Simultaneous capillary gas chromatographic profiling of medium-and long-chain fatty acid methyl esters with split injection. Correction for injection-related discrimination by the 'bracketing' method.

Split injection-related discrimination can be a source of inaccuracy and imprecision in quantitative capillary gas chromatographic profiling methods for compounds with relatively big differences in boiling points, such as the methyl esters of medium- and long-chain fatty acids prepared from biological materials. We systematically investigated a standard containing equal masses of saturated fatty acid methyl esters, with chain lengths from C5 to C26, under different injection conditions, including injection temperature, sample volume and split ratio. Day-to-day performance was studied under one set of conditions. Normalized peak areas, reciprocal response factors, using either C17 or C23 as an internal standard, and 'bracketed' reciprocal response factors (peak area of each analyte divided by half the sum of the peak areas of two adjacent esters were calculated. In all experiments the bracketed reciprocal response factors were found to be closest to unity with the lowest coefficients of variation.