Source: Laboratory of Jeff Salacup - University of Massachusetts Amherst

The product of an organic solvent extraction, a total lipid extract (TLE), is often a complex mixture of hundreds, if not thousands, of different compounds. The researcher is often only interested in a handful of compounds or, if interested in many, may need to remove unwanted constituents that are"in the way" or co-eluting. For example, the concentrations of individual compounds in a sample are often determined on a gas chromatograph coupled to a flame-ionizing detector (GC-FID), because the relationship between FID response (in pA) and the amount of compound in a sample (e.g., ng/µL) is both linear and sensitive. The GC portion of the instrument separates different compounds in a sample based on their boiling point, chemical structure, and affinity with a solid phase that can change according to application. The result is a chromatogram (Figure 1), showing the separation of different chemical constituents in time, as well as their relative concentration (calculated as the area under the curve). However, sometimes more than one compound elutes off the GC at a time (Figure 1). In this case, sample purification is required before compounds can be confidently quantified.
 

Figure 1. A chromatogram showing the separation of different chemical constituents over time and their relative concentration (area under the curve). Co-eluting and separated peaks are shown.