Method of separating compound(s) from mixture(s)

Abstract

The invention relates to methods of separating compounds of interest from mixtures using high performance liquid chromatography (HPLC). An initial separation of a sample of a mixture is performed to predict a retention time and elution time for the compound of interest on a preparative scale HPLC column and, optionally, to verify that an expected compound of interest is present in the mixture. Preparative scale HPLC is then used to separate and collect at least a portion of the compound from the mixture using the predicted elution time of the compound.

What is claimed is:  
     
         1 . A method of separating a compound of interest from a mixture, the method comprising the steps of: 
 (a) providing a mixture containing a compound of interest;    (b) subjecting a portion of the mixture to a separation using thin layer chromatography to determine an Rf value for the compound of interest;    (c) predicting an elution time of the compound of interest on a preparative scale HPLC column using the determined Rf value for the compound of interest;    (d) subjecting all or a portion of the remaining mixture to a preparative scale HPLC system comprising a preparative scale HPLC column; and    (e) collecting at least a portion of the compound of interest using the predicted elution time.    
     
     
         2 . The method of  claim 1  wherein the step of predicting an elution time for the compound of interest comprises: 
 (1) predicting a retention time of the compound of interest from the preparative scale HPLC column using a predetermined correlation function between Rf value from the TLC and retention time on the preparative scale HPLC column along with the determined Rf value of the compound of interest; and  
 (2) selecting a window of time around the predicted retention time within which the compound is expected to elute.  
 
     
     
         3 . The method of  claim 1  wherein step (b) comprises: 
 (1) subjecting a portion of the mixture to a separation using thin layer chromatography to produce one or more spots or zones;  
 (2) analyzing the one or more spots or zones using a mass spectrometer to determine the spot or zone containing the compound of interest; and  
 (3) determining an Rf value for the compound of interest.  
 
     
     
         4 . The method of  claim 1  wherein an artificial neural network is used to predict the elution time in step (c).  
     
     
         5 . The method of  claim 1  wherein a dynamic correlation function is used to predict the elution time in step (c).