CN1304025C - Active part and effective component of Four Drug Decoction - Google Patents
Active part and effective component of Four Drug Decoction Download PDFInfo
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- CN1304025C CN1304025C CN 03123521 CN03123521A CN1304025C CN 1304025 C CN1304025 C CN 1304025C CN 03123521 CN03123521 CN 03123521 CN 03123521 A CN03123521 A CN 03123521A CN 1304025 C CN1304025 C CN 1304025C
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Abstract
The present invention discloses an active site of the decoction of four medicines, which is characterized in that the active site is composed of a part B and a part C. The present invention has the active site that medicinal materials of the decoction of four medicines are decocted for one to three times, and are filtered to obtain water decocted liquid; the water decocted liquid is concentrated in a rotary evaporation mode in the temperature which is lower than 50 to 60 DEG C., and is precipitated via 20% of ethanol to obtain precipitate; supernatant liquid is go on to be precipitated via 40% of ethanol to obtain precipitate; supernatant liquid is go on to be precipitated via 60% of ethanol to obtain precipitate; 725 ml of supernatant liquid is go on to be precipitated via 80% of ethanol to obtain precipitate. Supernatant liquid is concentrated into extract without the scent of ethanol; the extract is dissolved in ethyl acetate, and is extracted via the ethyl acetate; ethyl acetate liquid is merged, and a solvent is recovered to obtain extract; the residual liquid is extracted via butanol; butanol liquid is merged, and a solvent is recovered to obtain extract B; the residual liquid is concentrated until the residual liquid foams, and extract C is obtained.
Description
Invention field
The present invention relates to a kind of Chinese patent medicine active site, particularly relate to SIWU TANG active site and effective ingredient.
Background technology
The material base that SIWU TANG plays a role is its active chemical, due to the population effect that its effect of preventing and curing diseases is its number of chemical composition.But it is very weak to the SIWU TANG The Chemical Constituents at present, to fundamentally study the composition of prescription rule of SIWU TANG compound recipe, disclose, explain the scientific meaning of SIWU TANG composition principle from cell, molecular level, the material base research of compatibility Study on Mechanism and SIWU TANG compound recipe generation curative effect must be combined closely from theoretical level.But, SIWU TANG is extremely complicated as its effective ingredient of Chinese medicine compound, even its each single medicine of distinguishing the flavor of all can be used as a little compound recipe, therefore, we have at first carried out the separation of chemical position according to the size of polar difference and molecular weight to SIWU TANG decocting liquid, and carry out activity with indexs such as peripheral hemogram, the cultivations of bone marrow hematogenesis CFU-GM colony and follow the trail of; The methods such as efficient thin layer chromatography (HPTLC) and high performance liquid chromatography (HPLC) technology of using are simultaneously carried out preliminary analysis to the composition at each position and are identified.
Summary of the invention
The object of the present invention is to provide the SIWU TANG active site.
SIWU TANG active site of the present invention is characterized in that being made up of B and C two parts, and described effective site is to be decocted 1-3 time by the SIWU TANG medical material, filters, and gets decocting liquid; Decocting liquid concentrates in 20-70 ℃ of rotary evaporation, use the 15-25% ethanol precipitation, must precipitate, supernatant is proceeded the 35-45% ethanol precipitation, must precipitate, supernatant is proceeded the 55-65% ethanol precipitation, must precipitate supernatant and proceed the 75-85% ethanol precipitation, must precipitate, supernatant concentration to do not have ethanol distinguish the flavor of extractum, add dissolving and carry out ethyl acetate extraction, combined ethyl acetate liquid, reclaim solvent and get extract, remaining water liquid carries out the butanols extraction approximately, merges n-butyl alcohol liquid, reclaims solvent and gets extract B, remaining water liquid is concentrated into foaming approximately and can not continues, and gets extract C.
