CN100574775C - A kind of Herba Rabdosiae Lophanthoidis extract and its production and application - Google Patents
A kind of Herba Rabdosiae Lophanthoidis extract and its production and application Download PDFInfo
- Publication number
- CN100574775C CN100574775C CN200710026843A CN200710026843A CN100574775C CN 100574775 C CN100574775 C CN 100574775C CN 200710026843 A CN200710026843 A CN 200710026843A CN 200710026843 A CN200710026843 A CN 200710026843A CN 100574775 C CN100574775 C CN 100574775C
- Authority
- CN
- China
- Prior art keywords
- extract
- herba rabdosiae
- rabdosiae lophanthoidis
- preparation
- ursolic
- 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.)
- Expired - Fee Related
Links
Images
Landscapes
- Medicines Containing Plant Substances (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention provides a kind of Herba Rabdosiae Lophanthoidis extract, it is characterized in that containing 1.58～2.63% oleanolic acid, 4.87～6.17% ursolic acids, 3.16～4.47%2 Alpha-hydroxy ursolic acids, 3.24～4.28%2 α, 19 alpha-dihydroxy-ursolic acids.This extract is prepared by following method: Herba Rabdosiae Lophanthoidis is soaked in the water, and the normal pressure heating extraction is passed through macroporous adsorbent resin with water extract then, uses ethanol elution, and the collection concentration of alcohol is 60～95% eluent, last concentrating under reduced pressure, spray drying.The present invention utilizes the effective ingredient in the purification with macroreticular resin technology extraction Herba Rabdosiae Lophanthoidis, the heat-sensitive ingredients oleanolic acid in the crude drug, ursolic acid, 2 and 2 α have been kept as much as possible, 19 alpha-dihydroxy-ursolic acids make extract keep the natural feature of Herba Rabdosiae Lophanthoidis well.
Description
Technical field
The present invention relates to the field of Chinese medicines, be specifically related to Chinese medicine extract, particularly Herba Rabdosiae Lophanthoidis extract.
Background technology
Herba Rabdosiae Lophanthoidis is the dry herb of Labiatae Rabdosia plant Rabdosia lophanthoides Isodon lophanthoides (Benthan) H.Hara and mutation isodon lophanthoides Isodon lophanthoides var.gerardianus (Benthan) H.Hara thereof, has clearing away heat-damp and promoting diuresis, jaundice eliminating, the effect of cool the blood dissipate blood stasis, medical herbs commonly used for China's southern area treatment hepatitis B, clinical practice is extensive, determined curative effect, existing more is the nourishing the liver hepatoprotective Chinese medicine preparation and the health product of primary raw material with it at present, as XIAOYAN LIDAN PIAN, the Herba Rabdosiae Lophanthoidis oral liquid, ten flavor Herba Rabdosiae Lophanthoidiss etc.But these kind ubiquity preparation process fall behind, and dose is big, take shortcomings such as inconvenience and effect be not ideal enough.At present, China's medical industry level of development is backward relatively, new technique and new technology are owing to reasons such as idea and objective condition are difficult to popularize, with the Chinese crude drug is the medicine of feedstock production treatment hepatitis B, it mainly is the active component that adopts in decocting cooking method and the decoction and alcohol sedimentation technique extraction raw material, then the medicinal liquid that obtains is directly used or is processed into required dosage form, medium and small polar fraction of the active component of crude drug such as oleanolic acid (Oleanic acid), ursolic acid (Ursolicacid), 2 (2 α-hydroxyl Ursolic acid), 2 α, 19 alpha-dihydroxy-ursolic acid (2 α, 19 α-dihydroxylUrsolic acid) etc. triterpenes components then be difficult to keep or yield on the low side, influenced the drug effect of Herba Rabdosiae Lophanthoidis extract hepatoprotective.
Summary of the invention
The purpose of this invention is to provide a kind of Herba Rabdosiae Lophanthoidis extract.
Another object of the present invention provides described preparation method of extract.
A further object of the present invention provides the application of described extract in the preparation hepatitis B medicament.
The present invention realizes that the technical scheme of above-mentioned purpose is: a kind of Herba Rabdosiae Lophanthoidis extract, it is characterized in that containing the following material of mass percent: oleanolic acid 1.58～2.63%, ursolic acid 4.87～6.17%, 2 3.16～4.47%, 2 α, 19 alpha-dihydroxy-ursolic acids 3.24～4.28%.
The preparation method of Herba Rabdosiae Lophanthoidis extract of the present invention is:
(1) get the Herba Rabdosiae Lophanthoidis segment, add 8～15 times in water at every turn, 100 ℃ of heating extraction are 1.0～1.5 hours under normal pressure, repeat to extract 2～3 times;
(2) aqueous extract is concentrated into every milliliter and contains crude drug 0.5～1.0g, then under 5～20 ℃, 5000～10000rpm ultracentrifugation is got supernatant, with macroporous adsorptive resins on the flow velocity of 0.5～3BV/h (times column volume per hour), wait to adsorb saturated after, with the flow velocity eluting of 5～10BV water with 0.5～3BV/h, then with the flow velocity eluting of 10%～95% ethanol with 0.5～3BV/h, the collection concentration of alcohol is 60～95% eluent earlier, concentrate, spray drying promptly.
Above-mentioned preparation method, wherein the concentration of ethanol elution is preferably 60～95%, and the best is 80%;
Above-mentioned preparation method, wherein ultracentrifugation speed the best is 8000rpm, and last column flow rate the best is 1.5BV/h, and elution flow rate the best is 1BV/h, and eluting is 80% with concentration of alcohol the best.
