterized in that the preparation method further includes walking as follows
It is rapid:
(h) the Chinese toon extract sterling is lyophilized, obtains Chinese toon extract sterling powder.
4. the preparation method of Chinese toon extract as described in claim 1,2 or 3, which is characterized in that
In step (a), concentration of alcohol used be 60-80%, preferably 70%；And/or
In step (a), the temperature of the ultrasonic wave assisted extraction is 65-75 DEG C, preferably 70 DEG C；And/or
In step (a), the power of the ultrasonic wave assisted extraction is 200-400W, preferably 320W；And/or
In step (a), the time of the ultrasonic wave assisted extraction is 30-120min, preferably 60min, 45min；And/or
In step (a), the extraction solid-liquid ratio of the ultrasonic wave assisted extraction is 1:10-50, preferably 1:30 or 1:25；And/or
In step (a), ultrasonic wave assisted extraction Chinese toon two or more times, extract by the Chinese toon being used in mixed way in ethanol
Mixed liquor；And/or
In step (b), the speed of the centrifugation is 3500-4500r/min, preferably 4000r/min；And/or
In step (b), the time of the centrifugation is 10-30min, preferably 20min.
5. the preparation method of Chinese toon extract as described in Claims 2 or 3, which is characterized in that
In step (c), the weight ratio of Chinese toon raw material used in water used and step (a) is 1:2-10, preferably 1:5；And/or
In step (d), the macroreticular resin is AB-8 macroreticular resin；And/or
Before executing step (d), the small fashionable dress column of 12-36 is impregnated with 90-98% ethyl alcohol, then uses 90-98% ethanol washing, most
After be washed with water to no alcohol taste, it is preferable that impregnate 24 small fashionable dress columns with 95% ethyl alcohol and be finally washed with water then with 95% ethanol washing
To no alcohol taste；And/or
It is in step (d), the Chinese toona fruit crude extract dilution is excessively described big with the speed of the preferred 8mL/min of 5-10mL/min
Hole resin column.
6. the preparation method of Chinese toon extract as described in Claims 2 or 3, which is characterized in that
In step (f), concentration of alcohol used be 60-80%, preferably 70%.
7. the preparation method of Chinese toon extract as claimed in claim 3, which is characterized in that
Before step (h), the Chinese toon extract sterling is freezed into 5-10 hours, preferably 8 hours in -80 DEG C of refrigerators.
8. the preparation method of Chinese toon extract as claimed in claim 3, which is characterized in that
Step (h) obtains institute for the Chinese toon extract sterling to be placed in freeze drier after preferably 24 hours 12-36 hours
State Chinese toon extract powder.
9. the preparation method of Chinese toon extract as described in claim 1, which is characterized in that
The Chinese toon is selected from Chinese toona fruit.
10. the preparation method of Chinese toon extract as described in claim 1, which is characterized in that
The Chinese toon uses powder.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110302248A (en) * | 2019-06-25 | 2019-10-08 | 阜阳师范学院 | The method for extracting Chinese toon seed general flavone |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104861084A (en) * | 2015-06-15 | 2015-08-26 | 潍坊医学院 | Cedrela sinensis polysaccharide extracting method |
| CN105131096A (en) * | 2015-08-10 | 2015-12-09 | 潍坊医学院 | Fructus toonae sinensis protein extraction method |
| CN105919025A (en) * | 2016-05-03 | 2016-09-07 | 王胜 | Vegetable salt containing Chinese mahogany seed extract and preparation method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104861084A (en) * | 2015-06-15 | 2015-08-26 | 潍坊医学院 | Cedrela sinensis polysaccharide extracting method |
| CN105131096A (en) * | 2015-08-10 | 2015-12-09 | 潍坊医学院 | Fructus toonae sinensis protein extraction method |
| CN105919025A (en) * | 2016-05-03 | 2016-09-07 | 王胜 | Vegetable salt containing Chinese mahogany seed extract and preparation method thereof |
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| 邢莎莎: "《万方数据库》", 2 November 2012 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110302248A (en) * | 2019-06-25 | 2019-10-08 | 阜阳师范学院 | The method for extracting Chinese toon seed general flavone |
| CN110302248B (en) * | 2019-06-25 | 2021-10-26 | 阜阳师范学院 | Method for extracting total flavonoids from Chinese toon seeds |
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