CN101411739B - Method for preparing Caesalpinia sappan water extract - Google Patents
Method for preparing Caesalpinia sappan water extract Download PDFInfo
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- CN101411739B CN101411739B CN2008100796720A CN200810079672A CN101411739B CN 101411739 B CN101411739 B CN 101411739B CN 2008100796720 A CN2008100796720 A CN 2008100796720A CN 200810079672 A CN200810079672 A CN 200810079672A CN 101411739 B CN101411739 B CN 101411739B
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Abstract
The invention relates to a method for preparing water extract by taking traditional Chinese medicines as raw materials, in particular to a method for preparing sapanwood water extract and application of the sapanwood water extract. The method solves the problem of development and utilization of the sapanwood traditional Chinese medicine. The method comprises the following steps: the sapanwood traditional Chinese medicine is weighed and added with 5 times of water for decoction, and supernatant is collected; the concentration of the supernatant is concentrated to between 1 and 2 grams per milliliter; the supernatant is subjected to centrifugation and ultrafiltration, and filtrate is collected, added with proteins (Albumin) for full reaction, subjected to ultrafiltration, and collected; and the sapanwood water extract is obtained after adjustment of the pH value and sterilization. The method has the characteristics: firstly, the traditional Chinese medicine extract is prepared by means of ultrafiltration, so that effective ingredients in the sapanwood can be intercepted to the maximum degree and loss of the effective ingredients can be reduced; and secondly, by adoption of protein purification, trace impurities in the solution can be removed, the effective ingredients of the extract are more concentrated and the effect is obvious. Moreover, the sapanwood water extract has obvious effect on treating ascites tumor and leukemia.
Description
Technical field
The present invention relates to a kind of is the method for feedstock production aqueous extract with Chinese medicine, is specially a kind of Caesalpinia sappan water extract preparation method and application thereof.
Background technology
Lignum Sappan (another name: Lignum Sappan, Soviet Union's a tree, used in making timber for boats, the Soviet side, lignum sappan, palm fibre wood, Ramulus et Folium Bischofiae Javanicae, red bavin) is the dry duramen of pulse family Semen Caesalpiniae platymiscium Lignum Sappan Caesalpinia sappan L..The four seasons all receive, but with autumn for well.Cut down and get trunk or thick branch, prune crust and sapwood Rhizoma Atractylodis macrocephalae, purplish red or reddish yellow heartwood in the middle of getting is put the ventilation and is dried in the shade and get final product.
Lignum Sappan is distributed in ground such as Guangxi, Guangdong, Taiwan, Guizhou, Yunnan, Sichuan.Medicinal Lignum Sappan contains brazilin (brasilin), brasilein multiple compositions such as (brasilein).
Ancient Times in China has many argumentations to the clinical practice of Lignum Sappan.The Tang Materia Medica record: main removing blood stasis, puerperal, the blood feeling of distension and oppression was desired the dead." Haiyao Bencao, Oversea Materia Medica " record: main asthenia blood addiction gas is stopped up stagnant; The lochia uneasiness is stirred pain in the abdomen; And blockage of main and collateral channels, men and women's apoplexy, trismus and inability to speak.Should carefully grind the fragrant smalls tool for weighing medicine in ancient China of nipple, wine is fried in shallow oil the Soviet side and is gone dregs to be taken after mixing with liquid, and the upright evil thing of telling is recovered." Japan hanako materia medica " record: control married woman's vim and vigour cardiac and abdominal pain, month uncomfortable and postpartum debility of time.The damage blood stasis is pounced in evacuation of pus pain relieving, subduing inflammation, woman's aphonia, and blood is kept silent, and shoot is divided in dysentery with red and white feces and back." medical center bun will " record: the reinforcing the heart dissipating blood stasis, remove the wind heat of the absurd work of blood system." medication heart method " record: Lignum Sappan, go wind and Radix Saposhnikoviae with using.The Compendium of Material Medica record: Lignum Sappan has the effect of " removing blood stasis, evacuation of pus pain relieving "." book on Chinese herbal medicine is looked for the truth " record: Lignum Sappan, function have similar Flos Carthami, few with then can and blood, many with then can removing blood stasis.But the Flos Carthami slightly warm in nature and, then the cold nature cold is also for this.So all cause of disease the exterior and the interior wind rises, and that hyperamization stagnates is not all right, and the puerperal blood faints distension is dead with (desires), and blood pain abdominal mass due to blood stasis, through diseases such as the heap soil or fertilizer over and around the roots of holding one's breath, carbuncle, injury from falling down, all should phase disease close and recuperate under medical treatment with his medicine.
