CN1450074A - Process for extracting coptis total alkaloid and new use thereof - Google Patents
Process for extracting coptis total alkaloid and new use thereof Download PDFInfo
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- CN1450074A CN1450074A CN 02103880 CN02103880A CN1450074A CN 1450074 A CN1450074 A CN 1450074A CN 02103880 CN02103880 CN 02103880 CN 02103880 A CN02103880 A CN 02103880A CN 1450074 A CN1450074 A CN 1450074A
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Abstract
The process for extracting total alkaloid of coptis root includes the following steps: soaking coptis root in water and making ultrasonic wave treatment, under the condition of soaking in warm water making extraction of total alkaloid, reduced pressure concentrating obtained extract, then using concentrated hydrochloric acid to regulate pH value of concentrated extract, and adding sodium chloride and standing still, then utilizing suction filtration or centrifugal drying to collect the precipitate in the extract, drying precipitate, pulverizing dried precipitate so as to obtain the invented coptis total alkaloid hydrochloride finished product. Said invention also discloses new application of coptis total alkaloid in preparation of medicine with the action of reducing blood sugar and enhancing sensitivity of insulin.
Description
Technical field
The present invention relates to a kind of extraction process and new purposes thereof of Chinese medicine, particularly relate to a kind of extraction process and new purposes thereof of Rhizoma Coptidis total alkaloids.
Background technology
At present, coptis medicinal material is used to extract Rhizoma Coptidis total alkaloids, and the complete processing of employing is a reflux extraction, as ethanol water refluxing extraction or water refluxing extraction.Substantially comprise immersion, refluxing extraction, extracting solution is concentrated into the medicinal extract shape, directly oven dry pulverizing or elder generation form the alkaloid crystal of hydrochloride and dry processes such as pulverizing then, traditional extracting solution concentrate drying is that extracting solution is concentrated into dried cream must contain Rhizoma Coptidis total alkaloids then through drying and crushing finished product, perhaps extracting solution being concentrated into the medicinal extract shape drips concentrated hydrochloric acid then or is dissolved in and forms the alkaloid salt hydrochlorate in the dilute hydrochloric acid of small amount of thermal, form crystallization in the process of cooling, collect crystalline deposit thing and oven dry by filtering, this type of complete processing gained finished product is impure more, need to improve the purity of finished product by recrystallization, yet recrystallization causes the loss of Rhizoma Coptidis total alkaloids again inevitably, use ethanol to use solvent in addition as extracting, production cost is bigger, and need the problem of consideration equipment and secure context, also do not have simple and easy effective Rhizoma Coptidis total alkaloids extracting method at present.Diabetes have become one of principal disease of harm health of people.Over nearly 50 years, worldwide, the morbidity of diabetes B increases sharply.In the diabetes B pathogenesis, body is an important factor to insulin resistant.Phase late 1980s, after famous ' insulin resistance syndrome ' notion proposes, promoted the process of the research and the control of diabetes greatly.Dependency to diseases such as diabetes, cardiovascular and ephrosis has also had deep understanding simultaneously.Therefore, surplus in the of nearly 10 year over, insulin resistant and insulin resistance syndrome, the medical science forward position focus of always being paid close attention to.Meanwhile, the world of medicine's exploitation and use euglycemic agent improve diabetes B people's insulin resistant state, correct unusual carbohydrate metabolism, blood sugar is descended, thereby alleviate the toxicity of hyperglycemia to B cell, help the recovery of its function.
Technology contents
It is a kind of direct, simple and easy, efficient that the object of the invention is to provide, cheaply coptis chinensis total alkaloid extracting technique; The object of the invention is to provide a kind of Rhizoma Coptidis total alkaloids to have new purposes in the medicine of insulin-sensitizing effect in preparation; Another object of the present invention is to provide the new purposes of a kind of Rhizoma Coptidis total alkaloids in the medicine of preparation prevention and treatment diabetes.
