CN101255182B - Extraction separating method for bupleurum root saponin b2 - Google Patents
Extraction separating method for bupleurum root saponin b2 Download PDFInfo
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Abstract
The invention relates to a separation method for abstracting effective ingredient of Chinese traditional medicine, more particularly a separation method for abstracting bupleurum saponin b<2> which is the effective ingredient of bupleurum, wherein the method comprises purifying the total extract obtained by abstracting bupleurum by macroporous resin to obtain the crude product of bupleurum saponin b<2>, and preparing bupleurum saponin b<2> pure product by column chromatography and recrystallization or both processes of the bupleurum saponin b<2> crude product. The method is capable of effectively separating the Chinese traditional medicine to obtain bupleurum saponin b<2> monomer, is suitable for industrial production, and provides a foundation for better controlling the quality of the effective ingredient of bupleurum.
Description
Technical field
The extraction and separation method of pharmaceutically active ingredient is specifically related to the extraction and separation method about effective constituent in the bupleurum Chinese in the invention relates to.
Background technology
Saikoside b
2Being a kind of secondary saikoside, is to be transformed by primary saikoside d, is that the oxygen ether ring instability between 28 easily ruptures owing to C-13 in the saikoside d structure specifically, generates Δ
11,13 (18)The secondary saikoside b of-olea diene type
2Document shows: after 1) the radix bupleuri medicinal material boiled through traditional decocting, saikoside d all changed saikoside b into
2(Yukio Ogihara, MasakiAburada, Sho-Saiko-To:Scientific Evaluation and Clinical Applications, 1 edition (July 24,2003), CRC press, page 126); 2) contain in the compound preparation of radix bupleuri and all can detect a large amount of saikoside b
2(Li Jun, Shi Renbing, Liu Bin etc., the different compatibilities of Sini San are to saikoside a, b
2And the influence of the fried amount of bupleurum total saponin, Beijing University of Chinese Medicine's journal, 2007,30 (2): 115-120).As seen, saikoside b
2Being to contain saikoside constituents important in the bupleurum preparation, is the important indicator that control contains the bupleurum preparation quality.Therefore, in real work, need the high saikoside b of extraction separation purity
2Product in contrast.Particularly, how to simplify saikoside b containing in the middle of the drug development of radix bupleuri
2Extraction process, improve separation purity, be one of key link of preparation process research process.Although saikoside b in the existing bibliographical information radix bupleuri
2The extraction preparation method (Zhang Zhi is bright etc. for Chen Xikui, Zhang Ruyi, the research of saponin component in the black radix bupleuri, journal of Beijing Medical University, 1992,2 (3): 222-224), but, this method be utilize the silica gel medium pressure post repeatedly chromatographic separation obtain, requiring has certain device.Because extraction process route complexity in the past to the equipment requirements strictness, is not suitable for large-scale industrialization production.
Summary of the invention
The object of the present invention is to provide a kind of technology of extraction separation bupleurum Chinese effective constituent, i.e. extraction separation bupleurum Chinese effective constituent saikoside b
2Novel process.
Processing method of the present invention comprises the steps:
A: get the dregs of a decoction after radix bupleuri or radix bupleuri have extracted volatile oil, the suitable pulverizing adds solvent and extracts, and extracting solution concentrates and reclaims solvent, gets general extractive;
B: general extractive adds water and suitably disperses, last macroporous resin adsorption post, earlier with the removal of impurities of alkali aqueous solution wash-out, water is eluted to neutrality again, next use 10%～40% ethanol/water solution wash-out removal of impurities, with 40%～100% ethanol/water solution wash-out and collect this part elutriant, concentrate and reclaim solvent at last, saikoside b
2Crude product;
C: get saikoside b
2Crude product is admixed 2～10 times of weight part silica gel, volatilizes solvent, must mix sample silica gel; Other got 20～80 times of weight part silica gel activatings 2 hours, and wet method dress post will be mixed sample silica gel and add capital, with the mixed solvent wash-out of chloroform-methanol-water, collect elutriant, and the thin layer inspection is known, and merges and contains saikoside b
2Stream part, reclaim solvent and get saikoside b
2Elaboration;
Get saikoside b
2Elaboration, recrystallization in the mixed solvent of acetone-water or chloroform-methanol-water filters, drying, saikoside b
2Pure product.
