CN101628907B - Fluorescein sodium refining method - Google Patents
Fluorescein sodium refining method Download PDFInfo
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- CN101628907B CN101628907B CN2009100414434A CN200910041443A CN101628907B CN 101628907 B CN101628907 B CN 101628907B CN 2009100414434 A CN2009100414434 A CN 2009100414434A CN 200910041443 A CN200910041443 A CN 200910041443A CN 101628907 B CN101628907 B CN 101628907B
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Abstract
A fluorescein sodium refining method relates to a method for preparing high purity fluorescein sodium by using reagent grade fluorescein sodium as raw material, comprising the following steps: taking reagent grade fluorescein sodium, adding polar solvent, then adding activated carbon, refluxing for 10-60min, filtrating, taking filtrate to add hydrochloric acid while stirring, reducing the temperature to room temperature, standing to ensure that fluorescein sodium is fully precipitated, filtrating, washing the precipitate with polar solvent, then washing with pure water, placing the fluorescein in water, adjusting the pH value to 8.5-9.0 with sodium hydroxide or sodium carbonate, and filtrating to obtain more than 99.5wt% of fluorescent sodium. The method has simple operation and is applicable to industrialized batch production; the whole process is a one-step acidic-basic refining process and the amount of wastewater is less, thus facilitating environmental protection; the method can effectively improve the purity of the product and the clarity of the solution, thus facilitating the quality control of the injection product and reducing the risk of clinical medication.
Description
Technical field
The present invention relates to a kind of process for purification that only contains the heterocyclic organic compounds of Sauerstoffatom, specifically is to be the method for the highly purified uranine of feedstock production pharmaceutical grade with the SILVER REAGENT uranine.
Background technology
The chemistry of uranine is called 9-(adjacent carboxyl phenyl)-6-hydroxyl-3H-xanthene-3-ketone disodium salt, and English is called fluorescein sodium, can be used for the mensuration of cornea damage, optical fundus blood vessel angiographic diagnosis and blood circulation time.The uranine of highly purified pharmaceutical grade is used to prepare the injection that radiography is used.Meet the pharmacopeia requirement though be used to prepare the uranine raw material of injection at present, the weight content of uranine can reach 98.5%, but can find to still have more a plurality of impurity to exist with high-efficient liquid phase chromatogram technique analysis, because of there being impurity to disturb, when using spectrophotometry, even concentration dilution is 1.0% o'clock, all greater than 4.0, instrument can't accurate reading in its optical density value of 600nm wavelength place; For the uranine injection liquid that is contained in 10% in ampoule or 20% concentration, under natural light, to inspect, liquid color is profound, can't carry out the visible foreign matters inspection.Major impurity is a phthalate anhydride in the uranine according to the literature; Resorcinol; 6-hydroxyl-7-sulfonic group-9-(adjacent carboxyl phenyl)-3H-xanthene-3-ketone, 6-hydroxyl-9-(adjacent ethoxy carbonyl phenyl)-3H-xanthene-3-ketone or 2-(2,4-dihydroxy-benzene formyl radical) phenylformic acid etc.Because the existence of these impurity, its uranine injection liquid is injected into human body also can bring certain side effect, as anaphylaxis such as nausea and vomiting, skin urticaria, coughs, can produce anaphylactic shock when serious, can't guarantee the safety of medication.Because these impurity and uranine structural similitude, the lock out operation difficulty is bigger, impurity is soluble in water under alkaline condition, at this moment, activated carbon is obviously low to its absorption, and these impurity and fluorescein co-precipitation under acidic conditions removed these impurity with traditional method to prepare highly purified uranine is what be difficult to realize so those skilled in the art think.Organic preparation chemistry handbook (in) the P280 page or leaf, introduced with the method for uranine recrystallization and purified.Because uranine has the red