At present many to the achievement in research of Lignum Sappan, for example the patent No. is the new purposes of 20041000047593 disclosed a kind of Lignum Sappan extract, this patent provides a kind of preparation method of Lignum Sappan extract, main feature is to use alcohol reflux, and points out that this extract can treat the under-supply caused disease of cerebral blood flow.The patent No. 2004100441119 discloses the patent application of a kind of Lignum Sappan ethyl acetate extract and effect thereof, and pointed out that this extract can significantly eliminate the rejection that organ transplantation produces, this extract also is to have utilized some traditional extracting method, and for example decocting method, supersound extract method, percolation, supercritical are quenched and followed the example of etc.The relevant article for the research of Lignum Sappan medical function is also a lot.
Summary of the invention
The present invention provides a kind of Caesalpinia sappan water extract preparation method and application thereof for the problem for the development and use that solve the Chinese medicine Lignum Sappan.
The present invention is realized that by following technical scheme a kind of Caesalpinia sappan water extract preparation method may further comprise the steps:
(1), title Chinese medicine Lignum Sappan adds 4-7 times of water, decoction 30-60min, collection supernatant; Data with optimum efficiency decoct 40min for adding 5 times of water,
(2), repeat above-mentioned (1) step, merge supernatant, centrifugal to supernatant, getting the supernatant concentration that obtains after centrifugal again is 1-2g/ml to concentration; Optimum data is 1g/ml,
(3), the concentrated solution that obtains after last step concentrated is centrifugal, divide secondary to carry out ultrafiltration, the aperture: relative molecular weight is respectively 6k and 3k, collects filtrate, merges.This method is than water extract-alcohol precipitation, or Chinese medicine traditional extraction process such as alcohol extracting-water precipitating can keep effective ingredient morely.
(4), in the filtrate that merged, add the protein (Albumin) of filtrate quality 0.1-0.5%, at 50-60 ℃, fully reaction under the 10-20min condition.Its main purpose is to eliminate residual trace impurity in the aqueous extract.Optimum data is 0.2% protein (Albumin), at 56 ℃, and fully reaction under the 15min condition.
(5), ultrafiltration, the aperture: relative molecular weight is respectively 3k, collects filtrate.
(6), adjust pH, the sterilization, obtain Caesalpinia sappan water extract.
The Caesalpinia sappan water extract that the method for the invention obtains is remarkable to treatment ascites tumor and leukemia effect, sees Table 1 and Fig. 1, below add up for therapeutic effect,
Table 1
Experiment mice H22 hepatic ascites tumor adopts abdominal cavity inoculation, 1X10
6Individual cell/only, respectively through vein and intraperitoneal administration, every kind of approach is with 50 behind the 24h, 100mgkg-1d-1, dosage, successive administration 7d.The normal saline of matched group injection equal volume is observed and is respectively organized the tumor-bearing mice life span, and repeated experiments 2 times is observed 60d, surpasses 60d and presses 60d calculating.Calculate the increase in life span of medication group respectively.Its increase in life span of abdominal cavity treatment ascites tumor mice can reach 193.4% as seen from Table 1.
K562 (people's red pulp leukemia cell line) suspension is adjusted into 1 * 10
6Individual cell/ml adds in 24 well culture plates 1000ul/ hole.Lignum Sappan extracting solution with variable concentrations joins in the cell suspension again, the 100ul/ hole.The every hole of matched group adds the 100ul normal saline, and mixing is put 37 ℃ of people, 5%CO
2Cultivate 36h in the incubator.Collecting cell adds that an amount of PBS of people is centrifugal to be washed 1 time.Add 5ul rhAnnexinV-PE in the sample cell, mixing is placed 20min in the room temperature lucifuge, adds that an amount of PRS of people is centrifugal to be washed 1 time again, abandons supernatant.Add PBS liquid 500ul in the precipitation, mixing.1h in-flow cell instrument is measured.Result as schematically shown in Figure 1 shows that Caesalpinia sappan water extract can kill 99.6% cancerous cell in 36 hours.