In the aforesaid method in the steps A radix bupleuri be the root of the arbitrary plant of umbelliferae Bupleurum, or commercially available radix bupleuri medicine materical crude slice also comprises the various processed products through concocting; The dregs of a decoction that radix bupleuri has extracted volatile oil are the dregs of a decoction of radix bupleuri after through wet distillation or supercritical extraction; Extracting solvent is water or any one alcoholic solvent, perhaps the mixed solvent formed by a certain percentage of these solvents; Extracting mode is decoction, reflux, supersound extraction, microwave extraction, high pressure extract.
The method of steps A is in the aforesaid method: get the dregs of a decoction 25 weight parts after radix bupleuri medicinal material or radix bupleuri have extracted volatile oil, adding concentration is 10～90% ethanol 150～300 parts by volume, heating and refluxing extraction 0.5～2 hour, filter, the dregs of a decoction successively extract 2～5 times with same method, united extraction liquid, the dregs of a decoction discard; Extracting solution reclaims ethanol, and getting thick paste is general extractive 2.5～3.5 weight parts.
The preferred method of steps A is in the aforesaid method: get the dregs of a decoction 25 weight parts after radix bupleuri medicinal material or radix bupleuri have extracted volatile oil, adding concentration is 30% ethanol 250 parts by volume, heating and refluxing extraction 1.5 hours, filter, the dregs of a decoction successively extract 3 times with same method, united extraction liquid, and the dregs of a decoction discard; Extracting solution reclaims ethanol, and getting thick paste is general extractive 3.3 weight parts.
In the aforesaid method among the step B macroporous adsorbent resin be in nonpolar, low-pole or the middle polarity any one; Alkali aqueous solution is that the aqueous solution by alkali or salt constituents constitutes, and its concentration is 1～5%.
The method of step B is in the aforesaid method: get general extractive 2.5～3.5 weight parts, add the water dilution of medicinal material amount 3～5 parts by volume, go up any one macroporous adsorptive resins in nonpolar, low-pole or the middle polarity, resin demand is 6～10 times of parts by volume of medicinal material amount, blade diameter length ratio is 1: 5～8, absorption flow velocity 2～4BV/h; Earlier with 1～5% NaOH, KOH or NaCO
3Wash-out 7～15BV, flow velocity 2～5BV/h, water is eluted to effluent liquid for neutral again, follows the ethanol elution 4～8BV with 0～40%, flow velocity 2～5BV/h; Use ethanol elution 5～10BV of 40～100% at last, flow velocity 2～5BV/h collects this part elutriant, reclaims solvent, gets saikoside b
2Crude product 1.0～1.6 weight parts.
The preferred method of step B is in the aforesaid method: get general extractive 3.3 weight parts, add the water dilution of medicinal material amount 4 parts by volume, and last AB-8 type macroporous adsorptive resins, amount of resin is 8 parts by volume of medicinal material amount, blade diameter length ratio is 1: 6, absorption flow velocity 2BV/h; Earlier with 2% NaOH wash-out 9BV, flow velocity 2BV/h, water is eluted to effluent liquid for neutral again, follows the ethanol elution 5BV with 30%, flow velocity 3BV/h; Use 80% ethanol elution 8BV at last, flow velocity 3BV/h collects this part elutriant, reclaims solvent, gets saikoside b
2Crude product 1.5 weight parts.
The method of step C is in the aforesaid method: get saikoside b
2Crude product is admixed 4～6 times of weight parts, 100～200 order silica gel, volatilizes solvent, must mix sample silica gel; Other gets 100～200 order silica gel, 30～60 weight parts, activates 2 hours, wet method dress post, blade diameter length ratio is 1: 5～10, will mix sample silica gel and add capital, with 10～5: 0.01～5: the mixed solvent wash-out of 0.01～0.5 chloroform-methanol-water, collect elutriant, the thin layer inspection is known, and merges and contains saikoside b
2Stream part, reclaim solvent and get saikoside b
2Elaboration;
Get saikoside b
2Elaboration, in 10～0.01: 0.01～10 acetone-water and 10～1: 0.01～9: recrystallization in the mixed solvent of 0.01～0.9 chloroform-methanol-water, filter, drying, saikoside b
2Pure product.
