CN102028764A - Ternate buttercup root anti-tuberculosis valid target and preparation method and application thereof - Google Patents
Ternate buttercup root anti-tuberculosis valid target and preparation method and application thereof Download PDFInfo
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- radix ranunculi
- ranunculi ternati
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Abstract
The invention discloses a ternate buttercup root anti-tuberculosis valid target and a preparation method and application thereof. The valid target mainly contains 3-[4-O-beta-D-glucosyl]-phenyl]-2-acrylic acid, 4-[Formyl-5-(methoxymethyl)-1H-pyrrol-1-yl] butanoate and 5-hydroxymethyl furfural. The valid target is formed by extracting ternate buttercup root and enriching macroporous resin. The quality standard of the valid target is established by using the HPLC (High Performance Liquid Chromatography), wherein the content of the 3-[4-O-beta-D-glucosyl]-phenyl]-2-acrylic acid is 10-15%, the content of the 5-hydroxymethyl furfural is 10-15%, and the content of the 4-[Formyl-5-(methoxymethyl)-1H-pyrrol-1-yl] butanoate is 70-80%. The valid target can be used for preparing medicines for resisting tubercle bacillus and drug-resistant tubercle bacillus, and can be combined with the existing clinical medicines for treating tuberculosis, thereby enhancing the curative effect and lowering the recurrence rate.
Description
Technical field
The present invention relates to Chinese medicine, relate in particular to antiphthisic effective site of a kind of Radix Ranunculi Ternati and its production and use.
Background technology
Tuberculosis (TB) is current global range to one of infectious disease of the most threatening property of the mankind, in recent years, because drug resistance, anti-multiple medicines (multidrugresistanttuberculosis, MDR one TB) generation of mycobacterium tuberculosis and propagation and tuberculosis merge the HIV infection, make that once being controlled at very low-level tuberculosis epidemic situation revives.Long than the easier generation drug resistance of other treatment of diseases, treatment cycle, be the big main cause of tuberculotherapy difficulty.At present, the integrated use bigeminy compound preparation that the international public praises highly is (as Rifinah, it is peaceful to defend lung) and three compound preparations (as Rifater, defend the lung spy), adopt the therapeutic scheme of 2+4, still need 6 months treatment time at least, and need drug combination, more increased the consumption figure and the toxicity of antituberculotics.Therefore, develop that novel antituberculotics shortens treatment cycle, the toxicity and the control drug resistance that reduce chemotherapeutics still is difficult point, the emphasis of tuberculosis prevention and treatment work.
Radix Ranunculi Ternati be the little Herba Ranunculi Japonici of Ranunculaceae ranunculus (
Ranunculus ternatusThunb.) dry tuber.Effect with heat-clearing and toxic substances removing, eliminating stagnation repercussive, treatment pulmonary tuberculosis, lymphoid tuberculosis, pharyngolaryngitis etc. have remarkable result.The chemical constitution study of system shows that Radix Ranunculi Ternati contains sterol, γ-ketone-δ-Wu Neizhi, 5 hydroxymethyl furfural, 4-[Formyl-5-(methoxymethyl)-1H-pyrrol-1-yl] multiple chemical compound such as butanoate, we instruct by pharmacologically active, track the antiphthisic effective site of Radix Ranunculi Ternati, the isolating means of employing system confirm that this effective site mainly contains 3-[4-O-β-D-glucosyl group]-phenyl]-2-acrylic acid, 4-[Formyl-5-(methoxymethyl)-1H-pyrrol-1-yl] butanoate, three kinds of compositions of 5 hydroxymethyl furfural; And by in the body, external evaluating drug effect confirms that this effective site all has significant therapeutical effect to Mycobacterium tuberculosis type strain, drug resistance mycobacterium tuberculosis strain, can be developed to antiphthisic new Chinese medicine, and is significant to clinical treatment lungy.
Summary of the invention
The purpose of this invention is to provide antiphthisic effective site of a kind of Radix Ranunculi Ternati and its production and use.
