CN102643258A - Method for separating and purifying chemical components in Herba Gueldenstaedtia - Google Patents
Method for separating and purifying chemical components in Herba Gueldenstaedtia Download PDFInfo
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Abstract
The invention discloses a method for separating and purifying such five compounds as 7,2'-dihydroxy-4'-methyl-isoflavone, onocerin, maackiain and the like. The invention separates 7,2'-dihydroxy-4'-methyl-isoflavone, maackiain, onocerin, onocol-7-O-beta-D-glucoside and adenosin from Herba Gueldenstaedtia for the first time; and the purities of the compounds are respectively higher than 98%. By using the low-price Herba Gueldenstaedtia as the raw material, the method disclosed by the invention can greatly lower the cost of the raw material, provides a new extraction source for the five compounds, and also provides a new method for preparing various standard compounds.
Description
Technical field
The invention provides the separation purification method of chemical ingredients in the cold medicine of skin.
Background technology
Onocol is claimed Formononetin again, and neochanin is given birth to the peaceful B of former buddhist, the pula brain, and Daidezin 4 ′ – methyl ethers are flavones ingredient.Modern study shows that onocol has estrogen-like effects, and antioxygenation can obviously promote the non-specific immunity system, and anticancer cytosis also has reducing blood-fat, effects such as diuresis.Onocol derives from red clover, the thorny restharrow at first, and the later stage is found gradually that this compounds also can separate and obtains from other plants such as the Radix Astragali, Flower of Beltleaf Primrose.Tan Rihong etc. are raw material with Resorcinol and homoanisic acid, the synthetic onocol (Tan Rihong, etc., the separation of onocol and study on the synthesis, chemical research and application, 200,921 5 phases of volume) that obtained.Onocol-7-O-β-D-glucoside is one of common glycosides derivatives of onocol, is more common in the plants such as the Radix Astragali Ren Xinyi etc.; Through onocol, 1-bromo-2,3,4; 6-O-is tetra-acetylated-and α-D-Glucopyranose carries out condensation reaction and obtains onocol-7-O-β-D-glucoside (Ren Xinyi is etc., the study on the synthesis of onocol-7-O-β-D-glucoside; The Tianjin chemical industry, 2006 20 2 phases of volume).
Maackiain is claimed horse cacaine, Maackia amurensis Rupr et Maxim English, Albizzia kalkora element again, is flavones ingredient.Modern study shows that maackiain has anti-microbial effect, and antitumor action is inhibited to the cAMP phosphodiesterase.At present, maackiain is still mostly is from plants such as Koryo Chinese scholartree, root of subprostrate sophora, to separate to obtain.
Adenosine, promptly 9-β-D-ribofuranosyl VITAMIN B4 is a kind of important medicine intermediate, utilizes adenosine to can be made into many medicines such as tetracycline, vidarabine etc.At present, mostly the preparation of adenosine is with chemical synthesis is main, as being that starting raw material carries out the synthetic of adenosine by inosine.
7,2 '-dihydroxyl-4 '-methyl-NOVASOY 400, be isoflavonoid, have certain biological activity, isoflavonoid can be used for the treatment of cardiovascular disorder more.At present also rarely seen 7,2 '-dihydroxyl-4 '-separation and purification and the compound method of methyl-NOVASOY 400.
Above-claimed cpd all has a good application prospect, and in order to reduce the production cost of these compounds, minimizing is depended on unduly existing resource, and the researchist is also seeking more raw materials source and preparation method.
The cold medicine of skin is the traditional herbal medicine of the peaceful river valley treatment of Sichuan Panxi Diqu flu; The local common people are called " skin typhoid fever ", " Flaccid Coelogyne " again; Derive from the dry herb of Yunnan, pulse family Gueldenstaedtia verna platymiscium river Gueldenstaedtia verna Gueldenstaedtia delavayi Franch.; All herbal medicine, the four seasons can adopt; Cold in nature, flavor is arduous, clearing heat and detoxicating, analgesia." skin tremble with fear medicine " speech beginning is shown in " Xichang herbal medicine ", claims that in this book it derives from the herb of Papilionaceae Gueldenstaedtia verna platymiscium health Yunnan Gueldenstaedtia verna Amblytropsis delavayi (Fr.) C.Y.Wu.Health Yunnan Gueldenstaedtia verna is Yunnan, the river Gueldenstaedtia verna Gueldenstaedtia delavayi Franch. that " Chinese Plants will " is recorded.But the cold medicine of skin is not same plant with " Chinese medicine voluminous dictionary " and the contained Gueldenstaedtia verna Gueldenstaedtia multiflora Bgun. of traditional Chinese medical science universities and colleges Chinese materia medica teaching material.
