Abstract:
A novel process for the purification of poloxamers which contain aldehyde impurities, and particularly for the purification of the poloxamers named poloxamer 188 and poloxamer 407, which process includes dissolving the poloxamer(s) in a solvent which solvent may optionally contain an acid or a mixture of acids; if the solvent in which the poloxamer(s) has been dissolved is free from acid, acid is added; whereafter the aldehydes are removed.

Description:
CROSS REFERENCE TO RELATED APPLICATIONS 
     The present application represents U.S. national stage of international application PCT/SE00/01305 with an international filing date of Jun. 19, 2000. The international application claims priority to Swedish application 9902364-0, filed on Jun. 21, 1999. 
    
    
     FIELD OF THE INVENTION 
     The present invention relates to a novel process for the purification of poloxamers which contain aldehyde impurities, and particularly for the purification of the poloxamers named poloxamer 188 and poloxamer 407. 
     BACKGROUND OF THE INVENTION 
     Poloxamers are synthetic block copolymers of hydrophilic ethylene oxide chains and hydrophobic propylene oxide chains, having the formula HO—[C 2 H 4 O] a —[C 3 H 6 O] b —[C 2 H 4 O] a —H, wherein a and b represent the number of hydrophilic and hydrophobic chains respectively. 
     The poloxamers can be used as surfactants, emulsifying agents, solubilizing agents or wetting agents in different compositions. WO97/38675 discloses a composition comprising one or more local anaesthetics, water and one or more surfactants, preferably poloxamer(s). Since the poloxamers have both hydrophobic and hydrophilic domains they can, in combination with for example a local anaesthetic, provide a composition having thermoreversible gelling properties. 
     Commercially available poloxamers often contain relatively high amounts of aldehydes. These aldehydes may react with the active compound, for example a local anaesthetic, and form non-desirable compounds which have to be separated from the final product. This also means that a higher amount of the active compound is required, since some of the active compound is reacting with the aldehydes and thus is lost during the purification process. 
     Thus, there is a need for an improved purification process in order to reduce the concentration of aldehydes in poloxamers. 
     OUTLINE OF THE INVENTION 
     The present invention is directed to a novel process for the purification of poloxamers, which contain aldehyde impurities, whereby 
     (i) the poloxamer(s) is dissolved in a solvent, which solvent may optionally contain an acid or a mixture of acids, 
     (ii) if the solvent in which the poloxamer(s) has been dissolved is free from acid, acid is added; whereafter 
     (iii) the aldehydes are removed. 
     The process according to the present invention is suitable for the purification of poloxamers containing aldehyde impurities, said poloxamers having the general formula HO—[C 2 H 4 O] a —[C 3 H 6 O] b —[C 2 H 4 O] a —H, wherein a and b represent the number of hydrophilic and hydrophobic chains. The process according to the present invention is particularly suitable for the purification of Lutrol F68®, which also has the name poloxamer 188 and wherein a=80 and b=27, or Lutrol F127®,which also has the name poloxamer 407 and wherein a=101 and b=56, the definitions being in accordance with USP (1995) HF18, p.2279. The process may also be used for the purification of a mixture of poloxamers, and specifically a mixture of poloxamer 188 and poloxamer 407, as well as for the purification of other polymers containing monomers such as ethylene oxide, propylene oxide or similar, said polymers containing aldehyde impurities. 
     The solvent used for dissolving the poloxamer may be water, an alcohol such as methanol. ethanol or propanol, or any other suitable solvent. Preferably water is used as a solvent. 
     The temperature of the solvent or the mixture of solvent and poloxamer is not essential, however, the poloxamer is dissolved more quickly at a temperature below room temperature. A preferred solvent temperature is from 0 to +10° C. 
     The concentration of poloxamer in the poloxamer solution is preferably 5-21% by weight. 
     More preferred the concentration is 15% by weight. 
     The acid(s) added to the poloxamer solution may be any acid(s), but preferably one or a mixture of two or more of hydrochloric acid, acetic acid or citric acid. More preferably the acid is one or a mixture of hydrochloric acid or acetic acid, but most preferred is to use acetic acid alone. pH for the solution after having added the acid should be from 1-7, preferably from 1-5. The amount of added acid is dependent on the desired pH. 
     The acid or acids may be added to the solvent before or after the poloxamer is added to the solvent. When the poloxamer and the acid have been added to the solvent, and the poloxamer is dissolved, the acid needs some time to interact, whereafter the aldehydes are removed from the solution by drying. The drying may be evaporation performed by boiling the solution, whereby the aldehydes are vaporized, drying the solution on a fluid bed, spray drying, freeze drying, vacuum evaporation or letting a stream of gas, such as helium or nitrogen, pass through the solution, spray crystallisation, spray granulation or any other method of drying known to the skilled person. Preferably the aldehydes are evaporated by drying the solution on a fluid bed and at a temperature as of &lt;47° C. for the poloxamer. A higher temperature provides a quicker drying of the solution, but the temperature may however not be too high since the poloxamers may melt at higher temperatures. 
     The process according to present invention is particularly suitable for the removal of formaldehyde, acetaldehyde and propionaldehyde from poloxamers. Other aldehyde impurities may however also be removed by using the process according to the present invention. The present process is particularly suitable for the removal of acetaldehyde and propionaldehyde from poloxamers. 
     Also within the scope of the invention is a poloxamer purified from aldehydes according to the process described above. 
     DETAILED DESCRIPTION OF THE INVENTION 
     The reduction of aldehydes in the following Examples has been determined according to the analytical method described below. 
     Analytical Method 
     The sample is dissolved and diluted in water. Any aldehydes present in the poloxamers will react with 2,4-dinitrophenylhydrazine or 3-methyl-2-benzothiazolonehydrazone at room temperature to form derivatives. The derivatives are extracted with cyclohexane followed by evaporation of the organic phase. The residual is dissolved in the mobile phase and the derivates are separated using liquid chromatography and quantified by standard addition or external standard calibration. 
    