Extract of the present invention adopts the purification by macroporous resin technology to extract Herba Rabdosiae Lophanthoidis and obtains, this method does not need at high temperature to carry out for a long time, heat-sensitive ingredients in the Herba Rabdosiae Lophanthoidis in the active skull cap components such as oleanolic acid, ursolic acid, 2,2 α, 19 alpha-dihydroxy-ursolic acids are difficult for decomposing destruction, have kept the natural feature of Herba Rabdosiae Lophanthoidis well.
Extract of the present invention can be used for preparing the medicine for the treatment of hepatitis B, and this medicine can be enteric coated capsule, soft capsule, drop pill, dispersible tablet etc.
Described enteric coated capsule is made up of extract of the present invention and HPMC, micropowder silica gel and magnesium stearate.Its preparation method is, with Herba Rabdosiae Lophanthoidis extract and excipient HPMC and the micropowder silica gel mixed by 1: 1.5: 0.5, granulates earlier, and drying adds 0.2% the abundant mix homogeneously of magnesium stearate lubricant again, and the filling enteric capsule shell is made.
Described soft capsule system is made up of extract of the present invention and an amount of vegetable oil, 3%～5% Cera Flava and a small amount of wetting agent.Its preparation method is, earlier with the mixture of extract of the present invention and the adjuvant mixed by 1: 2.5, goes up the Zhanang machine again and suppresses and form.
Described drop pill is mixed with PEG4000 and PEG60000 by extract of the present invention and forms.Its preparation method is, earlier extract of the present invention and PEG4000 and PEG6000 pressed 1: 0.5: 2.5 mixed, and fusion again splashes in the coolant and makes.
Described dispersible tablet is mixed with HPMC, lactose and starch by extract of the present invention and forms.Its preparation method is earlier extract of the present invention and HPMC, lactose and starch to be pressed 1: 0.7: 0.5: 1 mixed, granulate, and drying, granulate adds 1% magnesium stearate mix homogeneously again, goes up the tablet machine compacting again and forms.
Extract of the present invention can be identified by the following method:
1, high performance liquid chromatography:
(1) chromatographic condition: Kromasil RP-C
18Chromatographic column (4.6mm * 250mm, 5 μ m); 0.1% acetic acid-acetonitrile-methanol (10: 5: 85); Flow velocity: 0.8ml/min; 25 ℃ of column temperatures; Detect wavelength: 210nm; Sample size: 5 μ l; Theoretical cam curve is by 2 α, and 19 alpha-dihydroxy-ursolic acids, 2, oleanolic acid, ursolic acid calculate and all should be not less than 5000.
(2) preparation of standard solution: precision takes by weighing 2 α, 19 alpha-dihydroxy-ursolic acids, 2, oleanolic acid, ursolic acid reference substance are an amount of, add dissolve with methanol and standardize solution, making concentration respectively is 2 α, 19 alpha-dihydroxy-ursolic acid 0.24mg/ml, 2 0.20mg/ml, oleanolic acid 0.16mg/ml, the solution of ursolic acid 0.28mg/ml, standby.Accurate each 1ml of above-mentioned standard solution that draws puts in the 5ml volumetric flask, and is fully mixed, makes the standard mixed solution, standby.
(3) above-mentioned standard mixed solution 1,2,4,6,8,10, the 20 μ l of the accurate respectively absorption of the standard curve and the range of linearity thereof, measure according to selected chromatographic condition sample introduction, the chromatogram of standard mixed solution is seen Fig. 1,2 α, the retention time of 19 alpha-dihydroxy-ursolic acids, 2, oleanolic acid, ursolic acid is respectively 13.08min, 14.19min, 18.35min, 19.31min; Carry out linear regression with each composition quality of peak area (Y) (X), regression equation is respectively: 2 α, 19 alpha-dihydroxy-ursolic acid Y=425.038X-32.4, γ=0.9998, the range of linearity 48～960ng; 2 Y=352.057X-19.9, γ=0.9999, the range of linearity 40～800ng; Oleanolic acid Y=513.2X+11.715, γ=0.9996, the range of linearity 32～640ng; Ursolic acid Y=764.5X+14.724, γ=0.9998, the range of linearity 56～1120ng; γ is a linear coefficient.
(4) the sample determination precision takes by weighing each 100mg of extract sample of the present invention, adds methanol constant volume to 5ml, ultrasonicly makes its dissolving, before the sample introduction with 0.20 μ m filtering with microporous membrane.The accurate 10 μ l that draw measure according to selected chromatographic condition sample introduction, according to the quality X of each composition of regression equation calculation of standard mixed solution gained.
Below further specify beneficial effect of the present invention by concrete experiment:
1, extract of the present invention is to carbon tetrachloride (CCl
4) due to the protective effect of rat acute hepatic injury
60 rats are divided into normal group, model group, high, medium and low dosed administration group, 6 groups of positive controls at random.The administration group is pressed 200mgkg
-1, 100mgkg
-1, 50mgkg
-1Dosage is irritated stomach, every day 1 time, continuous 10 days with the extract of following preparation example 1 gained; Positive controls is pressed 45mgkg
-1Bifendate, every day 1 time, continuous 10 days.Administration the 1st, 4,7,10 days (totally 4 times), model group and administration group are pressed 2mLkg
-1Lumbar injection 10%CCl
4Modeling.The 11st day, behind the last administration 24h, get blood, measure serum glutamic pyruvic transminase (ALT), glutamic oxaloacetic transaminase, GOT (AST), alkali phosphatase (ALP) and total bilirubin (T-Bil), survey simultaneously that body weight, liver are heavy, spleen is heavy and thymus is heavy, ask organ coefficient, and get hepatic tissue and make pathology and observe.
