CN1085674C - Safflower total uranidin, its preparation and application - Google Patents
Safflower total uranidin, its preparation and application Download PDFInfo
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- CN1085674C CN1085674C CN99123713A CN99123713A CN1085674C CN 1085674 C CN1085674 C CN 1085674C CN 99123713 A CN99123713 A CN 99123713A CN 99123713 A CN99123713 A CN 99123713A CN 1085674 C CN1085674 C CN 1085674C
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Abstract
The present invention relates to total safflower yellow extracted from a safflower as a Chinese medicinal material, a preparation method for the total safflower yellow, the new application of the total safflower yellow to the field of medical preparation, and medicines and health-care products prepared from the total safflower yellow. The total safflower yellow has the functions of resisting thrombosis and myocardial ischemia, inhibiting platelet aggregation, removing free radicals and inhibiting platelet activation factors, and can treat or prevent various blood circulation dysfunctional diseases of cardiovascular and cerebrovascular diseases, etc.
Description
The present invention relates to a kind of total composition that from the activating blood herbs safflower, prepares and preparation method thereof, the new purposes of this total composition and the medicine and the Halth-care composition of composition thereof.
Chinese medicine safflower is the dried floral of feverfew Carthamus tinctorius L., is a promoting blood flow and remove blood stasis drug commonly used, can be used for coronary heart disease, many disturbance of blood circulation treatment of diseases such as stenocardia.Safflower is used as medicine with decocting liquid clinically, carthamin yellow is the main water soluble component of safflower, be one mix the phenyl styryl ketone glucoside [referring to, Gao Qiming, the pharmacological research overview of Chinese medicine safflower, combination of Chinese tradiational and Western medicine magazine, 1984,4 (12): 758-760.] the Flos Carthami total flavochromes crude product be the food color (GB2760-86) that allow to use of a country [referring to Zhou Liguo, edible natural pigment and extract and use.158-160. Jinan, Shandong science tech publishing house .], studies show that the many-sided cardiovascular pharmacological effect of its tool, but anti-freezing, promote fibrinolytic, antithrombotic forms, microcirculation improvement etc. [referring to. Gao Qiming, the pharmacological research overview of Chinese medicine safflower, combination of Chinese tradiational and Western medicine magazine, 1984,4 (12): 758-760.].The preparation method of this crude product Flos Carthami total flavochromes is a water extraction and alcohol precipitation method, because of its without method removal of impurity such as chromatographies, its thin layer collection of illustrative plates is that a hangover colour band from the initial point to the forward position is [referring to Tianjin institute of Pharmaceutical Industry etc., the pharmacological research of the research of safflower (two) carthamin yellow and isolate thereof. western modern medicine magazine .1980:9 (1) 2-8], point out its purity lower.Flos Carthami total flavochromes only has crude product that water extraction and alcohol precipitation method makes at present as food color production, does not still have Flos Carthami total flavochromes medicine or health-care preparation.
The invention provides a kind of Flos Carthami total flavochromes, this Flos Carthami total flavochromes can make with following method: be raw material with the safflower, add an amount of polar solvent lixiviate, the filtering dregs of a decoction merge vat liquor, concentrate and obtain concentrated solution; The concentrated solution that obtains through above-mentioned steps is carried out column chromatography, remove behind the impurity again with another polarity elutriant wash-out Flos Carthami total flavochromes with polarity elutriant wash-out; Or adsorb extracting concentrated solution with the chromatography carrier, directly washing Flos Carthami total flavochromes and make impurity absorption on the chromatography carrier with the polarity elutriant, the Flos Carthami total flavochromes elutriant reclaims solvent, and concentrate drying is promptly.
The present invention also provides a kind of preparation method of Flos Carthami total flavochromes, and this method comprises the steps: with the safflower to be raw material, adds an amount of polar solvent lixiviate, and the filtering dregs of a decoction merge vat liquor, concentrates and obtains extracting concentrated solution; The extraction concentrated solution that above-mentioned steps is obtained carries out column chromatography, removes behind the impurity again with another polarity elutriant wash-out Flos Carthami total flavochromes with polarity elutriant wash-out; Or adsorb extracting concentrated solution with the chromatography carrier, directly washing Flos Carthami total flavochromes and make impurity absorption on the chromatography carrier with the polarity elutriant, the Flos Carthami total flavochromes elutriant reclaims solvent, and concentrate drying is promptly.
In a preparation method's of the present invention preferred embodiment, described extraction is to extract 2-4 time with polar solvent, and each 1-3 days, extraction was 5-50 times of safflower crude drug amount with the polar solvent consumption.
In another preferred embodiment of preparation method of the present invention, describedly be used for deimpurity safflower and extract concentrated solution concentration and be equivalent to 0.5-10 gram crude drug/milliliter.
In another preferred embodiment of preparation method of the present invention, described column chromatography impurities removing method is to remove safflower with the polyamide column chromatography method to extract impurity in the concentrated solution, and its safflower extracts concentrated solution and used Silon is 1 than by volume: 1-30.
In another preferred embodiment of preparation method of the present invention, described column chromatography carrier impurities removing method is to extract impurity in the concentrated solution with the silica gel adsorption safflower, and its safflower extracts concentrated solution and used silica gel powder is 1 than by volume: 1-30.
In another preferred embodiment of preparation method of the present invention, described column chromatography carrier impurities removing method is with the Flos Carthami total flavochromes in the macroporous resin adsorption safflower extraction concentrated solution, with elutriant wash-out impurity, and its safflower extracts concentrated solution and big pore resin is 1 than by volume: 1-30.
In another preferred embodiment of preparation method of the present invention, described polarity elutriant is that water or concentration are the ethanol of 20-100 weight percent, methyl alcohol or aqueous propanol solution.
In another preferred embodiment of preparation method of the present invention, described polarity elutriant is that water or concentration are the ethanol of 40-95 weight percent, methyl alcohol or aqueous propanol solution.