    
     THE INVENTION WILL NOW BE DESCRIBED MORE IN DETAIL BY THE FOLLOWING NON-LIMITING EXAMPLES. 
     EXAMPLE 1 
     Purging with Helium 
     HCl was added to water to a concentration of 0.3 mol/l. Poloxamer 407 was dissolved in the HCl solution to a concentration of 5% by weight (w/w). The solution was purged with helium for 2 hours and kept at room temperature over night. The solution was then purged with helium for an additional 3 hours. The solution was kept at room temperature and helium was added at a flow of 1 bubble/second. The amount of aldehydes was reduced by 80%. 
     EXAMPLE 2 
     Evaporation by Using Nitrogen 
     HCl was added to water to a concentration of 0.3 mol/l. Poloxamer 407 was dissolved in the HCl solution to a concentration of 5% by weight (w/w). The solution was kept at room temperature for 3 hours. 80% of the solution was evaporated by using nitrogen flow. 0.3 mol/l HCl solution was added to the initial volume. The amount of aldehydes was reduced by 95%. 
     EXAMPLE 3 
     Vacuum Evaporation 
     5.5% w/w Poloxamer 188 and 15.5% w/w poloxamer 407 were dissolved in water. Citric acid was added to pH 2.7 whereafter the solution was kept at room temperature for at least 24 hours. Then 5% by weight of the solution was evaporated by using a rotavapour. The concentration of acetaldehyde was decreased from 135 ppm to 30 ppm and the concentration of propionaldehyde was decreased from 429 ppm to &lt;10 ppm. 
     EXAMPLE 4 
     Fluid Bed Drying—Hydrochloric Acid 
     Poloxamer 407 was dissolved in water to a concentration of 15% w/w, whereafter hydrochloric acid was added to pH 2. The solution was kept in a refrigerator for 6 days. The solution was dried by using fluid bed drying. The concentration of acetaldehyde was decreased from 337 ppm to 26 ppm and the propionaldehyde level was decreased from 1.35*10 3  ppm to 76 ppm. 
     EXAMPLE 5 
     Fluid Bed Drying—Citric Acid 
     18.22 g Citric acid,having a concentration of 50%, was added to 2000 g of water. Poloxamer 407 was dissolved in the citric acid solution to a concentration of 15% w/w. The solution was kept in a refrigerator for 6 days. The solution was dried in a fluid bed. The concentration of acetaldehyde and propionaldehyde was decreased from 358 ppm to 5 ppm and from 1.39*10 3  ppm to &lt;6 ppm respectively. 
     EXAMPLE 6 
     Fluid Bed Drying—Acetic Acid (Poloxamer 188) 
     120 g Concentrated acetic acid was added to 2000 g of water. Poloxamer 188 was dissolved in the acetic acid solution to a concentration of 15% w/w. The solution was kept in a refrigerator for 6 days. The solution was dried in a fluid bed. The concentration of acetaldehyde was decreased from 186 ppm to 8 ppm and the concentration of propionaldehyde from 297 ppm to &lt;6 ppm. 
     EXAMPLE 7 
     Fluid Bed Drying—Acetic Acid (Poloxamer 407) 
     120 g Concentrated acetic acid was added to 2000 g of water. Poloxamer 407 was dissolved in the acetic acid solution to a concentration of 15% w/w. The solution was kept in a refrigerator for 6 days. The solution was dried in a fluid bed, whereby the concentration of acetaldehyde was decreased from 358 ppm to 12 ppm and the concentration of propionaldehyde from 1.39*10 3  ppm to 9 ppm. 
     EXAMPLES 8-10 
     Purification of Poloxamer 188 in Full Scale 
     The poloxamer was dissolved in water, whereafter acetic acid was added. After at least 24 hours interaction, the solution was dried using a fluid bed dryer. In order to speed up the process, purified poloxamer was used as starting material in the fluid bed dryer. The first batch was however produced without purified poloxamer as starting material. In the following examples material from the previous batch was used as starting material. 
     Equipment and Process Parameters 
     