The present invention discloses a kind of SIWU TANG active site, and its feature is characterized in that mainly containing paeoniflorin 9.7%, fructose 16.5%, glucose 10.1%, sucrose 22.3%.The preparation method of this effective site is: the SIWU TANG medical material decocts 1-3 time, filters, and gets decocting liquid; Decocting liquid concentrates in 20-70 ℃ of rotary evaporation, use the 15-25% ethanol precipitation, must precipitate, supernatant is proceeded the 35-45% ethanol precipitation, must precipitate, supernatant is proceeded the 55-65% ethanol precipitation, must precipitate supernatant and proceed the 75-85% ethanol precipitation, must precipitate, supernatant concentration to do not have ethanol distinguish the flavor of extractum, add dissolving and carry out ethyl acetate extraction, combined ethyl acetate liquid reclaims solvent and gets extract, and remaining water liquid carries out the butanols extraction approximately, merge n-butyl alcohol liquid, reclaiming solvent must this active site.
Press practice of pharmacy, preparation of pharmaceutical compositions of the present invention can be become the various clinical pharmaceutical dosage form, comprise the dosage form of oral formulations or parenterai administration.Said oral formulations is selected from a kind of in the middle of the tablet, capsule, pill, granule, suspensoid, drop pill, oral liquid; Said parenterai administration dosage form is selected from a kind of in the middle of injection, aerosol, suppository or the subcutaneous administration dosage form.Medicine of the present invention also can add conventional drug excipient, as solvent, disintegrating agent, correctives, antiseptic, coloring agent etc.Paeoniflorin has blood tonification effect.
Indexs such as all hemogram, bone marrow hematogenesis CFU-GM colony cultivation are carried out activity tracking beyond the chemical position that following experimental example has separated with SIWU TANG decocting liquid; The methods such as efficient thin layer chromatography (HPTLC) and high performance liquid chromatography (HPLC) technology of using are simultaneously carried out preliminary analysis to the composition of active site and are identified.
Following experimental example further specifies the present invention.
The separation of experimental example 1, SIWU TANG active component
For the first time separate at the position: SIWU TANG through decocting, filtration, concentrate, be mixed with 100% medicinal liquid (being that every ml medicinal liquid contains crude drug 1g), progressively add ethanol then to 20%-80%, get 20% pure hypostasis (A), 80% pure hypostasis (B), 80% precipitate with ethanol supernatant extract (C) respectively.Carry out preliminary activity analysis (seeing Table 1) with A, B, C three parts.
Table 1 SIWU TANG and for the first time the position separator to the influence of syndrome of deficiency of blood mice peripheral blood leucocyte (x ± s)
| According to the back time | Number of animals | Group | ||||
| Matched group | The SIWU TANG group | The A group | The B group | The C group | ||
| 0 3 5 7 10 13 | 10 10 10 10 10 10 | 13.5±5.0 1.81±0.2 2.38±1.0 Δ 3.94±1.0 3.21±0.5 Δ 3.62±1.1 | 13.4±1.7 1.74±0.3 3.49±1.0* 4.40±2.0 4.29±1.3* 4.16±0.5 | 15.1±3.9 1.62±0.2 2.91±1.3 5.17±1.8 3.49±0.9 3.45±0.2 Δ | 15.4±2.9 2.04±0.4 3.29±0.9* 3.97±1.6 4.54±1.8* 4.51±1.3 | 14.0±1.9 1.69±0.3 3.06±1.0 5.55±1.5* 4.15±0.9* 3.80±0.6 |
Annotate: * and model group compare, * P＜0.05, * * P＜0.01;
ΔCompare with the SIWU TANG group,
ΔP＜0.05,
The Δ ΔP＜0.01.
The activity of results suggest B, C two parts and SIWU TANG quite after, carrying out second time position separates: SIWU TANG through decocting, filtration, concentrate, be mixed with 100% medicinal liquid (being that every ml medicinal liquid contains crude drug 1g), progressively add ethanol 20-80%, obtain 20-40%, 40-60%, 60-80% alcohol hypostasis (B1), (B2), (B3) respectively; The precipitate with ethanol supernatant concentration is removed ethanol, carries through ethyl acetate, n-butyl alcohol collection successively then, and it is (C1), (C2) that the recovery solvent gets extract, and the surplus water partial concentration gets extract and is (C3), further carries out activity rating.