The invention provides a kind of Flos Carthami total flavochromes and relate to tissue ischemia particularly myocardial ischemia and cerebral ischemia, the application due to thrombosis and platelet activating factor and the free radical in the medicine of damage disease in preparation prevention and treatment.
The invention provides a kind of Flos Carthami total flavochromes and relate to tissue ischemia particularly myocardial ischemia and cerebral ischemia, the application due to thrombosis and platelet activating factor and the free radical in the healthcare products of damage disease in preparation prevention and treatment.
The invention provides the medicine of disturbance of blood circulation diseases such as a kind of treatment and prevention cardiovascular and cerebrovascular diseases, wherein contain the Flos Carthami total flavochromes and the pharmaceutically useful carrier of treatment cardio-cerebrovascular diseases significant quantity.
The invention provides the healthcare products of disturbance of blood circulation diseases such as a kind of treatment and prevention cardiovascular and cerebrovascular diseases, wherein contain the Flos Carthami total flavochromes and the pharmaceutically useful carrier of treatment cardiovascular and cerebrovascular diseases significant quantity.
The present invention is used for preparation treatment and prevention relates to tissue ischemia particularly myocardial ischemia and cerebral ischemia, and the disturbance of blood circulation disease of thrombosis and tool platelet activating factor and radical damage is the medicine and the Halth-care composition of thrombotic diseases and myocardial ischemia disease particularly.It is Flos Carthami total flavochromes and pharmaceutically-acceptable excipients that this composition contains activeconstituents, and carrier or thinner are made into medicine or health-care preparation with ordinary method.
During the preparation said composition, Flos Carthami total flavochromes can be mixed or use carrier embedding, this carrier with carrier can be capsule or other loading forms.Carrier can be solid, semisolid or liquid substance, and it is as the carrier of Flos Carthami total flavochromes, vehicle or medium.Said composition can be a tablet, pill, suppository, powder, spirit, suspension, emulsion, solution, syrup, various ways such as injection.
Variety carrier, vehicle or thinner can be used for this composition.As lactose, glucose, sucrose, sorbyl alcohol, N.F,USP MANNITOL, starch, polyoxyethylene glycol, gelatin etc.Also can add lubricant, wetting agent, correctives, suspension agent, sanitas etc. in the said preparation.Any method preparation that composition of the present invention can use this area to adopt usually, to reach after patient's use, activeconstituents is brought into play the purpose of maximum drug effect.
But this medicine and Halth-care composition oral administration, intravenous injection, intramuscular injection, through skin or subcutaneous injection administration, various administrations such as per rectum.But single or multiple uses continuously, and the pharmaceutical composition using dosage then should generally should be controlled at 0.001-1.0 gram Flos Carthami total flavochromes/kg body weight sky according to patient's particular case decision.
The present invention takes water to carry, and the polyamide column chromatography elution method makes the higher Flos Carthami total flavochromes of a purity after the silica gel adsorption or the macroreticular resin absorbing method removal of impurity, and these methods are not appeared in the newspapers as yet in the application that the safflower composition extracts in the process for preparing.They can remove plurality of impurities effectively, obviously improve the purity of Flos Carthami total flavochromes, three kinds of methods prepare all visible 6-8 fluorescence spot of thin layer collection of illustrative plates of the Flos Carthami total flavochromes of gained, the crude product of water extraction and alcohol precipitation method gained thin layer fluorescence pattern under similarity condition then is the serious colour band of a hangover, rarely seen 2-3 blear spot (seeing accompanying drawing) on it points out Flos Carthami total flavochromes purity of the present invention to be height than crude product.And this method is easy and simple to handle, is easy to grasp, and can be used as the rough purification method that Flos Carthami total flavochromes preparation and safflower water soluble component detect.It is reported the anti-freezing of carthamin yellow tool, promote fibrinolytic, antithrombotic forms, cardiovascular pharmacological effects such as microcirculation improvement, experimental result shows, the Flos Carthami total flavochromes tool that the present invention makes is removed free radical, the antiplatelet incitant, anticoagulant, antithrombotic form, and effect such as resist myocardial ischemia, therefore it can be used for many disturbance of blood circulation treatment of diseases and prevention, be particularly useful for cerebral thrombosis, cerebral ischemia and stenocardia, the treatment and the prevention of tool thrombosis such as myocardial infarction and ischemia illness.
The following drug effect of the Flos Carthami total flavochromes that the present invention makes (abbreviation flavine) tool (used flavine is the preparation of embodiment 4 methods in the following experiment):
(1) flavine suppresses the effect of the platelet aggregation that platelet activating factor brings out
New zealand rabbit, body weight 3-4 kilogram, male and female are not limit.Chronolog type platelet aggregation instrument, platelet activating factor (PAF) is a Sigma company product, other reagent are homemade analytical pure.Get 10 milliliters of tame rabbit ear arteria comitans nervi mediani blood (ACD: whole blood=anti-freezing in 1: 6), 500 rev/mins centrifugal 5 minutes, get platelet rich plasma, 2000 rev/mins centrifugal 5 minutes thrombocyte, no calcium tyrode's solution washed twice with pH6.5, wash at every turn back 2000 rev/mins centrifugal 5 minutes, abandon supernatant, add pH7.4 and contain 0.25% bovine serum albumin 50mM Tris tyrode's solution (TTBSA) and adjust PC to 2 * 10
8/ milliliter is rabbit washing platelet suspension (WRP).Get 0.18 milliliter of WRP, add 0.01 milliliter of Flos Carthami total flavochromes solution after 37 degree (the mentioned temperature scale of this paper is ceslius scale) preheatings, add after 0.01 milliliter of platelet activating factor with turbidimetry record curve of platelet aggregation.The blank pipe replaces flavine solution with distilled water.Compare with flavine pipe platelet aggregation rate and blank pipe platelet aggregation rate, calculate the anticoagulant rate and weigh its drug effect.
