Assay
Place about 100 mg of Stearic Acid in a small conical flask fitted with a suitable reflux attachment. Place about 50 mg of
USP Stearic Acid RS and about 50 mg of
USP Palmitic Acid RS in a similar flask. Treat each flask as follows. Add 5.0 mL of a solution prepared by dissolving 14 g of boron trifluoride in methanol to make 100 mL, swirl to mix, and reflux for 15 minutes or until the solid is dissolved. Cool, transfer the reaction mixture with the aid of 10 mL of chromatographic solvent hexane to a 60-mL separator, and add 10 mL of water and 10 mL of saturated sodium chloride solution. Shake, allow to separate, then drain and discard the lower, aqueous layer. Pass the hexane layer through 6 g of anhydrous sodium sulfate (previously washed with chromatographic solvent hexane) into a suitable flask. Using a syringe fitted with a suitable needle, introduce a 1-µL to 2-µL portion of the assay preparation (which contains the Stearic Acid) into a suitable gas chromatograph equipped with a flame-ionization detector. The column preferably is of glass, 1.5 m in length and 3 mm in inside diameter, and it is packed with 15% G4 on support S1A. The carrier gas is helium, passed through a bed of molecular sieve for drying, if necessary. The temperatures of the port and the detector are maintained at 210
, and the column temperature is maintained at 165
.
System suitability
In a suitable chromatogram, the resolution factor,
R (see
Chromatography 621), is not less than 2.0 between the peaks from methyl palmitate and methyl stearate (located by comparison with the chromatogram of the standard preparation), and five replicate injections of a single sample show a coefficient of variation of not more than 1.5% in the percentage of methyl stearate and methyl palmitate, respectively. Measure the peak areas of the fatty acid esters in the chromatogram, and determine the percentage of C
18H
36O
2 in the portion of Stearic Acid taken by the formula:
100(A / B),
in which
A is the area due to the methyl stearate peak, and
B is the sum of the areas of all of the fatty acid ester peaks in the chromatogram. Similarly, determine the percentage of C
16H
32O
2.