Determination of cholesterol in fat and muscle of pig by HPLC and capillary gas chromatography with solvent venting injection

Abstract

AbstractTwo methods for determination of cholesterol in fat and muscle of pig were evaluated: extraction with chloroform:methanol (2:1, v/v) followed by saponification (method 1) and direct saponification (method 2). HPLC and GC were used to determine cholesterol concentrations. GC analysis was performed with a capillary column of 100 μm using a PTV injector in the modes of cold split and solvent venting. Cholesterol was analyzed without derivatization. Both methods of extraction did not present significant differences (p > 0.01). Sample analysis by GC with solvent venting injection and HPLC showed the lowest % r.s.d. but GC in the cold split mode allowed to obtain a shorter analysis time. Cholesterol concentrations obtained by HPLC were not statistically different from the results obtained by GC with solvent venting injection and were slightly lower than those previously reported. Cholesterol concentrations in fat and muscle tissues respectively ranged from 52 to 77 mg/100 g and from 55 to 65 mg/100 g.