Table 2 SIWU TANG and B position separator thereof to the influence of syndrome of deficiency of blood mice peripheral blood leucocyte (x ± s, n=10)
| The irradiation back time (my god) | Group | ||||
| Model group | The SIWU TANG group | The B1 group | The B2 group | The B3 group | |
| 0 3 5 7 10 13 | 14.22±3.0 1.28±0.2 2.18±0.3 ΔΔ 4.52±0.6 ΔΔ 3.90±0.8 Δ 3.70±0.8 Δ | 14.59±2.5 1.42±0.4 2.78±0.4** 6.42±1.3** 5.24±1.5* 4.74±1.2* | 15.61±2.6 1.12±0.3 2.56±0.8 4.62±1.5 Δ 3.68±1.1 Δ 4.08±1.0 | 13.37±2.1 1.10±0.2 Δ 2.44±0.6 4.96±0.9 ΔΔ 4.40±1.2 4.10±1.1 | 14.66±1.9 1.75±0.4** 2.58±0.5* 4.34±0.8 ΔΔ 3.94±0.8 Δ 3.84±1.4 |
Annotate: compare with model group,
*P＜0.05,
*P＜0.01;
ΔCompare with the SIWU TANG group,
ΔP＜0.05,
The Δ ΔP＜0.01
Table 3 SIWU TANG and C position separator thereof to the influence of syndrome of deficiency of blood mice peripheral blood leucocyte (x ± s, n=10)
| The irradiation back time (my god) | The SIWU TANG group | ||||
| Model group | Group | The C1 group | The C2 group | The C3 group | |
| 0 3 5 7 10 13 | 14.22±3.0 1.28±0.2 2.18±0.3 ΔΔ 4.52±0.6 ΔΔ 3.90±0.8 Δ 3.70±0.8 Δ | 14.59±2.5 1.42±0.4 2.78±0.4** 6.42±1.3** 5.24±1.5* 4.74±1.2* | 13.49±3.1 1.29±0.3 2.32±0.5 Δ 5.22±1.0 Δ 3.67±1.2 Δ 4.26±1.1 | 15.37±3.9 1.51±0.4 2.54±0.4* 5.46±1.2* 3.70±1.8 5.10±1.1** | 13.86±1.6 1.29±0.2 3.00±0.4** 5.72±1.4* 4.38±1.3 4.74±1.1* |
Annotate: compare with model group,
*P＜0.05,
*P＜0.01;
ΔCompare with the SIWU TANG group,
ΔP＜0.05,
The Δ ΔP＜0.01
Experimental result shows, is that index is carried out the activity tracking to the SIWU TANG extract with peripheral blood leucocyte, bone marrow hematogenesis CFU-GM, points out 80% pure hypostasis (B) and 80% precipitate with ethanol supernatant (C) activity stronger; Active chase experiment prompting to the two further separation and Extraction thing: B3, C2, C3 position activity are stronger relatively, leukocyte count in the radiation damage syndrome of deficiency of blood mice peripheral blood that all can raise.But the blood tonification effect of B1, B2 and B3 is all not as good as B.
The influence that table 4 SIWU TANG and extract thereof are bred syndrome of deficiency of blood mice hemopoietic progenitor cell (x ± s, n=10)
| Group | CFU-GM (individual/10 5/MNC) | CFU-E (individual/10 5/MNC) | BFU-E (individual/10 5/MNC) | CFU-mix (individual/10 5/MNC) |
| Normal control model contrast Siwu Tang group C1 group C2 group C3 group B group Paeoniflorin group | 84.2±5.5*** 34.8±3.9 61.3±7.1*** 12.6±4.0*** 46.5±4.8** 43.2±2.6** 44.4±7.2* 64.0±10.5*** | 45.8±8.9*** 13.5±4.2 23.8±5.1* 9.3±2.8 18.5±4.0 16.5±4.2 27.5±12.1 27.5±8.2* | 39.8±8.0*** 11.3±2.2 47.0±10.2*** 10.3±2.6* 15.0±3.7 14.5±2.1 37.5±13.5** 22.0±5.4* | 30.0±8.7** 5.0±1.8 12.5±2.5** 8.0±0.8* 13.3±4.0** 5.5±1.7 34.0±14.9** 33.3±7.5*** |
Annotate: * and model are compared * P＜0.05, * * P＜0.01, * * * P＜0.001
External hemopoietic progenitor cell colony cultivation results follows up the activity of enriching blood of having verified SIWU TANG separation and Extraction thing, and finds that first paeoniflorin has blood tonification effect.