The method of step C is preferably in the aforesaid method: get saikoside b
2Crude product 1.0～1.6 weight parts are admixed 3～5 times of weight parts, 100～200 order silica gel, volatilize solvent, must mix sample silica gel; Other gets 100～200 order silica gel, 40～50 weight parts, 100～105 ℃ activate 2 hours, wet method dress post, blade diameter length ratio is 1: 5～10, to mix sample silica gel and add capital, with 10～5: 0.01～5: the mixed solvent wash-out of 0.01～0.5 chloroform-methanol-water, collect elutriant, the thin layer inspection is known, and merges and contains saikoside b
2Stream part, reclaim solvent and get saikoside b
2Elaboration 0.08～0.16 weight part;
Get saikoside b
2Elaboration 0.8～0.16 weight part, in 10～0.01: 0.01～10 acetone-water or 10～1: 0.01～9: recrystallization in the mixed solvent of 0.01～0.9 chloroform-methanol-water, filter, drying, the saikoside b of 0.05～0.11 weight part
2Pure product.
The further preferred method of step C is in the aforesaid method: get saikoside b
2Crude product 1.5 weight parts are admixed 4 times of weight parts, 100～200 order silica gel, volatilize solvent, must mix sample silica gel; Other gets 100～200 order silica gel, 45 weight parts, and 100～105 ℃ activate 2 hours, wet method dress post, blade diameter length ratio 1: 7 will be mixed sample silica gel and add capital, with the mixed solvent wash-out of 9: 2: 0.2 chloroform-methanol-water, collect elutriant, the thin layer inspection is known, and merges and contains saikoside b
2Stream part, reclaim solvent and get saikoside b
2Elaboration 0.14 weight part.
Get saikoside b
2Elaboration 0.14 weight part, recrystallization in the mixed solvent of 6: 4 acetone-waters or 5: 1: 0.1 chloroform-methanol-water filters, drying, the saikoside b of 0.11 weight part
2Pure product.
Adopt present method to extract preparation radix bupleuri effective constituent saikoside b
2, the extraction process route is simple, and extraction separation purity improves, saikoside b
2Yield improve, be suitable for large-scale industrialization production.
Following experimental example is used to further specify but is not limited to the present invention.
Experimental example 1: saikoside b
2Purity test
Check purity with high performance liquid chromatography.Instrument: Waters 600 high performance liquid chromatographs.Chromatographic column: Hypersil-C18 chromatographic column, 250mm * 4.6mm, 5 μ m (spy of Dalian Erie).Moving phase: acetonitrile-water (30～50: 70～50); Flow velocity: 1.0～1.2mL/min; Column temperature: room temperature; Detect wavelength: 254nm; Sample size: 10 μ l; Retention time (RT): saikoside b
2Be 20～30min; Color atlas record time: 50min.Area normalization method is surveyed purity, saikoside b
2Purity be 98.3%.
Experimental example 2: saikoside b
2Structure determination
Fusing point Yanaco type micro melting point apparatus; Ultraviolet (UV) spectrum day island proper Tianjin UV-260 type ultraviolet-visible pectrophotometer; Infrared (IR) spectrum Perkin-Elmer 983G type Instrument measuring, the KBr compressing tablet; Proton nmr spectra and carbon spectrum (
1HNMR and
13HNMR) use the MERCURY-400 nmr determination; Mass spectrum (MS) is measured with JES-D300 type mass spectrograph.
The saikoside b of above-mentioned process for purification gained
2Pure product are: white granular crystallization, 235～237 ℃ of fusing points.UVλmax(nm)：244，252，262。IRυmax(KBr，cm
-1)：3416(OH)，1640(C＝C)。MS(m/z)：804(M+Na+H)，781(M+1)。
1H-NMR (δ, C
5D
5N): 0.87,0.91,0.98,0.99,1.04,1.67 (each 3H, s, 6 * CH3) (being 6 angular methyl(group) signals in the structure), 6.71 (1H, d, J=10.4Hz, H-11), 5.72 (1H, d, J=10.4Hz, H-12), 5.38 (1H, d, J=7.8Hz, glc H-1), (4.98 1H, d, J=7.8Hz, fuc H-1), 1.43 (3H, d, J=6.2Hz, fuc CH
3).