The antiphthisic effective site of Radix Ranunculi Ternati contains 3-[4-O-β-D-glucosyl group]-phenyl]-2-acrylic acid, 4-[Formyl-5-(methoxymethyl)-1H-pyrrol-1-yl] butanoate, three kinds of constituents of 5 hydroxymethyl furfural.
The content of three kinds of constituents of described effective site adopts the HPLC method to measure, wherein, 3-[4-O-β-D-glucosyl group]-phenyl]-the acrylic acid quality percentage composition of 2-is 10%～15%, the quality percentage composition of 5 hydroxymethyl furfural is 10%～15%, 4-[Formyl-5-(methoxymethyl)-1H-pyrrol-1-yl] the quality percentage composition of butanoate is 70%～80%.
Described effective site adopts ethanol extraction to make in conjunction with the method for purification by macroporous resin by Radix Ranunculi Ternati.
The preparation method of the antiphthisic effective site of Radix Ranunculi Ternati is: 1 weight portion Radix Ranunculi Ternati adds the water or the aquiferous ethanol of 5～15 weight portions, adopt and reflux, ultrasonic, merceration or diafiltration method are extracted 2～4 times, each extraction time 1～3 h, filter, be concentrated into 0.25～2g crude drug/mL concentration, filter or centrifugalize, supernatant separates by the macroporous resin detached dowel, weight ratio by crude drug supernatant applied sample amount and dried macroporous resin is 1:3～10, last sample flow velocity 1～6 BV/h, blade diameter length ratio is 1:2～1:10, the macroporous resin behind the last sample, adopt water successively, the aquiferous ethanol eluting, wherein, 2～8 column volumes of water elution, 2～8 column volumes of ethanol elution, elution flow rate 2～8BV/h, collect ethanol elution, concentrate, oven dry, drying under reduced pressure, infrared drying or microwave drying, pulverize, obtain the antiphthisic effective site of Radix Ranunculi Ternati.
The mass percent concentration of described aquiferous ethanol is 30%～80%.
Described weight ratio by crude drug supernatant applied sample amount and dried macroporous resin is 1:5～10, last sample flow velocity 1～4 BV/h, blade diameter length ratio is 1:5～1:8, macroporous resin behind the last sample adopts water, aquiferous ethanol eluting, wherein successively, 3～5 column volumes of water elution, 3～5 column volumes of aquiferous ethanol eluting, elution flow rate is 2～4BV/h.
The antiphthisic effective site of Radix Ranunculi Ternati is used for preparation treatment tuberculosis.
The Radix Ranunculi Ternati effective site of the present invention's preparation, adopt minimum doubling dilution to measure the experiment of Tuberculosis in vitro pyrenomycetes, the result shows that it all has good inhibition effect to Mycobacterium tuberculosis type strain H37Rv, the Mycobacterium tuberculosis of anti-multiple medicines the (MDR-TB), full drug resistance Mycobacterium tuberculosis (XDR-TB), and minimal inhibitory concentration is 2.5mg/mL.
Tuberculosis experiment confirm in the Radix Ranunculi Ternati effective site prepared by this method, body, it has significant therapeutical effect to lotus tubercule bacillus mice, can reduce the lotus bacterium number of mouse infection organ, prolongs life span.
Tuberculosis experiment confirm in the Radix Ranunculi Ternati effective site prepared by this method, body is united the lotus bacterium number that use can significantly reduce lotus tulase mouse infection organ with isoniazid, prolongs life span.
Radix Ranunculi Ternati effective site prepared by this method behind interpolation adjuvant or the excipient, can be made pharmaceutical dosage forms such as capsule, tablet, granule, becomes the treatment tuberculosis.
Description of drawings
Fig. 1 is that the high performance liquid chromatogram (HPLC-DAD) of Radix Ranunculi Ternati tuberculosis effective site is analyzed collection of illustrative plates.
The specific embodiment
Following experimental example is used to further specify but is not limited to the present invention.
The antiphthisic effective site of Radix Ranunculi Ternati contains 3-[4-O-β-D-glucosyl group]-phenyl]-2-acrylic acid, 4-[Formyl-5-(methoxymethyl)-1H-pyrrol-1-yl] butanoate, three kinds of constituents of 5 hydroxymethyl furfural.
