Chromatographic system (see Chromatography 621)
Prepare as directed for
Fatty Acid Composition under
Fats and Fixed Oils 401. Chromatograph the
Standard preparation, and record the peak responses as directed for
Procedure: the relative retention times are about 0.9 for methyl palmitate and 1.0 for methyl stearate; the resolution,
R, between methyl stearate and methyl palmitate is not less than 3.0.
Procedure
Separately inject equal volumes (about 1 µL) of the
Assay preparation and the
Standard preparation into the chromatograph, record the chromatograms, identify the methyl palmitate peak in the chromatogram obtained from the
Assay preparation by comparing the retention times of the peaks in that chromatogram with those in the chromatogram obtained from the
Standard preparation, and measure the responses for all the peaks, excluding the solvent peak. Calculate the percentage of C
16H
32O
2 in the portion of Palmitic Acid taken by the formula:
100A/B,
in which
A is the methyl palmitate peak response; and
B is the sum of the responses of all the peaks in the chromatogram except the solvent peak. Similarly calculate the percentage of stearic acid in the portion of Palmitic Acid taken.