So far, less to the research report of the cold medicine of skin both at home and abroad, the document record is also detailed inadequately.In the research of the cold medicine treatment flu to skin, He Dezhao sums up and has verified Yi Autonomous Prefecture of Liangshan, Sichuan Province use among the people " skin tremble with fear medicine "---be the experience of health Yunnan Gueldenstaedtia verna treatment flu, add the dried orange peel treatment with the cold medicine of skin and meeting the flu diagnosis and patient's 72 examples of catching a cold; Observe its clinical efficacy, result's 46 examples of fully recovering account for 63.89%; Produce effects 21 examples account for 29.17%, effective 5 examples; Account for 6.94% (He Dezhao; State, Liangshan Mountain traditional herbal medicine " skin tremble with fear medicine " adds dried orange peel treatment flu 72 examples, Chengdu University of Traditional Chinese Medicine's journal, 2004 27 3 phases of volume).
At present, also do not see the report of each chemical ingredients in the cold medicine of skin, do not see the method for from the cold medicine of skin, separating above-claimed cpd yet.
Summary of the invention
The object of the present invention is to provide the method for the multiple compound of separation and purification from the cold medicine of skin.
The invention provides 7,2 '-dihydroxyl-4 '-separation purification method of methyl-NOVASOY 400, it comprises following operation steps:
(1) get the cold medicine of skin, extracting in water, the water extract is used ethyl acetate extraction after concentrating, and gets the ethyl acetate extraction layer, behind the recovery solvent, gets the cold medicine ethyl acetate extract of skin;
(2) get the cold medicine ethyl acetate extract of skin, last silicagel column adopts the gradient elution mode; After methylene chloride-methanol=1:0V/V wash-out removal of impurities, use methylene chloride-methanol=20:1V/V wash-out again, collect 48 parts of elutriants; Every part is 1/4 column volume, promptly gets the A1-48 flow point;
(3) merge the A1-35 flow point, after concentrating, last silicagel column with sherwood oil-acetone=6:1V/V wash-out, is collected elutriant, and again through the VISOSE column chromatography, washed with methanol is collected 25 parts of elutriants altogether, and every part is 1/85 column volume, promptly gets the B1-25 flow point;
(4) get the B3-7 flow point, reclaim solvent after, with recrystallizing methanol, obtain 7,2 '-dihydroxyl-4 '-methyl-NOVASOY 400.
Further, the said silicagel column of step (2) is the column chromatography silica gel post, and wherein, silica gel is the 200-300 order;
The described silicagel column of step (3) is a thin layer silica gel G post, and the VISOSE post is a Sephadex G-25 post.
Further, the blade diameter length ratio of the said silicagel column of step (2) is 1:7-9, is preferably 1:8.
The blade diameter length ratio of the described silicagel column of step (3) is 1:13-15, is preferably 1:14, and the blade diameter length ratio of VISOSE post is 1:28-32, is preferably 1:30-31.
The present invention also provides the separation purification method of maackiain, and it comprises following operation steps:
Get the B1-25 flow point of aforementioned preparation, merge the B10-21 flow point, behind the recovery solvent,, obtain maackiain with recrystallizing methanol.
The present invention also provides the separation purification method of onocol, and it comprises following operation steps:
Get the A1-48 flow point of aforementioned preparation, merge the A36-41 flow point, after concentrating; Last silicagel column after methylene chloride-methanol=20:1V/V wash-out removal of impurities, carries out wash-out with methylene chloride-methanol=16:1V/V again; Collect elutriant; Reclaim solvent, crystallization and filtration, filter solids, obtain onocol through the ETHYLE ACETATE purifying.