The present invention seeks to be achieved through the following technical solutions: the coptis is broken earlier, the water logging bubble also carries out ultrasonication simultaneously then, water-soaking time is 30～240 minutes, make the water content of the coptis reach 100%～300%, the coptis behind the water logging bubble is soaked in temperature and carries out the total alkaloids extraction under the condition, extracting temperature is 70 ℃～98 ℃, the solvent water yield is 5～16 times of raw medicinal herbs weight during extraction, extraction time is 30～180 minutes, extraction time is 1～4 time, the gained extracting solution carries out concentrating under reduced pressure, vacuum tightness when concentrating is 0.04～0.08MPa, the volume of the water of concentrated extracting solution to 2～10 times raw medicinal herbs weight, regulate pH value to 1～5 of concentrated extracting solution then with concentrated hydrochloric acid, adding sodium-chlor again dissolves while hot, the sodium-chlor add-on is the weight of the water of 4%～16% concentration extraction liquid measure, concentrated extracting solution left standstill 4～24 hours, then by the throw out in suction filtration or the centrifuge dripping collection extracting solution, throw out is dried under 50 ℃～80 ℃ temperature, and dried throw out is pulverized can obtain Rhizoma Coptidis total alkaloids hydrochloride finished product.
Extraction process of the present invention is fit to the extraction of Rhizoma Coptidis total alkaloids, technology of the present invention has been used ultrasonication in immersion process, extraction process adopts temperature immersion formulation, extracting solution carries out appropriate concentrating under reduced pressure, it is to precipitate by forming the alkaloid salt hydrochlorate and form crystallization under salting out that Rhizoma Coptidis total alkaloids is separated from extracting solution, the crystalline deposit thing is collected with the method for suction filtration or centrifuge dripping, described ultrasonication is located in the water logging bubble process, it directly influences coptis medicinal material and soaks into speed and degree, and then influence the extraction effect of total alkaloids, adopt temperature immersion extraction method can simplify production unit and production operation greatly, the temperature immersion is extracted and also can be avoided impurity too much in the coptis to be brought in the extracting solution in addition, extracting solution is carried out appropriateness to be concentrated with the hydrochloric acid adjust pH again and adds sodium-chlor with the solubleness of further reduction Rhizoma Coptidis alkaloid hydrochloride in extracting solution, the few Rhizoma Coptidis total alkaloids content height of gained throw out impurity, and shorten the spissated time of extracting solution, concentration time shortens and can reduce in the Rhizoma Coptidis alkaloid some composition because of the long degree of decomposing of being heated, adopt suction filtration or centrifuge dripping to remove from throw out very effectively and be dissolved with the debris of sodium-chlor, thereby guaranteed the purity and the quality of Rhizoma Coptidis total alkaloids extract.Rhizoma Coptidis total alkaloids is identified: the thin layer chromatography qualitative analysis
With thin layer chromatography the throw out sample is carried out qualitative analysis, select silica gel g thin-layer plate for use, with benzene-ethyl acetate-methyl alcohol-Virahol-strong ammonia solution is developping agent, the ratio of test solution is 6: 3: 1.5: 1.5: 0.5, comprise berberine hydrochloride with reference substance in the experiment, Jatrorrhizine chloride and palmatine hydrochloride, solvent is a methyl alcohol, the methanol solution that is dissolved with throw out sample and contrast crystalline substance drops on the thin layer plate respectively, first balance 15 minutes in strong aqua, in developping agent, launch then, launch to finish the back and take out airing, inspect under the ultraviolet lamp of 365nm, it is reference that the result shows with the reference substance, berberine hydrochloride in the throw out, Jatrorrhizine chloride and palmatine hydrochloride all exist.The high performance liquid phase quantitative analysis:
