Microcolumns for the separation of analytes from a sample in the millisecond time scale

Abstract

The present invention generally relates to a microcolumn capable of separating an analyte from a sample in the millisecond time domain. The microcolumn is capable of such rapid separation by employing small column volumes that can tolerate medium to high flow rates. The invention also relates to a method of loading a microcolumn capable of separating an analyte from a sample in the millisecond time domain.

What is claimed is:  
     
         1 . A microcolumn for separating an analyte from a sample, the microcolumn comprising: 
 (a) an active layer, where the active layer is introduced into the microcolumn by a plurality of injections such that the active layer is capable of separating the analyte from the sample within the millisecond time domain; and    (b) an inert layer.    
     
     
         2 . The microcolumn of  claim 1  wherein said microcolumn has an internal diameter of about 50 microns to about 2 centimeters and a length of about 0.2 millimeters to about 2 centimeters.  
     
     
         3 . The microcolumn of  claim 1  wherein said microcolumn has an internal diameter of about 0.5 to about 2.1 millimeters and a length of about 1 millimeter to about 2 centimeters.  
     
     
         4 . The microcolumn of  claim 1  wherein the active layer is from about 10 microns to about 1.1 millimeters in length.  
     
     
         5 . The microcolumn of  claim 1  wherein the active layer is not less than 60 microns in length.  
     
     
         6 . The microcolumn of  claim 1  wherein the inert layer comprises particles selected from the group of materials consisting of diol-bonded silica, diol-bonded glass beads, agarose beads, hydroxylated perfusion media, and glycol coated perfusion media.  
     
     
         7 . The microcolumn of  claim 6  wherein the material comprises diol-bonded silica.  
     
     
         8 . The microcolumn of  claim 6  wherein the inert layer and active layer are formed from different materials.  
     
     
         9 . The microcolumn of  claim 6  wherein said microcolumn has the capacity to separate the analyte from the sample in about 1 to about 500 milliseconds after injection of the sample into the microcolumn.  
     
     
         10 . The microcolumn of  claim 6  wherein said microcolumn has the capacity to separate the analyte from the sample in about 5 to about 100 milliseconds after injection of the sample into the microcolumn.