Further, said silicagel column is a thin layer silica gel G post, and its blade diameter length ratio is 1:15-18, is preferably 16-17.
The present invention also provides the separation purification method of onocol-7-O-β-D-glucoside, and it comprises following operation steps:
A, get the cold medicine of skin, extracting in water, the water extract use ethyl acetate extraction after concentrating, and gets the ethyl acetate extraction layer, reclaim solvent after, the skin medicine ethyl acetate extract of trembling with fear;
B, get the cold medicine ethyl acetate extract of skin, last silicagel column, employing gradient elution mode; Earlier with after methylene chloride-methanol=1:0V/V wash-out removal of impurities; With the methylene chloride-methanol=20:1V/V wash-out of 12 times of column volumes, use methylene chloride-methanol=15:1V/V wash-out more earlier, 62 parts of the elutriants of collection methylene chloride-methanol=15:1; Every part is 1/4 column volume, promptly gets the C1-62 flow point;
C, get the C14-31 flow point, after concentrating, last silicagel column is after the removal of impurities of methylene chloride-methanol 12:1V/V wash-out; Carry out wash-out with 10:1V/V again, collect elutriant, filter; Filtrating is through the VISOSE column chromatography, and washed with methanol is collected meoh eluate; After reclaiming solvent, recrystallizing methanol obtains onocol-7-O-β-D-glucoside.
Further, the said silicagel column of step B is the column chromatography silica gel post, and blade diameter length ratio is 1:7-9, and wherein, silica gel is the 200-300 order;
The described silicagel column of step C is a thin layer silica gel G post, and blade diameter length ratio is 1:12-14; The VISOSE post is a Sephadex G-25 post, and blade diameter length ratio is 1:22-24.
The present invention also provides the separation purification method of adenosine, and it comprises following operation steps:
1., get the cold medicine of skin, extracting in water, the water extract use ethyl acetate extraction after concentrating, and gets the ethyl acetate extraction layer, reclaim solvent after, the skin medicine ethyl acetate extract of trembling with fear;
2., get the cold medicine ethyl acetate extract of skin, last silicagel column, employing gradient elution mode; After methylene chloride-methanol=1:0V/V wash-out removal of impurities; With the methylene chloride-methanol=20:1V/V wash-out of 12 times of column volumes, use methylene chloride-methanol=15:1V/V wash-out more earlier, 62 parts of the elutriants of collection methylene chloride-methanol=15:1; Every part is 1/4 column volume, promptly gets the C1-62 flow point; Use methylene chloride-methanol=7:1V/V wash-out again, collect 10 parts of elutriants, every part is 1/4 column volume, promptly gets the D1-10 flow point;
3., merge C53-62 flow point, D1-10 flow point, after concentrating, last silicagel column, with methylene chloride-methanol=10:1V/V wash-out, the collection elutriant behind the recovery solvent, through re-crystallizing in ethyl acetate, obtains adenosine.
Further, the 2. said silicagel column of step is the column chromatography silica gel post, and blade diameter length ratio is 1:7-9, and wherein, silica gel is the 200-300 order;
The 3. said silicagel column of step is a thin layer silica gel G post, and blade diameter length ratio is 1:12-14.
The present invention has isolated 7 first from the cold medicine of skin; 2 '-dihydroxyl-4 '-methyl-NOVASOY 400, maackiain, onocol, onocol-7-O-β-D-glucoside and 5 kinds of compounds of adenosine; The gained compound purity is all more than 98%, and the inventive method is raw material with the cold medicine of cheap skin, can significantly reduce raw materials cost; For above-mentioned 5 kinds of compounds provide new extraction source, also novel method is provided for the preparation of all cpds standard substance.