From table 1 and 2 as can be known, Herba Rabdosiae Lophanthoidis can significantly reduce the Rats with Acute Liver Injury Serum ALT, AST, and ALP level and T-Bil content have inhibitory action to increase of liver weight and atrophy of thymus gland.
Table 1 extract of the present invention is to CCl
4Due to the Rats with Acute Liver Injury Serum ALT, AST, ALP, the influence of T-Bil (x ± s, n=10)
| Group | Dosage/mgkg -1 | ALT/U·L -1 | AST/U·L -1 | ALP/U·L -1 | T-Bil/μmol·L -1 |
| Normal control | - | 35.72±5.58 | 149.78±13.81 | 176.80±49.54 | 1.302±0.308 |
| Model | - | 136.92±71.22 2) | 364.15±135.01 2) | 245.86±32.35 1) | 2.000±0.277 2) |
| High dose administration group | 200 | 76.32±27.26 | 254.27±97.14 | 185.36± 49.213 3) | 1.701±0.338 |
| Middle dosed administration group | 100 | 75.12±21.52 3) | 216.68±97.06 3) | 184.63±53.41 3) | 1.656±0.195 3) |
| Low dosage administration group | 50 | 66.50±24.66 3) | 237.61±58.90 3) | 184.45±39.42 3) | 1.644±0.192 4) |
| Positive controls | 45 | 56.21±13.83 3) | 226.09±88.84 3) | 208.13±48.54 | 1.561±0.258 4) |
Annotate: compare with the normal control group
1)P＜0.05,
2)P＜0.01; Compare with model group
3)P＜0.05,
4)P＜0.01.(table 2 together)
This extract of table 2 (ILVG) is to CCl
4Due to the Rats with Acute Liver Injury organ coefficient influence (x ± s, n=10)
| Group | Dosage/mgkg -1 | Liver coefficient/g100g -1 | Spleen coefficient/mgg -1 | Thymus coefficient/mgg -1 |
| Normal control | - | 3.613±0.312 | 2.529±0.293 | 1.685±0.432 |
| Model | - | 4.164±0.359 2) | 2.297±0.711 | 1.049±0.475 2) |
| High dose administration group | 200 | 3.627±0.313 4) | 2.584±0.487 | 1.525±0.344 3) |
| Middle dosed administration group | 100 | 3.639±0.316 4) | 2.164±0.215 | 1.695±0.734 3) |
| Low dosage administration group | 50 | 3.795±0.177 3) | 2.534±0.691 | 1.518±0.272 3) |
| Positive controls | 45 | 3.929±0.560 | 2.424±0.326 | 1.260±0.244 |
2, extract of the present invention is to the protective effect of rat acute hepatic injury due to the D-galactosamine
60 rats are divided into normal control group, model group, high, normal, basic dosed administration group, 6 groups of positive controls at random.The administration group is pressed 200mgkg
-1, 100mgkg
-1, 50mgkg
-1Dosage is with following preparation example 2 extract obtained filling stomaches; Positive controls is pressed 45mgkg
-1Bifendate is irritated stomach; Totally 7 days.In the 6th day afternoon of gastric infusion, model group and administration group are pressed 550mgkg
-1Body weight lumbar injection D-Gal once.The 11st day, behind the last administration 24h (lumbar injection 36h), measure the serum biochemistry index, and survey body weight and liver is heavy, spleen is heavy, thymus is heavy, ask organ coefficient, cut liver simultaneously and detect hepatic glycogen, get hepatic tissue and make pathology and observe.The results are shown in Table 3,4 and 5.
This extract of table 3 is to the influence of Rats with Acute Liver Injury Serum ALT, AST, ALP, TBA and T-Bil due to the D-Gal
| Group | Dosage (mgk g -1) | ALT (u·L -1) | AST (u·L -1) | ALP (u·L -1) | TBA (mol·L -1) | T-Bil (μmol·L -1) |
| The normal control group | - | 45.47± 6.59 | 185.05± 14.42 | 239.04± 36.66 | 18.26± 13.84 | 1.098± 0.352 |
| Model group | - | 1823.91± 755.95## | 2238.57± 1409.91## | 371.53± 126.21# | 237.75± 63.77## | 18.066± 9.759## |
| High dose administration group | 200 | 557.26± 971.98** | 542.68± 443.57** | 248.72± 90.10* | 96.59± 79.27** | 6.762± 6.460** |
| Middle dosed administration group | 100 | 445.83± 563.17** | 570.93± 544.82* | 257.38± 119.02 | 73.75± 39.55** | 6.834± 6.904* |
| Low dosage administration group | 20 | 326.94± 358.34** | 578.01± 429.82** | 255.68± 81.25* | 83.81± 91.62** | 6.111± 7.740** |
| Positive controls | 45 | 204.09± 203.70** | 443.45± 329.83** | 247.74± 45.01* | 105.35± 99.61** | 7.091± 9.174* |
Annotate: compare with the normal control group: #P＜0.05 ##P＜0.01; Compare with model group: * P＜0.05 * * P＜0.01 (table 4,5 together)
This extract of table 4 (ILVG) is to the influence of Rats with Acute Liver Injury serum T P, ALB, A/G and hepatic glycogen due to the D-Gal
| Group | Dosage (mgkg -1) | TP (g·L -1) | ALB (g·L -1) | A (g) | Hepatic glycogen (mgg -1) |
| The normal control group | - | 65.89±2.85 | 36.08±1.24 | 1.214±0.073 | 14.38±14.3 |
| Model group | - | 53.90±3.49## | 32.61±2.80## | 1.532± 0.120## | 3.18±2.40# |
| High dose administration group | 200 | 62.17±5.85* | 36.04±1.68** | 1.459±0.210 | 9.07±5.85* |
| Middle dosed administration group | 100 | 60.86±4.55* | 35.26±1.89* | 1.428±0.158 | 7.54±4.66* |
| Low dosage administration group | 20 | 61.85± 4.50* | 35.12±1.48* | 1.455±0.208 | 9.47±7.71* |