variant of two kinds of variants-be insoluble to alcohol ether and is dissolved in methyl alcohol and the yellow variant of alcoholic acid, yellow variant is heated and easily changes stable red variant into.When the preparation fluorescein, what the low temperature acidifying was separated out is yellow variant, and this kind variant dissolves in methyl alcohol or ethanol.But the yellow variant of separating out from water contains large quantity of moisture, is not easy to filter, and also is not easy drying, and in heat drying or heating for dissolving process, yellow variant is to change undissolved red variant into, and is very difficult in the actually operating, and productive rate is low, the waste water generation is big.The CN101270124A patent disclosure a kind of novel method for preparing high purity fluorescein and uranine; it is with fluorescein or uranine and acetylation reagent reaction generation di-acetyl fluorescein; thereby utilize the different chemical polarity of impurity and di-acetyl fluorescein; impurity is separated; obtain highly purified uranine through saponification, acidifying, salify again, the weight content of uranine can reach more than 99.2%.Although this method is feasible,, the di-acetyl fluorescein heating or meeting under the fiery condition, can decomposite the radiotoxicity material as a kind of radiotoxicity material, has stronger hazardness.Simultaneously, processing step is many and loaded down with trivial details, the cost height.So we seek a kind of simple and feasible method in order to solve the highly purified medicinal uranine of preparation.
Summary of the invention
The objective of the invention is to prepare the medicinal high-purity fluorescein sodium of weight content more than 99.5% with SILVER REAGENT uranine (weight content is 90.0～95.0%) with simple and feasible method.
To achieve the object of the present invention, the technical solution adopted in the present invention is that weight content is 90.0～95.0% uranine, the polar solvent that adds 2～12 times of volumes of uranine weight is a solvent, 1～10% of adding uranine weight activated carbon after reflux to uranine dissolves, continue 10～60 minutes after-filtration that reflux, get filtrate and add the calculated amount hydrochloric acid of (uranine of 100g adds the concentrated hydrochloric acid of 56ml), stir, and make it be cooled to room temperature, place, uranine is separated out fully, filter, filtrate is washed till the chlorion (Cl≤0.35%) up to specification with pure water after with the polar solvent washing precipitation, fluorescein is placed the water of 2～5 times of amounts, regulate pH value to 8.5～9.0 with sodium hydroxide or sodium bicarbonate, filter, filtrate is condensed into thick liquid, dry below 105 ℃, promptly getting weight content is more than 99.5%, and foreign matter content is below 0.5%, and 600nm place absorbancy is less than 0.5 uranine when being made into 10% (W/V) solution.
Said polar solvent has many, in the present invention preferably water, dehydrated alcohol, 50～99% ethanolic soln, methyl alcohol, dimethyl formamide, acetone, propyl carbinol or chloroform: the isopropylcarbinol volume ratio is 1: 1 a solution.
The measuring method of uranine weight content among the present invention: adopt high performance liquid chromatography
Chromatographic condition and system suitability octadecylsilane chemically bonded silica are weighting agent; The sodium dihydrogen phosphate volume ratio of methyl alcohol: 0.1mol/ml is that 50: 50 solution is made moving phase; The detection wavelength is 235nm; Flow velocity is 1ml/ minute.Number of theoretical plate calculates by the uranine peak should be not less than 2000, and the resolution of uranine peak and adjacent impurity peaks should meet the requirements.
The preparation of reference liquid: precision takes by weighing a certain amount of di-acetyl fluorescein (USP DiacetylfluoresceinRS) (the anhydrous di-acetyl fluorescein of 110.7mg is equivalent to the uranine of 100mg), be dissolved in the ethanol of 10ml, put in the 100ml volumetric flask, the sodium hydroxide solution that adds 2ml 2.5mol, rotation heating is 20 minutes in the boiling water steam, puts coldly, is diluted with water to scale, mixing is made the reference substance solution that every 1ml contains uranine 1mg.