13C-NMR (δ, C
5D
5N): the parent nucleus signal: 38.4 (C-1), 26.1 (C-2), 81.6 (C-3), 43.8 (C-4), 47.3 (C-5), 18.9 (C-6), 32.5 (C-7), 41.2 (C-8), 54.0 (C-9), 36.7 (C-10), 126.3 (C-11), 126.3 (C-12), 136.2 (C-13), 42.0 (C-14), 32.6 (C-15), 67.9 (C-16), 45.4 (C-17), 133.1 (C-18), 39.1 (C-19), 32.7 (C-20), 35.6 (C-21), 24.6 (C-22), 64.7 (C-23), 13.1 (C-24), 18.4 (C-25), 17.4 (C-26), 21.8 (C-27), 64.7 (C-28), 25.1 (C-29), 32.3 (C-30); Furan sugar signal: 106.0 (C-1 '), 71.8 (C-2 '), 85.5 (C-3 '), 71.9 (C-4 '), 71.1 (C-5 '), 17.2 (C-6 '); Glucose signals: 106.7 (C-1 "), 75.9 (C-2 "), 78.7 (C-3 "), 72.2 (C-4 "), 78.5 (C-5 "), 62.9 (C-6 ").
Above spectral data is consistent with bibliographical information, and structure is as follows:
Experimental example 3 macroporous adsorbent resin enrichment saikoside b
2The experiment of crude product
Get radix bupleuri medicinal material 2.5kg, adding concentration is 30% ethanol 25L, and heating and refluxing extraction 1.5h filters, and the dregs of a decoction successively extract 3 times with same method, last united extraction liquid, and the dregs of a decoction discard, and extracting solution reclaims ethanol, gets thick paste 0.33kg.
Get above-mentioned thick paste 0.33kg, add water 10L dilution, last AB-8 macroporous adsorbent resin, amount of resin 20L (volume in the water), the resin column blade diameter length ratio is 1: 6, absorption flow velocity 2BV/h, earlier with 2% NaOH wash-out 9BV, flow velocity 2BV/h, water is eluted to effluent liquid and is neutrality again, follow ethanol elution 5BV, flow velocity 3BV/h with 30%; Use 80% ethanol elution 8BV at last, flow velocity 3BV/h collects this part elutriant, reclaims solvent, gets saikoside b
2Crude product 0.15kg.
Experimental example 4 column chromatography methods and recrystallization method coupling prepare saikoside b
2The experiment of pure product
Saikoside b
2The method that adopts column chromatography and recrystallization coupling for preparing of pure product prepares.The sorbent material that uses during column chromatography is silica gel G (100～105 ℃ activate 2 hours for Haiyang Chemical Plant, Qingdao's production, 4.5kg).Get above-mentioned saikoside crude product 0.15kg, admix in 0.6kg 100～200 purpose silica gel, volatilize solvent, must mix sample silica gel, sample on the wet method (column diameter height ratio 1: 7).With the mixed solvent wash-out of 9: 2: 0.2 chloroform-methanol-water, collect elutriant, every part of 300mL collects 80 parts altogether, every part be evaporated to small volume after, check with thin-layer chromatography.Thin-layer chromatography condition: reverse phase silica gel precoated plate; Developping agent: 40% methanol/water solution.Saikoside b in the 31st～40 part wherein
2Content higher, with its merging, decompressing and extracting gets saikoside b
2Elaboration white powder 0.014kg.
Get saikoside b
2Elaboration 0.014kg, with small amount of methanol dissolving, recrystallization in the mixed solvent of 5: 1: 0.1 chloroform-methanol-water, filter, drying, the saikoside b of 0.011kg
2Pure product are the white granular crystallization.
Embodiment 1:Saikoside b
2The preparation of pure product
Get radix bupleuri medicinal material 2.5kg, adding concentration is 30% ethanol 20L, and heating and refluxing extraction 2h filters, and the dregs of a decoction successively extract 3 times with same method, last united extraction liquid, and the dregs of a decoction discard, and extracting solution reclaims ethanol, gets thick paste 0.31kg.