| Positive controls | 45 | 61.42±3.97** | 35.32± 1.32* | 1.431±0.197 | 6.90±6.35 |
This extract of table 5 (ILVG) is to the influence of Rats with Acute Liver Injury organ coefficient due to the D-Gal
| Group | Dosage (mgkg -1) | Liver coefficient (%) | Spleen coefficient (‰) | Thymus coefficient (‰) |
| Normal control | - | 3.263±0.497 | 2.627±0.347 | 2.038±0.331 |
| Model | - | 3.988±0.390## | 3.090±0.406# | 1.515±0.284## |
| High dose administration group | 200 | 3.667±0.204 | 2.933±0.656 | 1.814±0.280* |
| Middle dosed administration group | 100 | 3.642±0.163* | 3.057±0.424 | 1.834±0.286* |
| Low dosage administration group | 50 | 3.566±0.316* | 3.021±0.416 | 1.819±0.165* |
| Positive controls | 45 | 3.861±0.449 | 2.859±0.438 | 1.612±0.302 |
More than the table result shows, extract of the present invention can significantly reduce Rats with Acute Liver Injury Serum ALT, AST, ALP level and TBA, T-Bil content, the increase of hepatic glycogen in TP, ALB and the hepatic tissue has inhibitory action to increase of liver weight and atrophy of thymus gland in the promotion serum.
3, extract of the present invention (ILVG) is to the protective effect of canavaline (ConA) induced mice immunity hepatic injury
Adopt mouse tail vein injection canavaline (ConA) to cause liver injury model, 72 of NIH mices are divided into 6 groups at random, i.e. normal control group, model group, high, medium and low dosed administration group, and the bifendate positive controls, 12 every group, the administration group is pressed 240mgkg
-1, 120mgkg
-1, 60mgkg
-1Dosage is with following preparation example 3 extract obtained gastric infusions; Positive controls oral bifendate 45mgkg
-1, every day 1 time, continuous 5 days.4h model group and the disposable tail vein injection ConA of administration treated animal 20mg/kg after the last administration.Fasting be can't help plucking behind the water 8h eyeball and is got blood and put to death, and separation of serum is measured glutamate pyruvate transaminase (ALT), glutamic oxaloacetic transaminase, GOT (AST) in the serum, surveys body weight simultaneously, liver is heavy, spleen is heavy and thymus is heavy, asks organ coefficient.Fix with 4% formalin solution with position liver (leftlobe of liver), and get hepatic tissue and do the pathology observation.The results are shown in Table 6,7.
This extract of table 6 (ILVG) is to immunologic liver injury due to the canavaline (ConA)
Mice serum ALT, and the influence of AST (x ± s, n=12)
| Group | Dosage/mgkg -1 | ALT/U·L -1 | AST/U·L -1 |
| Normal control | - | 24.31±8.42 | 21.49±9.61 |
| Model | - | 220.28±53.82 2) | 156.94±48.68 2) |
| High dose administration group | 60 | 179.17±46.80 | 118.95±41.87 |
| Middle dosed administration group | 120 | 110.26±59.58 3) | 91.53±49.49 3) |
| Low dosage administration group | 240 | 56.21±42.15 3) | 39.74±25.43 3) |
| Positive controls | 45 | 133.88±49.58 3) | 91.62±65.97 3) |
Annotate: compare with the normal control group
1)P＜0.05,
2)P＜0.01; Compare with model group
3)P＜0.05,
4)P＜0.01 (table 7 together)
The result of table 6 shows that all apparently higher than normal group (P＜0.05, or P＜0.01), the high, medium and low dosage group of extract of the present invention can significantly reduce ALT value (P＜0.05) for model group rat blood serum ALT, AST, in, low dose group can significantly reduce ALT, AST value.
This extract of table 7 (ILVG) to the influence of hepatic injury mice organ coefficient due to the canavaline (ConA) (x ± s, n=12)
| Group | Dosage/mgkg -1 | Liver coefficient/mg10g -1 | Spleen coefficient/mg10g -1 | Thymus coefficient/mg10g -1 |
| Normal control | - | 511.91±41.72 | 40.48±6.54 | 37.25±10.26 |
| Model | - | 593.94±33.45 2) | 94.46±16.48 2) | 24.28±6.40 1) |
| High dose administration group | 60 | 568.65±38.41 | 81.95±15.46 | 28.85±5.54 3) |
| Middle dosed administration group | 120 | 560.06±36.51 3) | 78.99±17.70 3) | 29.66±4.74 4) |
| Low dosage administration group | 240 | 538.92±57.5 4) | 69.51±19.12 4) | 31.50±4.26 4) |
| Positive controls | 45 | 565.35±36.31 | 79.73±16.15 3) | 26.40±4.07 |
Table 7 is the result show, model group Mouse Liver coefficient is apparently higher than normal group (P＜0.01), and the thymus coefficient is starkly lower than normal group (P＜0.01), illustrates that the model group liver increases, atrophy of thymus gland.The high, medium and low dosage group of extract of the present invention all has the increase of the liver of inhibition, atrophy of thymus gland effect, significant difference (P＜0.05, or P＜0.01) is arranged, and the bifendate group is not seen this effect.