Experimental example 2: the preliminary analysis at SIWU TANG compound active position is identified
One, HPTLC shows that SIWU TANG B active part mainly is made up of paeoniflorin, monosaccharide, disaccharide (being likely sucrose) and sees Fig. 1,2.
Two, HPLC shows that the SIWU TANG n-butyl alcohol partly contains paeoniflorin and sees Fig. 3
Three, ESIMS shows that the SIWU TANG n-butyl alcohol partly contains paeoniflorin and sees Fig. 4,5,6
Four, TLC shows that the monosaccharide of SIWU TANG n-butyl alcohol part is mainly fructose and glucose is seen Fig. 7
Experimental example 3: the assay of SIWU TANG n-butyl alcohol part paeoniflorin, fructose, glucose, sucrose is measured (method slightly) through HPLC, in the SIWU TANG n-butyl alcohol part extractum, contains: paeoniflorin 9.7%, fructose 16.5%, glucose 10.1%, sucrose 22.3%.
Description of drawings
Fig. 1,2 SIWU TANG B active part HPTLC figure
Fig. 3: SIWU TANG n-butyl alcohol section H PLC figure
Fig. 4,5,6: SIWU TANG n-butyl alcohol part ESIMS figure
Fig. 7: SIWU TANG n-butyl alcohol part TLC figure
The following example all can be realized the effect of above-mentioned experimental example.
The preparation method of embodiment 1 SIWU TANG n-butanol extract (B) and active site C:
SIWU TANG medical material 600g, first fries in shallow oil and adds water 3000ml, soaks 1h, fries in shallow oil 30min, three layers of filtered through gauze, second fries in shallow oil and adds water 2100ml, fries in shallow oil three layers of filtered through gauze of 20min, decocting liquid 3700ml.Decocting liquid≤60 a ℃ rotary evaporation is concentrated into 600ml, 20% ethanol precipitation, must precipitate (G part, 14.91g), supernatant 685ml proceed 40% ethanol precipitation must precipitate (the F part, 58.5g), supernatant 500ml, proceed 60% ethanol precipitation, must precipitate (E, 15.47g), supernatant 725ml proceeds 80% ethanol precipitation, must precipitate (D, 59.33g), supernatant 1320ml, be concentrated into no ethanol distinguish the flavor of extractum (86.2g), add water 300ml dissolve the about 500ml of water liquid, carry out ethyl acetate extraction (500ml, 500ml, 300ml), combined ethyl acetate liquid, reclaim solvent and get extract A (1.934g), the about 500ml of remaining water liquid carries out n-butanol extraction (500ml, 400ml, 300ml), merge n-butyl alcohol liquid, reclaim solvent and get extract B (11.5g), the about 500ml of remaining water liquid is concentrated into foaming and can not continues, and gets extract C (74.8g).
The preparation of experimental example 2 capsules
Effective site B 500g of the present invention, C 500g, medical starch 1000g, mix homogeneously, the capsule of packing into No. 1, every 0.2g, each oral 2-3 grain, twice of every day.
The preparation of embodiment 3 tablets
Effective site B 200g of the present invention, C 800g, medical starch 400g, dextrin 100g, mix homogeneously is used an amount of alcohol granulation, through the pelletizing machine granulate, tabletting, every 0.25g, oral, each 2, twice of every day.
The preparation of embodiment 4 granules
Effective site B 15g of the present invention, C 85g,, Icing Sugar 100g, mix homogeneously is used an amount of alcohol granulation, and drying, granulate, packing are promptly.Oral, a 5g, twice on the one.