       
         
               
               
               
             
           
               
                   
                   
               
             
             
               
                   
                 Fluid Bed Dryer: 
                 Glatt GVW 32″ 
               
               
                   
                 Filter Shaking: 
                 2015 
               
               
                   
                 Spray Nozzle: 
                 1 * 6 head, central, 1.8 mm 
               
               
                   
                 Spray Nozzle Position: 
                 4 v down 
               
               
                   
                 Pump: 
                 Membrane-Pump 
               
               
                   
                   
               
             
          
         
       
     
     
       
         
               
               
               
             
           
               
                   
                   
               
               
                   
                 Process Parameter 
                 Poloxamer 188 (Examples 8-10) 
               
               
                   
                   
               
             
             
               
                   
                 Water 
                 135 kg 
               
               
                   
                 Acetic Acid 
                 15 kg 
               
               
                   
                 Poloxamer 
                 100 kg 
               
               
                   
                 PH 
                 &lt;2.8 
               
               
                   
                 Interaction Time 
                 &gt;24 h &lt; 1 week 
               
               
                   
                 Starting Material 
                 100 kg 
               
               
                   
                 Inlet Air Flow 
                 4000-5000 m 3   
               
               
                   
                 Inlet Air Temperature 
                 48-52° C. 
               
               
                   
                 Product Temperature 
                 &lt;47° C. 
               
               
                   
                 Spray Pressure 
                 3.8-4.2 bar 
               
               
                   
                 Spray Flow 
                 500-600 g/min 
               
               
                   
                   
               
             
          
         
       
     
     Analytical Results 
     3 Batches of Poloxamer 188 were purified according to the process with the process parameters given above. The levels of aldehydes are presented in the table below. 
     