The result
PAF concentration flavine concentration (g/L) anticoagulant rate (%)
2nM 0.0 0.0
0.21 28.0
0.42 32.9
0.85 60.0
1.69 79.4
The result shows that flavine suppresses the tangible dose-effect relationship of rabbit washing platelet gathering tool that platelet activating factor is brought out.The flavine that other embodiment method makes is identical with The above results trend according to this law experiment gained data.
(2) flavine suppresses the effect that thrombocyte that platelet activating factor brings out discharges serotonin (5-TH)
The WRP that above method makes, get 0.8 milliliter of WRP, add flavine solution, 37 ℃ of preheatings add 2000 rev/mins of 37 ℃ of reactions of 10 microlitre platelet activating factors 4 minutes after 2 minutes centrifugal 10 minutes, get supernatant liquor and add the TTBSA0.7 milliliter for 0.3 milliliter, it is centrifugal 20 minutes to add 3500 rev/mins of 0.3 milliliter of 100% Tricholroacetic Acids again, get 0.5 milliliter of supernatant liquor, add 2 milliliters of 0.05% o-phthalaldehyde(OPA)/HCl solution, 100 ℃ of heating in water bath 10 minutes, add after the water-cooled behind 2 milliliters of chloroform mixings 3500 rev/mins centrifugal 20 minutes, get 1 milliliter of supernatant measure 5-TH fluorescence (Ex360nm, Em475nm).The negative control pipe replaces flavine solution with distilled water, is reagent blank to replace the WRP pipe with concentration flavine TTBSA diluent.Each reaction tubes measured value all deduct the reagent blank value record 5-TH fluorescent value F (Ex360nm, Em475nm).
The result
PAF flavine concentration (g/L) F (Ex360nm, Em475nm)
2nM 0 354.7
0.103 329.6
0.207 309.0
0.414 263.8
0.828 234.6
The result shows that flavine suppresses the tangible dose-effect relationship of 5-TH release tool that platelet activating factor causes.The flavine that other embodiment method makes is identical with The above results trend according to this law experiment gained data.
(3) flavine suppresses the effect that mouse capillary permeability that platelet activating factor brings out increases
Kunming mouse, male and female are not limit, body weight 20-40 gram, random packet, behind the mouse peritoneal injection flavine solution 30 minutes, 2 milliliters/kilogram of tail vein injection 3% Evan's Blue normal saline solutions, behind the tail vein injection 30 minutes in mouse one rear flank leg quadriceps muscle of thigh injection PAF 0.25% bovine serum albumin normal saline solution to cause the local inflammation reaction, inject with volume physiological saline in same animal opposite side back leg quadriceps muscle of thigh, inject and put to death animal in back 30 minutes, get two back leg muscles respectively and shred, add 2 milliliters of methane amides after every in the leg muscle fragment, 56 ℃ were extracted 24 hours, get supernatant liquor after centrifugal 20 minutes for 3000 rev/mins, is blank in 620nm with every animal physiological brine side back leg extracting solution, measures offside back leg extracting solution absorbancy, weighs the change of mouse capillary permeability with these data.Not damaged blank group is with physiologic saline for substitute PAF, the negative blank of physiologic saline for substitute flavine solution.
The result
In the blank group PAF group low dose group dosage group high dose group PAF dosage-191nM * 0.05ml with left with left with left flavine dosage--0.05g/kg 0.07g/kg 0.100g/kgn 12 12 12 12 12A,620 0.080 ± 0.045 0.456 ± 0.156▲ ▲0.342 ± 0.129* 0.322 ± 0.135* 0.222 ± 0.071*
Compare with the blank group ▲ ▲ * P＜0.05_ is compared with the PAF group in P＜0.01
The above results shows, the flavine abdominal injection, and that can suppress obviously that PAF causes that the local capillary permeability of mouse increases makes the tangible dose-effect relationship of apparatus.The flavine that other embodiment method makes is identical with The above results trend according to this law experiment gained data.
(4) flavine is removed the free radical effect
Detect the effect that flavine is removed hydroxy radical qiao with phenanthroline iron oxidation style, contain phenanthroline 0.75mmol/L in the reaction system, FeSO
40.75mmol/L, 150mmol/L pH7.4 phosphate buffered saline buffer, H
2O
20.01%, add behind the flavine and other reagent add H
2O
2, 37 ℃ of insulations were surveyed A536nm after 60 minutes.With the negative contrast of distilled water, the damage pipe does not add H
2O
2And antioxidant.Being the colorimetric blank with concentration flavine diluent.Hydroxy radical qiao clearance rate meter d algorithm
D=[(A
536 dosings-A
536 damages)/(A
536 not damages-A
536 damages)] * 100%
The result
Flavine concentration (g/L) hydroxy radical qiao clearance rate (%)
0.0 0.0
10.1 53.9
11.0 60.8
11.8 70.0
12.6 68.9
14.3 74.7
The result shows, what flavine was removed hydroxy radical qiao makes the tangible dose-effect relationship of apparatus.The flavine that other embodiment method makes is identical with The above results trend according to this law experiment gained data.
(5) the thrombotic effect of flavine mouse
Male mouse of kunming, body weight 18-22 gram is divided into five groups at random by the body weight equilibrium, i.e. normal control group, the acetylsalicylic acid group, flavine is low, in, high dose group.Fasting is 12 hours before the experiment, be positioned over mouse in the fixer successively in 30 minutes behind the gastric infusion, make its afterbody vertical, after cut off at distance tail point 1.5mm place, pick up counting immediately and, write down the hemorrhage termination time rapidly with 1.5cm place under 37 ℃ of physiological saline liquid levels of its tail point immersion
The result
Grouping dosage (g/kg) bleeding time (second)
Physiological saline----203 ± 169
Acetylsalicylic acid 0.20 500 ± 186
* *
Flavine low dosage 0.10 334 ± 249
*
Dosage 0.26 398 ± 201 in the flavine
*
Flavine high dosage 0.62 461 ± 193
* *
Compare * P＜0.05, * * P＜0.01, * * * P＜0.001 with the physiological saline group
The result shows that the flavine that the present invention makes suppresses the thrombotic tangible dose-effect relationship of apparatus of doing of mouse, and the flavine that other embodiment method makes is identical with The above results trend according to this law experiment gained data.