Description of drawings
The extraction flow process of Fig. 1 ethyl acetate extract
The separation of Fig. 2 ethyl acetate extract, purge process
Embodiment
The extraction separation and the structure of the cold medicine chemical ingredients of embodiment 1 skin are identified
1 instrument, reagent
Superconduction high resolution NMR spectrometer: Bruker Avance 600 probe:13C-1H DUL TE:300K
High-resolution mass spectrometer: Micromass Zabspec
Column chromatography filler: silica gel H (200～300 order), Haiyang Chemical Plant, Qingdao;
Chromatographic silica gel version: HPTLC plate, Haiyang Chemical Plant, Qingdao;
Thin-layer chromatography (TLC) silica gel: silica gel G, silica gel G F-254, Haiyang Chemical Plant, Qingdao;
Chromatographic solvent system: sherwood oil: acetone (different ratios); Chloroform: acetone (different ratios); Methylene dichloride: methyl alcohol (different ratios); Methylene dichloride: ETHYLE ACETATE (different ratios);
TLC developer: 10% sulfuric acid ethanol liquid; Iodine vapor; Uv lamp (254nm, 365nm);
Solvent for use is analytical pure, is produced by the Long Huagongshijichang of Chengdu section;
2 vegetable materials
The cold medicine herb of skin picks up from casual arm town, Mianning County, state, the Liangshan Mountain in April, 2008, identifies through professor Jiang Guihua of pharmaceutical college of Chengdu University of Traditional Chinese Medicine, is the dry herb of Yunnan, pulse family Gueldenstaedtia verna platymiscium river Gueldenstaedtia verna Gueldenstaedtia delavayi Franch..Sheet is existing in pharmaceutical college of Chengdu University of Traditional Chinese Medicine sample shop.
The preparation of 3 water extracts
The dry herb 12kg of the cold medicine of skin is cut into segment, decocting 3 times (10 times of water gagings of medicinal material/inferior, 30min/ time), and collecting decoction is concentrated into 30L.
The extraction of 4 ethyl acetate extracts
Water extract gradation (each 2000ml) places separating funnel (5000ml); With 2000ml/ extracted several times of ETHYLE ACETATE, the combined ethyl acetate extraction liquid is with the Rotary Evaporators concentrating under reduced pressure; Decompression and solvent recovery to medicinal extract shape; Be transferred in the furnace pot of constant weight, put again that (50 ℃) dry by the fire to dry-powdered in the vacuum drying apparatus, weigh the heavy 64.1g of ethyl acetate extract dry powder.The dry powder cryopreservation is treated the column chromatography for separation use.
It extracts flow process and sees Fig. 1.
The sepn process of 5 ethyl acetate extracts
Ethyl acetate extract (64.1g), with the silica gel column chromatography initial gross separation, 2500g (200-300 order) silica gel dress post (diameter 10cm, length 80cm); Use methylene chloride-methanol (1:0,20:1,15:1 successively; 10:1,7:1) gradient elution is collected elutriant; Collect 169 parts of flow points altogether, wherein, methylene chloride-methanol 1:0 is that 1-49 part, methylene chloride-methanol 20:1 are that 50-97 part, methylene chloride-methanol 15:1 are that 98-159 part, methylene chloride-methanol 7:1 are 160-169 part; Every part is 500ml, detects through TLC to merge identical flow point, is able to down flow point altogether;
(1) collects 50-84 flow point (18.4g), after the thin layer silica gel G is mixed appearance, through silica gel in atmosphere pressure post (diameter 2.5cm, length 35cm repeatedly; The thin layer silica gel G) flash chromatography, sherwood oil-acetone 6:1 carries out wash-out, collects elutriant, again through sephadex column Sephadex G-25 (50-100 order) 2.6*80cm chromatography; With washed with methanol, every 5ml collects a, collects 25 parts altogether, merges the 3-7 flow point; After reclaiming solvent,, promptly get compound IV (12mg) with recrystallizing methanol; Merge the 10-21 flow point, behind the recovery solvent,, promptly get compound V (20.4mg) with recrystallizing methanol;
(2) collect 85-90 flow point (4.9g), after the thin layer silica gel G is mixed appearance, through silica gel in atmosphere pressure post (diameter 1.5cm repeatedly; Length 25cm; The thin layer silica gel G) flash chromatography carries out gradient elution with methylene chloride-methanol 20:1,16:1 successively, collects the 16:1 elutriant; Reclaim solvent, crystallization and filtration, filter solids and obtain compound VIII (25mg) through the ETHYLE ACETATE purifying.
