CN1935176B - Preparation method of safflower extract containing total safflower yellow - Google Patents

Abstract

The present invention relates to a preparation method of safflower extract, comprising: leaching and extracting safflower with at least 2 times the water temperature to obtain extract I; Elute with alcohol, collect the eluted part of alcohol, recover the eluent until the alcohol content is below 25%, and obtain eluent II; ultrafiltrate through a ceramic membrane to obtain ultrafiltrate III; recover the solvent from the ultrafiltrate under reduced pressure to a thick paste obtained after drying; the extract is characterized by the total safflower yellow content>80%, the hydroxyl safflower yellow A content>20%, and the kaempferol-3-O-rutinoside content>0.6%.

Description

Preparation method of safflower extract containing total safflower yellow

technical field

The present invention relates to a preparation method of safflower extract, specifically, the safflower extract containing total safflower yellow pigment and its preparation method, as well as the pharmaceutical composition containing the extract and its application.

Background technique

The traditional Chinese medicine safflower is the dried flower of Carthamus tinctorius L., a plant of Compositae. It is a traditional medicine for promoting blood circulation and removing blood stasis. It has a good curative effect on many blood circulation disorders and diseases. At the same time, the total safflower yellow is widely used as a food coloring because of its good coloring ability and strong stability. Since the 1970s, the research on the chemical components of the traditional Chinese medicine safflower has been very in-depth, and its active ingredients have been determined to be water-soluble total safflower yellow, and its main effective monomer components are hydroxy safflower yellow A and so on. There are many reports on the production and preparation methods of total safflower yellow pigment. The early commonly used method is mainly water extraction and alcohol precipitation, and the extract is obtained after drying the extract [Zhou Yufang et al., Extraction of active components of safflower, Journal of Xinjiang Institute of Technology, 1997 , 18(4):270-272]; in recent years, it has been extracted with water or dilute alcohol or heated, and the extract is purified by polyamide or macroporous adsorption resin, and the extract is obtained after drying [see CN 1101424C; CN 1085674C]. The above two patents use polar solvent extraction, the extract is simply treated and then separated and purified with an adsorbent to absorb the active ingredients, and the extract is obtained after drying. This method improves the quality of the safflower extract to a certain extent, but due to The composition of the active ingredients and the amount of impurities in the extract prepared by this method are not clear, resulting in poor stability and safety of the preparation developed by it, especially not suitable for injections. With the clinical application of traditional Chinese medicine injections There are many reports of adverse reactions caused by traditional Chinese medicine injections [Mo Binbin et al., Statistical analysis of literature on adverse reactions of traditional Chinese medicine injections, Zhongnan Pharmacy, 2003, 1(3): 184-185], most of which are due to the use of traditional The extraction process and the degree of refinement are insufficient, so its drug safety is a concern and concern in the industry. How to develop a mature preparation process, so that the active ingredients and content of safflower extract are clear, high in purity, and less in impurities, and at the same time, polymer macromolecules and pyrogens can be removed, and the safflower extract containing total safflower yellow pigment can be improved. The stability of the preparation of the extract, so that it can be directly used in injections, thereby ensuring the safety of safflower preparations, especially injections, has become a problem that people in the industry are working hard to solve.

Contents of the invention

The present inventor obtained the active ingredient of safflower extract through a large number of experimental studies, that is, an extract containing total safflower yellow pigment (also known as safflower yellow pigment). The product is obtained through refining, wherein the content of total safflower yellow is very high, which solves the defect of low active ingredients in the prior art, achieves the purpose of clarifying the composition of the active ingredients of safflower, and improves its content, thereby improving the Bioavailability, increases the therapeutic effect.

The object of the present invention is to provide the preparation method of the above-mentioned safflower extract containing total safflower yellow pigment, which method includes the extraction and refining process, especially the technology of adopting HPD600 type or HP-20 type macroporous adsorption resin and ceramic membrane ultrafiltration , so that the content of active ingredients in the safflower extract is high and the impurities are less, which is more conducive to providing its application in preparations.

The purpose of the present invention is also to provide a safflower extract, which is obtained by conventional extraction and separation and refined by ceramic membrane ultrafiltration. The extract has clear components and is more suitable for direct use in the injection of safflower extract.