Get above-mentioned thick paste 0.31kg, add water 10L dilution, last AB-8 macroporous adsorbent resin, amount of resin 20L (volume in the water), the resin column blade diameter length ratio is 1: 6, absorption flow velocity 2BV/h, earlier with 2% NaOH wash-out 10BV, flow velocity 2BV/h, water is eluted to effluent liquid and is neutrality again, follow ethanol elution 8BV, flow velocity 3BV/h with 25%; Use 90% ethanol elution 6BV at last, flow velocity 3BV/h collects this part elutriant, reclaims solvent, gets saikoside b
2Crude product 0.14kg.
Saikoside b
2The method that adopts column chromatography and recrystallization coupling for preparing of pure product prepares.The sorbent material that uses during column chromatography is silica gel G (100～105 ℃ activate 2 hours for Haiyang Chemical Plant, Qingdao's production, 4.5kg).Get above-mentioned saikoside crude product 0.14kg, admix in 0.55kg 160～200 purpose silica gel, volatilize solvent, must mix sample silica gel, sample on the wet method (column diameter height ratio 1: 6).With the mixed solvent wash-out of 7: 2: 0.2 chloroform-methanol-water, collect elutriant, every part of 250mL collects 80 parts altogether, every part be evaporated to small volume after, check with thin-layer chromatography.Thin-layer chromatography condition: reverse phase silica gel precoated plate; Developping agent: 50% methanol/water solution.Saikoside b in the 25th～35 part wherein
2Content higher, with its merging, decompressing and extracting gets saikoside b
2Elaboration white powder 0.013kg.
Get saikoside b
2Elaboration 0.013kg, with small amount of methanol dissolving, recrystallization in the mixed solvent of 6: 4 acetone-waters, filter, drying, the saikoside b of 0.009kg
2Pure product are the white granular crystallization.
Embodiment 2:Saikoside b
2The preparation of pure product
Get radix bupleuri medicinal material 2.5kg, adding concentration is 20% ethanol 30L, and heating and refluxing extraction 2h filters, and the dregs of a decoction successively extract 4 times with same method, last united extraction liquid, and the dregs of a decoction discard, and extracting solution reclaims ethanol, gets thick paste 0.35kg.
Get above-mentioned thick paste 0.35kg, add water 9L dilution, last AB-8 macroporous adsorbent resin, amount of resin 22.5L (volume in the water), the resin column blade diameter length ratio is 1: 7, absorption flow velocity 2BV/h, earlier with 3% NaOH wash-out 9BV, flow velocity 3BV/h, water is eluted to effluent liquid and is neutrality again, follow ethanol elution 5BV, flow velocity 2BV/h with 35%; Use 70% ethanol elution 10BV at last, flow velocity 2BV/h collects this part elutriant, reclaims solvent, gets saikoside b
2Crude product 0.11kg.
Saikoside b
2The method that adopts column chromatography and recrystallization coupling for preparing of pure product prepares.The sorbent material that uses during column chromatography is silica gel G (100～105 ℃ activate 2 hours for Haiyang Chemical Plant, Qingdao's production, 3.5kg).Get above-mentioned saikoside crude product 0.11kg, admix in 0.44kg 100～200 purpose silica gel, volatilize solvent, must mix sample silica gel, sample on the wet method (column diameter height ratio 1: 5).With the mixed solvent wash-out of 6: 1: 0.1 chloroform-methanol-water, collect elutriant, every part of 300mL, every part be evaporated to small volume after, check with thin-layer chromatography.Thin-layer chromatography condition: reverse phase silica gel precoated plate; Developping agent: 50% methanol/water solution.Saikoside b in the 20th～26 part wherein
2Content higher, with its merging, decompressing and extracting gets saikoside b
2Elaboration white powder 0.010kg.
Get saikoside b
2Elaboration 0.010kg, with small amount of methanol dissolving, recrystallization in the mixed solvent of 5: 1: 0.1 chloroform-methanol-water, filter, drying, the saikoside b of 0.006kg
2Pure product are the white granular crystallization.
Embodiment 3:Saikoside b
2The preparation of pure product
Get the radix bupleuri dregs of a decoction 2.5kg that has carried behind the volatile oil, add concentration and be 50% ethanol 30L, heating and refluxing extraction 1.5h filters, and the dregs of a decoction successively extract 3 times with same method, last united extraction liquid, the dregs of a decoction discard, extracting solution reclaims ethanol, thick paste 0.26kg.