(3) collect 111-128 flow point (6.3g), after the thin layer silica gel G is mixed appearance, through silica gel in atmosphere pressure post (diameter 2cm, length 25cm; The thin layer silica gel G) flash chromatography carries out gradient elution with methylene chloride-methanol 12:1,10:1 successively, merges the 10:1 elutriant, filters; Filtrating is through sephadex column Sephadex G-25 (50-100 order) 2.6*60cm chromatography, and methanol-eluted fractions is collected elutriant; After reclaiming solvent, recrystallizing methanol obtains compound IX (15mg).
(4) collect 150-169 flow point (6.7g), after the thin layer silica gel G is mixed appearance, through silica gel in atmosphere pressure post (diameter 1.5cm; Length 20cm; The thin layer silica gel G) flash chromatography, methylene chloride-methanol 10:1 carries out wash-out, collects elutriant; After reclaiming solvent, after re-crystallizing in ethyl acetate, obtain compound X (20mg).
Its separation and purification process is seen Fig. 2.
2.6 the structure of compound is identified
Through the application of systematic solvent extraction and plurality of color spectral technology, separate to obtain 5 compounds from the ethyl acetate extract of the cold medicine herb of skin.Through physics and chemistry constant measuring and spectroscopic analysis, identify the structure of 5 compounds temporarily.
Compound IV white needle (methyl alcohol), mp195～197 ℃;
(c 0.10, methyl alcohol).Iron trichloride-Tripotassium iron hexacyanide reaction is positive.1H?NMR:δ：3.56(1H，m，H-3)，3.63（3H，s，OCH3-4′），3.78（1H，t，J=10.26Hz，H-2a），4.29（1H，dd，J=10.98,5.16Hz，H-2a），5.62（1H，d，J=7.02Hz，H-4），6.37（1H，d，J=2.4Hz，H-8），6.56（1H，dd，J=2.22,8.04Hz，H-5′），6.67（1H，d，J=2.22Hz，H-3′），6.86（1H，d，J=2.22Hz，H-8），6.94（1H，dd，J=8.10,2.58Hz，H-6），7.20（1H，d，J=8.40Hz，H-6′）,7.58（1H，d，J=8.40Hz，H-5）。13C?NMR：δ162.6（C-4′），δ162.4（C-2′），δ161.5（C-7），δ158.4（C-9），δ133.8（C-5），δ126.3（C-,6′），δ121.0（C-1′），δ112.9（C-10），δ111.9（C-6），δ107.5（C-5′），δ105.2（C-8），δ98.2（C-3′），δ80.2（C-4）,δ67.7（C-2），δ41.0（C-3）,δ56.3（C-OCH3）。Through HSQC, HHCOSY, DEPT spectrum its nuclear magnetic data has been carried out accurate ownership, be accredited as 7,2 '-dihydroxy-4 '-methoxy-isoflavanol (7,2 '-dihydroxyl-4 '-methyl-NOVASOY 400).Its structure is seen formula 1.Through measuring, the purity of this compound is 98.10%.
The structure of formula 1 compound IV
The colourless needle of compound V (methyl alcohol, pyridine); The FeCl3 coupling reaction is positive, and uv lamp (254nm) is red-purple blackening, mp180～181 ℃ ［ 23 ］ down; 1H NMR: δ 7.53 (1H, d, J=8.82Hz, H-1), 6.90 (1H, dd, J=8.46,2.16Hz, H-2), 6.84 (1H, s, H-7); δ 6.83 (1H, d, J=2.22Hz, H-4), 6.64 (1H, s, H-10), 5.91 (1H, d, J=1.14Hz ,-OCH2O-aH); 5.88 (1H, d, J=1.14Hz ,-OCH2O-bH), 5.57 (1H, d, J=7.32Hz; H-11a, 4.25 (1H, dd, J=11.04,4.74Hz, H-6 β), 3.77 (1H; T, J=10.62Hz, H-6 α), 3.48 (1H, m, H-6a) .13C NMR: δ 40.5 (C-6a), 66.5 (C-6); 79.0 (C-11a), 93.7 (C-10), 101.5 (OCH2-O), 104.0 (C-4), 105.3 (C-7), 110.7 (C-2), 111.8 (C-11b); 118.7 (C-6b), 132.5 (C-1), 141.9 (C-8), 148.3 (C-9), 154.7 (C-10a), 157.3 (C-4a), 160.4 (C-3).MS:m/z：307[M+Na],m/z：285[M+H]。Infer that according to 1H NMR and 13C NMR signal this compound is a known compound, promptly maackiain (the horse cacaine, maackiain).Its structure is seen formula 2.Through measuring, the purity of this compound is 99.95%.