With high-efficient liquid phase technique the throw out sample is carried out qualitative and quantitative analysis, the SODIUM PHOSPHATE, MONOBASIC of selecting acetonitrile and 0.02mol/L for use be organic and inorganic flow mutually, the ratio of two-phase is 36: 64, flow rate of mobile phase is 0.85ml/min, sample size is 20 μ l, comprise berberine hydrochloride with the contrast crystalline substance in the experiment, Jatrorrhizine chloride and palmatine hydrochloride, selecting deionized water for use is the solvent of throw out sample and reference substance, throw out sample feeding concentration is set in 0.05mg/ml, detect wavelength set at 345nm, with reference substance high performance liquid phase collection of illustrative plates is contrast, useful technology of institute is extracted berberine hydrochloride in the gained throw out, Jatrorrhizine chloride and palmatine hydrochloride all exist, make the standard regression line with known sample introduction concentration of reference substance and corresponding high performance liquid phase collection of illustrative plates absorption peak area, one cover extraction process orthogonal test gained throw out sample is carried out the high performance liquid phase atlas analysis, calculate that content of berberine hydrochloride reaches 55%～74% in the throw out, about root alkali of hydrochloric acid and palmatine hydrochloride content reach 5.3%～6.8% and 5.5%～8.5% respectively, see Table 1 and table 2.Table 1 reference substance and throw out sample feeding concentration and high performance liquid chromatography absorption peak area
The calculated value of three kinds of hydrochloric acid alkaloid salt concentration and percentage composition in the table 2. throw out sample
| Numbering | Berberine hydrochloride | Jatrorrhizine chloride | Palmatine hydrochloride | |||
| Concentration mg/ml | Peak area | Concentration mg/ml | Peak area | Concentration mg/ml | Peak area | |
| Reference substance | ||||||
| 1 | ????0.04 | ???51.25082 | ???0.000612 | ???1.11604 | ???0.00196 | ???2.256885 |
| 2 | ????0.015 | ???17.49666 | ???0.00102 | ???1.30146 | ???0.002548 | ???2.58995 |
| 3 | ????0.02 | ???21.82086 | ???0.00153 | ???2.041665 | ???0.00294 | ???2.925785 |
| 4 | ????0.03 | ???34.59527 | ???0.00204 | ???2.53755 | ???0.00098 | ???1.144535 |
| 5 | ????0.035 | ???39.44524 | ???0.00255 | ???3.296285 | ???0.001568 | ???1.668731 |
| The throw out sample | ||||||
| 1??(0.04992mg/ml) | ??36.085 | ???3.5323 | ???3.1647 | |||
| 2??(0.04849mg/ml) | ??34.079 | ???3.2031 | ???3.1271 | |||
| 3??(0.04954mg/ml) | ??38.203 | ???3.8564 | ???3.6603 | |||
| 4??(0.04954mg/ml) | ??31.697 | ???3.3796 | ???2.7697 | |||
| 5??(0.04992mg/ml) | ??37.109 | ???3.3891 | ???3.4157 | |||
| 6??(0.04946mg/ml) | ??38.001 | ???3.8148 | ???3.6852 | |||
| 7??(0.04949mg/ml) | ??40.433 | ???3.8272 | ???3.7006 | |||
| 8??(0.04947mg/ml) | ??32.838 | ???3.5409 | ???3.1457 | |||
| 9??(0.04946mg/ml) | ??44.410 | ???4.1484 | ???4.1273 | |||
| Numbering | Berberine hydrochloride | Jatrorrhizine chloride | Palmatine hydrochloride | ||||||
| Peak area | Concentration mg/ml | Percentage composition % | Peak area | Concentration mg/ml | Percentage composition % | Peak area | Concentration mg/ml | Percentage composition % | |
| ??1 | ??36.085 | ??0.0304 | ???60.9 | ??3.5323 | ??0.0028 | ????5.7 | ??3.1647 | ??0.0032 | ????6.3 |
| ??2 | ??34.079 | ??0.0289 | ???59.6 | ??3.2031 | ??0.0026 | ????5.3 | ??3.1271 | ??0.0031 | ????6.4 |
| ??3 | ??38.203 | ??0.0321 | ???64.8 | ??3.8564 | ??0.0031 | ????6.3 | ??3.6603 | ??0.0037 | ????7.5 |
| ??4 | ??31.697 | ??0.0271 | ???54.7 | ??3.3796 | ??0.0027 | ????5.5 | ??2.7697 | ??0.0027 | ????5.5 |
