CN103494860A - Method for preparing lithospermum extract and application of lithospermum extract - Google Patents
Method for preparing lithospermum extract and application of lithospermum extract Download PDFInfo
- Publication number
- CN103494860A CN103494860A CN201310470068.1A CN201310470068A CN103494860A CN 103494860 A CN103494860 A CN 103494860A CN 201310470068 A CN201310470068 A CN 201310470068A CN 103494860 A CN103494860 A CN 103494860A
- Authority
- CN
- China
- Prior art keywords
- extraction
- radix arnebiae
- radix
- extract
- arnebiae extract
- 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
Abstract
The invention discloses a method for preparing a lithospermum extract and application of the lithospermum extract, and relates to the field of medicines. The invention aims at disclosing the method for preparing the lithospermum extract and application of the lithospermum extract to preparing a medicine for treating tumors. The method comprises the following steps: 1, preparing lithospermum powder; 2, extracting effective parts mainly being naphthoquinone compounds of the lithospermum powder obtained in the step 1 by utilizing a CO2 supercritical extraction technology, thus obtaining the wine-colored thick-cream lithospermum extract. The application of the lithospermum extract is the application of the lithospermum extract in preparing the medicine for treating tumors. According to the invention, gynecologic tumors and digestive tract cell lines are emphatically researched and the research result verifies that the effective parts of lithospermum naphthoquinone have exact biological activity for inhibiting growth and inducing apotosis to tumor cell lines, thereby providing a scientific experimental basis for developing hard lithospermum and the effective parts of lithospermum naphthoquinone into a new anticancer drug with a remarkable curative effect and specific targets.
Description
Technical field
The present invention relates to field of medicaments, be specifically related to preparation method and the application thereof of Radix Arnebiae extract.
Background technology
Tumor disease has become one of archenemy that threatens human health and life, and its M & M is all rising year by year.Mortality rate is only second to the cardiovascular diseases, is the mankind's second largest killer.How by effective measures, to prevent and treat malignant tumor has become the top priority of China down to whole world disease preventing and treating, is also particularly medical domain key subjects anxious to be resolved of the whole society.But, due to the difficulty of early diagnosis of cancer, a considerable amount of cancer patients will rely on Drug therapy, therefore, the research and development of antitumor drug are the key areas that biomedical science develops rapidly with application.
Cervical cancer is one of modal woman's sexual reproduction road malignant tumor, in China's cervical cancer, accounts for first of female genital tract malignant tumor.Not only in the female sex organ carcinoma, account for first place but also be the most common carcinoma in the various malignant tumor of women.Ovarian cancer is that a class is difficult to by the cancer of early discovery, is one of common malignant tumor of female sex organ, accounts for more than 4% of gynecological's cancerous protuberance, and its mortality rate occupies the gynecological tumor first place, and sickness rate accounts for the 3rd, inferior to cervical cancer and carcinoma of corpus uteri.The sickness rate of ovarian cancer constantly rises in recent years, very harmful to women's health.In the last thirty years, the treatment of gynecological's carcinoma has obtained considerable progress, but the curative effect of ovarian cancer there is no obvious improvement only.Cancer of pancreas is the digestive tract kinds of tumor that a kind of grade malignancy is very high, and its pathogenesis is still not clear, and in malignant tumor, the M & M of cancer of pancreas is all higher.Cancer of pancreas has the biological behaviour of high malignancy, and clinical process is dangerous, and within 5 years, survival rate is very low, more than 60, plants in malignant tumor, and prognosis is the poorest.
TCM on Tumor is compared its unique advantage is arranged with the western medical treatment tumor, and Chinese medicine has very abundant theory and clinical experience on tumor diagnosis and treatment.Chinese medicine is due to complicated component, the comprehensive function of, too many levels multi-level by bringing into play, many target spots and antitumor.The traditional Chinese medical science takes full advantage of the Chinese medicine multiple target effect and carries out integrally-regulated from Overall View to body in conjunction with theory of Chinese medical science on treatment tumor thinking.Chinese medicine has good research and development prospect, finding antitumor effective ingredient treatment malignant tumor from Chinese herbal medicine is more and more accepted by numerous scholars and patient at present, and oneself becomes Therapeutic Method commonly used, it is one of effective means in Multimodality Therapy of Malignant Tumors.
Summary of the invention
The objective of the invention is to disclose a kind of preparation method and the application in the medicine of preparation treatment tumor thereof of Radix Arnebiae extract.
The preparation method of a kind of Radix Arnebiae extract of the present invention, carry out according to the following steps:
One, prepare Arnebia Root: Radix Arnebiae (Radix Lithospermi) is placed under the condition that temperature is 40 ℃～50 ℃ and dries, obtain dry Radix Arnebiae (Radix Lithospermi), then be ground into powder, cross sieve No. 1, retain under sieve, obtain Arnebia Root;
Two, utilize CO
2the Arnebia Root that supercritical extraction technique obtains step 1 carries out take the extraction of naphthoquinone constituents as main effective site: the Arnebia Root that step 1 is obtained is packed in extraction kettle, extraction kettle is heated to 40 ℃～50 ℃, extraction-container and is heated to 40 ℃～50 ℃, basin and is heated to 30 ℃～40 ℃, then open CO
2bottle, control CO
2flow is 20L/h～30L/h, by high-pressure pump, system is pressurizeed, when the extracting pressure of the extraction kettle parsing pressure that is 15MPa～20MPa, extraction-container is 4MPa～8MPa, start cycling extraction, and maintenance extraction kettle constant temperature and pressure, after extraction 2h～3h, from the extraction-container discharge hole for discharge, obtain wine-colored stiff paste shape Radix Arnebiae extract.
The application of a kind of Radix Arnebiae extract of the present invention is the application of Radix Arnebiae extract in the medicine of preparation treatment tumor.