The structure of formula 2 compound V
Compound IX white, needle-shaped crystals (methyl alcohol) is soluble in methyl alcohol, mp217～219 ℃ .1H NMR: δ: 8.42 (1H, s, H-2), 8.04 (1H, d, J=8.76Hz, H-5); 7.52 (2H, d, J=8.40Hz, H-2 ', 6 '), 7.23 (1H, s, H-8), 7.13 (1H; D, J=8.82Hz, H-6), 6.98 (2H, d, J=8.40Hz, H-3 ', 5 '); 5.09 (1H, d, J=6.96Hz, glc H-1 "), 3.30 (3H, s, OCH3). δ 4.5 ~ 5.5: on the sugar-OH, on δ 3 ~ 4 sugar-H.13C?NMR：δ175.1（C-4），δ161.9（C-7），δ159.5（C-4′），δ157.5（C-9），δ154.1（C-2），δ130.5（C-2′,6′），δ127.4（C-5），δ124.5（C-1′），δ123.9（C-3），δ119.0（C-10），δ116.1（C-6），δ114.1（C-3′5′），δ103.9（C-8），δ100.5（C-1"），δ77.7（C-3"），δ77.0（C-5"），δ73.6（C-2"），δ70.1（C-4"），δ61.1（C-6"），δ55.6（C-OCH3）。1H NMR and 13C NMR data all with 4 '-methoxyl group NOVASOY 400-7-O-β-D-glucoside, i.e. onocol-7-O-β-D-glucoside (basically identical of formononetin-7-O-β-D-glucoside).Its structure is seen formula 3.Through measuring, the purity of this compound is 98.92%.
The structure of formula 3 compound IX
Compound VIII white needle (methyl alcohol), mp257～258 ℃; Iron trichloride-Tripotassium iron hexacyanide coupling reaction is positive.
1H NMR: δ: 8.30 (1H, s, H-2), 7.95 (1H, dd, J=8.76,2.22Hz, H-5); 7.49 (2H, d, J=8.40Hz, H-2 ', 6 '), 6.96 (1H, d, J=8.76Hz; H-3 ', 5 '), 6.92 (1H, dd, J=8.76,2.22Hz, H-6); 6.85 (2H, d, J=2.22Hz, H-8), 3.77 (3H, s, OCH3) .13C NMR: δ 175.1 (C-4); δ 163.0 (C-4 '), δ 159.4 (C-7), δ 157.9 (C-9), δ 153.8 (C-3), δ 130.5 (C-2 ', 6 '), δ 127.8 (C-5); δ 124.7 (C-1 '), δ 123.6 (C-2), δ 117.1 (C-10), δ 115.6 (C-6), δ 114.1 (C-3 ' 5 '), δ 102.6 (C-8) .MS:m/z:291 [M+Na], m/z:269 [M+H]. is onocol with its nuclear magnetic data with contrast authenticating compound VIII.Its structure is seen formula 4.Through measuring, the purity of this compound is 99.94%.
The structure of formula 4 compound VIII
Compound X white needle; Be dissolved in methyl-sulphoxide; 1H NMR: δ: 8.32 (1H, s, H-8), 8.12 (1H, s, H-2), 7.30 (2H, s; H-NH2), 5.86 (1H, d, J=6.24Hz, H-1 '), 5.00 ~ 5.50 (3H, OH-H), 4.59 (1H; M, H-2 '), 4.12 (1H, m, H-3 '), 3.94 (1H, m; H-4 '), 3.65 (1H, m, H-5 '), 3.53 (1H, m, H-5 ').13C?NMR：δ156.6（C-6），δ152.8（C-2），δ149.6（C-4），δ140.4（C-8），δ119.8（C-5），δ88.4（C-1′），δ86.4（C-4′），δ73.9（C-2′），δ71.1（C-3′），δ62.2（C-5′）。MS：m/z：255[M+Na],m/z：237[M+H]。Confirm that the compound X is 9-(β-D-ribofuranosyl)-Adenosine (adenosine).Its structure is seen formula 5.Through measuring, the purity of this compound is 98.75%.