(6) flavine suppresses the effect that ADP brings out platelet aggregation
New zealand rabbit, body weight 3-4 kilogram, male and female are not limit.Instrument: Chronolog type platelet aggregation instrument, ADP are BM company product, and other reagent are homemade analytical pure.Get tame rabbit ear arteria comitans nervi mediani blood (3.8% Sodium Citrate: whole blood=anti-freezing in 1: 9), 500 rev/mins centrifugal 5 minutes, platelet rich plasma (PRP), centrifugal 10 minutes of 3000 rev/mins of subnatants platelet poor plasma (PPP), adjusting the PRP PC with PPP is 2 * 10
8/ milliliter is got 0.165 milliliter of PRP, adds 0.03 milliliter of flavine solution after 37 degree (the mentioned temperature scale of this paper is ceslius scale) preheatings, adds behind 0.005 milliliter of 1g/L ADP with turbidimetry record curve of platelet aggregation.The blank pipe replaces flavine solution with distilled water.Compare with flavine pipe platelet aggregation rate and blank pipe platelet aggregation rate, calculate the anticoagulant rate.
The result
Drug dose anticoagulant rate (%)
H
2O - 0.0
Acetylsalicylic acid 4.8mmol/L 52.4
Flavine 1.5g/L 14.1
Flavine 2.2g/L 27.8
Flavine 3.3g/L 42.9
Flavine 5.0g/L 56.9
Flavine 7.5g/L 66.9
The result shows that the flavine that the present invention makes suppresses the tangible dose-effect relationship of rabbit platelet aggregation tool that ADP brings out, and the flavine that other embodiment method makes is identical with The above results trend according to this law experiment gained data.
(7) flavine suppresses the effect of the rat heart muscle ischemic that Pituitrin brings out
The male wistar rat, body weight 180-220 gram, screen animal according to laxative remedy before the experiment: rats by intraperitoneal injection urethane 1-1.2g/kg anesthesia, quiet notes Pituitrin 0.5 or 1.0u/kg, select the T ripple to rise more than 0.1 millivolt or descend to surpass 50% responsive animal and be divided into three groups at random, be the physiological saline group, pannonit group and Flos Carthami total uranidin group is carried out pharmacodynamic experiment after 4-6 hour.(quiet notes 0.75u/kg person is 7 to 30 minutes intravenous injection Pituitrins in every group of 15 rats of each experimental group behind medicaments for resisting myocardial ischemia or the physiological saline abdominal injection, quiet notes 1.0u/kg person is 8), before the intravenous injection Pituitrin and quiet notes back 5,10,15,20,30,45s, 1,2,3,5,10,15,20min writes down the II lead electrocardiogram, with behind the quiet notes Pituitrin before the injection T ripple rise more than 1 millivolt or descend that to surpass 50% be that the medicine function of resisting myocardial ischemia is invalid, 1 millivolt of T ripple rising less than or decline less than 50% are that drug effect is effective.
The result
The invalid routine number of the n grouping effective routine number of dosage (mg/kg)
15 physiological saline groups-5 10
15 pannonit groups 5 12
*3
15 flavine groups 500 12
*3
Compare * P＜0.05 with the physiological saline group
The result shows that flavine tool that the present invention makes suppresses the effect of the rat heart muscle ischemic that Pituitrin brings out, and the flavine that other embodiment method makes is identical with The above results trend according to this law experiment gained data.
(8) flavine suppresses the effect of the rat heart muscle ischemic that Racemic isoproterenol brings out
The male wistar rat, body weight 240-260 gram, being divided into two groups at random is physiological saline group and Flos Carthami total uranidin group, abdominal injection urethane 1g/kg anesthesia, 30 minutes pneumoretroperitoneums of Flos Carthami total flavochromes or physiological saline abdominal injection injection Racemic isoproterenol 0.05mg/kg gives behind Flos Carthami total flavochromes or the physiological saline 12 hours first and 24 hours the same manners are respectively given Flos Carthami total flavochromes or physiological saline once.First abdomen annotate different third kidney after 24 hours again with method injection with the dosage Racemic isoproterenol, inject 3min II lead electrocardiogram behind different third kidney with last before different third kidney of record initial injection.Choose five J points calculating J/R values in the electrocardiogram(ECG, average as this point data, carry out own control to reaching last before different third kidney to J/R value behind different third kidney first with every animal, calculate and give different third kidney preceding absolute value that reaches last to the difference of J/R value behind different third kidney first, relatively with the t method of inspection.Giving the variation of different third kidney front and back J/R value, is that myocardial ischemia is obvious with the person of being changed significantly, and changes apparent author and is suppressed for ischemic.
The result
N grouping dosage (g/kg) J/R difference
8 physiological saline groups-0.146 ± 0.126
*
8 Flos Carthami total flavochromes groups 0.75 0.175 ± 0.203
*To comparing P＜0.05 before different third kidney with after giving different third kidney.
The result shows that the Flos Carthami total flavochromes abdominal injection can obviously suppress the rat heart muscle ischemic that different third kidney brings out.The flavine that other embodiment method makes is identical with The above results trend according to this law experiment gained data.
