Method for measuring the content of a botulinum toxin in a formulation

Abstract

A method of measuring the concentration of a bioactive agent is disclosed.

1 . A method of determining the concentration of a bioactive agent in a pharmaceutical formulation comprising a bioactive agent, comprising:
 determining a correlation between the bioactive agent and a marker compound in the formulation by deriving a ratio of the bioactive agent to the marker compound in the formulation,   measuring the concentration of the marker compound in the formulation, and   determining the concentration of the bioactive agent using the concentration of the marker compound and the ratio of the bioactive agent to the marker compound,   wherein the bioactive agent is a botulinum toxin, and   wherein the marker compound is sodium ion or chloride ion.   
   
   
       2 . The method of  claim 1 , wherein the marker compound is detected by inductively-coupled plasma atomic emission spectroscopy, high-performance liquid chromatography, fluorescence spectroscopy, liquid-chromatography/mass-spectrometry, ultraviolet/visible spectroscopy, infrared spectroscopy, electrochemical methods using; sodium ion selective electrode, or chloride ion selective electrode. 
   
   
       3 . The method of  claim 1 , wherein the bioactive agent is  Clostridium Botulinum  toxin A. 
   
   
       4 . The method of  claim 2 , wherein the bioactive agent is  Clostridium Botulinum  toxin A. 
   
   
       5 . The method of  claim 1 , wherein the formulation is a liquid formulation. 
   
   
       6 . The method of  claim 2 , wherein the formulation is a liquid formulation. 
   
   
       7 . A method of providing cosmetic treatment using a botulinum toxin, comprising:
 determining the content of the botulinum toxin in a pharmaceutical formulation comprising the botulinum toxin,   providing an amount of botulinum toxin according to the content determined according to the determining step, and   administering to a user the amount of the botulinum toxin in the formulation,   wherein determining comprises
 determining a correlation between the botulinum toxin and a marker compound in the formulation by deriving a ratio of the bioactive agent to the marker compound in the formulation, 
 measuring the concentration of the marker compound in the formulation, and 
 determining the concentration of the botulinum toxin using the concentration of the marker compound and the ratio of the bioactive agent to the marker compound; and 
   wherein the marker compound is selected from the group consisting of albumin, and human serum albumin.   
   
   
       8 . The method of  claim 7 , wherein the marker compound is detected by inductively-coupled plasma atomic emission spectroscopy, high-performance liquid chromatography, fluorescence spectroscopy, liquid-chromatography/mass-spectrometry, ultraviolet/visible spectroscopy, infrared spectroscopy, electrochemical methods using; sodium ion selective electrode, or chloride ion selective electrode. 
   
   
       9 . The method of  claim 7 , wherein the botulinum toxin is  Clostridium Botulinum  A 
   
   
       10 . The method of  claim 8 , wherein the botulinum toxin is  Clostridium Botulinum  A 
   
   
       11 . The method of  claim 7 , wherein the formulation is a liquid formulation. 
   
   
       12 . The method of  claim 8 , wherein the formulation is a liquid formulation. 
   
   
       13 . A method of providing cosmetic treatment using a botulinum toxin, comprising:
 determining the content of the botulinum toxin in a pharmaceutical formulation comprising the botulinum toxin,   providing an amount of botulinum toxin according to the content determined according to the determining step, and   administering to a user the amount of the botulinum toxin in the formulation,   wherein determining comprises
 determining a correlation between the botulinum toxin and a marker compound in the formulation by deriving a ratio of the bioactive agent to the marker compound in the formulation, 
 measuring the concentration of the marker compound in the formulation, and 
 determining the concentration of the botulinum toxin using the concentration of the marker compound and the ratio of the bioactive agent to the marker compound; and 
   wherein the marker compound is sodium ion or chloride ion.   
   
   
       14 . The method of  claim 13 , wherein the marker compound is detected by inductively-coupled plasma atomic emission spectroscopy, high-performance liquid chromatography, fluorescence spectroscopy, liquid-chromatography/mass-spectrometry, ultraviolet/visible spectroscopy, infrared spectroscopy, electrochemical methods using; sodium ion selective electrode, or chloride ion selective electrode. 
   
   
       15 . The method of  claim 13 , wherein the botulinum toxin is  Clostridium Botulinum  A 
   
   
       16 . The method of  claim 14 , wherein the botulinum toxin is  Clostridium Botulinum  A 
   
   
       17 . The method of  claim 13 , wherein the formulation is a liquid formulation. 
   
   
       18 . The method of  claim 14 , wherein the formulation is a liquid formulation.