The structure of formula 5 compound X
The present invention has isolated 7 first from the cold medicine of skin; 2 '-dihydroxyl-4 '-methyl-NOVASOY 400, maackiain, onocol, onocol-7-O-β-D-glucoside and 5 kinds of compounds of adenosine; The gained compound purity is all more than 98%, and the inventive method is raw material with the cold medicine of cheap skin, can significantly reduce raw materials cost; For above-mentioned 5 kinds of compounds provide new extraction source, also novel method is provided for the preparation of all cpds standard substance.
Claims (10)
1.7,2 '-dihydroxyl-4 '-separation purification method of methyl-NOVASOY 400, it is characterized in that: it comprises following operation steps:
(1) get the cold medicine of skin, extracting in water, the water extract is used ethyl acetate extraction after concentrating, and gets the ethyl acetate extraction layer, behind the recovery solvent, gets the cold medicine ethyl acetate extract of skin;
(2) get the cold medicine ethyl acetate extract of skin, last silicagel column adopts the gradient elution mode; After methylene chloride-methanol=1:0V/V wash-out removal of impurities, use methylene chloride-methanol=20:1V/V wash-out again, collect 48 parts of elutriants; Every part is 1/4 column volume, promptly gets the A1-48 flow point;
(3) merge the A1-35 flow point, after concentrating, last silicagel column with sherwood oil-acetone=6:1V/V wash-out, is collected elutriant, and again through the VISOSE column chromatography, washed with methanol is collected 25 parts of elutriants altogether, and every part is 1/85 column volume, promptly gets the B1-25 flow point;
(4) get the B3-7 flow point, reclaim solvent after, with recrystallizing methanol, obtain 7,2 '-dihydroxyl-4 '-methyl-NOVASOY 400.
2. separation purification method according to claim 1 is characterized in that: the said silicagel column of step (2) is the column chromatography silica gel post, and wherein, silica gel is the 200-300 order;
The described silicagel column of step (3) is a thin layer silica gel G post, and the VISOSE post is a Sephadex G-25 post.
3. separation purification method according to claim 2 is characterized in that: the blade diameter length ratio of the said silicagel column of step (2) is 1:7-9;
The blade diameter length ratio of the described silicagel column of step (3) is 1:13-15, and the blade diameter length ratio of VISOSE post is 1:28-32.
4. the separation purification method of maackiain, it is characterized in that: it comprises following operation steps:
The weighting profit requires the B1-25 flow point of 1 preparation, merges the B10-21 flow point, behind the recovery solvent, with recrystallizing methanol, obtains maackiain.
5. the separation purification method of onocol, it is characterized in that: it comprises following operation steps:
The weighting profit requires the A1-48 flow point of 1 preparation, merges the A36-41 flow point, after concentrating; Last silicagel column after methylene chloride-methanol=20:1V/V wash-out removal of impurities, carries out wash-out with methylene chloride-methanol=16:1V/V again; Collect elutriant; Reclaim solvent, crystallization and filtration, filter solids, obtain onocol through the ETHYLE ACETATE purifying.
6. the separation purification method of onocol according to claim 5, it is characterized in that: said silicagel column is a thin layer silica gel G post, and its blade diameter length ratio is 1:15-18.