4, pathological observation
The liver tissue lesions classification with reference to " gastroenterology and hepatopathy magazine " (Lang Zhenwei waits the classifications of .CHB patient's difference, the expression study [J] of TGF-β 1 in the hepatic tissue by stages for Wang Xinxin, Sun Lin. gastroenterology and hepatopathy magazine, 2004,13 (3): 291.).It is that the center is radial arrangement with the central vein that light microscopic is observed the normal group hepatocyte down, the sinus hepaticus no abnormality seen.Obvious pathological changes all appears in the whole murine liver tissue of model group, most of swelling of liver cell in the lobule, and the Cytoplasm puffing, the balloon sample that is that has becomes, visible tangible white spotty necrosis and focal necrosis, the visible massive inflammatory cells infiltrated of necrosis region; Portal area lymphocyte and mononuclear cell inflammatory infiltration are particularly evident, and the visible red cell is heaped in the sinus hepaticus.As seen extract group of the present invention, bifendate group part hepatic tissue are dispersed in spotty necrosis and focal necrosis, and the mouse liver injury degree all obviously alleviates than model group, and inflammatory cell infiltration significantly reduces.Illustrate that extract of the present invention has the certain protection effect to rat acute hepatic injury due to the canavaline (Con A).Adopt the Kruskal-Wallis method to test, the pathological change difference of hepatic necrosis, inflammatory cell infiltration has significance meaning (extract group: He=34.84,28.33 between demonstration group as a result, P＜0.01), adopt the Nemenyi method to compare in twos, the result shows that the hepatic necrosis of extract group of the present invention, bifendate group, inflammatory cell infiltration pathological change all are starkly lower than model group, difference has significance meaning (P＜0.05 or P＜0.01), sees Table 8.
Table 8 Herba Rabdosiae Lophanthoidis causes the influence of mouse liver injury pathological change to Con A
Description of drawings
Fig. 1 is the high-efficient liquid phase chromatogram of standard mixed solution, and wherein a is 2 α, 19 alpha-dihydroxy-ursolic acids, and b is a 2, and c is an oleanolic acid, and d is a ursolic acid.
Fig. 2 is the extract obtained high-efficient liquid phase chromatogram of embodiment 1, and wherein a is 2 α, 19 alpha-dihydroxy-ursolic acids, and b is a 2, and c is an oleanolic acid, and d is a ursolic acid.
The specific embodiment
Preparation example
Example 1
Get the Herba Rabdosiae Lophanthoidis 2000g of fresh dried, cut into 1～2cm segment.
Get the Herba Rabdosiae Lophanthoidis segment, add 12 times in water and soaked 12 hours, in 100 ℃ of following heating extraction 1.5 hours, filtered while hot, medicinal residues add 10 times in water again, heating extraction 1 hour, filtered while hot.Merge ethanol extract twice; Aqueous extract is concentrated into every milliliter contains crude drug 0.90g, under 5～20 ℃ with the 5000rpm ultracentrifugation, get supernatant and be added on the AB-8 type macroporous adsorbent resin of having handled well (production of Tianjin Chemical Plant of Nankai Univ.), with macroporous adsorptive resins on the flow velocity of 2BV/h (times column volume per hour), wait to adsorb saturated after, earlier with the flow velocity eluting of 6BV water with 1.5BV/h, again successively with 5BV 60% ethanol, 10BV80% ethanol flow velocity eluting with 1BV/h, collection concentration is 80% ethanol elution, concentrate spray drying.
Precision takes by weighing said extracted matter sample 100mg, adds methanol constant volume to 5ml, ultrasonicly makes its dissolving, with 0.20 μ m filtering with microporous membrane.The accurate 10 μ l that draw carry out high-performance liquid chromatogram determination, chromatographic condition: Kromasil RP-C
18Chromatographic column (4.6mm * 250mm, 5 μ m); 0.1% acetic acid-acetonitrile-methanol (10: 5: 85); Flow velocity: 0.8ml/min; 25 ℃ of column temperatures; Detect wavelength: 210nm; Sample size: 5 μ l; Theoretical cam curve is not less than 5000.The results are shown in Figure 2, at Rt is that 13.08min, 14.19min, 18.35min, 19.31min place have four peaks, reference standard mixed solution chromatogram (Fig. 1), these four peaks are respectively 2 α as can be known, 19 alpha-dihydroxy-ursolic acids, 2, oleanolic acid, ursolic acid, each composition of regression equation calculation according to standard mixed solution gained is 2 α, 19 alpha-dihydroxy-ursolic acids 4.07%, 2 3.68%, oleanolic acid 2.63%, ursolic acid 5.62%.
Example 2
Get the Herba Rabdosiae Lophanthoidis 1000g of fresh dried, cut into 1～2cm segment.
Get the Herba Rabdosiae Lophanthoidis segment, add 15 times in water and soaked 12 hours, in 100 ℃ of following heating extraction 1.0 hours, filtered while hot, medicinal residues add 10 times in water again, heating extraction 0.5 hour, filtered while hot.Merge ethanol extract twice; Aqueous extract is concentrated into every milliliter contains crude drug 0.74g, under 5～20 ℃ with the 8000rpm ultracentrifugation, get supernatant and be added on the AB-8 type macroporous adsorbent resin of having handled well (production of Tianjin Chemical Plant of Nankai Univ.), with macroporous adsorptive resins on the flow velocity of 3BV/h (times column volume per hour), wait to adsorb saturated after, earlier with the flow velocity eluting of 6BV water with 0.8BV/h, again successively with 3BV 60% ethanol, 6BV80% ethanol flow velocity eluting with 1BV/h, collection concentration is 80% ethanol elution, concentrate spray drying; Contain 3.24% 2 α in extract obtained, 19 alpha-dihydroxy-ursolic acids, 4.47% 2,1.58% oleanolic acid and 5.34% ursolic acid.