Advantage of the present invention:
One, the invention provides a kind of preparation method of Radix Arnebiae extract, take Radix Arnebiae (Radix Lithospermi) as raw material, utilize the carbon dioxide supercritical fluid extraction technology to carry out take to it efficient enrichment of carrying out that naphthoquinone constituents is main effective site, extraction, purifying process, supercritical fluid extraction is compared with traditional chemical solvent extraction method, its superiority is residual without chemical solvent, pollution-free, avoid the extract high temperature pyrolysis, activity and the technique of protection biological active substances are simple, extraction efficiency is high and purity is high, is conducive to commercial production;
Two, the present invention adopts four factor one or three horizontal quadrature designs, take content and extracted amount as index, uses CO
2supercritical liquid extraction technique is optimized selection to the extraction conditions of the naphthoquinone constituents in Chinese Drug Zicao, show that the optimum extraction condition of Naphthoquinone in Zicao constituents is: extracting pressure 17MPa, 40 ℃ of extraction temperature, parsing pressure 5MPa, 40 ℃ of resolution temperatures, flow 25L/h; The methodological study of measuring total naphthoquinone constituents content in extract with shikonin through spectrophotography shows, the relative standard deviation of its precision, stability, repeatability and average recovery all is less than 3%, illustrate that the method accurately, reliably; The HPLC content assaying method that application has been set up is measured 4 kinds of main constituents such as acetylshikonins in effective part group, and as calculated, the total content of four kinds of compositions is 58.42 ± 6.15%, accounts for 87.30 ± 3.22% of total naphthoquinone content;
Three, the present invention confirms its anti-tumor activity effect by the external activity screening experiment, and result of study shows, Radix Arnebiae (Radix Lithospermi) effective site can be induced the kinds of tumor cells apoptosis; Radix Arnebiae (Radix Lithospermi) effective site can suppress the Growth of Cells of human cervical cancer 1 squamous cell carcinoma Siha, Proliferation of Human Ovarian Cell Skov3, human pancreatic cancer cell Bxpc3, human cervical carcinoma cell Hela, Proliferation of Human Ovarian Cell Ho-8910, human liver cancer cell HepG2, and be dosage and time dependence, show that Radix Arnebiae (Radix Lithospermi) effective site all has stronger cytotoxic activity to kinds of tumors, the 48h Radix Arnebiae extract all is less than 0.9ug/ml to the IC50 value of six kinds of cells; Pharmacological research finds that Radix Arnebiae (Radix Lithospermi) effective site has antitumor action, proves that its growth to 6 kinds of tumor cells of people all has inhibitory action, and is obvious dose-effect relationship, is the experimental basis of having explored efficient and economic natural antitumor drug provision.
Four, to take emphatically gynecological tumor and digestive tract cell strain be object of study in the present invention, confirm that Gronwell naphthaquinone effective site has the biological activity of definite inhibition growth inducing apoptosis to tumor cell line, for gromwell root and naphthoquinone effective kind part thereof are developed to curative effect brilliance, target spot clear and definite PTS the experiment basis of science is provided, for the clinical treatment of gynecological cancer and alimentary tract cancer adopts rational scheme that the experimental data of theoretical foundation and science is provided.
The accompanying drawing explanation
Fig. 1 is Radix Arnebiae extract effect Siha cell growth curve, in Fig. 1-▲-Siha cell growth curve while being Radix Arnebiae extract effect 24h, in Fig. 1-◆-Siha cell growth curve while being Radix Arnebiae extract effect 48h, in Fig. 1-■-Siha cell growth curve while being Radix Arnebiae extract effect 72h;
Fig. 2 is Radix Arnebiae extract effect skov3 cell growth curve, in Fig. 2-▲-skov3 cell growth curve while being Radix Arnebiae extract effect 24h, in Fig. 2-◆-skov3 cell growth curve while being Radix Arnebiae extract effect 48h, in Fig. 2-■-skov3 cell growth curve while being Radix Arnebiae extract effect 72h;
Fig. 3 is Radix Arnebiae extract effect Bxpc3 cell growth curve, in Fig. 3-▲-Bxpc3 cell growth curve while being Radix Arnebiae extract effect 24h, in Fig. 3-◆-Bxpc3 cell growth curve while being Radix Arnebiae extract effect 48h, in Fig. 3-■-Bxpc3 cell growth curve while being Radix Arnebiae extract effect 72h;
Fig. 4 is Radix Arnebiae extract effect Ho-8910 cell growth curve, in Fig. 4-▲-Ho-8910 cell growth curve while being Radix Arnebiae extract effect 24h, in Fig. 4-◆-Ho-8910 cell growth curve while being Radix Arnebiae extract effect 48h, in Fig. 4-■-Ho-8910 cell growth curve while being Radix Arnebiae extract effect 72h;
Fig. 5 is Radix Arnebiae extract effect HepG2 cell growth curve, in Fig. 5-▲-HepG2 cell growth curve while being Radix Arnebiae extract effect 24h, in Fig. 5-◆-HepG2 cell growth curve while being Radix Arnebiae extract effect 48h, in Fig. 5-■-HepG2 cell growth curve while being Radix Arnebiae extract effect 72h;
Fig. 6 is Radix Arnebiae extract effect Hela cell growth curve, in Fig. 6-▲-Hela cell growth curve while being Radix Arnebiae extract effect 24h, in Fig. 6-◆-Hela cell growth curve while being Radix Arnebiae extract effect 48h, in Fig. 6-■-Hela cell growth curve while being Radix Arnebiae extract effect 72h;
Fig. 7 be 48h various dose Radix Arnebiae extract to six kinds of Tumoricidal action figure, the in Fig. 7 is that 48h concentration is that 0.075 μ g/mL Radix Arnebiae extract is to six kinds of Tumoricidal action figure, in Fig. 7
for 48h concentration be 0.15 μ g/mL Radix Arnebiae extract to six kinds of Tumoricidal action figure, the ■ in Fig. 7 is that 48h concentration is that 0.75 μ g/mL Radix Arnebiae extract is to six kinds of Tumoricidal action figure;
Fig. 8 is normal Siha cellular morphology figure;
Fig. 9 is the Siha cellular morphology figure after Radix Arnebiae extract is processed;
Figure 10 is normal Skov3 cellular morphology figure;
Figure 11 is the Skov3 cellular morphology figure after Radix Arnebiae extract is processed;
Figure 12 is normal Bxpc3 cellular morphology figure;
Figure 13 is the Bxpc3 cellular morphology figure after Radix Arnebiae extract is processed.
The specific embodiment
The specific embodiment one: the preparation method of a kind of Radix Arnebiae extract of present embodiment, carry out according to the following steps:
One, prepare Arnebia Root: Radix Arnebiae (Radix Lithospermi) is placed under the condition that temperature is 40 ℃～50 ℃ and dries, obtain dry Radix Arnebiae (Radix Lithospermi), then be ground into powder, cross No. 1 sieve (2000 ± 70 μ m, 10 orders), retain under sieve, obtain Arnebia Root;
Two, utilize CO
2the Arnebia Root that supercritical extraction technique obtains step 1 carries out take the extraction of naphthoquinone constituents as main effective site: the Arnebia Root that step 1 is obtained is packed in extraction kettle, extraction kettle is heated to 40 ℃～50 ℃, extraction-container and is heated to 40 ℃～50 ℃, basin and is heated to 30 ℃～40 ℃, then open CO
2bottle, control CO
2flow is 20L/h～30L/h, by high-pressure pump, system is pressurizeed, when the extracting pressure of the extraction kettle parsing pressure that is 15MPa～20MPa, extraction-container is 4MPa～8MPa, start cycling extraction, and maintenance extraction kettle constant temperature and pressure, after extraction 2h～3h, from the extraction-container discharge hole for discharge, obtain wine-colored stiff paste shape Radix Arnebiae extract.