The preparation of sample: precision takes by weighing the uranine 100mg that is dried to constant weight, puts in the volumetric flask of 100ml, and thin up is made need testing solution to scale.
Measuring method: get each 10 μ l of reference substance solution and need testing solution, inject liquid chromatograph, the record color atlas is by the content of uranine in the external standard method calculating trial-product.
The detection method of related substance among the present invention: adopt high performance liquid chromatography
Chromatographic condition and system suitability with octyl silane group silica gel be packed column (5 μ m, 4.6mm * 250mm), column temperature is 30 ℃, mobile phase A is potassium primary phosphate 1.22g, is dissolved in water and is diluted to 1000ml, regulates pH to 2.0 with phosphoric acid; Mobile phase B is an acetonitrile; The detection wavelength is 220nm; According to the form below carries out linear gradient elution.
| Time (minute) | Mobile phase A (%) | Mobile phase B (%) |
| 0 | 85 | 15 |
| 25 | 20 | 80 |
| 34 | 20 | 80 |
| 35 | 85 | 15 |
| 45 | 85 | 15 |
The preparation of need testing solution: it is an amount of to get uranine, adds acetonitrile: water volume ratio is that 15: 85 solution is made the solution that contains the 0.5mg uranine among every 1ml as solvent, as need testing solution.
The preparation of contrast solution and reference substance solution: precision is measured need testing solution 1ml and is put in the 100ml measuring bottle, uses solvent acetonitrile: water volume ratio be 15: 85 solution dilution to scale, solution in contrast.Other respectively gets phthalic acid (impurity A) and Resorcinol (impurity B) reference substance is an amount of, the accurate title, decide, add acetonitrile: water volume ratio is that 15: 85 solution is made the solution that contains 2.5 μ g among every 1ml, as phthalic acid reference substance solution and Resorcinol reference substance solution.
Measuring method: respectively get reference substance solution 20 μ l, inject liquid chromatograph, regulate detection sensitivity, make the peak height of principal constituent chromatographic peak be about 20% of full range; Precision is measured each 20 μ l of need testing solution, contrast solution and reference substance solution again, injects liquid chromatograph respectively, the record color atlas.The resolution of main peak and adjacent each impurity peaks should meet the requirements in the need testing solution color atlas.In the need testing solution color atlas if any with the reference substance solution color atlas in impurity A and the corresponding peak of B, its peak area must not be greater than the area of impurity A and B in the reference substance solution color atlas; If any other impurity peaks, single impurity peak area must not be greater than 0.5 times of contrast solution main peak area (be equivalent to test solution weight content 0.5%), other impurity peak area and must not be greater than contrast solution main peak area (be equivalent to test solution weight content 1.0%).Ignore less than the impurity peaks of 0.05 times of contrast solution main peak area in the need testing solution color atlas.
Description of drawings
The HPLC color atlas of the uranine content before accompanying drawing 1 is made with extra care
The HPLC color atlas of accompanying drawing 3 Resorcinols
The HPLC color atlas of accompanying drawing 4 phthalic acids
The HPLC color atlas of accompanying drawing 5 refining preceding uranine related substances
The HPLC color atlas of the uranine content after accompanying drawing 6 is made with extra care
The HPLC color atlas of the uranine related substance after accompanying drawing 7 is made with extra care
Among the figure, the 1st, the peak of uranine, the 2nd, impurity peaks, the 3rd, the peak of Resorcinol, the 4th, the peak of phthalic acid.
Advantage of the present invention is:
1. simple to operate, easily realize industrialized mass production;
2. whole technology is an acid-alkali refining operation, and wastewater flow rate is few, is beneficial to environmental protection;
3, method of the present invention can improve the purity of product and the clarity of solution thereof effectively, is convenient to the risk of the quality control and the reduction clinical application of injection product.
Set forth technical scheme of the present invention in conjunction with the accompanying drawings below by specific embodiment.