| ??5 | ??37.109 | ??0.0312 | ???62.5 | ??3.3891 | ??0.0027 | ????5.4 | ??3.4157 | ??0.0034 | ????6.9 |
| ??6 | ??38.001 | ??0.0319 | ???64.5 | ??3.8148 | ??0.0031 | ????6.2 | ??3.6852 | ??0.0037 | ????7.5 |
| ??7 | ??40.433 | ??0.0338 | ???68.3 | ??3.8272 | ??0.0031 | ????6.2 | ??3.7006 | ??0.0037 | ????7.6 |
| ??8 | ??32.838 | ??0.0279 | ???56.4 | ??3.5409 | ??0.0028 | ????5.7 | ??3.1457 | ??0.0031 | ????6.3 |
| ??9 | ??44.410 | ??0.0368 | ???74.4 | ??4.1484 | ??0.0034 | ????6.8 | ??4.1273 | ??0.0042 | ????8.5 |
Experimental example 1, to the influence of tetraoxypyrimidine (Alloxan) hyperglycemia mouse blood sugar
The formation of alloxan hyperglycemia mouse: kunming mice, male, body weight 23-25g, tail vein injection alloxan 100mg/Kg.Predict blood sugar (glucose oxidase method) behind the 72h.Select the above person of 11.1mmol/L for use, be divided into 6 groups, be respectively control group, total alkali 25,50,100,200mg/kg and known hypoglycemic agents N1,N1-Dimethylbiguanide 200mg/kg group.Every day, gastric infusion was 1 time, in administration the 7th day, got blood and surveyed blood sugar.
Total alkali has stronger reduction effect to the blood sugar of Alloxan hyperglycemia mouse, and dose-effect relationship is preferably arranged, and the effect of total alkali 200mg/kg group obviously is better than N1,N1-Dimethylbiguanide 200mg/kg group, sees table 3 and Fig. 1 for details.
Table 3. total alkali is to the influence of Alloxan hyperglycemia mouse blood sugar
Experimental example 2. total alkalis are to the insulin tolerance of insulin resistant MSG mouse and the influence of glucose tolerance:
| Group | Dosage (mg/kg) | Blood sugar (mg/dl) | Blood sugar decline % |
| Control group | ????- | ????448.2±38.0 | |
| ????25 | ????409.8±93.8 | ?????8.6 | |
| Total alkali | ????50 | ????371.2±92.1 * | ?????17.2 |
| ????100 | ????328.7±94.1 ** | ?????26.7 | |
| ????200 | ????219.4±81.0 ** | ?????51.0 | |
| N1,N1-Dimethylbiguanide | ????200 | ????304.2±89.3 ** | ?????32.1 |
The formation of MSG mouse: newborn ICR mouse: be born and rose in 1st: subcutaneous injection every day L-Sodium Glutamate (MSG) 4g/Kg (by body weight), continuous 7 days, control group injection equivalent physiological saline.Ablactation after the 21st day, MSG group and control group are all by the male and female sub-cage rearing.Because MSG damage hypothalamus brings out hyperglycemia, hyperinsulinemia and the insulin resistant of animal.The MSG mouse is divided into 3 groups: control group, total alkali 100mg/kg and known euglycemic agent rosiglitazone 2.The 5mg/kg group.Every day, gastric infusion was 1 time, carried out the insulin tolerance experiment on the 7th day in administration: carried out the glucose tolerance experiment on the 12nd day in administration.
Table 4 and Fig. 2 explanation, total alkali has obviously improved the reactivity of insulin resistant MSG mouse to Regular Insulin, and subcutaneous injection Regular Insulin 0.4IU/kg measures 0,40 and 90 minute blood sugar, and area (AUC) all is starkly lower than contrast under the calculating blood glucose curve
Table 4. total alkali is to the influence of the plain tolerance of insulin resistant MSG mouse islets
Compare with control group,
*P＜0.05
| Group | Dosage (mg/kg) | Body weight (g) | Blood sugar (mg/dl) | ????????AUC ?????(mg/dl.hr) | ||
| ???????0｀ | ???????40｀ | ??????90｀ | ||||
| Contrast | ???- | ???52.1±7.9 | ???159.1±71.9 | ???129.5±69.5 | ???137.0±60.9 | ???207.2±73.9 |
| Rosiglitazone | ???2.5 | ???51.8±5.9 | ???176.8±116.2 | ???98.6±45.6 | ???127.0±42.9 | ???185.8±79.8 |
| Total alkali | ???100 | ???47.7±7.4 | ???117.0±47.5 | ???66.3±26.9 * | ???92.3±55.3 | ???127.2±40.1 * |
Table 5 and Fig. 3 show, total alkali has obviously improved the experiment of insulin resistant MSG mouse glucose tolerance, and oral administration gavage glucose 2g/kg measures that area (AUC) all is starkly lower than control group under 0,30 and 120 minute blood sugar and the blood glucose curve.