The content of three kinds of constituents of described effective site adopts the HPLC method to measure, wherein, 3-[4-O-β-D-glucosyl group]-phenyl]-the acrylic acid quality percentage composition of 2-is 10%～15%, the quality percentage composition of 5 hydroxymethyl furfural is 10%～15%, 4-[Formyl-5-(methoxymethyl)-1H-pyrrol-1-yl] the quality percentage composition of butanoate is 70%～80%.
Described effective site adopts ethanol extraction to make in conjunction with the method for purification by macroporous resin by Radix Ranunculi Ternati.
The preparation method of the antiphthisic effective site of Radix Ranunculi Ternati is: 1 weight portion Radix Ranunculi Ternati adds the water or the aquiferous ethanol of 5～15 weight portions, adopt and reflux, ultrasonic, merceration or diafiltration method are extracted 2～4 times, each extraction time 1～3 h, filter, be concentrated into 0.25～2g crude drug/mL concentration, filter or centrifugalize, supernatant separates by the macroporous resin detached dowel, weight ratio by crude drug supernatant applied sample amount and dried macroporous resin is 1:3～10, last sample flow velocity 1～6 BV/h, blade diameter length ratio is 1:2～1:10, the macroporous resin behind the last sample, adopt water successively, the aquiferous ethanol eluting, wherein, 2～8 column volumes of water elution, 2～8 column volumes of ethanol elution, elution flow rate 2～8BV/h, collect ethanol elution, concentrate, oven dry, drying under reduced pressure, infrared drying or microwave drying, pulverize, obtain the antiphthisic effective site of Radix Ranunculi Ternati.
The mass percent concentration of described aquiferous ethanol is 30%～80%.
Described weight ratio by crude drug supernatant applied sample amount and dried macroporous resin is 1:5～10, last sample flow velocity 1～4 BV/h, blade diameter length ratio is 1:5～1:8, macroporous resin behind the last sample adopts water, aquiferous ethanol eluting, wherein successively, 3～5 column volumes of water elution, 3～5 column volumes of aquiferous ethanol eluting, elution flow rate is 2～4BV/h.
The antiphthisic effective site of Radix Ranunculi Ternati is used for preparation treatment tuberculosis.
Embodiment 1
1 weight portion Radix Ranunculi Ternati adds the water or the aquiferous ethanol of 5 weight portions, adopts reflux, extract, 2 times, each extraction time 1 h, filter, be concentrated into 0.25g crude drug/mL concentration, filter, supernatant separates by the macroporous resin detached dowel, weight ratio by crude drug supernatant applied sample amount and dried macroporous resin is 1:3, last sample flow velocity 1 BV/h, blade diameter length ratio is 1:2, the macroporous resin behind the last sample, adopt water successively, the aquiferous ethanol eluting, wherein, 2 column volumes of water elution, 2 column volumes of ethanol elution, elution flow rate 2BV/h, collect ethanol elution, concentrate, oven dry, drying under reduced pressure, infrared drying or microwave drying, pulverize, obtain the antiphthisic effective site of Radix Ranunculi Ternati.
Embodiment 2
1 weight portion Radix Ranunculi Ternati adds the water or the aquiferous ethanol of 15 weight portions, adopts supersound extraction 2～4 times, each extraction times 3 h, filter, be concentrated into 2g crude drug/mL concentration, centrifugalize, supernatant separates by the macroporous resin detached dowel, weight ratio by crude drug supernatant applied sample amount and dried macroporous resin is 1:10, last sample flow velocity 6 BV/h, blade diameter length ratio is 1:10, the macroporous resin behind the last sample, adopt water successively, the aquiferous ethanol eluting, wherein, 8 column volumes of water elution, 8 column volumes of ethanol elution, elution flow rate 8BV/h, collect ethanol elution, concentrate, oven dry, drying under reduced pressure, infrared drying or microwave drying, pulverize, obtain the antiphthisic effective site of Radix Ranunculi Ternati.