Embodiment
Embodiment 1
Take by weighing uranine 100g, (uranine is to be provided by Tianjin reagent one factory, weight content is 95.0% (seeing accompanying drawing 1), (see accompanying drawing 2～5 through the HPLC analysis, foreign matter content is 7.1%,) adding 100% ethanol 500ml, reflux adds the activated carbon of 5g to dissolving, continue the 15 minutes after-filtration that reflux, get the hydrochloric acid (36.5%) that filtrate adds 56ml, stir, and make it be cooled to room temperature, placed 18 hours, filter, filter cake washs to chlorion after with the 200ml methanol wash up to specification only with purified water, fluorescein is placed the water of 200ml, add caustic lye of soda (concentration 40%) and regulate pH to 8.7, solution concentration 105 ℃ of dryings, promptly gets the uranine of weight 75g to thick liquid, with high effective liquid chromatography for measuring content is 99.8% (seeing accompanying drawing 6), refining yield is 75%, foreign matter content 0.25% (seeing accompanying drawing 7), 10% solution 600nm absorbancy 0.377.
Take by weighing uranine 100g (with the raw material of embodiment 1), add 50% ethanol 250ml, reflux is to dissolving, the activated carbon that adds 10g, continue the 30 minutes after-filtration that reflux, get the hydrochloric acid (36.5%) of filtrate Dropwise 5 6ml, stir, and make it be cooled to room temperature, placed 16 hours, filter, filter cake washs to chlorion is up to specification with purified water after with washing with acetone and ends, and fluorescein is placed the water of 200ml, add caustic lye of soda (concentration 40%) and regulate pH to 8.8, solution concentration 105 ℃ of dryings, promptly gets the uranine of weight 70g to thick liquid, with high effective liquid chromatography for measuring content is 99.7%, refining yield is 70%, foreign matter content 0.18%, 10% solution 600nm absorbancy 0.379.
Take by weighing uranine 100g, (with the raw material of embodiment 1) adds water 200ml, stirring and dissolving, the activated carbon that adds 10g, 30 minutes after-filtration of reflux, get the hydrochloric acid (36.5%) of filtrate Dropwise 5 6ml, stir, and make it be cooled to room temperature, placed 20 hours, filter, filter cake washs to chlorion is up to specification with purified water after with washing with alcohol and ends, and fluorescein is placed the water of 150ml, add caustic lye of soda (concentration 40%) and regulate pH to 8.75, solution concentration 105 ℃ of dryings, promptly gets the uranine of weight 60g to thick liquid, with high effective liquid chromatography for measuring content is 99.6%, refining yield is 60%, foreign matter content 0.32%, 10% solution 600nm absorbancy 0.495.
Take by weighing uranine 100g, (with the raw material of embodiment 1) adds methyl alcohol 1000ml, reflux is to dissolving, the activated carbon that adds 5g, continue the 30 minutes after-filtration that reflux, get the hydrochloric acid (36.5%) of filtrate Dropwise 5 6ml, stir, and make it be cooled to room temperature, placed 20 hours, filter, filter cake washs to chlorion is up to specification with purified water with dimethyl formamide washing back and ends, and fluorescein is placed the water of 150ml, add caustic lye of soda (concentration 40%) and regulate pH to 8.75, solution concentration 105 ℃ of dryings, promptly gets the uranine of weight 58.5g to thick liquid, with high effective liquid chromatography for measuring content is 99.8%, refining yield is 58.5%, foreign matter content 0%, 10% solution 600nm absorbancy 0.475.
Take by weighing uranine 100g, add acetone 1000ml, reflux adds the activated carbon of 8g to dissolving, continue the 30 minutes after-filtration that reflux, get the hydrochloric acid (36.5%) of filtrate Dropwise 5 6ml, stir, and make it be cooled to room temperature, placed 20 hours, filter, filter cake washs to chlorion is up to specification with purified water after with washing with alcohol and ends, and fluorescein is placed the water of 200ml, add caustic lye of soda (concentration 40%) and regulate pH to 8.70, solution concentration 105 ℃ of dryings, promptly gets the uranine of weight 53g to thick liquid, with high effective liquid chromatography for measuring content is 99.6%, refining yield is 53%, foreign matter content 0.42%, 10% solution 600nm absorbancy 0.468.