Example 3
Get the Herba Rabdosiae Lophanthoidis 2000g of fresh dried, cut into 1～2cm segment.
Get the Herba Rabdosiae Lophanthoidis segment, add 12 times in water and soaked 12 hours, in 100 ℃ of following heating extraction 1.5 hours, filtered while hot, medicinal residues add 10 times in water again, heating extraction 1 hour, filtered while hot.Merge ethanol extract twice; Aqueous extract is concentrated into every milliliter contains crude drug 0.90g, under 5～20 ℃ with the 5000rpm ultracentrifugation, get supernatant and be added on the AB-8 type macroporous adsorbent resin of having handled well (production of Tianjin Chemical Plant of Nankai Univ.), with macroporous adsorptive resins on the flow velocity of 2BV/h (times column volume per hour), wait to adsorb saturated after, earlier with the flow velocity eluting of 6BV water with 1.5BV/h, again successively with 5BV 60% ethanol, 8BV80% ethanol flow velocity eluting with 1BV/h, collection concentration is 80% ethanol elution, concentrate spray drying; Contain 3.81% 2 α in extract obtained, 19 alpha-dihydroxy-ursolic acids, 3.16% 2,2.19% oleanolic acid and 4.87% ursolic acid.
Example 4
Get the Herba Rabdosiae Lophanthoidis 2000g of fresh dried, cut into 1～2cm segment.
Get the Herba Rabdosiae Lophanthoidis segment, add 15 times in water and soaked 12 hours, in 100 ℃ of following heating extraction 1.5 hours, filtered while hot, medicinal residues add 10 times in water again, heating extraction 1 hour, filtered while hot.Merge ethanol extract twice; Aqueous extract is concentrated into every milliliter contains crude drug 0.88g, under 5～20 ℃ with the 8000rpm ultracentrifugation, get supernatant and be added on the AB-8 type macroporous adsorbent resin of having handled well (production of Tianjin Chemical Plant of Nankai Univ.), with macroporous adsorptive resins on the flow velocity of 2BV/h (times column volume per hour), wait to adsorb saturated after, earlier with the flow velocity eluting of 8BV water with 1.2BV/h, again successively with 3BV 60% ethanol, 8BV80% ethanol flow velocity eluting with 1BV/h, collection concentration is 80% ethanol elution, concentrate spray drying; Contain 4.28% 2 α in extract obtained, 1% alpha-dihydroxy-ursolic acid, 3.53% 2,1.98% oleanolic acid and 5.28% ursolic acid.
Example 5
Get the Herba Rabdosiae Lophanthoidis 3000g of fresh dried, cut into 1～2cm segment.
Get the Herba Rabdosiae Lophanthoidis segment, add 15 times in water and soaked 12 hours, in 100 ℃ of following heating extraction 1 hour, filtered while hot, medicinal residues add 10 times in water again, heating extraction 1 hour, filtered while hot.Merge ethanol extract twice; Aqueous extract is concentrated into every milliliter contains crude drug 0.85g, under 5～20 ℃ with the 8000rpm ultracentrifugation, get supernatant and be added on the AB-8 type macroporous adsorbent resin of having handled well (production of Tianjin Chemical Plant of Nankai Univ.), with macroporous adsorptive resins on the flow velocity of 2BV/h (times column volume per hour), wait to adsorb saturated after, earlier with the flow velocity eluting of 6BV water with 1.2BV/h, again successively with 4BV 60% ethanol, 8BV80% ethanol flow velocity eluting with 1BV/h, collection concentration is 80% ethanol elution, concentrate spray drying; Contain 4.16% 2 α in extract obtained, 19 alpha-dihydroxy-ursolic acids, 4.38% 2,2.54% oleanolic acid and 6.17% ursolic acid.
Application examples
Example 1
With preparation example 1 extract obtained 82.6g with an amount of ethanol dilution after, progressively increase according to equivalent that respectively excipient HPMC and micropowder silica gel are even by 1: 1.5: 0.5 mixed for method, cross No. 6 sieve series and become granule, under 60 ℃ of temperature dry 3 hours.Get 0
#Jejunum colloidal sol capsule.Fill above dried particles respectively, per 1 capsules 0.5g is coated with one deck mucialga of arabic gummy in the Nang Kou place, and cover sealing capsule behind dry gauze wiping capsule, is loaded in the airtight brown bottle, promptly.The consumption of said preparation is every day twice, and is each 4, oral.
Example 2
1 extract obtained 72.2g mixes with 120g Oleum Arachidis hypogaeae semen with preparation example, adds 10g Cera Flava and 1.25g tween 80, and the rotary Zhanang of reuse machine is pressed into 500 soft capsules (0.5g/ grain).The consumption of said preparation is every day twice, and is each 4, oral.
Example 3
2 extract obtained 41.3g mix with soybean oil 63g with preparation example, add 5.0g Cera Flava and 0.8g tween 80, and the rotary Zhanang of reuse machine is pressed into 300 soft capsules (0.5g/ grain).The consumption of said preparation is every day twice, and is each 4, oral.
Example 4
With preparation example 3 extract obtained 78.6g with an amount of ethanol dilution after again with starch 200g mixing, cross 120 mesh sieves, in 60 ℃ of oven dry 3 hours down, encapsulated (0.5g/ grain).The consumption of said preparation is every day twice, and is each 4, oral.