The comparison of the carthamin yellow (being called for short commercially available flavine) that Flos Carthami total flavochromes that the present invention makes (being called for short flavine of the present invention) and commercially available water extraction and alcohol precipitation method make:
(1) flavine of the present invention and commercially available flavine suppress the comparison of the platelet aggregation effect that platelet activating factor brings out
According to this specification sheets aforementioned " effect of the platelet aggregation that flavine inhibition platelet activating factor is brought out " method, prepare WRP in comparing two kinds of flavine effects with once testing with same animal blood, the result is as follows: the commercially available flavine concentration of PAF concentration flavine concentration of the present invention (g/L) (g/L) anticoagulant rate (%)
1nM 0.0 0.0 0.0
5.0 - 16.1
10.0 - 38.7
20.0 - 98.4
10.0 15.9
20.0 20.3
The above results shows that the Flos Carthami total flavochromes drug effect that the present invention makes is apparently higher than commercially available flavine.
(2) flavine acute toxicity test of the present invention
1) gastric infusion experiment
Get 20 of kunming mices, male and female half and half, mouse body weight 18-22 gram is observed the animal situation after the administration, observed record dead animal number of elements continuously seven days.
Gastric infusion group result:
N=20 flavine dosage of the present invention (g/kg) animal dead number (only)
10 0
The result shows, flavine of the present invention is irritated stomach dosage when reaching 10g/kg, none death of mouse.
2) abdominal injection experiment: get 80 of kunming mices, be divided into four groups at random, 20 every group, male and female half and half, mouse body weight 18-22 gram is observed the animal situation after the administration, observed continuously seven days, calculates medium lethal dose (LD50) with each group dead animal rate.
Intraperitoneal administration group result:
N=20 flavine dosage of the present invention (g/kg) animal dead number (only)
8.46 16
7.80 11
7.20 10
6.64 2
Calculate flavine abdominal injection LD50=7.54 ± 0.24g/kg of the present invention
According to the literature, commercially available flavine abdominal injection LD50=5.49g/kg (95% fiducial limit: 5.11-5.59g/kg), gastric infusion LD50=5.53g/kg (95% fiducial limit: 5.39-5.69g/kg), [referring to Huang Zhengliang etc.: the toxicological study of carthamin yellow, Gansu medicine, 1983, (4): 22-24] thus The above results show that the Flos Carthami total flavochromes toxicity that the present invention makes is starkly lower than commercially available flavine.
(3) flavine of the present invention and commercially available flavine thin-layer chromatography result's comparison
See accompanying drawing, tomographic results shows that the more commercially available flavine of Flos Carthami total flavochromes purity that the present invention makes obviously improves.
Embodiment
Following examples are in order to illustrate in greater detail implementation method of the present invention, rather than in order to limit the present invention.Embodiment 1 polymeric amide chromatography legal system is equipped with Flos Carthami total flavochromes:
Get 100 gram safflower crude drug medicinal powder, add 0.5 kg of water room temperature at every turn and soaked 48 hours, stir frequently therebetween, extract twice altogether, merge the water extract twice, remove by filter the dregs of a decoction, the 50-90 degree is evaporated to 200 milliliters and promptly gets the safflower water extracting liquid.Get above-mentioned safflower water extracting liquid, last column volume is 3000 milliliters a polyamide column, be eluted to elutriant Molish reaction and ninhydrin reaction feminine gender with distilled water, get final product to elutriant is colourless with the carthamin yellow that adsorbs on the 40 weight percent ethanol elution polyamide columns.Above-mentioned elutriant reclaims ethanol, and 50-90 degree concentrating under reduced pressure is drying to obtain Flos Carthami total flavochromes.Embodiment 2 polymeric amide chromatography legal systems are equipped with Flos Carthami total flavochromes
Get 100 gram safflower crude drug medicinal powder, add 0.5 kilogram of 20 weight percent methyl alcohol room temperature at every turn and soaked 48 hours, stir frequently therebetween, extract twice altogether, merge extracted twice liquid, remove by filter the dregs of a decoction, reclaim solvent, the 50-90 degree is evaporated to 200 milliliters and promptly gets the safflower water extracting liquid.Get above-mentioned safflower water extracting liquid, last column volume is 3000 milliliters a polyamide column, be eluted to elutriant Molish reaction and ninhydrin reaction feminine gender with distilled water, get final product to elutriant is colourless with the carthamin yellow that adsorbs on the 95 weight percent ethanol elution polyamide columns.Above-mentioned elutriant reclaims ethanol, and 50-90 degree concentrating under reduced pressure is drying to obtain Flos Carthami total flavochromes.Embodiment 3 polymeric amide chromatography legal systems are equipped with Flos Carthami total flavochromes:
Get 100 gram safflower crude drug medicinal powder, add 3000 milliliter of 20 weight percent ethanol room temperature at every turn and soaked 48 hours, stir frequently therebetween, extract twice altogether, merge extracted twice liquid, remove by filter the dregs of a decoction, reclaim ethanol, the 50-90 degree is evaporated to 40 milliliters and promptly gets safflower extraction concentrated solution.Get above-mentioned safflower and extract concentrated solution, last column volume is 1200 milliliters a polyamide column, be eluted to elutriant Molish reaction and ninhydrin reaction feminine gender with distilled water, get final product to elutriant is colourless with the carthamin yellow that adsorbs on the 75 weight percent ethanol elution polyamide columns.Above-mentioned elutriant reclaims ethanol, and 50-90 degree concentrating under reduced pressure is drying to obtain Flos Carthami total flavochromes.Embodiment 4 silica gel adsorptions prepare Flos Carthami total flavochromes