The invention provides a preparation method of safflower extract, which comprises: immersing Chinese herbal medicine safflower with at least 2 times the water temperature to obtain an extract; Concentrated alcohol (concentration is about 20-50%) is eluted, the alcohol eluted part is collected, and the eluate is recovered until the alcohol content is below 20%; ultrafiltration is performed through a ceramic membrane, and the ultrafiltrate is dried.

The advantage of using warm extraction of safflower medicinal materials in the method of the present invention is that some active ingredients contained in safflower are incompletely extracted during cold soaking (cold water immersion extraction method), and may be oxidized or decomposed under the situation of heating and refluxing extraction. However, the active ingredients are destroyed, and the screening experiment of initial extraction also proves that the effect of warm extraction is better than that of cold soaking and hot extraction. The extract obtained by 2-14 times of water temperature extraction not only improves the production of active ingredients with the help of the HPD600 or HP-20 macroporous adsorption resin and ultrafiltration membrane (ceramic membrane) separation technology suitable for safflower extract after screening and optimization rate and content, and effectively remove polymeric macromolecules and pyrogens in the safflower extract, so that it can be safely and effectively used in injection formulations.

In the above method, the extraction temperature of warm immersion is 40-90°C, preferably 50-80°C, most preferably 65-75°C; wherein the separation step includes first washing with 1-2 column bed volumes, and the diameter-to-height ratio of the column 1:6-1:10, to remove water-soluble impurities such as polysaccharides, discard the water, and then elute with low concentration (20%-50%, preferably 30%) alcohol for 3-5 column bed volumes, collect the eluted The liquid is recovered until the alcohol concentration is lower than 20%; the molecular weight cut-off of the ceramic membrane is preferably 10,000, and if the yield and other factors are considered comprehensively, it can also be 60-50,000, preferably 1-30,000, so as to improve the purity and yield, and more effectively remove polymeric macromolecules and pyrogens in the safflower extract.

There are many types of membrane ultrafiltration used in the field of extraction and purification of traditional Chinese medicine, such as polysulfone organic membranes, polyamide membranes, cellulose acetate membranes, ceramic membranes, tetrafluoroethylene membranes, etc., as long as they limit the same molecular weight cut-off range in theory Ultrafiltration membrane, its ultrafiltration effect should be the same. However, during the practical operation of the refined ultrafiltration of the safflower extract, the applicant found that for the preparation of injection preparations containing total safflower yellow, the ultrafiltration effect of the ceramic membrane is the best, and the ultrafiltration effect of the membrane is the best. Stability, acid and alkali resistance, permeability and other indicators have good performance. For filtering the viscous traditional Chinese medicine extract with high content of total safflower yellow pigment, the corresponding loss is greatly reduced, and the operability is strong. It is superior to the effect achieved by other membrane ultrafiltration. The comparative experimental results for the same amount of medicinal materials and the same molecular weight cut-off are as follows:

The present invention also screens the type and model of the adsorption resin, and the specific screening method is: take different types of resins such as HPD600, HPD100, Dianion HP-20, and 860021 type, and first perform pretreatment, get 1L of adsorption resin to pack, and set aside Put the safflower extract (wherein the content of hydroxysafflower flavin A is 7.628mg/ml) on the column, check whether it is saturated by TLC, first wash with water, then elute with different concentrations of ethanol, and collect the residual liquid of the column respectively , water washing solution, and ethanol eluting solution, measure the content of hydroxysafflower yellow A in it, and calculate the relevant characteristic parameters, the results are shown in Table 7.

Table 7 Characteristic parameters of different types of resin adsorption purification

From the results in Table 7, it can be seen that with hydroxysafflower yellow A as the index component, the safflower extract has different adsorption and elution effects on different resins, among which the HPD100 resin has a smaller amount than the column; HPD600, HP-20 , 860021 type resin has larger specific loading capacity and specific adsorption capacity, but when 860021 is eluted with 30% ethanol, the specific elution capacity is small, that is, the sample loss is serious. HP-20 and HPD 600 resins have relatively large specific loading capacity and large specific elution capacity, and have good adsorption and elution performance. Therefore, compared with the viscous safflower extract of the extract, HP-20 and HPD 600 resins are preferably used for separation and purification, more preferably HPD 600 resins.

Through experiments, it is determined that the loading conditions are as follows:

Diameter-to-height ratio: 1:6-1:10, preferably 1:8

Adsorption temperature: room temperature (<35°C)

Medicinal solution concentration: equivalent to 80-130g crude drug/liter, preferably 110g/L.

