Validasi metode analisis kolesterol menggunakan HPLC–ELSD dengan matriks sampel telur ayam

Abstract

A high-performance liquid chomatography (HPLC) method for the separation and quantification of cholesterol using an evaporative light-scattering detector (ELSD) is described. Detection of cholesterol was achieved in 1,50 until 1,53 minutes using cholesterol standard. A simplified binary mobile phase mixture of hexane and isopropanol (90:10) was used to separate cholesterol. Detection was accomplished with an ELSD, with the following settings: evaporation temperature 50 ºC, air pressure 2,2 bars, flow rate of mobile phase 2 mL/min and grain 7. A calibration curve was obtained from 0 to 5000 μg/g sample, the linear equation of cholesterol is y = 35,58x - 3140, with a correlation coefficient of 0,997. Instrument detection limit (IDL) was 1,07 μg/mL and limit of quantitation (LOQ) was 3,56 μg/mL. Recovery by spikin cholesterol standard in egg sample at low concentration was 122,13%, medium concentration was 108,23%, and high concentration was 44,71%. Methode detection limit (MDL) value was 2,30 μg/g sample. With intra reproducibility value was 0,04%.