Table 5. total alkali is to the influence of insulin resistant MSG mouse sugar tolerance
Compare with control group,
*P＜0.05
| Group | Dosage (mg/kg) | Body weight (g) | Blood sugar (mg/dl) | ???????AUC ????(mg/dl.hr) | ||
| ????????0｀ | ???????30｀ | ???????120｀ | ||||
| Control group | ????- | ??52.0±7.9 | ????111.9±24.8 | ???120.5±91.7 | ??127.9±32.4 | ???324.4±119.6 |
| Rosiglitazone | ???2.5 | ??51.5±5.9 | ????118.7±58.7 | ???188.1±86.3 | ??129.2±50.6 | ???314.7±132.9 |
| Total alkali | ???100 | ??44.6±8.4 | ????85.0±20.0 * | ???119.2±38.3 * | ??84.7±20.3 ** | ???204.0±53.9 * |
Conclusion:
Above-mentioned experimental result explanation, total alkali can obviously reduce the blood sugar of hyperglycemia mouse, is dose-effect relationship, and to the mouse of insulin resistance, total alkali can obviously improve its unusual Regular Insulin and sugar tolerance: the characteristics with euglycemic agent.In addition, finding in the experiment in the past that the euglycemic agent pioglitazone that has gone on the market increased the weight of animals, clinical also have a similar results, is considered to side effect, and total alkali does not have the body weight of increasing effect.
Description of drawings:
Fig. 1 total alkali is to the influence of Alloxan hyperglycemia mouse blood sugar;
Fig. 2 total alkali is to the influence of the plain tolerance of insulin resistant MSG mouse islets;
Fig. 3 total alkali is to the influence of MSG mouse glucose tolerance.
Embodiment:
When steeping, the broken water logging of the coptis carries out the 20kHz ultrasonication, water-soaking time is 120 minutes, make the water content of the coptis reach 230%, the coptis behind the water logging bubble is soaked in temperature and carries out the total alkaloids extraction under the condition, extracting temperature is 82 ℃, the solvent water yield is 10 times of raw medicinal herbs weight during extraction, extraction time is 60 minutes, extraction time is 2 times, the gained extracting solution carries out concentrating under reduced pressure, vacuum tightness when concentrating is 0.04～0.08MPa, the volume of the water of concentrated extracting solution to 3 times raw medicinal herbs weight: the pH value to 2 of regulating concentrated extracting solution then with concentrated hydrochloric acid, add sodium-chlor again, the sodium-chlor add-on is the weight of the water of 14% concentration extraction liquid measure, and concentrated extracting solution left standstill 12 hours, then by the throw out in suction filtration or the centrifuge dripping collection extracting solution, throw out is dried under 60 ℃ temperature, and dried throw out is pulverized can obtain Rhizoma Coptidis total alkaloids hydrochloride finished product
Claims (5)
1, the extraction process of Rhizoma Coptidis total alkaloids, it is characterized in that this technology is: ultrasonication is also carried out in coptis elder generation fragmentation water logging bubble then simultaneously, water-soaking time is 30～240 minutes, make the water content of the coptis reach 100%～300%, the coptis behind the water logging bubble is soaked in temperature and carries out the total alkaloids extraction under the condition, extracting temperature is 70 ℃～98 ℃, the solvent water yield is 5～16 times of raw medicinal herbs weight during extraction, extraction time is 30～180 minutes, extraction time is 1～4 time, the gained extracting solution carries out concentrating under reduced pressure, vacuum tightness when concentrating is 0.04～0.08MPa, the volume of the water of concentrated extracting solution to 2～10 times raw medicinal herbs weight, regulate pH value to 1～5 of concentrated extracting solution then with concentrated hydrochloric acid, adding sodium-chlor again dissolves while hot, the sodium-chlor add-on is the weight of the water of 4%～16% concentration extraction liquid measure, concentrated extracting solution left standstill 4～24 hours, then by the throw out in suction filtration or the centrifuge dripping collection extracting solution, throw out is dried under 50 ℃～80 ℃ temperature, and dried throw out is pulverized can obtain Rhizoma Coptidis total alkaloids hydrochloride finished product.