Get 100 gram safflower crude drug medicinal powder, add 0.5 kg of water room temperature at every turn and soaked 48 hours, stir frequently therebetween, extract twice altogether, merge the water extract twice, remove by filter the dregs of a decoction, the 50-90 degree is evaporated to 200 milliliters and promptly gets the safflower water extracting liquid.Get above-mentioned safflower and extract concentrated solution and mix thoroughly with silica-gel powder, promptly gets the Flos Carthami total flavochromes elutriant with the 2000 milliliter of 40 above-mentioned silica gel of weight percent ethanol elution after the 60 degree decompression oven dry with 1000 gram column chromatographies.Get the Flos Carthami total flavochromes elutriant, reclaim ethanol, promptly get Flos Carthami total flavochromes behind the 50-90 degree concentrating under reduced pressure evaporate to dryness.Embodiment 5 silica gel adsorptions prepare Flos Carthami total flavochromes
Get 100 gram safflower crude drug medicinal powder, add 0.5 kilogram of 20 weight percent methyl alcohol room temperature at every turn and soaked 48 hours, stir frequently therebetween, extract twice altogether, merge extracted twice liquid, remove by filter the dregs of a decoction, reclaim solvent, the 50-90 degree is evaporated to 200 milliliters and promptly gets the safflower water extracting liquid.Get above-mentioned safflower and extract concentrated solution and mix thoroughly with silica-gel powder, promptly gets the Flos Carthami total flavochromes elutriant with the 2000 milliliter of 95 above-mentioned silica gel of weight percent ethanol elution after the 60 degree decompression oven dry with 1000 gram column chromatographies.Get the Flos Carthami total flavochromes elutriant, reclaim ethanol, promptly get Flos Carthami total flavochromes behind the 50-90 degree concentrating under reduced pressure evaporate to dryness.Embodiment 6 silica gel adsorptions prepare Flos Carthami total flavochromes
Get 100 gram safflower crude drug medicinal powder, add 3000 milliliter of 20 weight percent ethanol room temperature at every turn and soaked 48 hours, stir frequently therebetween, extract twice altogether, merge extracted twice liquid, remove by filter the dregs of a decoction, reclaim ethanol, the 50-90 degree is evaporated to 40 milliliters and promptly gets safflower extraction concentrated solution.Get above-mentioned safflower and extract concentrated solution and mix thoroughly with silica-gel powder, promptly gets the Flos Carthami total flavochromes elutriant with the 2000 milliliter of 75 above-mentioned silica gel of weight percent ethanol elution after the 60 degree decompression oven dry with 1200 gram column chromatographies.Get the Flos Carthami total flavochromes elutriant, reclaim ethanol, promptly get Flos Carthami total flavochromes behind the 50-90 degree concentrating under reduced pressure evaporate to dryness.Embodiment 7 macroreticular resin absorbing methods prepare Flos Carthami total flavochromes:
Get 100 gram safflower crude drug medicinal powder, add 0.5 kg of water room temperature at every turn and soaked 48 hours, stir frequently therebetween, extract twice altogether, merge the water extract twice, remove by filter the dregs of a decoction, the 50-90 degree is evaporated to 200 milliliters and promptly gets the safflower water extracting liquid.Get above-mentioned safflower water extracting liquid, with volume be that 3000 milliliters macroporous resin is mixed thoroughly, after the 60 degree decompression oven dry, be eluted to elutriant Molish reaction and ninhydrin reaction feminine gender repeatedly with distilled water, get final product to elutriant is colourless with the carthamin yellow that adsorbs on the 40 weight percent ethanol elution macroporous resins.Above-mentioned elutriant reclaims ethanol, and 50-90 degree concentrating under reduced pressure is drying to obtain Flos Carthami total flavochromes.Embodiment 8 macroreticular resin absorbing methods prepare Flos Carthami total flavochromes
Get 100 gram safflower crude drug medicinal powder, add 0.5 kilogram of 20 weight percent methyl alcohol room temperature at every turn and soaked 48 hours, stir frequently therebetween, extract twice altogether, merge extracted twice liquid, remove by filter the dregs of a decoction, reclaim solvent, the 50-90 degree is evaporated to 200 milliliters and promptly gets the safflower water extracting liquid.Get above-mentioned safflower water extracting liquid, with volume be that 3000 milliliters macroporous resin is mixed thoroughly, after the 60 degree decompression oven dry, be eluted to elutriant Molish reaction and ninhydrin reaction feminine gender repeatedly with distilled water, get final product to elutriant is colourless with the carthamin yellow that adsorbs on the 95 weight percent ethanol elution macroporous resins.Above-mentioned elutriant reclaims ethanol, and 50-90 degree concentrating under reduced pressure is drying to obtain Flos Carthami total flavochromes.Embodiment 9 macroreticular resin absorbing methods prepare Flos Carthami total flavochromes
Get 100 gram safflower crude drug medicinal powder, add 3000 milliliter of 20 weight percent ethanol room temperature at every turn and soaked 48 hours, stir frequently therebetween, extract twice altogether, merge extracted twice liquid, remove by filter the dregs of a decoction, reclaim ethanol, the 50-90 degree is evaporated to 40 milliliters and promptly gets safflower extraction concentrated solution.Get above-mentioned safflower and extract concentrated solution, with volume be that 1200 milliliters macroporous resin is mixed thoroughly, after the 60 degree decompression oven dry, be eluted to elutriant Molish reaction and ninhydrin reaction feminine gender repeatedly with distilled water, get final product to elutriant is colourless with the carthamin yellow that adsorbs on the 75 weight percent ethanol elution macroporous resins, above-mentioned elutriant reclaims ethanol, and 50-90 degree concentrating under reduced pressure is drying to obtain Flos Carthami total flavochromes.Embodiment 10 macroreticular resin absorbing methods prepare Flos Carthami total flavochromes:
Get 100 gram safflower crude drug medicinal powder, add 0.5 kg of water room temperature at every turn and soaked 48 hours, stir frequently therebetween, extract twice altogether, merge the water extract twice, remove by filter the dregs of a decoction, the 50-90 degree is evaporated to 200 milliliters and promptly gets the safflower water extracting liquid.Get above-mentioned safflower water extracting liquid, last column volume is 3000 milliliters a macroporous resin column, be eluted to elutriant Molish reaction and ninhydrin reaction feminine gender with distilled water, get final product to elutriant is colourless with the carthamin yellow that adsorbs on the 40 weight percent ethanol elution macroporous resin column.Above-mentioned elutriant reclaims ethanol, and 50-90 degree concentrating under reduced pressure is drying to obtain Flos Carthami total flavochromes.Embodiment 11 macroreticular resin absorbing methods prepare Flos Carthami total flavochromes
Get 100 gram safflower crude drug medicinal powder, add 0.5 kilogram of 20 weight percent methyl alcohol room temperature at every turn and soaked 48 hours, stir frequently therebetween, extract twice altogether, merge extracted twice liquid, remove by filter the dregs of a decoction, reclaim solvent, the 50-90 degree is evaporated to 200 milliliters and promptly gets the safflower water extracting liquid.Get above-mentioned safflower water extracting liquid, last column volume is 3000 milliliters a macroporous resin column, be eluted to elutriant Molish reaction and ninhydrin reaction feminine gender with distilled water, get final product to elutriant is colourless with the carthamin yellow that adsorbs on the 95 weight percent ethanol elution macroporous resin column.Above-mentioned elutriant reclaims ethanol, and 50-90 degree concentrating under reduced pressure is drying to obtain Flos Carthami total flavochromes.Embodiment 12 macroreticular resin absorbing methods prepare Flos Carthami total flavochromes
Get 100 gram safflower crude drug medicinal powder, add 3000 milliliter of 20 weight percent ethanol room temperature at every turn and soaked 48 hours, stir frequently therebetween, extract twice altogether, merge extracted twice liquid, remove by filter the dregs of a decoction, reclaim ethanol, the 50-90 degree is evaporated to 40 milliliters and promptly gets safflower extraction concentrated solution.Get above-mentioned safflower and extract concentrated solution, last column volume is 1200 milliliters a macroporous resin column, be eluted to elutriant Molish reaction and ninhydrin reaction feminine gender with distilled water, get final product to elutriant is colourless with the carthamin yellow that adsorbs on the 75 weight percent ethanol elution macroporous resin column.Above-mentioned elutriant reclaims ethanol, and 50-90 degree concentrating under reduced pressure is drying to obtain Flos Carthami total flavochromes.
Make Flos Carthami total flavochromes with the inventive method and make spendable preparation by following examples method.Example of formulations 1 Flos Carthami total flavochromes oral hard capsule agent
Flos Carthami total flavochromes 100 grams
Dry starch 100 grams
1000 hard capsule of packing into after said components is mixed promptly get Flos Carthami total flavochromes oral hard capsule agent formulation embodiment 2 Flos Carthami total flavochromes injections
Flos Carthami total flavochromes 50 grams
Water for injection is an amount of
1000 milliliters of full doses
Get 50 gram Flos Carthami total flavochromess and add to 1000 milliliters according to a conventional method through can after with the dissolving of an amount of water for injection, step such as seal and make the Flos Carthami total flavochromes injection.Example of formulations 3 Flos Carthami total flavochromes suppositorys
Flos Carthami total flavochromes 500 grams
Semi-synthetic fatty acid glycerine fat is an amount of
Make 1000
Get the heating of semi-synthetic fatty acid glycerine fat and make and melt, suitably the cooling back adds 500 gram carthamin yellow mixings, the system bolt, and cooling promptly gets Flos Carthami total flavochromes suppository.Example of formulations 4 Flos Carthami total flavochromes oral liquids
Flos Carthami total flavochromes 0.5 gram
10 milliliters of distilled water
According to a conventional method through can, step such as seal and produce and promptly get the Flos Carthami total flavochromes oral liquid after the Flos Carthami total flavochromes dissolving.Example of formulations 5 Flos Carthami total flavochromes tablets
Flos Carthami total flavochromes 500 grams
Magnesium Stearate 5 grams
Starch 20 grams
Make 1000
Flos Carthami total flavochromes, behind Magnesium Stearate and the starch mixing according to a conventional method through compressing tablet, dressing, technologies such as bottling promptly get the Flos Carthami total flavochromes tablet.
In addition, in not running counter to spirit of the present invention, can carry out various changes and modifications to given embodiment.By the alternate embodiments that those skilled in the art in the art recognize, they also are included in the scope of claim.
Claims (13)
1. Flos Carthami total flavochromes, this Flos Carthami total flavochromes can prepare with following method
(1) with the safflower be raw material, add an amount of polar solvent lixiviate, the filtering dregs of a decoction merge vat liquor, concentrate and obtain extracting concentrated solution;
(2) the extraction concentrated solution that above-mentioned steps is obtained carries out column chromatography, removes behind the impurity again with another polarity elutriant wash-out Flos Carthami total flavochromes with polarity elutriant wash-out; Or adsorb extracting concentrated solution with the chromatography carrier, directly wash Flos Carthami total flavochromes and make impurity absorption on the chromatography carrier with the polarity elutriant;
(3) collect the Flos Carthami total flavochromes elutriant of removing impurity, reclaim solvent, concentrate drying promptly.
2. the preparation method of a Flos Carthami total flavochromes, this method comprises the step of following order
(1) with the safflower be raw material, add an amount of polar solvent lixiviate, the filtering dregs of a decoction merge vat liquor, concentrate and obtain extracting concentrated solution;
(2) the extraction concentrated solution that above-mentioned steps is obtained carries out column chromatography, removes behind the impurity again with another elutriant wash-out Flos Carthami total flavochromes with polarity elutriant wash-out; Or adsorb extracting concentrated solution with the chromatography carrier, directly wash Flos Carthami total flavochromes and make impurity absorption on the chromatography carrier with the polarity elutriant;
(3) collect the Flos Carthami total flavochromes elutriant of removing impurity, reclaim solvent, concentrate drying promptly.