The above-mentioned extracted and refined extract can be dried to obtain the solid extract prepared by the method of the present invention. The drying method can be any conventional drying method, preferably spray drying.

Compared with the preparation method disclosed in the prior art, the preparation method of the present invention shows obvious differences in the active ingredients contained in the obtained extract due to the different specific schemes adopted in the separation and purification steps, such as those prepared in patents 01131268.8 and 03157479.3 The extract has been refined by column chromatography for many times, the content of hydroxysafflor yellow A is greater than 70, and other yellow pigment compounds are removed as impurities, and the research results of the present invention show that the content of hydroxy safflower yellow A If the content is too high, the curative effect and activity are not as good as the index of total safflower yellow pigment. Therefore, the extract prepared by the method of the present invention has better clinical application value.

The innovation of the present invention is that by applying specific macroporous adsorption resin and ceramic membrane separation technology, the content of total safflower yellow in the safflower extract is improved, and at the same time, the preparation (especially the injection type) containing the extract The stability is greatly improved, the polymeric macromolecules and pyrogens in the plant extract are effectively removed, and the injection can be directly prepared, and the safety of the preparation is effectively guaranteed.

The present invention also provides a safflower extract, which is obtained by the above preparation method. After testing, the total safflower yellow content in the extract is at least 60%, preferably more than 80% (taken as hydroxyl red Anthocyanin A), wherein the content of hydroxysafflower yellow A is at least 15%, preferably more than 20%.

Since the active ingredient total safflower yellow contained in the extract of the present invention is a mixture, the effective compounds contained in it include hydroxysafflower yellow A, hydroxyl safflower yellow B, kaempferol- 3-O-rutinoside, quercetin-3-O-glucoside, etc., so far there is no literature publicly reporting what is the reasonable composition of the active ingredients of the traditional Chinese medicine safflower in the treatment of cardiovascular and cerebrovascular diseases, and what is the optimal treatment amount, so it is bound to It leads to defective characteristics such as difficult to grasp the dose-effect relationship and unstable curative effect, and its clinical experiment effect is not very satisfactory, especially for traditional Chinese medicine injection dosage forms, it is easier to grasp the quality of the preparation with exact composition and accurate content. The applicant found that the exact curative effect compound in the extract obtained by the preparation method of the present invention is (but not only) the therapeutically effective amount of hydroxysafflor flavin A, even other components other than hydroxy safflower flavin A (such as kaempferol-3-O-rutinoside, quercetin-3-O-glucoside) also play a role in advancing the treatment, so the present invention has carried out a large number of experimental tests, and the results confirm that the total A safflower extract with a safflower yellow content of at least 60%, in which hydroxysafflor yellow A is at least 15%, and a kaempferol-3-O-rutinoside content of at least 0.8% has a therapeutic effect of The best, significantly better than the safflower extract prepared by the method of the reference literature.

The method for determining the content of the above-mentioned total safflower yellow pigment is an ultraviolet absorptiometry, and the operation steps are:

Accurately weigh an appropriate amount of hydroxysafflower flavin A reference substance, and add water to make a reference solution. When measuring, take an appropriate amount of the extract prepared by the above-mentioned method, accurately weigh it, add water to dissolve as the test solution, and use water as a blank according to spectrophotometry (Appendix VA Spectrophotometry of Chinese Pharmacopoeia 2000 Edition). Measure the absorbance of the reference substance and the sample at a wavelength of 332nm, and calculate the amount equivalent to hydroxysafflower yellow A in the test solution according to the external standard method, to obtain final product.

The content determination of hydroxysafflor yellow A adopts high-performance liquid chromatography to determine, and the specific operation steps are as follows:

Octadecylsilane-bonded silica gel is used as filler; mobile phase A: 0.05 mol/L sodium dihydrogen phosphate solution (phosphoric acid adjusts the pH value to 4.0). Mobile Phase B: Acetonitrile. The flow rate is 1.0ml/min, and the detection wavelength is 403nm. The separation degree of hydroxysafflor yellow A and other impurity peaks should meet the requirements, and the number of theoretical plates should not be less than 3000 based on the calculation of hydroxy safflower yellow A peak.

Preparation of the reference substance solution: Accurately weigh an appropriate amount of the reference substance hydroxysafflower flavin A, dissolve it in water and dilute to an appropriate concentration as the reference substance solution.

Preparation of the test solution: take an appropriate amount of the extract, dissolve it in water and dilute it to an appropriate concentration.

