Validation of Analytical Method for Quantification of Egg Cholesterol Using Reversed Phase-High Performance Liquid Chromatography-Multiwavelength Detector

Y. L. Maslukhah (Graduate Student of Department of Food Science and Technology, Faculty of Agricultural Technology, IPB University (Bogor Agricultural University))
D. N. Faridah (Department of Food Science and Technology, Faculty of Agricultural Technology, IPB University (Bogor Agricultural University))
H. N. Lioe (Department of Food Science and Technology, Faculty of Agricultural Technology, IPB University (Bogor Agricultural University))

Tropical Animal Science Journal
Vol. 42 No. 3 (2019): Tropical Animal Science Journal

In this research, analytical method of cholesterol content in eggs by Reversed Phase-High Performance Liquid Chromatography-Multiwavelength Detector (RP-HPLC-MWD) was validated. Our experiment validated the modified method of AOAC 994.10:2012 to get a more simple and efficient analytical method of cholesterol content. The sample was saponified using 10% KOH concentration for 15 min at 80 °C, then this analytical method was validated. RP-HPLC-MWD condition was at 100% MeOH as a mobile phase, flow rate of 1.0 mL/min, detection UV at 205 nm, cholesterol was detected at 10.38±0.13 min. As a result, the coefficients of determination for instrument and method linearities reached 0.9991 and 0.9912, respectively. The limits of detection and quantification of RP-HPLC-MWD instrument were found at 5 and 10 μg/mL, respectively, while the method-detection limit and quantification limit were 250 and 500 μg/g sample, respectively. Recovery values for the cholesterol analysis ranged from 98.62% to 112.26%, with a precision of 1.05%‒3.90%. Additionally, intralab reproducibility was known to reach 3.27%. This validated method can be applied for the analysis of cholesterol in various eggs available in the market.