Example 5
2 extract 83.1g mix with 40g PEG4000 and 200g PEG6000 with preparation example, and fusion splashes into and promptly gets drop pill (50mg/ grain) in the coolant methyl-silicone oil.The consumption of said preparation is every day twice, and each 3.0g is oral.
Example 6
3 extract 98.7g mix with 60g PEG4000 and 300g PEG6000 with preparation example, and fusion splashes into and promptly gets drop pill (50mg/ grain) in the coolant methyl-silicone oil.The consumption of said preparation is every day twice, and each 3.0g is oral.
Example 7
4 extract 110.6g mix with 45g PEG4000 and 200g PEG6000 with preparation example, and fusion splashes into and promptly gets drop pill (50mg/ grain) in the coolant methyl-silicone oil.The consumption of said preparation is every day twice, and each 3.0g is oral.
Example 8
With the beta-cyclodextrin inclusion compound of preparation example 3 extracts and 1.5 times of amounts, add 60g starch, mix, after 80 mesh sieves 2 times, put in the blender, the adding distilled water is an amount of, stirs the system soft material 10 minutes, cross 16 mesh sieves and granulate, wet grain is dry in 60 ℃ of air dry ovens, and dry granular is crossed 16 mesh sieve granulate.Add carboxymethyl starch sodium 10g and magnesium stearate 2g, mix homogeneously, tabletting, the bag film-coat gets tablet.The consumption of said preparation is every day three times, and is each 2, oral.
Example 9
With the beta-cyclodextrin inclusion compound of preparation example 2 extracts and 1.5 times of amounts, add 80g starch, mix, after 80 mesh sieves 2 times, put in the blender, the adding distilled water is an amount of, stirs the system soft material 10 minutes, cross 16 mesh sieves and granulate, wet grain is dry in 60 ℃ of air dry ovens, and dry granular is crossed 16 mesh sieve granulate.Add carboxymethyl starch sodium 12g and magnesium stearate 2g, mix homogeneously, tabletting, the bag film-coat gets tablet.The consumption of said preparation is every day three times, and is each 2, oral.
Example 10
With the beta-cyclodextrin inclusion compound of preparation example 5 extracts and 1.5 times of amounts, add 100g starch, mix, after 80 mesh sieves 2 times, put in the blender, the adding distilled water is an amount of, stirs the system soft material 10 minutes, cross 16 mesh sieves and granulate, wet grain is dry in 60 ℃ of air dry ovens, and dry granular is crossed 16 mesh sieve granulate.Add carboxymethyl starch sodium 15g and magnesium stearate 2g, mix homogeneously, tabletting, the bag film-coat gets tablet.The consumption of said preparation is every day three times, and is each 2, oral.
Example 11
With the beta-cyclodextrin inclusion compound of preparation example 1 extract with 2 times of amounts, add 60g starch again, mix, after 80 mesh sieves 2 times, put in the blender, the adding distilled water is an amount of, stirs 10 minutes, the system soft material, cross 16 mesh sieves and granulate, wet grain is dry in 60 ℃ of air dry ovens, and dry granular is crossed 16 mesh sieve granulate, divide packing, promptly get granule.The consumption of said preparation is every day 3 times, each 1 bag (2g), and warm water is taken after mixing it with water.
Example 12
Preparation example 4 extracts and HPMC, lactose and starch were pressed 1: 0.7: 0.5: 1 mixed is even, granulate with 95% ethanol, 20 orders, dry 1 hour, 16 mesh sieve granulate add 1% magnesium stearate mix homogeneously again below 50 ℃, and compacting heavily is the tablet of 0.4g in flakes.Promptly get dispersible tablet.The consumption of said preparation is every day 2 times, and each 3, warm water is taken after mixing it with water.
Example 13
Preparation example 5 extracts and HPMC, lactose and starch were pressed 1: 0.9: 0.9: 1.2 mixed is even, granulate with 95% ethanol, 20 orders, dry 1 hour, 16 mesh sieve granulate add 1% magnesium stearate mix homogeneously again below 50 ℃, and compacting heavily is the tablet of 0.4g in flakes.Promptly get dispersible tablet.The consumption of said preparation is every day 2 times, and each 3, warm water is taken after mixing it with water.
Claims (9)
1, a kind of Herba Rabdosiae Lophanthoidis extract, it is characterized in that containing the following material of mass percent: oleanolic acid 1.58～2.63%, ursolic acid 4.87～6.17%, 2 3.16～4.47%, 2 α, 19 alpha-dihydroxy-ursolic acids 3.24～4.28%.
2, the preparation method of the described Herba Rabdosiae Lophanthoidis extract of claim 1, this preparation method is made up of following steps:
(1) get the Herba Rabdosiae Lophanthoidis segment, add 8～15 times in water at every turn, 100 ℃ of heating extraction are 1.0～1.5 hours under normal pressure, repeat to extract 2～3 times;
(2) aqueous extract is concentrated into every milliliter and contains crude drug 0.5～1.0g, with 5～20 ℃, 5000～10000rpm ultracentrifugation, get supernatant with macroporous adsorptive resins on the flow velocity of 0.5～3BV/h, wait to adsorb saturated after, with the flow velocity eluting of 5～10BV water with 0.5～3BV/h, then with the flow velocity eluting of 10%～95% ethanol with 0.5～3BV/h, the collection concentration of alcohol is 60～95% eluent earlier, concentrating under reduced pressure, spray drying.
3, according to the preparation method of the described Herba Rabdosiae Lophanthoidis extract of claim 2, the concentration of alcohol that it is characterized in that described eluent is 60～95%.
4, according to the preparation method of the described Herba Rabdosiae Lophanthoidis extract of claim 2, the concentration of alcohol that it is characterized in that described eluent is 80%.