Embodiment 5
SIWU TANG medical material 600g, first fries in shallow oil and adds water 3000ml, soaks 1h, fries in shallow oil 30min, three layers of filtered through gauze, second fries in shallow oil and adds water 2100ml, fries in shallow oil three layers of filtered through gauze of 20min, decocting liquid 3700ml; Decocting liquid≤60 a ℃ rotary evaporation is concentrated into 600ml, 20% ethanol precipitation must precipitate 14.91g, and supernatant 685ml proceeds 40% ethanol precipitation must precipitate 58.5g, supernatant 500ml, proceed 60% ethanol precipitation, must precipitate 15.47g, supernatant 725ml proceeds 80% ethanol precipitation, must precipitate 59.33g, supernatant 1320ml, be concentrated into no ethanol distinguish the flavor of extractum 86.2g, add water 300ml dissolve the about 500ml of water liquid, use 500ml respectively, 500ml, the 300ml ethyl acetate extracts, combined ethyl acetate liquid reclaims solvent and gets extract 1.934g, the about 500ml of remaining water liquid, use 500ml respectively, 400ml, the 300ml n-butyl alcohol extracts, and merges n-butyl alcohol liquid, and reclaiming solvent must this active site, mainly contain paeoniflorin 9.7%, fructose 16.5%, glucose 10.1%, sucrose 22.3%.Gained effective site adding Icing Sugar is an amount of, and mix homogeneously is used an amount of alcohol granulation, and drying, granulate, packing are promptly.Oral, a 5g, twice on the one.
Claims (7)
1, a kind of effective site of SIWU TANG is characterized in that being made up of B and C two parts, and B and C are prepared by following method, and the SIWU TANG medical material decocts 1-3 time, filters, and gets decocting liquid; Decocting liquid concentrates in 20-70 ℃ of rotary evaporation, use the 15-25% ethanol precipitation, must precipitate, supernatant is proceeded the 35-45% ethanol precipitation, must precipitate, supernatant is proceeded the 55-65% ethanol precipitation, must precipitate supernatant and proceed the 75-85% ethanol precipitation, must precipitate, supernatant concentration to do not have ethanol distinguish the flavor of extractum, add dissolving and carry out ethyl acetate extraction, combined ethyl acetate liquid, reclaim solvent and get extract, remaining water liquid carries out the butanols extraction, merges n-butyl alcohol liquid, reclaims solvent and gets extract B, remaining water liquid is concentrated into foaming and can not continues, and gets extract C.
2, the effective site of a kind of SIWU TANG as claimed in claim 1 is characterized in that SIWU TANG medical material 600g, and first fries in shallow oil and add water 3000ml, soak 1h, fry in shallow oil 30min, three layers of filtered through gauze, second fries in shallow oil and adds water 2100ml, fries in shallow oil three layers of filtered through gauze of 20min, decocting liquid 3700ml; Decocting liquid≤60 a ℃ rotary evaporation is concentrated into 600ml, 20% ethanol precipitation, must precipitate 14.91g, supernatant 685ml proceeds 40% ethanol precipitation must precipitate 58.5g, and supernatant 500ml proceeds 60% ethanol precipitation, must precipitate 15.47g, supernatant 725ml proceeds 80% ethanol precipitation, must precipitate 59.33g, supernatant 1320ml, be concentrated into no ethanol distinguish the flavor of extractum 86.2g, add water 300ml dissolve the about 500ml of water liquid, use 500ml respectively, 500ml, the 300ml ethyl acetate extracts, combined ethyl acetate liquid, reclaim solvent and get extract 1.934g, remaining water liquid 500ml uses 500ml respectively, 400ml, the 300ml n-butyl alcohol extracts, merge n-butyl alcohol liquid, reclaim solvent and get effective site B, remaining water liquid 500ml concentrates, and gets effective site C.
3, a kind of SIWU TANG effective site is characterized in that mainly containing paeoniflorin 9.7%, fructose 16.5%, glucose 10.1%, sucrose 22.3%;
This effective site is prepared by following method: the SIWU TANG medical material decocts 1-3 time, filters, and gets decocting liquid; Decocting liquid concentrates in 20-70 ℃ of rotary evaporation, use the 15-25% ethanol precipitation, must precipitate, supernatant is proceeded the 35-45% ethanol precipitation, must precipitate, supernatant is proceeded the 55-65% ethanol precipitation, must precipitate, and supernatant is proceeded the 75-85% ethanol precipitation, must precipitate, supernatant concentration to do not have ethanol distinguish the flavor of extractum, add dissolving and carry out ethyl acetate extraction, combined ethyl acetate liquid, reclaim solvent and get extract, remaining water liquid carries out the butanols extraction approximately, merges n-butyl alcohol liquid, and reclaiming solvent must this active site.