Embodiment 3
Get Radix Ranunculi Ternati medical material 2 kg after the pulverizing, add 6 times of amount 10% ethanol, heating and refluxing extraction 4 times, each 1h filters, and merge extractive liquid, is concentrated into 0.25 g crude drug/mL; Filter the back by D101 type macroporous resin, extracting solution is 1:5 by the ratio of Radix Ranunculi Ternati total glycosides content and dried resin, and last column flow rate 1BV/h, resin path height ratio are 1:3; Wash resin 3BV with water, elution flow rate is 2BV/h, and water elution liquid discards; With containing 30% aquiferous ethanol eluting 8BV, flow velocity is 2BV/h, collects ethanol elution; It is 1.1 that ethanol elution is evaporated to relative density, and drying under reduced pressure gets Radix Ranunculi Ternati effective site.The HPLC method records 3-[4-O-β-D-glucosyl group]-phenyl]-the acrylic acid content of 2-is 17%, 5 hydroxymethyl furfural is 35%, 4-[Formyl-5-(methoxymethyl)-1H-pyrrol-1-yl] and the content of butanoate is 40%.This effective site is pulverized, and adds 5% micropowder silica gel, and the capsule of packing into No. 1 promptly gets capsule.
Embodiment 4
With Radix Ranunculi Ternati decoction pieces 1kg, add 15 times of amount 80% ethanol, heating and refluxing extraction 4 times, each 3h filters, and merge extractive liquid, is concentrated into 2.0 g crude drug/mL; Centrifugal back is by the chromatographic column of AB-8 type macroporous resin, and extracting solution is 1:10 by the ratio of Radix Ranunculi Ternati total glycosides content and dried resin, and last column flow rate 4BV/h, resin path height ratio are 1:9; Wash resin 8BV with water, elution flow rate is 4BV/h, and water elution liquid discards; With containing 95% aquiferous ethanol eluting, flow velocity is 4BV/h, collects ethanol elution; It is 1.25 that ethanol elution is evaporated to relative density, and decompression or spray drying get Radix Ranunculi Ternati effective site.The HPLC method records 3-[4-O-β-D-glucosyl group]-phenyl]-the acrylic acid content of 2-is 11%, 5 hydroxymethyl furfural is 28%, 4-[Formyl-5-(methoxymethyl)-1H-pyrrol-1-yl] and the content of butanoate is 50%.This effective site is pulverized, and adds 5% dextrin, 1% magnesium stearate, adds 1% aspartame again, makes granule, promptly.
Embodiment 5
Get the Radix Ranunculi Ternati medical material 5kg after the pulverizing, add 8 times of amount 70% ethanol, heating and refluxing extraction 3 times, each 1h filters, and merge extractive liquid, is concentrated into 1.0 g crude drug/mL; Centrifugal (4000 rev/mins) back is by AB-8 type macroporous resin, and medicinal liquid is 1:5 by the ratio of Radix Ranunculi Ternati total glycosides content and dried resin, and last column flow rate 2BV/h, resin path height ratio are 1:5; Wash resin 3BV with water, elution flow rate is 4BV/h, and water elution liquid discards; With containing 70% water-ethanol eluting 3BV, flow velocity is 2BV/h, collects ethanol elution; It is that 1.1(50 ℃ of heat is surveyed that ethanol liquid is evaporated to relative density), drying under reduced pressure gets Radix Ranunculi Ternati effective site.The HPLC method records 3-[4-O-β-D-glucosyl group]-phenyl]-the acrylic acid content of 2-is 24%, 5 hydroxymethyl furfural is 34%, 4-[Formyl-5-(methoxymethyl)-1H-pyrrol-1-yl] and the content of butanoate is 47%.This effective site is pulverized, and adds 3% starch, 1% sucrose, and compacting promptly gets tablet in flakes.