Get above-mentioned thick paste 0.26kg, add water 11L dilution, last AB-8 macroporous adsorbent resin, amount of resin 19L (volume in the water), the resin column blade diameter length ratio is 1: 8, absorption flow velocity 3BV/h, earlier with 3% NaOH wash-out 10BV, flow velocity 3BV/h, water is eluted to effluent liquid and is neutrality again, follow ethanol elution 6BV, flow velocity 3BV/h with 30%; Use 80% ethanol elution 9BV at last, flow velocity 3BV/h collects this part elutriant, reclaims solvent, gets saikoside b
2Crude product 0.10kg.
Saikoside b
2The method that adopts column chromatography and recrystallization coupling for preparing of pure product prepares.The sorbent material that uses during column chromatography is silica gel G (100～105 ℃ activate 2 hours for Haiyang Chemical Plant, Qingdao's production, 3kg).Get above-mentioned saikoside crude product 0.10kg, admix in 0.50kg 100～200 purpose silica gel, volatilize solvent, must mix sample silica gel, sample on the wet method (column diameter height ratio 1: 6).With the mixed solvent wash-out of 9: 4: 0.4 chloroform-methanol-water, collect elutriant, every part of 300mL, every part be evaporated to small volume after, check with thin-layer chromatography.Thin-layer chromatography condition: reverse phase silica gel precoated plate; Developping agent: 45% methanol/water solution.Saikoside b in the 30th～40 part wherein
2Content higher, with its merging, decompressing and extracting gets saikoside b
2Elaboration white powder 0.009kg.
Get saikoside b
2Elaboration 0.009kg, with small amount of methanol dissolving, recrystallization in the mixed solvent of 7: 5 acetone-waters, filter, drying, the saikoside b of 0.006kg
2Pure product are the white granular crystallization.
Embodiment 4:Saikoside b
2The preparation of pure product
Get the radix bupleuri dregs of a decoction 2.5kg that has carried behind the volatile oil, add concentration and be 70% ethanol 25L, heating and refluxing extraction 1h filters, and the dregs of a decoction successively extract 4 times with same method, last united extraction liquid, the dregs of a decoction discard, extracting solution reclaims ethanol, thick paste 0.28kg.
Get above-mentioned thick paste 0.28kg, add water 11L dilution, last AB-8 macroporous adsorbent resin, amount of resin 20L (volume in the water), the resin column blade diameter length ratio is 1: 7, absorption flow velocity 3BV/h, earlier with 2% NaOH wash-out 11BV, flow velocity 3BV/h, water is eluted to effluent liquid and is neutrality again, follow ethanol elution 4BV, flow velocity 3BV/h with 40%; Use 85% ethanol elution 10BV at last, flow velocity 4BV/h collects this part elutriant, reclaims solvent, gets saikoside b
2Crude product 0.12kg.
Saikoside b
2The method that adopts column chromatography and recrystallization coupling for preparing of pure product prepares.The sorbent material that uses during column chromatography is silica gel G (100～105 ℃ activate 2 hours for Haiyang Chemical Plant, Qingdao's production, 3.6kg).Get above-mentioned saikoside crude product 0.12kg, admix in 0.48kg 100～200 purpose silica gel, volatilize solvent, must mix sample silica gel, sample on the wet method (column diameter height ratio 1: 7).With the mixed solvent wash-out of 9: 3: 0.3 chloroform-methanol-water, collect elutriant, every part of 250mL, every part be evaporated to small volume after, check with thin-layer chromatography.Thin-layer chromatography condition: reverse phase silica gel precoated plate; Developping agent: 55% methanol/water solution.Saikoside b in the 25th～35 part wherein
2Content higher, with its merging, decompressing and extracting gets saikoside b
2Elaboration white powder 0.010kg.
Get saikoside b
2Elaboration 0.010kg, with small amount of methanol dissolving, recrystallization in the mixed solvent of 5: 2: 0.2 chloroform-methanol-water, filter, drying, the saikoside b of 0.008kg
2Pure product are the white granular crystallization.