       
         
               
               
               
               
             
               
               
               
               
             
           
               
                   
               
               
                   
                 Formaldehyde 
                 Acetaldehyde 
                 Propionaldehyde 
               
               
                 Batch 
                 μg/g 
                 μg/g 
                 μg/g 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 Poloxamer 188 
                 3.8 
                 1.4 * 10 2   
                 2.8 * 10 2   
               
               
                 Example 8 
                 &lt;1.9 
                 &lt;20 
                 &lt;48 
               
               
                 Polaxamer 188 
               
               
                 Example 9 
                 &lt;1.9 
                 &lt;20 
                 &lt;48 
               
               
                 Polaxamer 188 
               
               
                 Example 10 
                 &lt;1.9 
                 &lt;20 
                 &lt;48 
               
               
                 Polaxamer 188 
               
               
                   
               
             
          
         
       
     
     EXAMPLES 11-13 
     Purification of Poloxamer 407 in Full Scale 
     The poloxamer was dissolved in water, whereafter acetic acid was added. After at least 24 hours interaction, the solution was dried using a fluid bed dryer. In order to speed up the process, purified poloxamer was used as starting material in the fluid bed dryer. The first batch was however produced without purified poloxamer as starting material. In the following examples material from the previous batch was used as starting material. 
     Equipment and Process Parameters 
     
       
         
               
               
               
             
           
               
                   
                   
               
             
             
               
                   
                 Fluid Bed Dryer: 
                 Glatt GVW 32″ 
               
               
                   
                 Filter Shaking: 
                 2015 
               
               
                   
                 Spray Nozzle: 
                 1 * 6 head, central, 1.8 mm 
               
               
                   
                 Spray Nozzle Position: 
                 4 v down 
               
               
                   
                 Pump: 
                 Membrane-Pump 
               
               
                   
                   
               
             
          
         
       
     
     
       
         
               
               
               
             
           
               
                   
                   
               
               
                   
                 Process Parameter 
                 Poloxamer 407 (Examples 11-13) 
               
               
                   
                   
               
             
             
               
                   
                 Water 
                 592.1 kg 
               
               
                   
                 Acetic Acid 
                 47.4 kg 
               
               
                   
                 Poloxamer 
                 150 kg 
               
               
                   
                 PH 
                 &lt;2.8 
               
               
                   
                 Interaction Time 
                 &gt;24 h &lt; 1 week 
               
               
                   
                 Starting Material 
                 50 kg 
               
               
                   
                 Inlet Air Flow 
                 4000-5000 m 3   
               
               
                   
                 Inlet Air Temperature 
                 48-52° C. 
               
               
                   
                 Product Temperature 
                 &lt;47° C. 
               
               
                   
                 Spray Pressure 
                 3.8-4.2 bar 
               
               
                   
                 Spray Flow 
                 700-900 g/min 
               
               
                   
                   
               
             
          
         
       
     
     Analytical Results 
     3 Batches of Poloxamer 407 were purified according to the process with the process parameters given above. The levels of aldehydes are presented in the table below. 
     
       
         
               
               
               
               
             
               
               
               
               
             
           
               
                   
               
               
                   
                 Formaldehyde 
                 Acetaldehyde 
                 Propionaldehyde 
               
               
                 Batch 
                 μg/g 
                 μg/g 
                 μg/g 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 Poloxamer 407 
                 5.2 
                 3.0 * 10 2   
                 1.3 * 10 3   
               
               
                 Example 11 
                 &lt;1.9 
                 &lt;20 
                 &lt;48 
               
               
                 Poloxamer 407 
               
               
                 Example 12 
                 2.1 
                 &lt;20 
                 &lt;48 
               
               
                 Poloxamer 407 
               
               
                 Example 13 
                 &lt;1.9 
                 &lt;20 
                 &lt;48 
               
               
                 Poloxamer 407 
               
               
                   
               
             
          
         
       
     
     The examples show that by using the process according to present invention the concentrations of aldehydes, specifically acetaldehyde and propionaldehyde. are reduced in both poloxamers 407 and 188. Further it has been shown that all the acids hydrocloric acid, acetic acid and citric acid can be used in the present process. Furthermore the examples also show that drying of the solution may be performed in many different ways, such as purging with helium, evaporation by using nitrogen, vacuum evaporation or fluid bed drying.