Embodiment 6
Radix Ranunculi Ternati pharmaceutical decocting piece 5kg adds 10 times of amount 50% ethanol, heating and refluxing extraction 2 times, and each 1.5h filters, and merge extractive liquid, is concentrated into 0.5g crude drug/mL; Filter the back by D201 type macroporous resin, medicinal liquid is 1:9 by the ratio of Radix Ranunculi Ternati total glycosides content and dried resin, and last column flow rate 3BV/h, resin path height ratio are 1:7; Wash resin 5BV with water, elution flow rate is 4BV/h, and water elution liquid discards; With containing 50% water-ethanol eluting 10BV, flow velocity is 4BV/h, collects ethanol elution; It is that 1.25(50 ℃ of heat is surveyed that ethanol liquid is evaporated to relative density), spray drying gets Radix Ranunculi Ternati effective site.The HPLC method records 3-[4-O-β-D-glucosyl group]-phenyl]-the acrylic acid content of 2-is 11%, 5 hydroxymethyl furfural is 28%, 4-[Formyl-5-(methoxymethyl)-1H-pyrrol-1-yl] and the content of butanoate is 50%.This effective site is pulverized, and adds 5% galactose, and compacting promptly gets tablet in flakes.
Embodiment 7
Radix Ranunculi Ternati pharmaceutical decocting piece 5kg adds 15 times of amount 30% ethanol, heating and refluxing extraction 3 times, and each 0.5h filters, and merge extractive liquid, is concentrated into 0.25g crude drug/mL; Centrifugal (8000 rev/mins) back is by HPD450 macroporous resin, and medicinal liquid is 1:6 by the ratio of Radix Ranunculi Ternati total glycosides content and dried resin, and last column flow rate 4BV/h, resin path height ratio are 1:9; Wash resin 2BV with water, elution flow rate is 3BV/h, and water elution liquid discards; With containing 50% water-ethanol eluting 5BV, flow velocity is 3BV/h, collects ethanol elution; It is that 1.2(50 ℃ of heat is surveyed that ethanol liquid is evaporated to relative density), spray drying gets Radix Ranunculi Ternati effective site.The HPLC method records 3-[4-O-β-D-glucosyl group]-phenyl]-the acrylic acid content of 2-is 3%, 5 hydroxymethyl furfural is 5%, 4-[Formyl-5-(methoxymethyl)-1H-pyrrol-1-yl] and the content of butanoate is 21%.This effective site is pulverized, and adds 5% magnesium stearate, and the capsule of packing into No. 2 promptly gets capsule.
Experimental example 1 high performance liquid chromatography detects the content of main component in the Radix Ranunculi Ternati effective site
Adopt constituent 4-[Formyl-5-(the methoxymethyl)-1H-pyrrol-1-yl of high effective liquid chromatography for measuring Radix Ranunculi Ternati effective site] butanoate, 5 hydroxymethyl furfural, 3-[4-O-β-D-glucosyl group]-phenyl]-the acrylic acid content of 2-.Instrument: Shimadzu 20A high performance liquid chromatograph-DAD UV-detector (Japan).Chromatographic column: TC RP-C
18Chromatographic column, 250mm * 4.6 mm(Aglient, USA).Mobile phase: methanol-0.4% acetic acid water (gradient elution).Elution program is: 0.01min-35min methanol 5%-19%, acetic acid water 95%-81%; 35min-45min methanol 19%-19%, acetic acid water 81%-81%.Flow velocity: 1.0 mL/min.Column temperature: 40 oC.Detect wavelength: 290 nm.Sample size: 10 μ L.Retention time (RT): 3-[4-O-β-D-glucosyl group]-phenyl]-2-acrylic acid (chemical compound 1) is 8-10 min, 5 hydroxymethyl furfural is 12-15 min, 4-[Formyl-5-(methoxymethyl)-1H-pyrrol-1-yl] butanoate is 39-42 min.Adopt external standard method 3-[4-O-β-D-glucosyl group]-phenyl]-2-acrylic acid, 5 hydroxymethyl furfural, 4-[Formyl-5-(methoxymethyl)-1H-pyrrol-1-yl] content of butanoate.
The experiment of experimental example 2 Radix Ranunculi Ternati effective site Tuberculosis in vitro pyrenomycetes
Material and method
1) culture medium: 7H9 fluid medium (containing 10%ADC, 0.03% agar).