5,, it is characterized in that described ultracentrifugation speed is 8000rpm according to the preparation method of the described Herba Rabdosiae Lophanthoidis extract of claim 2.
6,, it is characterized in that described upward column flow rate is 1.5BV/h according to the preparation method of the described Herba Rabdosiae Lophanthoidis extract of claim 2.
7,, it is characterized in that described eluting is 80% with concentration of alcohol the best according to the preparation method of the described Herba Rabdosiae Lophanthoidis extract of claim 2.
8, the application of the described Herba Rabdosiae Lophanthoidis extract of claim 1 in the medicine of preparation treatment hepatitis B.
9, medicine according to claim 8 is enteric coated capsule, soft capsule, drop pill or dispersible tablet.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200710026843A CN100574775C (en) | 2007-02-08 | 2007-02-08 | A kind of Herba Rabdosiae Lophanthoidis extract and its production and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200710026843A CN100574775C (en) | 2007-02-08 | 2007-02-08 | A kind of Herba Rabdosiae Lophanthoidis extract and its production and application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101011461A CN101011461A (en) | 2007-08-08 |
| CN100574775C true CN100574775C (en) | 2009-12-30 |
Family
ID=38699296
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN200710026843A Expired - Fee Related CN100574775C (en) | 2007-02-08 | 2007-02-08 | A kind of Herba Rabdosiae Lophanthoidis extract and its production and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN100574775C (en) |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103417620B (en) * | 2013-07-09 | 2015-02-11 | 赵晨阳 | Application of serrate rabdosia herb extract serving as STAT3 signal specific inhibiter in preparation of anti-tumor drug |
| CN105106222B (en) * | 2015-09-14 | 2020-05-08 | 新乡医学院 | Application of emetic acid in preparing medicament for treating or preventing Alzheimer disease caused by estrogen deficiency |
| CN106046076B (en) * | 2016-06-24 | 2018-11-16 | 广州白云山和记黄埔中药有限公司 | A method of preparing thin flower Rabdosia lophanthoides glycosides A and NSC 619661 simultaneously from Rabdosia plant |
| CN109395430A (en) * | 2018-11-21 | 2019-03-01 | 神威药业集团有限公司 | A kind of extracting method suitable for Traditional Chinese medicine water-soluble heat-sensitive ingredients |
| CN109331053A (en) * | 2018-11-21 | 2019-02-15 | 神威药业集团有限公司 | A kind of extracting method suitable for Traditional Chinese medicine water-soluble heat-sensitive ingredients |
-
2007
- 2007-02-08 CN CN200710026843A patent/CN100574775C/en not_active Expired - Fee Related
Non-Patent Citations (2)
| Title |
|---|
| 狭基线纹香茶菜（溪黄草）的化学成分与抗乙肝病毒作用研究. 胡英杰等.中草药，第36卷第11期. 2005 |
| 狭基线纹香茶菜(溪黄草)的化学成分与抗乙肝病毒作用研究. 胡英杰等.中草药，第36卷第11期. 2005 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101011461A (en) | 2007-08-08 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN112274620B (en) | Traditional Chinese medicine composition for treating novel coronavirus pneumonia, preparation method, detection method and application thereof | |
| CN100574775C (en) | A kind of Herba Rabdosiae Lophanthoidis extract and its production and application | |
| CN101396384A (en) | Asiatic centella extract and preparation methode thereof | |
| CN1985976A (en) | Abortion preventing Yimu preparation and its new preparing process | |
| CN101361812A (en) | Preparation method of rhubarb glycyrrhiza preparation | |
| CN1977889A (en) | Medicinal composition of astragalus, salvia miltrorrhiza and oxymatrine, and its preparing method | |
| WO2013155995A1 (en) | Red yeast and kudzu root pharmaceutical composition for regulating blood lipids and preparation method therefor | |
| CN102920964B (en) | Traditional Chinese medicine preparation for curing cough | |
| CN100384429C (en) | Capsule for treating cardiac and cerebral vascular diseases and its preparing method and application | |
| CN1985927A (en) | Medicine composition for treating hepatic disease mainly | |
| CN106266949A (en) | A kind of compound Chinese medicinal preparation and its preparation method and application | |
| CN101785799B (en) | Extract of total triterpenes in dracocephalum moldavica and preparation method and application thereof | |
| CN106309729B (en) | A kind of Chinese medicine composition adjusting chronic hepatitis immune function | |
| CN102370677A (en) | Three-flavor sandalwood preparation and preparation method thereof | |
| CN101549075B (en) | Traditional Chinese medicine composition for treating mammary gland hyperplasia and preparation method thereof | |
| CN101385846A (en) | Preparation method of Jiawei Xiaoyao power preparation | |
| CN1969963A (en) | Method for extracting medicament effective component for treating hepatitis and medicinal composition thereof | |
| CN101422522B (en) | Preparation method of traditional Chinese medicine preparation | |
| CN102406897B (en) | Xingnaojing solid medicinal composition and preparation method thereof | |
| CN101884700A (en) | Tea-containing Chinese medicinal composition | |
| CN1331522C (en) | Red tangerine peel medicine for treating sputum cough and its preparation method | |
| CN102309543A (en) | Compound salvia miltiorrhiza concentrated preparation and preparation and detection method thereof | |
| CN104189296A (en) | Traditional Chinese medicine oral solution for treating jaundice of pigs and preparation method thereof | |
| CN1977888B (en) | Medicinal composition of baicalin, ganoderma lucidum and salvia miltrorrhiza | |
| CN100358554C (en) | Chinese medicinal capsules for disinfecting and relieving toxicity and their preparation |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20091230 Termination date: 20210208 |