4, the preparation method of effective site as claimed in claim 3 is characterized in that SIWU TANG medical material 600g, and first fries in shallow oil and add water 3000ml, soak 1h, fry in shallow oil 30min, three layers of filtered through gauze, second fries in shallow oil and adds water 2100ml, fries in shallow oil three layers of filtered through gauze of 20min, decocting liquid 3700ml; Decocting liquid≤60 a ℃ rotary evaporation is concentrated into 600ml, 20% ethanol precipitation, must precipitate 14.91g, supernatant 685ml proceeds 40% ethanol precipitation must precipitate 58.5g, supernatant 500ml, proceed 60% ethanol precipitation, must precipitate 15.47g, supernatant 725ml proceeds 80% ethanol precipitation, must precipitate 59.33g, supernatant 1320ml, be concentrated into no ethanol distinguish the flavor of extractum 86.2g, add water 300ml dissolve water liquid 500ml, use 500ml respectively, 500ml, the 300ml ethyl acetate extracts, combined ethyl acetate liquid reclaims solvent and gets extract 1.934g, remaining water liquid 500ml, use 500ml respectively, 400ml, the 300ml n-butyl alcohol extracts, and merges n-butyl alcohol liquid, and reclaiming solvent must this active site.
5,, it is characterized in that also adding the excipient of pharmaceutically accepting as claim 1,2,3 or 4 described effective sites.
6, as claim 1,2,3 or 4 described effective sites, it is characterized in that making the dosage form of clinical acceptance.
7, effective site as claimed in claim 6 is characterized in that the dosage form of making is a kind of in the middle of tablet, capsule, pill, granule, suspensoid, drop pill, oral liquid, injection, aerosol, suppository or the subcutaneous administration dosage form.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 03123521 CN1304025C (en) | 2003-05-09 | 2003-05-09 | Active part and effective component of Four Drug Decoction |
| CNB2006101618153A CN100467026C (en) | 2003-05-09 | 2003-05-09 | Application of paeoniflorin for enriching the blood |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 03123521 CN1304025C (en) | 2003-05-09 | 2003-05-09 | Active part and effective component of Four Drug Decoction |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB2006101618153A Division CN100467026C (en) | 2003-05-09 | 2003-05-09 | Application of paeoniflorin for enriching the blood |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1548079A CN1548079A (en) | 2004-11-24 |
| CN1304025C true CN1304025C (en) | 2007-03-14 |
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| CN 03123521 Expired - Fee Related CN1304025C (en) | 2003-05-09 | 2003-05-09 | Active part and effective component of Four Drug Decoction |
| CNB2006101618153A Expired - Lifetime CN100467026C (en) | 2003-05-09 | 2003-05-09 | Application of paeoniflorin for enriching the blood |
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| CNB2006101618153A Expired - Lifetime CN100467026C (en) | 2003-05-09 | 2003-05-09 | Application of paeoniflorin for enriching the blood |
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Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101897714B (en) * | 2009-05-25 | 2012-11-28 | 北京卓越同创药物研究院 | Use of paeoniflorin |
| CN101984997B (en) * | 2010-10-11 | 2012-03-14 | 南京中医药大学 | Anti-tumor composition containing four-substance decoction extract and application thereof |
| CN105377280B (en) * | 2013-07-18 | 2019-09-03 | 北京中医药大学 | A kind of drug and preparation method thereof for enriching blood |
| CN104725449A (en) * | 2013-12-19 | 2015-06-24 | 北京农学院 | Paeoniflorin extracting method |
| CN114585368A (en) * | 2019-11-22 | 2022-06-03 | 张作光 | Application of albiflorin or paeoniflorin in preventing and/or treating renal anemia |
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2003
- 2003-05-09 CN CN 03123521 patent/CN1304025C/en not_active Expired - Fee Related
- 2003-05-09 CN CNB2006101618153A patent/CN100467026C/en not_active Expired - Lifetime
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| Publication number | Publication date |
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| CN100467026C (en) | 2009-03-11 |
| CN1548079A (en) | 2004-11-24 |
| CN101040864A (en) | 2007-09-26 |
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