3. the preparation method of claim 2 is characterized in that described extraction is to extract 2-4 time with polar solvent, and each 1-3 days, extraction was 5-50 times of safflower crude drug weight with an amount of polar solvent consumption.
4. the preparation method of claim 2, it is characterized in that extracting used polar solvent is water or ethanol, methyl alcohol and aqueous propanol solution.
5. the preparation method of claim 2 is characterized in that describedly being used for deimpurity safflower and extracting concentrated solution concentration and be equivalent to 0.1-10 gram crude drug/milliliter.
6. the preparation method of claim 2 is characterized in that removing safflower with the polyamide column chromatography method extracts impurity in the concentrated solution, and its safflower extracts concentrated solution and used Silon is 1 than by volume: 1-30.
7. the preparation method of claim 2 is characterized in that extracting impurity in the concentrated solution with the silica gel adsorption safflower, and its safflower extracts concentrated solution and used silica gel powder is 1 than by volume: 1-30.
8. the preparation method of claim 2 is characterized in that extracting active principle in the concentrated solution with the macroporous resin adsorption safflower, removes impurity, and its safflower to extract concentrated solution be 1 than by volume with used macroporous resin: 1-30.
9. the preparation method of claim 2 is characterized in that described polarity elutriant is that water or concentration are the ethanol of 20-100 weight percent, methyl alcohol or aqueous propanol solution.
10. the preparation method of claim 2 is characterized in that described polarity elutriant is that water or concentration are the ethanol of 40-95 weight percent, methyl alcohol or aqueous propanol solution.
11. medicine and healthcare products for the treatment of cardiovascular and cerebrovascular diseases, the Flos Carthami total flavochromes and the pharmaceutically useful carrier that it is characterized in that containing treatment or prevent the claim 1 of disturbance of blood circulation disease significant quantities such as cardiovascular and cerebrovascular diseases.
12. the medicine of claim 11 and healthcare products is characterized in that making spendable preparations such as capsule, injection, tablet, suppository and oral liquid.
13. the Flos Carthami total flavochromes of claim 1 relates to tissue ischemia particularly myocardial ischemia and cerebral ischemia in preparation prevention and treatment, the medicine of damage disease and the application in the healthcare products due to thrombosis and platelet activating factor and the free radical.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN99123713A CN1085674C (en) | 1998-11-24 | 1999-11-16 | Safflower total uranidin, its preparation and application |
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN981251218 | 1998-11-24 | ||
| CN 98125121 CN1264708A (en) | 1998-11-24 | 1998-11-24 | Total safflower yellow and its preparing process and application |
| CN99123713A CN1085674C (en) | 1998-11-24 | 1999-11-16 | Safflower total uranidin, its preparation and application |
Publications (2)
| Publication Number | Publication Date |
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| CN1272500A CN1272500A (en) | 2000-11-08 |
| CN1085674C true CN1085674C (en) | 2002-05-29 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN99123713A Expired - Lifetime CN1085674C (en) | 1998-11-24 | 1999-11-16 | Safflower total uranidin, its preparation and application |
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Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1300161C (en) * | 2003-09-22 | 2007-02-14 | 浙江永宁制药厂 | Yellow pigment of safflower preparation method and application |
| CN100418522C (en) * | 2004-01-08 | 2008-09-17 | 山东瑞阳制药有限公司 | High-dose hydroxy safflower yellow A or its medically-acceptable salt application for preparing medicine for cerebral apoplexy induced from being ischemic |
| CN1813709B (en) * | 2005-01-31 | 2012-04-18 | 浙江永宁药业股份有限公司 | Safflower yellow pigment dropping pills and its manufacturing method and application |
| CN1935176B (en) * | 2005-09-23 | 2010-07-14 | 北京星昊医药股份有限公司 | Method for preparing red flower extract containing total red flower uranidin |
| CN102657691A (en) * | 2012-06-02 | 2012-09-12 | 悦康药业集团安徽天然制药有限公司 | Extraction process of carthamin yellow |
| CN103446020B (en) * | 2013-09-02 | 2015-03-04 | 上海应用技术学院 | Cosmetics pigment composition containing safflower natural pigment extract and preparation method thereof |
| CN106189352B (en) * | 2016-07-19 | 2018-06-26 | 广州中大南沙科技创新产业园有限公司 | A kind of extracting method of carthamin yellow |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH09296124A (en) * | 1996-03-04 | 1997-11-18 | Toyo Ink Mfg Co Ltd | Separation of yellow safflower pigment and sugar from safflower extract |
| JPH10306224A (en) * | 1997-05-08 | 1998-11-17 | Toyo Ink Mfg Co Ltd | Production of safflower yellow pigment |
-
1999
- 1999-11-16 CN CN99123713A patent/CN1085674C/en not_active Expired - Lifetime
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH09296124A (en) * | 1996-03-04 | 1997-11-18 | Toyo Ink Mfg Co Ltd | Separation of yellow safflower pigment and sugar from safflower extract |
| JPH10306224A (en) * | 1997-05-08 | 1998-11-17 | Toyo Ink Mfg Co Ltd | Production of safflower yellow pigment |
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| CN1272500A (en) | 2000-11-08 |
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Assignee: Shanxi Huahui Kaide Pharmaceutical Co., Ltd. Assignor: Beijing-City Inst. of Cardiopulmonary Vascular Diseases Contract fulfillment period: 2009.8.27 to 2019.11.15 Contract record no.: 2009110000217 Denomination of invention: Total safflower yellow and its preparing process and application Granted publication date: 20020529 License type: Exclusive license Record date: 2009.10.9 |
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Free format text: EXCLUSIVE LICENSE; TIME LIMIT OF IMPLEMENTING CONTACT: 2009.8.27 TO 2019.11.15; CHANGE OF CONTRACT Name of requester: SHANXI HUAHUI KAIDE PHARMACEUTICAL CO.,LTD. Effective date: 20091009 |
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