2) medicine and processing thereof (test tube doubling dilution): Radix Ranunculi Ternati effective site, isoniazid (INH) dmso solution are made solution (storing solution) just, behind the circulation steam sterilization, make it be designed concentration in culture medium.
3) tulase and in culture medium concentration: experimental bacteria is Mycobacterium tuberculosis type strain H37Rv, MDR-TB, XDR-TB.Will be on modified Russell medium well-grown bacterial strain, wear into bacteria suspension with agate mortar, making its concentration in culture medium is 1 * 10
5CFU/mL.
4) cultivation is put 37 ℃ of cultivations with observation, is as the criterion with the result around cultivating.
The result
In the 7H9 culture medium, Radix Ranunculi Ternati effective site is 2.5 mg/mL at the minimal inhibitory concentration (MIC) of H37Rv, the Mycobacterium tuberculosis of anti-multiple medicines the (MDR-TB), full drug resistance Mycobacterium tuberculosis (XDR-TB), shows that Radix Ranunculi Ternati effective site all has good vitro inhibition effect to tulase type strain and persister.
Tuberculosis experiment in the experimental example 3 Radix Ranunculi Ternati effective site bodies
1) laboratory animal: SPF level C57BL/6 mice, age in 6-8 week, female and male mice half and half.Weigh labelling, random packet.
2) experiment reagent or material:
Tubercule bacillus H37Rv strain; Modified Russell medium (sodium glutamate, potassium dihydrogen phosphate, magnesium sulfate, citrate of magnesia, glycerol, distilled water, whole egg liquid, 2% peacock green, potato starch); Middlebrook 7H9 fluid medium and Middlebrook 7H11 solid medium; Normal saline; Phosphate buffered saline(PBS) (PBS); Formaldehyde.
3) medicine:
Supply the reagent thing: Radix Ranunculi Ternati effective site;
Chinese medicine contrast: Radix Ranunculi Ternati capsule (being commercially available clinical used treatment Chinese patent medicine lungy);
Western medicine contrast: isoniazid (being commercially available clinical used treatment tuberculosis).
4) be taken on the modified Russell medium 2 weeks of growth, and the culture of the mycobacterium hominis H37Rv of multiple poison in animal body, wear into the even bacteria suspension (sterile saline solution or PBS) of 1mg/ml with dismembyator.Behind the mouse anesthesia with tuberculosis model yet to be built, every above-mentioned bacteria suspension of mice collunarium 10 μ L.
5) with the mice random packet of modeling success, be respectively model group, negative control group, the high, medium and low dosage group of Radix Ranunculi Ternati effective site, Chinese medicine positive drug control group (Radix Ranunculi Ternati capsule), Western medicine positive drug control group (isoniazid), the high, medium and low dosage adding of Radix Ranunculi Ternati effective site isoniazid matched group (the high, medium and low dosage group of drug combination), irritate stomach every day and give the medicine or the distilled water of variable concentrations, continued medication for 8 weeks.
6) after experiment finishes, measure the lotus bacterium number of respectively organizing mouse infection organ-lung, spleen.
The result
To infecting the Cavia porcellus of tulase H37Rv, Radix Ranunculi Ternati effective site can significantly reduce the lotus bacterium number of lotus bacterium mouse lung, spleen, pathogenic index obviously reduces (P＜0.01), the slightly inferior and isoniazid of effect, Radix Ranunculi Ternati effective site and isoniazid share, pathogenic index further reduces (P＜0.001), and effect is better than isoniazid (table 1).
Table 1 Radix Ranunculi Ternati total glycosides and associating Rimactazid are to the exponential influence of Cavia porcellus organ disease
Claims (7)
1. antiphthisic effective site of Radix Ranunculi Ternati, it is characterized in that: effective site contains 3-[4-O-β-D-glucosyl group]-phenyl]-2-acrylic acid, 4-[Formyl-5-(methoxymethyl)-1H-pyrrol-1-yl] butanoate, three kinds of constituents of 5 hydroxymethyl furfural.