Above animal and cell experiment confirm that the Lignum Sappan extracting solution that the method for the invention obtains can be applied to preparation treatment ascites tumor and leukemic medicine, has DEVELOPMENT PROSPECT.
In sum, compare technical characterstic of the present invention with the method for existing traditional preparation process Lignum Sappan extracting solution: 1, adopt ultrafiltration to prepare Chinese medicine extraction liquid and can hold back the active ingredient in the Lignum Sappan to greatest extent and reduce loss of active ingredients.2, adopting the protein purification method can remove trace impurity in the solution, is that the effective ingredient of extract is more concentrated and effect is remarkable.
Description of drawings
Fig. 1 represents that variable concentrations Lignum Sappan liquid is to the K562 cytotoxicity
The specific embodiment
1 one kinds of Caesalpinia sappan water extract preparation methoies of embodiment may further comprise the steps:
(1), title Chinese medicine Lignum Sappan adds 5 times of waters for injection, decoction 40min, collection supernatant;
(2), repeat above-mentioned (1) step, merge supernatant, to the centrifugal 30min of supernatant 3500r, getting supernatant concentration again is 1g/ml to concentration;
(3), the centrifugal 30min of concentrated solution 3500r that obtains after last step concentrated, divide secondary to carry out ultrafiltration, the aperture: relative molecular weight is respectively 6k and 3k, collects filtrate, merges.This method is than water extract-alcohol precipitation, or Chinese medicine traditional extraction process such as alcohol extracting-water precipitating can keep effective ingredient morely.
(4), in the filtrate that merged, add 0.2% protein (Albumin), at 56 ℃, fully reaction under the 15min condition.Its main purpose is to eliminate residual trace impurity in the aqueous extract.
(5), ultrafiltration, the aperture: relative molecular weight is respectively 3k, collects filtrate.
(6), transfer pH6,100 ℃ of sterilizations obtain Caesalpinia sappan water extract.
(1), title Chinese medicine Lignum Sappan adds 4 times of waters for injection, decoction 30min, collection supernatant;
(2), repeat above-mentioned (1) step, merge supernatant, to the centrifugal 30min of supernatant 3500r, getting supernatant concentration again is 2g/ml to concentration;
(3), the centrifugal 30min of concentrated solution 3500r that obtains after last step concentrated, divide secondary to carry out ultrafiltration, the aperture: relative molecular weight is respectively 6k and 3k, collects filtrate, merges.This method is than water extract-alcohol precipitation, or Chinese medicine traditional extraction process such as alcohol extracting-water precipitating can keep effective ingredient morely.
(4), in the filtrate that merged, add 0.5% protein (Albumin), at 50 ℃, fully reaction under the 10min condition.Its main purpose is to eliminate residual trace impurity in the aqueous extract.
(5), ultrafiltration, the aperture: relative molecular weight is respectively 3k, collects filtrate.
(6), transfer pH6,100 ℃ of sterilizations obtain Caesalpinia sappan water extract.
(1), title Chinese medicine Lignum Sappan adds 7 times of waters for injection, decoction 60min, collection supernatant;
(2), repeat above-mentioned (1) step, merge supernatant, to the centrifugal 30min of supernatant 3500r, getting supernatant concentration again is 1.5g/ml to concentration;
(3), the centrifugal 30min of concentrated solution 3500r that obtains after last step concentrated, divide secondary to carry out ultrafiltration, the aperture: relative molecular weight is respectively 6k and 3k, collects filtrate, merges.This method is than water extract-alcohol precipitation, or Chinese medicine traditional extraction process such as alcohol extracting-water precipitating can keep effective ingredient morely.
(4), in the filtrate that merged, add 0.1% protein (Albumin), at 60 ℃, fully reaction under the 20min condition.Its main purpose is to eliminate residual trace impurity in the aqueous extract.
(5), ultrafiltration, the aperture: relative molecular weight is respectively 3k, collects filtrate.