The dry root that Radix Arnebiae (Radix Lithospermi) described in the present embodiment step 1 is comfrey Radix Arnebiae (Radix Lithospermi) (Lithospermum erythrorhizon sieb.et Zucc), also claim gromwell root; In Radix Arnebiae (Radix Lithospermi), main component is the naphthoquinone constituents, there is thermolability through evidence Naphthoquinone in Zicao constituents, shikonin the heating more than 60 ℃ color become atropurpureus by redness, with temperature rising pace of change, accelerate, thin layer chromatography checks that speckle disappears, and the prompting alkannin is destroyed.
Present embodiment provides a kind of preparation method of Radix Arnebiae extract; take Radix Arnebiae (Radix Lithospermi) as raw material; utilize the carbon dioxide supercritical fluid extraction technology to carry out take to it efficient enrichment of carrying out that naphthoquinone constituents is main effective site; extraction, purifying process; supercritical fluid extraction is compared with traditional chemical solvent extraction method; its superiority is residual without chemical solvent; pollution-free; avoid the extract high temperature pyrolysis; activity and the technique of protection biological active substances are simple; extraction efficiency is high and purity is high, is conducive to commercial production.
Present embodiment adopts four factor one or three horizontal quadrature designs, take content and extracted amount as index, uses CO
2supercritical liquid extraction technique is optimized selection to the extraction conditions of the naphthoquinone constituents in Chinese Drug Zicao, show that the optimum extraction condition of Naphthoquinone in Zicao constituents is: extracting pressure 17MPa, 40 ℃ of extraction temperature, parsing pressure 5MPa, 40 ℃ of resolution temperatures, flow 25L/h; The methodological study of measuring total naphthoquinone constituents content in extract with shikonin through spectrophotography shows, the relative standard deviation of its precision, stability, repeatability and average recovery all is less than 3%, illustrate that the method accurately, reliably; The HPLC content assaying method that application has been set up is measured 4 kinds of main constituents such as acetylshikonins in effective part group, and as calculated, the total content of four kinds of compositions is 58.42 ± 6.15%, accounts for 87.30 ± 3.22% of total naphthoquinone content;
Present embodiment confirms its anti-tumor activity effect by the external activity screening experiment, and result of study shows, Radix Arnebiae (Radix Lithospermi) effective site can be induced the kinds of tumor cells apoptosis; Radix Arnebiae (Radix Lithospermi) effective site can suppress the Growth of Cells of human cervical cancer 1 squamous cell carcinoma Siha, Proliferation of Human Ovarian Cell Skov3, human pancreatic cancer cell Bxpc3, human cervical carcinoma cell Hela, Proliferation of Human Ovarian Cell Ho-8910, human liver cancer cell HepG2, and be dosage and time dependence, show that Radix Arnebiae (Radix Lithospermi) effective site all has stronger cytotoxic activity to kinds of tumors, the 48h Radix Arnebiae extract all is less than 0.9ug/ml to the IC50 value of six kinds of cells; Pharmacological research finds that Radix Arnebiae (Radix Lithospermi) effective site has antitumor action, proves that its growth to 6 kinds of tumor cells of people all has inhibitory action, and is obvious dose-effect relationship, is the experimental basis of having explored efficient and economic natural antitumor drug provision.
It is object of study that present embodiment be take emphatically gynecological tumor and digestive tract cell strain, confirm that Gronwell naphthaquinone effective site has the biological activity of definite inhibition growth inducing apoptosis to tumor cell line, for gromwell root and naphthoquinone effective kind part thereof are developed to curative effect brilliance, target spot clear and definite PTS the experiment basis of science is provided, for the clinical treatment of gynecological cancer and alimentary tract cancer adopts rational scheme that the experimental data of theoretical foundation and science is provided.
The specific embodiment two: present embodiment is different from the specific embodiment one: the preparation method of Radix Arnebiae extract, carry out according to the following steps: one, prepare Arnebia Root: Radix Arnebiae (Radix Lithospermi) is placed under the condition that temperature is 45 ℃～50 ℃ and dries, obtain dry Radix Arnebiae (Radix Lithospermi), then be ground into powder, cross sieve No. 1, retain under sieve, obtain Arnebia Root; Two, utilize CO
2the Arnebia Root that supercritical extraction technique obtains step 1 carries out take the extraction of naphthoquinone constituents as main effective site: the Arnebia Root that step 1 is obtained is packed in extraction kettle, extraction kettle is heated to 40 ℃～45 ℃, extraction-container and is heated to 40 ℃～45 ℃, basin and is heated to 30 ℃～40 ℃, then open CO
2bottle, control CO
2flow is 20L/h～25L/h, by high-pressure pump, system is pressurizeed, when the extracting pressure of the extraction kettle parsing pressure that is 15MPa～18MPa, extraction-container is 4MPa～6MPa, start cycling extraction, and maintenance extraction kettle constant temperature and pressure, after extraction 2h～3h, from the extraction-container discharge hole for discharge, obtain wine-colored stiff paste shape Radix Arnebiae extract.Other is identical with the specific embodiment one.
The specific embodiment three: present embodiment is different from the specific embodiment one or two: the preparation method of Radix Arnebiae extract, carry out according to the following steps: one, prepare Arnebia Root: Radix Arnebiae (Radix Lithospermi) is placed under the condition that temperature is 45 ℃～48 ℃ and dries, obtain dry Radix Arnebiae (Radix Lithospermi), then be ground into powder, cross sieve No. 1, retain under sieve, obtain Arnebia Root; Two, utilize CO
2the Arnebia Root that supercritical extraction technique obtains step 1 carries out take the extraction of naphthoquinone constituents as main effective site: the Arnebia Root that step 1 is obtained is packed in extraction kettle, extraction kettle is heated to 40 ℃～42 ℃, extraction-container and is heated to 40 ℃～42 ℃, basin and is heated to 30 ℃～40 ℃, then open CO
2bottle, control CO
2flow is 22L/h～25L/h, by high-pressure pump, system is pressurizeed, when the extracting pressure of the extraction kettle parsing pressure that is 16MPa～17MPa, extraction-container is 5MPa～6MPa, start cycling extraction, and maintenance extraction kettle constant temperature and pressure, after extraction 2h～3h, from the extraction-container discharge hole for discharge, obtain wine-colored stiff paste shape Radix Arnebiae extract.Other is identical with the specific embodiment one or two.