Determination method: Accurately draw 20 μl each of the reference substance solution and the test solution respectively, inject it into a liquid chromatograph for determination, and calculate it by the peak area according to the external standard method.

The determination of kaempferol-3-O-rutinoside content is determined by high-performance liquid chromatography, and the operation steps are:

Octadecylsilane bonded silica gel is used as filler; mobile phase A: 0.05mol/L phosphoric acid. Mobile Phase B: Acetonitrile. The flow rate is 1.0ml/min, and the detection wavelength is 365nm. The separation degree of kaempferol-3-O-rutinoside and other impurity peaks should meet the requirements, and the number of theoretical plates should not be less than 3000, calculated according to the peak of kaempferol-3-O-rutinoside.

Preparation of the reference substance solution: Accurately weigh an appropriate amount of the reference substance kaempferol-3-O-rutinoside, add water to dissolve and dilute to an appropriate concentration as the reference substance solution.

Preparation of the test solution: take an appropriate amount of the extract, dissolve it in water and dilute it to an appropriate concentration.

Determination method: Accurately draw 20 μl each of the reference substance solution and the test solution respectively, inject it into a liquid chromatograph for determination, and calculate it by the peak area according to the external standard method.

The present invention also provides a pharmaceutical composition, which contains a clinically effective dose of safflower (safflower yellow) extract and conventional pharmaceutical auxiliary materials, and the clinically recommended effective dose of the medicine is 60-240 mg/day.

The above-mentioned pharmaceutical composition includes oral dosage forms and injection dosage forms, preferably injections, including large infusion solutions, freeze-dried powder injections and small water injections. Among the above drugs, as an auxiliary component used in conjunction with the active ingredient, at least one of mannitol, glucose, dextran or sodium chloride is recommended to be selected according to different specific injections. For example, for freeze-dried powder injections, conventional freeze-drying methods can be used, with water as the solvent, and the extract is added to an appropriate amount of scaffold to freeze-dry. It is recommended to use at least one of mannitol, glucose, and dextran as an auxiliary agent. The stent agent is composed together with the extract; wherein mannitol is preferably used as the stent agent. For drugs in the form of isotonic infusion preparations that can be used directly, at least one of mannitol, glucose, and sodium chloride can be selected as an auxiliary component; among them, sodium chloride is preferably used as an auxiliary component. When preparing small injection injections and large infusion preparations, antioxidants can be added, and antioxidants can be ethylenediaminetetraacetic acid, ethylenediaminetetraacetic acid disodium salt, ascorbic acid, sodium ascorbate, sodium thiosulfate and sodium metabisulfite one or more of them.

The present invention also provides the application of the pharmaceutical composition containing the safflower extract in the preparation of medicines for treating cardiovascular and cerebrovascular diseases.

The drug effect test result of safflower yellow pigment extract of the present invention is as follows:

(1) Study on anti-myocardial hypoxia effect of safflower yellow intravenous administration

1. Effects on the disappearance time of the tracheal ECG in mice:

60 test mice were randomly divided into 6 groups, 10 male and half males in each group. The test mice were anesthetized with urethane (1.2g/kg) intraperitoneally, fixed, and the front of the neck was surgically exposed and freed from the trachea. The MS·302 multimedia biological signal recording and analysis system was connected to the limbs of the mice through needle electrodes, and the electrocardiogram of the standard II lead was collected, and the tail vein was punctured for administration. The first group and the control group were injected with normal saline. The 2nd, 3rd, and 4th groups are safflower yellow pigment high, medium and low dosage groups, and the dosages are 120mg/kg, 60mg/kg, and 30mg/kg respectively; the 5th group is the safflower yellow pigment oral administration group, and the dosage is 60mg/kg, intragastric administration 30 minutes before closing the trachea. The administration volume of each group is 0.2ml/10g, the administration speed is 0.02ml/s, and the required concentration is prepared with 0.9% NaCl before the test. Immediately after the injection, the trachea was closed with an arteriole clip, and the time from closing the trachea to the disappearance of the electrocardiogram was recorded for each mouse in each group. The significance of the test data was processed by the t-test of paired data. The test results are shown in Table 1. In the selected dose range, the intravenous injection of safflower yellow significantly prolongs the disappearance time of electrocardiogram compared with the control group. There is a very significant difference; There was no significant difference between the trends and the control group.