(6), transfer pH6,100 ℃ of sterilizations obtain Caesalpinia sappan water extract.
Claims (3)
1. Caesalpinia sappan water extract preparation method is characterized in that: may further comprise the steps:
(1), title Chinese medicine Lignum Sappan adds 4-7 times of water, decoction 30-60min, collection supernatant;
(2), repeat above-mentioned (1) step, merge supernatant, centrifugal to supernatant, getting the supernatant concentration that obtains after centrifugal again is 1-2g/ml to concentration;
(3), the concentrated solution that obtains after last step concentrated is centrifugal, divide secondary to carry out ultrafiltration, the aperture: relative molecular weight is respectively 6k and 3k, collects filtrate, merges;
(4), in the filtrate that merged, add the albumin of filtrate quality 0.1-0.5%, at 50-60 ℃, react under the 10-20min condition;
(5), ultrafiltration, the aperture: relative molecular weight is respectively 3k, collects filtrate;
(6), adjust pH, the sterilization, obtain Caesalpinia sappan water extract.
2. Caesalpinia sappan water extract preparation method according to claim 1 is characterized in that: may further comprise the steps:
(1), title Chinese medicine Lignum Sappan adds 5 times of water, decoction 40min, collection supernatant;
(2), repeat above-mentioned (1) step, merge supernatant, centrifugal to supernatant, getting the supernatant concentration that obtains after centrifugal again is 1g/ml to concentration;
(3), the concentrated solution that obtains after last step concentrated is centrifugal, divide secondary to carry out ultrafiltration, the aperture: relative molecular weight is respectively 6k and 3k, collects filtrate, merges;
(4), in the filtrate that merged, add the albumin of filtrate quality 0.2%, at 56 ℃, react under the 15min condition;
(5), ultrafiltration, the aperture: relative molecular weight is respectively 3k, collects filtrate;
(6), adjust pH, the sterilization, obtain Caesalpinia sappan water extract.
3. Caesalpinia sappan water extract preparation method according to claim 1 and 2 is characterized in that: in step (2), to the centrifugal 30min of supernatant 3500r, to the centrifugal 30min of concentrated solution 3500r, transferring pH in step (6) is 6,100 ℃ of sterilizations in step (3).
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| CN101683380B (en) * | 2009-05-11 | 2011-08-24 | 山西省肿瘤医院 | Method for preparing irrigating solution with anti-tumor function |
| CN101904892B (en) * | 2010-05-22 | 2012-06-06 | 山西省肿瘤医院 | Preparation process for Sappan Wood extract perfusate and application thereof in treating bladder cancer |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1059847A (en) * | 1990-09-15 | 1992-04-01 | 安徽省亳州市神医制药厂 | Method for extracting traditional Chinese medicine components |
| CN1393250A (en) * | 2001-07-04 | 2003-01-29 | 周亚滨 | Aqueous extract of sapanwood for treating acute rejection reaction after organ transplantation |
| CN101019835A (en) * | 2007-03-09 | 2007-08-22 | 上海师范大学 | Lignum sappan and its prepn process and application |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1059847A (en) * | 1990-09-15 | 1992-04-01 | 安徽省亳州市神医制药厂 | Method for extracting traditional Chinese medicine components |
| CN1393250A (en) * | 2001-07-04 | 2003-01-29 | 周亚滨 | Aqueous extract of sapanwood for treating acute rejection reaction after organ transplantation |
| CN101019835A (en) * | 2007-03-09 | 2007-08-22 | 上海师范大学 | Lignum sappan and its prepn process and application |
Non-Patent Citations (4)
| Title |
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| 任连生等.苏木水提物抗癌作用机制的研究.《山西医药杂志》.2000,第29卷(第3期),201-203. * |
| 徐建国等.苏木提取液抑制肿瘤作用的研究.《肿瘤研究与临床》.2006,第18卷(第11期),726-727. * |
| 徐建国等.苏木煎液抗小鼠白血病作用的研究.《中国中药杂志》.1991,第16卷(第5期),306. * |
| 赵焕新等.苏木研究进展.《齐鲁药事》.2007,第26卷(第2期),102-105. * |
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