The specific embodiment four: present embodiment is different from one of specific embodiment one to three: the preparation method of Radix Arnebiae extract, carry out according to the following steps: one, prepare Arnebia Root: Radix Arnebiae (Radix Lithospermi) is placed under the condition that temperature is 45 ℃ and dries, obtain dry Radix Arnebiae (Radix Lithospermi), then be ground into powder, cross sieve No. 1, retain under sieve, obtain Arnebia Root; Two, utilize CO
2the Arnebia Root that supercritical extraction technique obtains step 1 carries out take the extraction of naphthoquinone constituents as main effective site: the Arnebia Root that step 1 is obtained is packed in extraction kettle, extraction kettle is heated to 40 ℃, extraction-container and is heated to 40 ℃, basin and is heated to 30 ℃, then open CO
2bottle, control CO
2flow is 25L/h, by high-pressure pump, system is pressurizeed, when the extracting pressure of the extraction kettle parsing pressure that is 17MPa, extraction-container is 5MPa, start cycling extraction, and maintenance extraction kettle constant temperature and pressure, after extraction 2h, from the extraction-container discharge hole for discharge, obtain wine-colored stiff paste shape Radix Arnebiae extract.Other is identical with one of specific embodiment one to three.
The specific embodiment five: the application of a kind of Radix Arnebiae extract of present embodiment is the application of Radix Arnebiae extract in the medicine of preparation treatment tumor.
The application of present embodiment Radix Arnebiae extract is Radix Arnebiae extract directly to be made to the medicine for the treatment of tumor.
Present embodiment confirms its anti-tumor activity effect by the external activity screening experiment, and result of study shows, Radix Arnebiae (Radix Lithospermi) effective site can be induced the kinds of tumor cells apoptosis; Radix Arnebiae (Radix Lithospermi) effective site can suppress the Growth of Cells of human cervical cancer 1 squamous cell carcinoma Siha, Proliferation of Human Ovarian Cell Skov3, human pancreatic cancer cell Bxpc3, human cervical carcinoma cell Hela, Proliferation of Human Ovarian Cell Ho-8910, human liver cancer cell HepG2, and be dosage and time dependence, show that Radix Arnebiae (Radix Lithospermi) effective site all has stronger cytotoxic activity to kinds of tumors, the 48h Radix Arnebiae extract all is less than 0.9ug/ml to the IC50 value of six kinds of cells; Pharmacological research finds that Radix Arnebiae (Radix Lithospermi) effective site has antitumor action, proves that its growth to 6 kinds of tumor cells of people all has inhibitory action, and is obvious dose-effect relationship, is the experimental basis of having explored efficient and economic natural antitumor drug provision.
Adopt following verification experimental verification effect of the present invention:
Test one: a kind of preparation method of Radix Arnebiae extract, carry out according to the following steps:
One, prepare Arnebia Root: Radix Arnebiae (Radix Lithospermi) is placed under the condition that temperature is 45 ℃ and dries, obtain dry Radix Arnebiae (Radix Lithospermi), then be ground into powder, cross sieve No. 1, retain under sieve, obtain Arnebia Root;
Two, utilize CO
2the Arnebia Root that supercritical extraction technique obtains step 1 carries out take the extraction of naphthoquinone constituents as main effective site: take Arnebia Root that 200 gram step 1 obtain and pack in the 1L extraction kettle, extraction kettle is heated to 40 ℃, extraction-container and is heated to 40 ℃, basin and is heated to 30 ℃, then open CO
2bottle, control CO
2flow is 25L/h, by high-pressure pump, system is pressurizeed, when the extracting pressure of the extraction kettle parsing pressure that is 17MPa, extraction-container is 5MPa, start cycling extraction, and maintenance extraction kettle constant temperature and pressure, after extraction 2h, from the extraction-container discharge hole for discharge, obtain wine-colored stiff paste shape Radix Arnebiae extract.
The dry root that Radix Arnebiae (Radix Lithospermi) described in this test procedure one is comfrey Radix Arnebiae (Radix Lithospermi) (Lithospermum erythrorhizon sieb.et Zucc), also claim gromwell root.
The comparison of extracting method:
Supersound extraction: take Arnebia Root (cross No. 1 sieve after the lower Arnebia Root of sieve) 100g, join in 95% ethanol supersound extraction 2 times, each 30min, after filtering, decompression and solvent recovery is to without alcohol, and drying, obtain Radix Arnebiae extract.
Merceration extracts: take Arnebia Root (cross No. 1 sieve after sieve under Arnebia Root) 100g, join in 95% ethanol merceration 36 hours, filter, filtrate decompression reclaims solvent to without alcohol, and drying, obtain Radix Arnebiae extract.
Reflux, extract: take Arnebia Root (cross No. 1 sieve after sieve under Arnebia Root) 100g, join in 95% ethanol of 800mL the 45min that refluxes, filter, the 95% alcohol reflux 30min that adds again 600mL, filter merging filtrate, reclaim solvent to nothing alcohol, drying, obtain Radix Arnebiae extract.
Percolation extracts: take Arnebia Root (crossing Arnebia Root under the sieve after sieving for No. 1) 100g, add 200mL95% ethanol moistening 6 hours, the medicated powder that moistening is good evenly adds percolator, get rid of in cylinder and flood 36 hours after air, flow velocity percolation with 3mL/10min, the reclaim under reduced pressure percolate is to nothing alcohol, and drying, obtain Radix Arnebiae extract.
Test one method extraction by the present invention: obtain Radix Arnebiae extract.
Supersound extraction, merceration extraction, reflux, extract,, percolation are extracted and tested one method by the present invention and extract each 10mg of Radix Arnebiae extract obtained, add respectively 95% dissolve with ethanol and be settled to 20mL, obtain 5 parts of Radix Arnebiae extract solution, ultraviolet-spectrophotography is measured trap at the wavelength place of 516nm, measure total naphthoquinone content, every part of triplicate the results are shown in Table 1:
The comparison of table 1 extracting method
As shown in Table 1, according to the comparison of different extracting method, test one extracting method by the present invention Radix Arnebiae (Radix Lithospermi) is extracted, total naphthoquinone content is the highest.
The investigation of temperature in leaching process:
Get three parts of Arnebia Roots, every part of 1g, 95% ethanol ultrasonic extraction, reclaim under reduced pressure extracting solution (recovered temperature is respectively 50 ℃, 60 ℃, 70 ℃), drying, obtain three parts of Radix Arnebiae extracts, gets three parts of each 10mg of Radix Arnebiae extract sample, respectively add 95% dissolve with ethanol and be settled to 10mL, obtain three parts of Radix Arnebiae extract solution, ultraviolet-spectrophotography is measured trap at the wavelength place of 516nm, measures total naphthoquinone content, every part of triplicate the results are shown in Table 2:
Table 2 extracts the investigation of temperature
As shown in Table 2, the affect highly significant of temperature on the naphthoquinone constituents, should not at high temperature extract, and extracts temperature and be controlled in 60 ℃.