Table 1 Effect of intravenous injection of safflower yellow on the disappearance time of electrocardiogram in mice with closed trachea

Each group compared with the control group ^* P<0.05 ^** P<0.01

2. Effects on the tolerance to hypoxia in mice:

Contain soda lime (Beipiao Mining Bureau Calcium Hydroxide Factory, 991001), seal the bottle mouth with vaseline, adjust the CY-2 oxygen measuring instrument, connect it to the airtight container, and record the oxygen content (%) in the container Change and record the death time (min) of the mice. The significance of the test data is processed with the t-test of the paired data. The test results are shown in Table 2. Within the selected dosage range, the intravenous injection of safflower yellow significantly reduces oxygen consumption and prolongs the survival time of mice. There were significant and very significant differences compared with the control group. The safflower yellow 60mg/kg dose group was given by intragastric administration, which had certain effects on reducing the oxygen consumption and prolonging the survival time of the tested mice, but there was no significant difference from the control group.

Table 2 The effect of intravenous injection of safflower yellow on the oxygen consumption (%) and survival time (min) of mice

Comparing each administration group with the control group ^* P<0.05 ^** P<0.01

(2) The effect of safflower yellow on anti-platelet aggregation and thrombosis

1. Effect on ADP-induced platelet aggregation:

60 male Wister rats were randomly divided into 6 groups, the control group was intravenously injected with 0.9% NaCl safflower yellow pigment, the doses were 80mg/kg, 40mg/kg, 20mg/kg, and the safflower yellow pigment was administered by intragastric administration The dosage of each group is 40mg/kg, and each group of medicines is prepared with 0.9% sodium chloride before administration, and the volume of intravenous injection is 5ml/kg, and the injection speed is 2.0ml/min. The volume of intragastric administration was 20ml/kg, administered once a day for 7 consecutive days, and 1 hour after the last administration, blood was collected from the retroorbital venous plexus and anticoagulated with 0.38% sodium citrate (1:9). Anticoagulant blood was centrifuged at 600 rpm for 10 minutes to prepare platelet-rich plasma (PRP) and the remaining plasma was prepared at 3000 rpm for 15 minutes to prepare platelet-poor plasma (PPP).

Preheat the PAM-3 dual-channel platelet aggregation instrument for 30 minutes. When the pointer of the machine thermometer points to 37°C, use a micropipette to transfer 200 μl of PPP into the clean cuvette, stir and heat for 5 minutes, adjust the PPP potentiometer to make the pointer of the recorder point to 20, take out the cuvette, and then take 200 μl of the same blood sample PRP, pour it into the turbidimetric cup, and heat it up to 37°C. Adjust the PRP potentiometer to make the pointer of the recorder point to 80. At this time, add 20 μl of newly prepared ADP working solution (2.2 μM) into the PRP, observe the change of the pointer of the recorder, trace the platelet aggregation curve, and calculate the platelet aggregation rate (%) , converted the platelet aggregation inhibition rate, and carried out comparative significance determination. The test results showed that the administration of the selected dose 7 days in advance had a significant inhibitory effect on the ADP-induced platelet aggregation rate, and there was a significant or very significant difference between each administration group and the control group. Significant differences. See Table 3 for details.

n＝10)Table 3 Effect of intravenous injection of safflower yellow on ADP-induced platelet aggregation in rats (

n=10)

Compared with the control group ^* P <0.05, ^** P <0.01

2. Effects on platelet-dependent platelet formation

36 rabbits (male body weight 2.0-3.0kg) were randomly divided into 6 groups, 6 in each group, the control group, and 3 dosage groups of safflower yellow intravenously injected, and the dosages were 44mg/kg, 22mg/kg, 11mg/kg; in the safflower yellow group, the dosage was 22mg/kg, and in the positive control drug compound Danshen injection group, the dosage was 1500mg/kg. The control group was intravenously injected with 0.9% NaCl, and each drug group was prepared with 0.9% NaCl before administration, and the volume of intravenous injection was 2ml/kg. The volume of intragastric administration is 5ml/kg. Administer once a day for 7 consecutive days. After the last administration, inject anesthesia with pentobarbital sodium at 30 mg/kg into the marginal ear vein.

The dorsal position of the tested animals was fixed, the front of the neck was skin-prepared, operated, and the common carotid artery was freed. A No. 1 surgical silk thread was inserted into one side of the common carotid artery with a small fine needle for about 3 cm. After being stored in the artery for 2 hours, the carotid artery was cut out. Common artery, take out thrombus, weigh wet weight after removing silk thread, take the thrombus wet weight as observation index, carry out intergroup comparison, significant measurement, test result shows, in the selected dosage range, administering 7 days in advance has the greatest effect on rabbit cervical total Arterial thrombosis has obvious inhibitory effect, and there are significant or very significant differences between each administration group and the control group, see Table 4 for details.