Embodiment 6
Uranine 100g, add dimethyl formamide 300ml, reflux is to dissolving, the activated carbon that adds 10g, continue the 30 minutes after-filtration that reflux, get the hydrochloric acid (36.5%) of filtrate Dropwise 5 6ml, stir, and make it be cooled to room temperature, placed 18 hours, filter, filter cake washs to chlorion is up to specification with purified water with dimethyl formamide washing back and ends, and fluorescein is placed the water of 200ml, add caustic lye of soda (concentration 40%) and regulate pH to 8.70, solution concentration 105 ℃ of dryings, promptly gets the uranine of weight 73g to thick liquid, with high effective liquid chromatography for measuring content is 99.9%, refining yield is 73%, foreign matter content 0.28%, 10% solution 600nm absorbancy 0.364.
Embodiment 7
Uranine 100g, add propyl carbinol 1200ml, reflux adds the activated carbon of 10g to dissolving, continue the 30 minutes after-filtration that reflux, get the hydrochloric acid (36.5%) of filtrate Dropwise 5 6ml, stir, and make it be cooled to room temperature, placed 18 hours, filter, filter cake washs to chlorion is up to specification with purified water after with washing with alcohol and ends, and fluorescein is placed the water of 200ml, add caustic lye of soda (concentration 40%) and regulate pH to 8.70, solution concentration 105 ℃ of dryings, promptly gets the uranine of weight 45g to thick liquid, with high effective liquid chromatography for measuring content is 99.7%, refining yield is 46%, foreign matter content 0.42%, 10% solution 600nm absorbancy 0.425.
Uranine 100g, add chloroform: isopropylcarbinol (1: 1) 1200ml, reflux is to dissolving, the activated carbon that adds 10g, continue the 30 minutes after-filtration that reflux, get the hydrochloric acid (36.5%) of filtrate Dropwise 5 6ml, stir, and make it be cooled to room temperature, placed 18 hours, filter, filter cake washs to chlorion is up to specification with purified water after with methanol wash and ends, and fluorescein is placed the water of 200ml, add caustic lye of soda (concentration 40%) and regulate pH to 8.70, solution concentration 105 ℃ of dryings, promptly gets the uranine of weight 43g to thick liquid, with high effective liquid chromatography for measuring content is 99.6%, refining yield is 43%, foreign matter content 0.48%, 10% solution 600nm absorbancy 0.451.
Claims (1)
1. the process for purification of a uranine, it is characterized in that getting weight content and be 90.0～95.0% uranine, the polar solvent that adds 2～12 times of volumes of uranine weight, 1～10% of adding uranine weight activated carbon after reflux to uranine dissolves, continue 10～60 minutes after-filtration that reflux, get filtrate and stir the hydrochloric acid that adds calculated amount down, add the concentrated hydrochloric acid of 56ml with the uranine of 100g, and make it be cooled to room temperature, place, fluorescein is separated out fully, filter, use ethanol, methyl alcohol, dimethyl formamide or washing with acetone post precipitation place fluorescein in the water of 2～5 times of amounts again with the pure water washing, regulate pH value to 8.5～9.0 with sodium hydroxide or yellow soda ash, filter, filtrate is condensed into thick liquid, drying, promptly get weight content at the uranine more than 99.5%
Described polar solvent is water, dehydrated alcohol, 50～99% ethanolic soln, methyl alcohol, dimethyl formamide, acetone, propyl carbinol or chloroform: the isopropylcarbinol volume ratio is 1: 1 a solution.
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