2. the antiphthisic effective site of a kind of Radix Ranunculi Ternati according to claim 1, it is characterized in that: the content of three kinds of constituents of described effective site adopts the HPLC method to measure, wherein, 3-[4-O-β-D-glucosyl group]-phenyl]-the acrylic acid quality percentage composition of 2-is 10%～15%, the quality percentage composition of 5 hydroxymethyl furfural is 10%～15%, 4-[Formyl-5-(methoxymethyl)-1H-pyrrol-1-yl] the quality percentage composition of butanoate is 70%～80%.
3. the antiphthisic effective site of a kind of Radix Ranunculi Ternati according to claim 1 is characterized in that: described effective site adopts ethanol extraction to make in conjunction with the method for purification by macroporous resin by Radix Ranunculi Ternati.
4. the preparation method of the antiphthisic effective site of Radix Ranunculi Ternati as claimed in claim 1, it is characterized in that: 1 weight portion Radix Ranunculi Ternati adds the water or the aquiferous ethanol of 5～15 weight portions, adopt and reflux, ultrasonic, merceration or diafiltration method are extracted 2～4 times, each extraction time 1～3 h, filter, be concentrated into 0.25～2g crude drug/mL concentration, filter or centrifugalize, supernatant separates by the macroporous resin detached dowel, is 1:3～10 by the weight ratio of crude drug supernatant applied sample amount and dried macroporous resin, last sample flow velocity 1～6 BV/h, blade diameter length ratio is 1:2～1:10, macroporous resin behind the last sample adopts water successively, the aquiferous ethanol eluting, wherein, 2～8 column volumes of water elution, 2～8 column volumes of ethanol elution, elution flow rate 2～8BV/h collects ethanol elution, concentrate, oven dry, drying under reduced pressure, infrared drying or microwave drying are pulverized, and obtain the antiphthisic effective site of Radix Ranunculi Ternati.
5. the preparation method of the antiphthisic effective site of a kind of Radix Ranunculi Ternati according to claim 4 is characterized in that: the mass percent concentration of described aquiferous ethanol is 30%～80%.
6. the preparation method of the antiphthisic effective site of a kind of Radix Ranunculi Ternati according to claim 4, it is characterized in that: described weight ratio by crude drug supernatant applied sample amount and dried macroporous resin is 1:5～10, last sample flow velocity 1～4 BV/h, blade diameter length ratio is 1:5～1:8, and the macroporous resin behind the last sample adopts water, aquiferous ethanol eluting successively, wherein, 3～5 column volumes of water elution, 3～5 column volumes of aquiferous ethanol eluting, elution flow rate is 2～4BV/h.
7. the purposes of the antiphthisic effective site of Radix Ranunculi Ternati as claimed in claim 1 is characterized in that: be used for preparation treatment tuberculosis.
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| CN103768168A (en) * | 2012-10-25 | 2014-05-07 | 奇复康药物研发(苏州)有限公司 | Extraction method for r ranunculus ternatus thunb tannin |
| CN106674163A (en) * | 2017-01-05 | 2017-05-17 | 贵州大学 | Anti-mycobacterium tuberculosis compound and preparation method and application thereof |
| CN110361457A (en) * | 2018-03-26 | 2019-10-22 | 北京哈三联科技有限责任公司 | The HPLC detection method of 5 hydroxymethyl furfural content in medical product |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103768167A (en) * | 2012-10-25 | 2014-05-07 | 奇复康药物研发(苏州)有限公司 | Extraction method for ranunculus ternatus thunb triterpene component |
| CN103768168A (en) * | 2012-10-25 | 2014-05-07 | 奇复康药物研发(苏州)有限公司 | Extraction method for r ranunculus ternatus thunb tannin |
| CN106674163A (en) * | 2017-01-05 | 2017-05-17 | 贵州大学 | Anti-mycobacterium tuberculosis compound and preparation method and application thereof |
| CN106674163B (en) * | 2017-01-05 | 2019-07-19 | 贵州大学 | A kind of anti-mycobacterium tuberculosis compound and the preparation method and application thereof |
| CN110361457A (en) * | 2018-03-26 | 2019-10-22 | 北京哈三联科技有限责任公司 | The HPLC detection method of 5 hydroxymethyl furfural content in medical product |
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