Table 4 The effect of intravenous administration of safflower yellow on the thrombosis of common carotid artery in rabbits ( mg)

Compared with the control group ^* P<0.05 ^** P<0.01

The extracts involved in the pharmacological experiments of the present invention were prepared according to the method of Example 1.

Detailed ways

The following examples are to illustrate the present invention in more detail, but not to limit the present invention.

Example 1 Preparation of safflower extract containing total safflower yellow by water extraction

Take 2000g of safflower medicinal material, add 10 times of water (the weight of the medicinal material), stir and extract twice at 70°C for 1 hour each time, filter, combine the secondary extracts, and cool to room temperature. According to the ratio of the amount of sample on the column to the amount of resin (1:10/W:V), pass HP-20 macroporous adsorption resin column chromatography, the diameter-to-height ratio of the column is 1:7, and use 1-2ml/cm ² / Min flow rate eluted 1 column bed volume and discarded, followed by 25% ethanol 1-2ml/cm ² /min flow rate 4 column bed volumes, 30% ethanol eluted part was the total safflower yellow pigment part, 70 The solvent is recovered under reduced pressure at ℃ to a concentrated solution with a specific gravity of about 1.05. The above solution is ultrafiltered through a ceramic membrane with a molecular weight cut-off of 8000, and the filtrate is collected and dried to obtain the total safflower yellow of the extract, which can be directly used to prepare injections.

After testing, the total safflower yellow content in the extract is 85%, the hydroxyl safflower yellow A content is 25%, and the kaempferol-3-O-rutinoside content is 0.7%.

Example 2 Preparation of safflower extract containing total safflower yellow by water extraction

Take 2000g of safflower medicinal material, add 8 times of water (medicinal material weight), stir and extract twice at 75°C for 1 hour each time, filter, combine the secondary extracts, and cool to room temperature. According to the ratio of the amount of sample on the column to the amount of resin (1:8/W:V), add HPD 600 macroporous adsorption resin column chromatography ^. Min flow rate eluted 1 column bed volume and discarded, followed by 40% ethanol 1-2ml/cm ² /min flow rate 4 column bed volumes, 40% ethanol eluted part was the total safflower yellow pigment part, 70 The solvent is recovered under reduced pressure at ℃ to a concentrated solution with a specific gravity of about 1.02. The above solution is ultrafiltered through a ceramic membrane with a molecular weight cut-off of 10,000. The filtrate is collected and dried to obtain the total safflower yellow extract.

After testing, the total safflower yellow content in the extract is 81%, the hydroxyl safflower yellow A content is 23%, and the kaempferol-3-O-rutinoside content is 0.65%.

Example 3 Preparation of safflower extract containing total safflower yellow pigment by dilute alcohol extraction

Get 2000g of safflower medicinal material, add 10 times of 30% ethanol (medicinal material weight), reflux extraction 2 times, each time for 1 hour, filter, merge the secondary extract, reclaim the extract to no alcohol, according to the amount of sample on the column and The ratio of the amount of resin (1:10/W:V), add HP-20 macroporous adsorption resin column chromatography, the diameter-to-height ratio of the column is 1:10, and eluted with water at a flow rate of 1-2ml/ ^cm2 /min1 The column bed volume was discarded, followed by 30% ethanol at a flow rate of 1-2ml/cm ² /min for 4 column bed volumes, the eluted part of 35% ethanol was the safflower yellow part, and the solvent was recovered under reduced pressure at 70°C until For a concentrated solution with a specific gravity of about 1.05, the above solution is ultrafiltered through an organic membrane with a molecular weight cut-off of 10,000, and the filtrate is collected and dried to obtain the safflower yellow extract.

After testing, the total safflower yellow content in the extract is 88%, the hydroxyl safflower yellow A content is 27%, and the kaempferol-3-O-rutinoside content is 0.72%.