2, the extraction process of Rhizoma Coptidis total alkaloids according to claim 1, it is characterized in that this technology is: carry out the 20kHz ultrasonication when the broken water logging of the coptis is steeped, water-soaking time is 120 minutes, make the water content of the coptis reach 230%, the coptis behind the water logging bubble is soaked in temperature and carries out the total alkaloids extraction under the condition, extracting temperature is 82 ℃, the solvent water yield is 10 times of raw medicinal herbs weight during extraction, extraction time is 60 minutes, extraction time is 2 times, the gained extracting solution carries out concentrating under reduced pressure, vacuum tightness when concentrating is 0.04～0.08MPa, the volume of the water of concentrated extracting solution to 3 times raw medicinal herbs weight: regulate the pH value to 2 of concentrated extracting solution then with concentrated hydrochloric acid, add sodium-chlor again, the sodium-chlor add-on is the weight of the water of 14% concentration extraction liquid measure, concentrated extracting solution left standstill 12 hours, by the throw out in suction filtration or the centrifuge dripping collection extracting solution, throw out is dried under 60 ℃ temperature then, and dried throw out is pulverized can obtain Rhizoma Coptidis total alkaloids hydrochloride finished product
3, the application of Rhizoma Coptidis total alkaloids in the preparation hypoglycemic drug.
4, Rhizoma Coptidis total alkaloids has application in the medicine of insulin-sensitizing effect in preparation.
5, Rhizoma Coptidis total alkaloids is in prevention and the application for the treatment of in the diabetes B.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1321995C (en) * | 2005-08-02 | 2007-06-20 | 山东轻工业学院 | Total coptis alkaloid preparation method |
| WO2007068143A1 (en) * | 2005-12-13 | 2007-06-21 | Lanqin Yang | Preparation of plant oil, crude protein and fiber by water extraction |
| CN101108214B (en) * | 2007-07-20 | 2010-05-19 | 湖南师范大学 | Method of separating and extracting natural base from coptis chinensis with latex membrane |
| CN101874831A (en) * | 2009-04-30 | 2010-11-03 | 成都中医药大学 | New application of coptis product processed by ginger juice |
| CN110613755A (en) * | 2019-09-26 | 2019-12-27 | 吉林省中药制剂工程研究中心有限公司 | Preparation method of rhizoma coptidis total alkaloids and application of rhizoma coptidis total alkaloids in treatment of diabetes |
-
2002
- 2002-04-05 CN CN 02103880 patent/CN1223601C/en not_active Expired - Fee Related
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1321995C (en) * | 2005-08-02 | 2007-06-20 | 山东轻工业学院 | Total coptis alkaloid preparation method |
| WO2007068143A1 (en) * | 2005-12-13 | 2007-06-21 | Lanqin Yang | Preparation of plant oil, crude protein and fiber by water extraction |
| CN101108214B (en) * | 2007-07-20 | 2010-05-19 | 湖南师范大学 | Method of separating and extracting natural base from coptis chinensis with latex membrane |
| CN101874831A (en) * | 2009-04-30 | 2010-11-03 | 成都中医药大学 | New application of coptis product processed by ginger juice |
| CN110613755A (en) * | 2019-09-26 | 2019-12-27 | 吉林省中药制剂工程研究中心有限公司 | Preparation method of rhizoma coptidis total alkaloids and application of rhizoma coptidis total alkaloids in treatment of diabetes |
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| CN1223601C (en) | 2005-10-19 |
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