The comparison of different technology conditions:
At CO
2in supercritical extraction process, the factor that affects extraction yield mainly contains extraction time, extracting pressure, extraction temperature, parsing pressure, resolution temperature, CO
2flows etc., this test is extracted Radix Arnebiae (Radix Lithospermi) medical material optimum process condition with regard to these factors and is determined.
1, the selection of extraction initial condition:
According to previous literature report about CO
2the research of supercritical extraction Radix Arnebiae (Radix Lithospermi) and trial test result, the initial extraction conditions that this test is set is extracting pressure 25MPa, 45 ℃ of extraction temperature are resolved pressure 4MPa, 45 ℃ of resolution temperatures.
2, determining of extraction time:
Take Radix Arnebiae (Radix Lithospermi) medicinal powder (crossing Arnebia Root under the sieve after sieving for No. 1) 200 grams, test one Radix Arnebiae (Radix Lithospermi) SFE-CO according to the present invention
2the basic procedure of method extraction process and extraction initial condition are proposed extraction, and collect once per half an hour, observes extracted amount, and result shows, receives three times later substantially without extract, and for guaranteeing extraction fully, this test determines that extraction time is 2 hours.
3, CO
2the selection of flow:
Take three parts of Radix Arnebiae (Radix Lithospermi) crude drug powder (crossing Arnebia Root under the sieve sieved for No. 1) 200 grams, test one Radix Arnebiae (Radix Lithospermi) SFE-CO according to the present invention
2the basic procedure of method extraction process and extraction initial condition are extracted, and extraction time is 2 hours, investigates best CO
2flow the results are shown in Table 3:
Table 3CO
2the investigation result of flow
Result shows, CO
2the content that flow is the 25L/h hydroxyl naphthoquinone total pigment is higher, so CO is chosen in this test
2flow be 25L/h.
4, orthogonal experiment Optimized Extraction condition:
According to above-mentioned monofactorial experimental result, through analysis-by-synthesis, slective extraction pressure (A), extraction temperature (B), parsing pressure (C), 4 principal elements of resolution temperature (D), each sets 3 levels, designs four factor three horizontal quadratures experiment L
9(3
4), extraction time is 2 hours, CO
2flow is 20L/h, and Radix Arnebiae (Radix Lithospermi) is carried out to CO
2the supercritical extraction factor level is in Table 4, and experimental result is in Table 5, and interpretation is in Table 6:
Table 4 supercritical CO
2fluid extraction L
9(3
4) factor level
Select L9 (3 according to table 4
4) orthogonal table, each factors combine and interpretation of result are in Table 5, and the content of effective site and extracted amount are the important indicators of investigating technological parameter, and we combine content and two indexs of extracted amount of naphthoquinone constituents technique have been done to the weighted analysis of variance, the results are shown in Table 6.
Table 5 extraction process L
9(3
4) orthogonal experiments
The analysis of table 6 orthogonal experiments
Table 7 analysis of variance table
F tables look-up
(0.05)(2,2)=19.00
Result shows, C
2d
2b
2a
1for optimised process, resolving pressure is 5MPa, resolution temperature is 40 ℃, extraction temperature is 40 ℃, extracting pressure is 17MPa (table 7). the results of analysis of variance shows to resolve pressure, these two factors of resolution temperature experimental result is had to the significance impact, and two of extracting pressure and extraction temperature are without obvious difference.
The technique repeated experiments:
For determining the stability of technique, we have carried out having carried out replica test 3 times by best technological parameter, in extracting pressure, are 17MPa, and extraction temperature is 40 ℃, resolve pressure 5MPa, and resolution temperature is 40 ℃, CO
2tested under the condition that flow is 25L/h, be the results are shown in Table 8:
Table 8 technology stability result of the test
Result shows, it is basicly stable that this tests definite process conditions.
As shown in the above, this experiment adopts four factor one or three horizontal quadrature designs, take content and extracted amount as index, uses CO
2supercritical liquid extraction technique is optimized selection to the extraction conditions of the naphthoquinone constituents in Chinese Drug Zicao, show that the optimum extraction condition of Naphthoquinone in Zicao constituents is: extracting pressure 17MPa, 40 ℃ of extraction temperature, parsing pressure 5MPa, 40 ℃ of resolution temperatures, flow 25L/h.The methodological study of measuring total naphthoquinone constituents content in extract with shikonin through ultraviolet spectrophotometry shows, the relative standard deviation of its precision, stability, repeatability and average recovery all is less than 3%, illustrate that the method accurately, reliably.
The assay of total naphthoquinone compound and 4 kinds of main constituents in Radix Arnebiae (Radix Lithospermi) effective site:
1., need testing solution preparation: get 5 parts, different batches Radix Arnebiae (Radix Lithospermi) effective site sample (Radix Arnebiae extract prepared by method of the present invention), every part of about 10mg, accurately weighed, put in volumetric flask, add 95% ethanol 10mL, standardize solution, cross pre-column and filter membrane (0.45 μ m), obtain 5 parts of test liquid samples;
2., total naphthoquinone compound assay: 5 parts of test liquid samples that 1. step made with ultraviolet visible spectrophotometry are measured absorbance A under the 516nm wavelength, and the specific absorbance (E1%1cm) of Shikonin (C16H16O5) of take is the total naphthoquinone compound content of 242 calculating.
3., main constituent assay: get 5 parts of test liquid samples difference injection liquid chromatographies that 1. 20 μ L steps make, every part of triplicate the results are shown in Table 9:
The effective site assay result of table 9 Radix Arnebiae extract
As shown in Table 9, Radix Arnebiae (Radix Lithospermi) is through CO
2the content of the effective site of Radix Arnebiae extract prepared by supercritical extraction process should be 63%～74%, has reached the specification requirement of effective part group.
The HPLC content assaying method that application has been set up is measured 4 kinds of main constituents such as acetylshikonins in effective part group, and as calculated, the total content of four kinds of compositions is 58.42 ± 6.15%, accounts for 87.30 ± 3.22% of total naphthoquinone content.
The effective site antitumor activity of Radix Arnebiae extract:
The screening Radix Arnebiae extract is to different cell strain cytotoxicity:
Get the different densities tumor cell, 1*10
5individual/hole, be inoculated in 96 well culture plates, after cultivation 24h, adds concentration to be respectively the Radix Arnebiae extract solution of 0.075 μ g/mL, 0.15 μ g/mL, 0.75 μ g/mL, continues cultivation 24,48h, 72h, with mtt assay, measures light absorption value, presses formula and calculate:
Suppression ratio %=(1-OD
the dosing group/ OD
matched group) * 100%
With average suppression ratio mapping, application SPSS13.0 computed in software IC50 value.
Described tumor cell is cervical cancer cell strain Siha, Hela; Ovarian Cancer Cells Skov3, Ho-8910; Pancreas cancer cell strain Bxpc3; Hepatoma cell strain HepG2; All purchased from Harbin Medical University's scientific experiment center.