Example 4 Preparation of (total) safflower yellow freeze-dried powder injection

Weigh 310g of dextran, dissolve in 4500ml of water for injection, then weigh 300g of safflower yellow extract prepared by the method of the present invention, and dissolve in the above solution. Adjust the pH to 7.0±0.2 with 20% sodium hydroxide solution. Add the prepared amount of 0.1% needles, heat and stir with activated carbon for 15 minutes, filter with suction, and add water for injection to the filtrate to 5000 ml. Fine filtration, intermediate inspection, filling, freeze-drying, capping, finished product inspection, packaging.

Example 5 Preparation of (total) safflower yellow freeze-dried powder injection

Weigh 300g of mannitol, dissolve in 4500ml of water for injection, then weigh 300g of safflower yellow extract prepared by the method of the present invention, and dissolve in the above solution. Adjust the pH to 7.0±0.2 with 20% sodium hydroxide solution. The solution was ultrafiltered with a ceramic membrane with a molecular weight cut-off of 10,000, and the ultrafiltration was performed twice with 8000ml of water. The ultrafiltrate is used for intermediate inspection, filling, freeze-drying, capping, finished product inspection, and packaging.

Embodiment 6 (total) preparation of safflower yellow pigment infusion

Take 320g of the safflower yellow extract prepared by the above method, add 834.7g of sodium chloride and 90L of water for injection, stir to dissolve, add an appropriate amount of 0.1mol/L NaOH, adjust the pH to about 7.0, and supplement water for injection to a sufficient amount , then add activated carbon for needles with a liquid volume of 0.15%, stir for 15 minutes, remove the carbon by suction filtration, after the content and pH value are qualified, then fine filter and subpackage, (105 ° C) autoclaving, to obtain.

Example 7 (Total) Preparation of safflower yellow pigment small water injection

Take 250g of the safflower yellow extract prepared by the above method, add 80.0g of sodium chloride and 10L of water for injection, stir to dissolve, add an appropriate amount of 0.1mol/L NaOH, adjust the pH to about 7.0, add 10g of antioxidant sulfur Sodium sulfite, add water for injection to a sufficient amount, then add activated carbon for needles with a liquid volume of 0.1%, stir for 15 minutes, and remove carbon by suction filtration. Sterilize by autoclaving.

The preferred embodiments of the present invention have been described above, but they are not intended to limit the present invention. Modifications and changes to the embodiments disclosed herein may be made by those skilled in the art without departing from the scope and spirit of the invention.

Claims (9)

1. preparation method that contains the Flos Carthami extract of total Carthamus yellow, comprising:

(1) Flos Carthami is obtained extracting solution I with at least 2 times of water temperature lixiviates, and it is 40-90 ℃ that described warm macerating extracts temperature;

(2) extracting solution concentrate concentrated solution, be 1 with blade diameter length ratio on the concentrated solution then: 6-1: 10 HPD600 type or HP-20 type macroporous adsorbent resin separate, described separation comprises earlier with 1-2 bed volume washing, discard water liquid, the pure eluting 3-5 bed volume of reuse concentration 20%-50%, collect pure eluting part, recovery eluent extremely pure content obtains eluent II below 20%;

(3) by ceramic membrane ultrafitration, obtain ultrafiltrate III, the molecular cut off of described ceramic membrane is 6000-5 ten thousand;

(4) ultrafiltrate III is 1.05-1.20 through being recycled to relative density, gets final product.

2. the described preparation method of claim 1, wherein, Flos Carthami is got with 2-14 times of water temperature lixiviate, obtains extracting solution I.

3. the described preparation method of claim 1, wherein the molecular cut off of the ceramic membrane described in the step (3) is 1-3 ten thousand.

4. the described preparation method of claim 1, wherein the alcohol used of the eluting described in the step (2) is 30% ethanol.

5. the described preparation method of claim 1, wherein the determining alcohol of the eluent II that obtains of step (2) is 5%-15%.

6. a Flos Carthami extract that contains total Carthamus yellow that makes with the described preparation method of claim 1 is characterized by total carthamus tinctorius yellow color content＞80%, hydroxyl safflor yellow A content＞20%, kaempferol-3-O-rutinoside content＞0.6%.

7. a pharmaceutical composition is characterized in that containing described Flos Carthami extract and the pharmacy acceptable auxiliary that contains total Carthamus yellow of the claim 6 for the treatment of effective dose.

8. the described pharmaceutical composition of claim 7, wherein this pharmaceutical composition is an injection.

9. the described pharmaceutical composition of claim 8, wherein said injection contains the described Flos Carthami extract 1-3 of claim 6 part, mannitol, dextran or sodium chloride 0.8-5 part.