Claims (6)
1. saikoside b
2Extraction and separation method, it is characterized in that comprising the steps:
A: get the dregs of a decoction after radix bupleuri or radix bupleuri have extracted volatile oil, the suitable pulverizing adds solvent and extracts, and extracting solution concentrates and reclaims solvent, gets general extractive;
B: general extractive adds water-dispersion, last macroporous resin adsorption post, earlier with the removal of impurities of alkali aqueous solution wash-out, water is eluted to neutrality again, next remove assorted with 10%～40% ethanol aqueous wash, with 40%～100% aqueous ethanolic solution wash-out and collect this part elutriant, concentrate and reclaim solvent at last, saikoside b
2Crude product;
C: get saikoside b
2Crude product is admixed 2～10 times of weight part silica gel, volatilizes solvent, must mix sample silica gel; Other got 20～80 times of weight part silica gel activatings 2 hours, and wet method dress post will be mixed sample silica gel and add capital, with the mixed solvent wash-out of chloroform-methanol-water, collect elutriant, and the thin layer inspection is known, and merges and contains saikoside b
2Stream part, reclaim solvent and get saikoside b
2Elaboration;
Get saikoside b
2Elaboration, recrystallization in the mixed solvent of acetone-water or chloroform-methanol-water filters, drying, saikoside b
2Pure product.
2. according to the extraction and separation method described in the claim 1, it is characterized in that: radix bupleuri is the root of the arbitrary plant of umbelliferae Bupleurum in the described steps A, or commercially available radix bupleuri medicine materical crude slice, also comprises the various processed products through concocting; It is the dregs of a decoction of radix bupleuri after through wet distillation or supercritical extraction that radix bupleuri has extracted the dregs of a decoction behind the volatile oil; Extracting solvent is water or any one alcoholic solvent, perhaps the mixed solvent formed by a certain percentage of these solvents; Extracting mode is any one in decoction, reflux, supersound extraction, microwave extraction, the high pressure extract.
3. according to the extraction and separation method described in the claim 1, it is characterized in that: macroporous adsorbent resin is any one in nonpolar, low-pole or the middle polarity among the described step B; Alkali aqueous solution is that the aqueous solution by alkali or strong base-weak acid salt constituents constitutes, and its concentration is 1～5%.
4. according to the extraction and separation method described in the claim 1, it is characterized in that: described step C is:
Get saikoside b
2Crude product is admixed 4～6 times of weight parts, 100～200 order silica gel, volatilizes solvent, must mix sample silica gel; Other gets 100～200 order silica gel, 30～60 weight parts, activates 2 hours, wet method dress post, blade diameter length ratio is 1: 5～10, will mix sample silica gel and add capital, with 10～5: 0.01～5: the mixed solvent wash-out of 0.01～0.5 chloroform-methanol-water, collect elutriant, the thin layer inspection is known, and merges and contains saikoside b
2Stream part, reclaim solvent and get saikoside b
2Elaboration;
Get saikoside b
2Elaboration, in 10～0.01: 0.01～10 acetone-water or 10～1: 0.01～9: recrystallization in the mixed solvent of 0.01～0.9 chloroform-methanol-water, filter, drying, saikoside b
2Pure product.
5. according to the extraction and separation method described in the claim 1, it is characterized in that: this method comprises the steps:
A: get the dregs of a decoction 25 weight parts after radix bupleuri medicinal material or radix bupleuri have extracted volatile oil, adding concentration is 10～90% ethanol 150～300 parts by volume, heating and refluxing extraction 0.5～2 hour, filter, the dregs of a decoction successively extract 2～5 times with same method, united extraction liquid, and the dregs of a decoction discard; Extracting solution reclaims ethanol, and getting thick paste is general extractive 2.5～3.5 weight parts;
B: get general extractive 2.5～3.5 weight parts, add the water dilution of 3～5 times of parts by volume of medicinal material amount, go up any one macroporous adsorptive resins in nonpolar, low-pole or the middle polarity, resin demand is 6～10 times of parts by volume of medicinal material amount, blade diameter length ratio is 1: 5～8, absorption flow velocity 2～4BV/h; Earlier with 1～5% NaOH, KOH or NaCO3 wash-out 7～15BV, flow velocity 2～5BV/h, water is eluted to effluent liquid for neutral again, follows the ethanol elution 4～8BV with 10～40%, flow velocity 2～5BV/h; Use ethanol elution 5～10BV of 40～100% at last, flow velocity 2～5BV/h, the ethanol elution part of collection 40～100% reclaims solvent, gets saikoside b
2Crude product 1.0～1.6 weight parts;
C: get saikoside b
2Crude product 1.0～1.6 weight parts are admixed 3～5 times of weight parts, 100～200 order silica gel, volatilize solvent, must mix sample silica gel; Other gets 100～200 order silica gel, 40～50 weight parts, 100～105 ℃ activate 2 hours, wet method dress post, blade diameter length ratio is 1: 5～10, to mix sample silica gel and add capital, with 10～5: 0.01～5: the mixed solvent wash-out of 0.01～0.5 chloroform-methanol-water, collect elutriant, the thin layer inspection is known, and merges and contains saikoside b
2Stream part, reclaim solvent and get saikoside b
2Elaboration 0.08～0.16 weight part;
Get saikoside b
2Elaboration 0.08～0.16 weight part, in 10～0.01: 0.01～10 acetone-water or 10～1: 0.01～9: recrystallization in the mixed solvent of 0.01～0.9 chloroform-methanol-water, filter, drying, the saikoside b of 0.05～0.11 weight part
2Pure product.