Fig. 1 is Radix Arnebiae extract effect Siha cell growth curve, in Fig. 1-▲-Siha cell growth curve while being Radix Arnebiae extract effect 24h, in Fig. 1-◆-Siha cell growth curve while being Radix Arnebiae extract effect 48h, in Fig. 1-■-Siha cell growth curve while being Radix Arnebiae extract effect 72h; Fig. 2 is Radix Arnebiae extract effect skov3 cell growth curve, in Fig. 2-▲-skov3 cell growth curve while being Radix Arnebiae extract effect 24h, in Fig. 2-◆-skov3 cell growth curve while being Radix Arnebiae extract effect 48h, in Fig. 2-■-skov3 cell growth curve while being Radix Arnebiae extract effect 72h; Fig. 3 is Radix Arnebiae extract effect Bxpc3 cell growth curve, in Fig. 3-▲-Bxpc3 cell growth curve while being Radix Arnebiae extract effect 24h, in Fig. 3-◆-Bxpc3 cell growth curve while being Radix Arnebiae extract effect 48h, in Fig. 3-■-Bxpc3 cell growth curve while being Radix Arnebiae extract effect 72h; Fig. 4 is Radix Arnebiae extract effect Ho-8910 cell growth curve, in Fig. 4-▲-Ho-8910 cell growth curve while being Radix Arnebiae extract effect 24h,
In Fig. 4-◆-Ho-8910 cell growth curve while being Radix Arnebiae extract effect 48h, in Fig. 4-■-Ho-8910 cell growth curve while being Radix Arnebiae extract effect 72h; Fig. 5 is Radix Arnebiae extract effect HepG2 cell growth curve, in Fig. 5-▲-HepG2 cell growth curve while being Radix Arnebiae extract effect 24h, in Fig. 5-◆-HepG2 cell growth curve while being Radix Arnebiae extract effect 48h, in Fig. 5-■-HepG2 cell growth curve while being Radix Arnebiae extract effect 72h; Fig. 6 is Radix Arnebiae extract effect Hela cell growth curve, in Fig. 6-▲-Hela cell growth curve while being Radix Arnebiae extract effect 24h, in Fig. 6-◆-Hela cell growth curve while being Radix Arnebiae extract effect 48h, in Fig. 6-■-Hela cell growth curve while being Radix Arnebiae extract effect 72h; Experimental result shows that Radix Arnebiae extract all has obvious inhibitory action (Fig4-1 to Fig4-6) to the growth of six kinds of people source cancerous cell, in the concentration range arranged, has obvious dosage and time dependence.Show that Radix Arnebiae extract all has certain cytotoxic activity to these cells.Radix Arnebiae extract all is less than 0.9ug/ml to the IC50 value of the 48h of six kinds of tumor cells.In these cells, Radix Arnebiae extract is the highest to the sensitivity of siha, Skov3 cell, and higher inhibitory action is arranged during 24h, presents significant inhibitory action when 48h.To in these two kinds of cytosis 72h along with the increase of dosage and time, inhibitory action strengthens gradually, shows that Radix Arnebiae extract presents significant time and dose dependent to the effect of siha, skov3 cell.To the inhibitory action major embodiment of siha, skov3 cell in early days; Radix Arnebiae extract and acetylshikonin are different to the inhibitory action of all the other cells and Skov3, siha cell, and its sensitivity is slightly poor, and inhibitory action is slower, but also can reach obvious inhibitory action when 48h.Result shows, Radix Arnebiae extract has strong active anticancer to tumor cell, and different cell strains is shown to different sensitivity, illustrates that the Radix Arnebiae extract active anticancer has certain specificity.
Various dose administration group at 48h to six kinds of cytotoxicity:
Fig. 7 be 48h various dose Radix Arnebiae extract to six kinds of Tumoricidal action figure, the in Fig. 7 is that 48h concentration is that 0.075 μ g/mL Radix Arnebiae extract is to six kinds of Tumoricidal action figure, in Fig. 7
for 48h concentration be 0.15 μ g/mL Radix Arnebiae extract to six kinds of Tumoricidal action figure, the ■ in Fig. 7 is that 48h concentration is that 0.75 μ g/mL Radix Arnebiae extract is to six kinds of Tumoricidal action figure; Observe six kinds of tumor cells survival rate under the effect of Radix Arnebiae extract with mtt assay and significantly descend, and be dose dependent.The inhibitory action sensitivity of Radix Arnebiae extract to Siha, Skov3, Ho-8910 cell proliferation.
Morphological observation:
With every hole 2 * 10
5individual cell density is inoculated in 6 orifice plates, after cultivating 24h, adds concentration to be respectively the Radix Arnebiae extract of 0.075 μ g/mL, 0.15 μ g/mL, 0.75 μ g/mL, respectively at 0,6,12, the 24h observation by light microscope.
Described cell is cervical cancer cell strain Siha, Hela; Ovarian Cancer Cells Skov3, Ho-8910; Pancreas cancer cell strain Bxpc3; Hepatoma cell strain HepG2; All purchased from Harbin Medical University's scientific experiment center.
Radix Arnebiae extract effect kinds of tumor cells metamorphosis:
Observe the normal group cell under inverted microscope and be the epitheliated type adherent growth, cell is extension, flat, the cell homogenizing is and transparent, and cell generally has 2-4 kernel, and nuclear membrane, kernel profile are obvious, the iuntercellular close structure, Growth of Cells is vigorous. and administration group cell outline strengthens, contrast increases, and becomes circle and floats, and cell contact fluffs, proliferation slows, in kytoplasm, granule increases.See shown in Fig. 8 to Figure 13, Fig. 8 is normal Siha cellular morphology figure; Fig. 9 is the Siha cellular morphology figure after Radix Arnebiae extract is processed; Figure 10 is normal Skov3 cellular morphology figure; Figure 11 is the Skov3 cellular morphology figure after Radix Arnebiae extract is processed; Figure 12 is normal Bxpc3 cellular morphology figure; Figure 13 is the Bxpc3 cellular morphology figure after Radix Arnebiae extract is processed.
Cultured cell has more fixing form and structure under normal cultivation conditions, under inverted microscope, can observe this form and structure.After Radix Arnebiae extract is processed, the variation of larger form and structure all appears in 6 kinds of tumor cells of experimental group.Show that Growth of Cells is suppressed, cell quantity reduces, cavity (6 kinds of cells have) occurs, cell shrinkage, form pleomorphic type increase the variations such as (8910, bxpc3) and projection shortening.The sign in early stage that causes cell death after Here it is drug effect, more necrocytosis feature appears in hepg2 and 8910 after drug treating, the cavity that siha, HeLa, bxpc3 cell occur is many than other cell, is the difference that medicine causes the application point difference of cell.The core shrinkage all appears in six kinds of cells, and apoptotic body appears in prompting, illustrates that Radix Arnebiae extract can cause six kinds of apoptotic generations.