7. the separation purification method of onocol-7-O-β-D-glucoside, it is characterized in that: it comprises following operation steps:
A, get the cold medicine of skin, extracting in water, the water extract use ethyl acetate extraction after concentrating, and gets the ethyl acetate extraction layer, reclaim solvent after, the skin medicine ethyl acetate extract of trembling with fear;
B, get the cold medicine ethyl acetate extract of skin, last silicagel column, employing gradient elution mode; Earlier with after methylene chloride-methanol=1:0V/V wash-out removal of impurities; With the methylene chloride-methanol=20:1V/V wash-out of 12 times of column volumes, use methylene chloride-methanol=15:1V/V wash-out more earlier, 62 parts of the elutriants of collection methylene chloride-methanol=15:1; Every part is 1/4 column volume, promptly gets the C1-62 flow point;
C, get the C14-31 flow point, after concentrating, last silicagel column is after the removal of impurities of methylene chloride-methanol 12:1V/V wash-out; Carry out wash-out with 10:1V/V again, collect elutriant, filter; Filtrating is through the VISOSE column chromatography, and washed with methanol is collected meoh eluate; After reclaiming solvent, recrystallizing methanol obtains onocol-7-O-β-D-glucoside.
8. the separation purification method of onocol according to claim 7-7-O-β-D-glucoside is characterized in that: the said silicagel column of step B is the column chromatography silica gel post, and blade diameter length ratio is 1:7-9, and wherein, silica gel is the 200-300 order;
The described silicagel column of step C is a thin layer silica gel G post, and blade diameter length ratio is 1:12-14; The VISOSE post is a Sephadex G-25 post, and blade diameter length ratio is 1:22-24.
9. the separation purification method of adenosine, it is characterized in that: it comprises following operation steps:
1., get the cold medicine of skin, extracting in water, the water extract use ethyl acetate extraction after concentrating, and gets the ethyl acetate extraction layer, reclaim solvent after, the skin medicine ethyl acetate extract of trembling with fear;
2., get the cold medicine ethyl acetate extract of skin, last silicagel column, employing gradient elution mode; After methylene chloride-methanol=1:0V/V wash-out removal of impurities; With the methylene chloride-methanol=20:1V/V wash-out of 12 times of column volumes, use methylene chloride-methanol=15:1V/V wash-out more earlier, 62 parts of the elutriants of collection methylene chloride-methanol=15:1; Every part is 1/4 column volume, promptly gets the C1-62 flow point; Use methylene chloride-methanol=7:1V/V wash-out again, collect 10 parts of elutriants, every part is 1/4 column volume, promptly gets the D1-10 flow point;
3., merge C53-62 flow point, D1-10 flow point, after concentrating, last silicagel column, with methylene chloride-methanol=10:1V/V wash-out, the collection elutriant behind the recovery solvent, through re-crystallizing in ethyl acetate, obtains adenosine.
10. the separation purification method of adenosine according to claim 9, it is characterized in that: the 2. said silicagel column of step is the column chromatography silica gel post, and blade diameter length ratio is 1:7-9, wherein, silica gel is the 200-300 order;
The 3. said silicagel column of step is a thin layer silica gel G post, and blade diameter length ratio is 1:12-14.
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| CN107296825A (en) * | 2017-08-25 | 2017-10-27 | 成都中医药大学 | A kind of skin is trembled with fear medicine medicinal extract or its pharmaceutical composition and preparation method thereof |
| CN107519232A (en) * | 2017-10-16 | 2017-12-29 | 铜仁职业技术学院 | One kind extraction Gueldenstaedtia verna extractive of general flavone and preparation method thereof |
| CN115300497A (en) * | 2021-05-08 | 2022-11-08 | 四川大学 | Application of Korean sophoricoside in preparing medicine for inhibiting tumor metastasis |
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| CN101690752B (en) * | 2008-06-06 | 2013-04-10 | 广西灵峰药业有限公司 | Quality control method of medicine composition |
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| Title |
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| 《成都中医药大学学报》 20040930 何德昭 凉山州民间草药"皮寒药"加陈皮治疗感冒72例 11-12 第27卷, 第3期 * |
| 何德昭: "凉山州民间草药"皮寒药"加陈皮治疗感冒72例", 《成都中医药大学学报》 * |
| 王军宪等: "米口袋的化学成分研究", 《西北植物学报》 * |
| 韩毅丽等: "米口袋化学成分的研究", 《山西中医学院学报》 * |
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| CN102641324B (en) | 2014-04-02 |
| CN102645497B (en) | 2013-08-28 |
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| CN102643258B (en) | 2014-03-19 |
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