Future potential of targeted component analysis by multidimensional liquid chromatography-mass spectrometry.
Multidimensional liquid chromatography (MDLC) may be used in either (i) the profiling mode where it is the objective to fractionate all components in a mixture or (ii) the targeted component mode in which it is the objective to determine specific analytes. This paper focuses on targeted component analysis from complex mixtures, addressing the critical operations of analyte selection and transport from the first to the second dimension. Although the physical operation of switching a component into the second dimension with computer controlled valving is simple, it is shown that changes in analyte retention time and peak width with column age and fouling are a serious problem. The analyte moves out of the preselected time window for valve switching and quantitation is compromised in the second dimension. It is proposed that a solution to the "drifting peak" phenomenon in targeted component analysis is to use binary mobility elution in the first dimension. Binary mobility refers to those systems, such as affinity chromatography, in which analyte mobility is generally either 0 or 1 relative to mobile phase velocity. Coupling these binary changes in analyte mobility in the first dimension with valve switching eliminates the "drifting peak" phenomenon. In addition, it is shown that a wide time window may be used in affinity separations without compromising the separation or accumulating contaminants. Several cases are described in which immunosorbents were used with reversed phase columns to provide quantitative targeted component analyses from complex mixtures.
Regnier
F
F
Department of Chemistry, Purdue University, West Lafayette, IN 47907-1393, USA.
Huang
G
G
D000818:Animals; D000906:Antibodies / Q000737:chemistry / Q000276:immunology; D000941:Antigens / Q000032:analysis / Q000302:isolation & purification; D002417:Cattle; D002853:Chromatography, Liquid / Q000379:methods*; D006454:Hemoglobins / Q000302:isolation & purification; D006801:Humans; D007328:Insulin / Q000276:immunology / Q000302:isolation & purification; D013058:Mass Spectrometry / Q000379:methods*; D004364:Pharmaceutical Preparations / Q000032:analysis*; D011506:Proteins / Q000032:analysis / Q000737:chemistry*
D000906:Antibodies; D000941:Antigens; D006454:Hemoglobins; D007328:Insulin; D004364:Pharmaceutical Preparations; D011506:Proteins