Experimental result shows, Radix Arnebiae extract and acetylshikonin can be induced the kinds of tumor cells apoptosis.Radix Arnebiae extract and acetylshikonin can suppress the Growth of Cells of human cervical cancer 1 squamous cell carcinoma Siha, Proliferation of Human Ovarian Cell Skov3, human pancreatic cancer cell Bxpc3, human cervical carcinoma cell Hela, Proliferation of Human Ovarian Cell Ho-8910, human liver cancer cell HepG2, and are dosage and time dependence.Show that Radix Arnebiae extract all has stronger cytotoxic activity to kinds of tumors, the 48h Radix Arnebiae extract all is less than 0.9ug/mL to the IC50 value of six kinds of cells.In six kinds of tumor cells, the highest to the sensitivity of Siha cell, the poorest to the sensitivity of HepG2.
This experiment Primary Study Radix Arnebiae (Radix Lithospermi) effective site anti-tumor activity in vitro, prove that its growth to 6 kinds of tumor cells of people all has inhibitory action, and be obvious dose-effect relationship, be the experimental basis of having explored efficient and economic natural antitumor drug provision.
Claims (5)
1. the preparation method of a Radix Arnebiae extract is characterized in that the preparation method of Radix Arnebiae extract is carried out according to the following steps:
One, prepare Arnebia Root: Radix Arnebiae (Radix Lithospermi) is placed under the condition that temperature is 40 ℃～50 ℃ and dries, obtain dry Radix Arnebiae (Radix Lithospermi), then be ground into powder, cross sieve No. 1, retain under sieve, obtain Arnebia Root;
Two, utilize CO
2the Arnebia Root that supercritical extraction technique obtains step 1 carries out take the extraction of naphthoquinone constituents as main effective site: the Arnebia Root that step 1 is obtained is packed in extraction kettle, extraction kettle is heated to 40 ℃～50 ℃, extraction-container and is heated to 40 ℃～50 ℃, basin and is heated to 30 ℃～40 ℃, then open CO
2bottle, control CO
2flow is 20L/h～30L/h, by high-pressure pump, system is pressurizeed, when the extracting pressure of the extraction kettle parsing pressure that is 15MPa～20MPa, extraction-container is 4MPa～8MPa, start cycling extraction, and maintenance extraction kettle constant temperature and pressure, after extraction 2h～3h, from the extraction-container discharge hole for discharge, obtain wine-colored stiff paste shape Radix Arnebiae extract.
2. the preparation method of a kind of Radix Arnebiae extract according to claim 1, the preparation method that it is characterized in that Radix Arnebiae extract is carried out according to the following steps: one, prepare Arnebia Root: Radix Arnebiae (Radix Lithospermi) is placed under the condition that temperature is 45 ℃～50 ℃ and dries, obtain dry Radix Arnebiae (Radix Lithospermi), then be ground into powder, cross sieve No. 1, retain under sieve, obtain Arnebia Root; Two, utilize CO
2the Arnebia Root that supercritical extraction technique obtains step 1 carries out take the extraction of naphthoquinone constituents as main effective site: the Arnebia Root that step 1 is obtained is packed in extraction kettle, extraction kettle is heated to 40 ℃～45 ℃, extraction-container and is heated to 40 ℃～45 ℃, basin and is heated to 30 ℃～40 ℃, then open CO
2bottle, control CO
2flow is 20L/h～25L/h, by high-pressure pump, system is pressurizeed, when the extracting pressure of the extraction kettle parsing pressure that is 15MPa～18MPa, extraction-container is 4MPa～6MPa, start cycling extraction, and maintenance extraction kettle constant temperature and pressure, after extraction 2h～3h, from the extraction-container discharge hole for discharge, obtain wine-colored stiff paste shape Radix Arnebiae extract.
3. the preparation method of a kind of Radix Arnebiae extract according to claim 1, the preparation method that it is characterized in that Radix Arnebiae extract is carried out according to the following steps: one, prepare Arnebia Root: Radix Arnebiae (Radix Lithospermi) is placed under the condition that temperature is 45 ℃～48 ℃ and dries, obtain dry Radix Arnebiae (Radix Lithospermi), then be ground into powder, cross sieve No. 1, retain under sieve, obtain Arnebia Root; Two, utilize CO
2the Arnebia Root that supercritical extraction technique obtains step 1 carries out take the extraction of naphthoquinone constituents as main effective site: the Arnebia Root that step 1 is obtained is packed in extraction kettle, extraction kettle is heated to 40 ℃～42 ℃, extraction-container and is heated to 40 ℃～42 ℃, basin and is heated to 30 ℃～40 ℃, then open CO
2bottle, control CO
2flow is 22L/h～25L/h, by high-pressure pump, system is pressurizeed, when the extracting pressure of the extraction kettle parsing pressure that is 16MPa～17MPa, extraction-container is 5MPa～6MPa, start cycling extraction, and maintenance extraction kettle constant temperature and pressure, after extraction 2h～3h, from the extraction-container discharge hole for discharge, obtain wine-colored stiff paste shape Radix Arnebiae extract.
4. the preparation method of a kind of Radix Arnebiae extract according to claim 1, the preparation method that it is characterized in that Radix Arnebiae extract is carried out according to the following steps: one, prepare Arnebia Root: Radix Arnebiae (Radix Lithospermi) is placed under the condition that temperature is 45 ℃ and dries, obtain dry Radix Arnebiae (Radix Lithospermi), then be ground into powder, cross sieve No. 1, retain under sieve, obtain Arnebia Root; Two, utilize CO
2the Arnebia Root that supercritical extraction technique obtains step 1 carries out take the extraction of naphthoquinone constituents as main effective site: the Arnebia Root that step 1 is obtained is packed in extraction kettle, extraction kettle is heated to 40 ℃, extraction-container and is heated to 40 ℃, basin and is heated to 30 ℃, then open CO
2bottle, control CO
2flow is 25L/h, by high-pressure pump, system is pressurizeed, when the extracting pressure of the extraction kettle parsing pressure that is 17MPa, extraction-container is 5MPa, start cycling extraction, and maintenance extraction kettle constant temperature and pressure, after extraction 2h, from the extraction-container discharge hole for discharge, obtain wine-colored stiff paste shape Radix Arnebiae extract.