6. according to the extraction and separation method described in the claim 1, it is characterized in that: this method comprises the steps:
A: get the dregs of a decoction 25 weight parts after radix bupleuri medicinal material or radix bupleuri have extracted volatile oil, adding concentration is 30% ethanol 250 parts by volume, and heating and refluxing extraction 1.5 hours is filtered, and the dregs of a decoction successively extract 3 times with same method, united extraction liquid, and the dregs of a decoction discard; Extracting solution reclaims ethanol, and getting thick paste is general extractive 3.3 weight parts;
B: get general extractive 3.3 weight parts, add the water dilution of 4 times of parts by volume of medicinal material amount, last AB-8 type macroporous adsorptive resins, resin demand are 8 times of parts by volume of medicinal material amount, and blade diameter length ratio is 1: 6, absorption flow velocity 2BV/h; Earlier with 2% NaOH wash-out 9BV, flow velocity 2BV/h, water is eluted to effluent liquid for neutral again, follows the ethanol elution 5BV with 30%, flow velocity 3BV/h; Use 80% ethanol elution 8BV at last, flow velocity 3BV/h collects this part elutriant, reclaims solvent, gets saikoside b
2Crude product 1.5 weight parts;
C: get saikoside b
2Crude product 1.5 weight parts are admixed 4 times of weight parts, 100～200 order silica gel, volatilize solvent, must mix sample silica gel; Other gets 100～200 order silica gel, 45 weight parts, and 100～105 ℃ activate 2 hours, wet method dress post, blade diameter length ratio 1: 7 will be mixed sample silica gel and add capital, with the mixed solvent wash-out of 9: 2: 0.2 chloroform-methanol-water, collect elutriant, the thin layer inspection is known, and merges and contains saikoside b
2Stream part, reclaim solvent and get saikoside b
2Elaboration 0.14 weight part;
Get saikoside b
2Elaboration 0.14 weight part, recrystallization in the mixed solvent of 6: 4 acetone-waters or 5: 1: 0.1 chloroform-methanol-water filters, drying, the saikoside b of 0.11 weight part
2Pure product.
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| CN106138132A (en) * | 2016-07-19 | 2016-11-23 | 广州中大南沙科技创新产业园有限公司 | A kind of extracting method of saikoside |
| CN106176851A (en) * | 2016-07-22 | 2016-12-07 | 南京正宽医药科技有限公司 | The process of preparing Chinese medicine extracting method of a kind of Radix Bupleuri decoction pieces and Radix Bupleuri compositions |
| CN107385894A (en) * | 2017-08-15 | 2017-11-24 | 苏州市纤维检验所 | A kind of plant base anti-UV fabric finishing agent and its preparation technology |
| CN109908192A (en) * | 2019-04-24 | 2019-06-21 | 漯河医学高等专科学校 | A kind of extracting method of saikoside |
| CN110623970A (en) * | 2019-10-12 | 2019-12-31 | 复旦大学 | Application of saikosaponin B2 in preparing medicine for preventing or treating renal fibrosis |
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| 陈喜奎等.黑柴胡中皂甙成分的研究.北京医科大学学报.1992,24(3),222-224. * |
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