5. the application of a kind of Radix Arnebiae extract that prepared by preparation method as claimed in claim 1, it is characterized in that the application of Radix Arnebiae extract in the medicine of preparation treatment tumor.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310470068.1A CN103494860A (en) | 2013-10-10 | 2013-10-10 | Method for preparing lithospermum extract and application of lithospermum extract |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310470068.1A CN103494860A (en) | 2013-10-10 | 2013-10-10 | Method for preparing lithospermum extract and application of lithospermum extract |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN103494860A true CN103494860A (en) | 2014-01-08 |
Family
ID=49860183
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310470068.1A Pending CN103494860A (en) | 2013-10-10 | 2013-10-10 | Method for preparing lithospermum extract and application of lithospermum extract |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103494860A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104774151A (en) * | 2015-01-30 | 2015-07-15 | 泰山医学院 | Preparation technology of Mount Taishan Radix Lithospermi naphthoquinone active monomers |
| CN107648296A (en) * | 2017-08-18 | 2018-02-02 | 合肥丰洁生物科技有限公司 | A kind of preparation method of Radix Arnebiae extract |
| CN111560289A (en) * | 2020-04-15 | 2020-08-21 | 青岛市妇女儿童医院（青岛市妇幼保健院、青岛市残疾儿童医疗康复中心、青岛市新生儿疾病筛查中心） | Method for preparing lithospermum oil by ultrasonic-assisted supercritical fluid extraction |
| CN113638245A (en) * | 2021-08-10 | 2021-11-12 | 宜兴中大纺织有限公司 | Plant dye one-bath method dyeing method of polyester-cotton blended yarn |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101234100A (en) * | 2008-02-04 | 2008-08-06 | 浙江大学 | Application of naphthoquinone compounds |
| CN101669981A (en) * | 2009-10-30 | 2010-03-17 | 黑龙江中医药大学 | Method for extracting hydroxyl naphthoquinone total pigment with supercritical carbon dioxide |
| CN101721452A (en) * | 2010-01-04 | 2010-06-09 | 新疆大学 | New process for improving utilization ratio of lithospermum |
| CN101919895A (en) * | 2010-03-15 | 2010-12-22 | 王喜军 | Traditional Chinese medicine extract with anti-HCV (hepatitis C virus) function |
| CN101683331B (en) * | 2008-09-27 | 2013-01-30 | 厦门大学 | Application of alkannin derivant |
-
2013
- 2013-10-10 CN CN201310470068.1A patent/CN103494860A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101234100A (en) * | 2008-02-04 | 2008-08-06 | 浙江大学 | Application of naphthoquinone compounds |
| CN101683331B (en) * | 2008-09-27 | 2013-01-30 | 厦门大学 | Application of alkannin derivant |
| CN101669981A (en) * | 2009-10-30 | 2010-03-17 | 黑龙江中医药大学 | Method for extracting hydroxyl naphthoquinone total pigment with supercritical carbon dioxide |
| CN101721452A (en) * | 2010-01-04 | 2010-06-09 | 新疆大学 | New process for improving utilization ratio of lithospermum |
| CN101919895A (en) * | 2010-03-15 | 2010-12-22 | 王喜军 | Traditional Chinese medicine extract with anti-HCV (hepatitis C virus) function |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104774151A (en) * | 2015-01-30 | 2015-07-15 | 泰山医学院 | Preparation technology of Mount Taishan Radix Lithospermi naphthoquinone active monomers |
| CN107648296A (en) * | 2017-08-18 | 2018-02-02 | 合肥丰洁生物科技有限公司 | A kind of preparation method of Radix Arnebiae extract |
| CN111560289A (en) * | 2020-04-15 | 2020-08-21 | 青岛市妇女儿童医院（青岛市妇幼保健院、青岛市残疾儿童医疗康复中心、青岛市新生儿疾病筛查中心） | Method for preparing lithospermum oil by ultrasonic-assisted supercritical fluid extraction |
| CN113638245A (en) * | 2021-08-10 | 2021-11-12 | 宜兴中大纺织有限公司 | Plant dye one-bath method dyeing method of polyester-cotton blended yarn |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101647842B (en) | Fructus Podophylli extract and application thereof | |
| CN102309655B (en) | Traditional Chinese medicine composition with effect of preventing and treating cancers and application of Chinese medicinal composition | |
| CN103768534A (en) | Traditional Chinese medicinal composition with anti-tumor activity | |
| Zeng et al. | Laminaria japonica polysaccharides effectively inhibited the growth of nasopharyngeal carcinoma cells in vivo and in vitro study | |
| CN103494860A (en) | Method for preparing lithospermum extract and application of lithospermum extract | |
| CN102370695B (en) | Qinglongyi active extract, its preparation method and its application | |
| CN108567820A (en) | A kind of Chinese medicine composition and preparation method thereof for oncotherapy | |
| CN101904974A (en) | Chinese medicinal composition for treating malignant tumor diseases and preparation method thereof | |
| CN101850032A (en) | Anti-tumor traditional Chinese medicine composition and preparation method and application thereof | |
| CN103933541B (en) | A kind of glutathion anti-cancer composition | |
| CN102858359B (en) | Medicinal composition comprising alcohol-soluble and water-insoluble licorice extract, pharmaceutical preparation, pharmaceutical application, therapeutic method, and preparative method thereof | |
| CN104857040A (en) | New purpose of embelia parviflora wall for preparing medicines for treating liver fibrosis | |
| CN101168008B (en) | Medicinal composition with tumor inhibition function and preparation method and application thereof | |
| CN108542927A (en) | The application of the obcordate aspidopterys stem of falling the heart and its extract in anti-tumor aspect | |
| CN111704535B (en) | 3-carbonyl malus spectabilis ketone, extraction method and application in preparing antitumor drugs | |
| CN101385749A (en) | Use of heaven tree extract in preparing antineoplastic medicine | |
| CN104069194A (en) | A traditional Chinese medicine composition having anticancer effects and a preparing method and uses thereof | |
| CN106074682A (en) | Total alkaloid of sophora alopecuroide extract and preparation method thereof and the application prepared in medicament for resisting cervical cancer | |
| CN103191267B (en) | Traditional Chinese medicine composition for treating liver cancers | |
| CN102114065B (en) | Longhairy antenoron herb and common threewingnut root Chinese medicinal composition with effect of preventing and treating cancer and application thereof | |
| CN102805829B (en) | Preparation method of rhizoma smilacis glabrae extract and applications thereof in tumor treatment | |
| CN106310116A (en) | Rhizoma Zedoariae oil, and application and pharmaceutical preparation thereof | |
| CN104529968B (en) | Anti-tumor diterpenoid compound, and pharmaceutical composition, preparation method and application thereof | |
| CN104257955A (en) | Anti-hepatoma traditional Chinese medicine mixture containing arisaema amurense and production method of anti-hepatoma traditional Chinese medicine mixture | |
| CN104825922A (en) | Traditional Chinese medicine composition with anti-colorectal cancer activity and preparation method and application of traditional Chinese medicine composition |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C05 | Deemed withdrawal (patent law before 1993) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20140108 |