Rapid determination of cholesterol in milk containing emulsified foods

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

“…Recovery of cholesterol analysis using HPLC-DAD (Bavisetty & Narayan, 2015;Albuquerque et al, 2016) ranging from 95.2%, 98%, to 111%-125%, respectively. The recovery of cholesterol analysis using HPLC-UV operated at fixed wavelength reached 84%-103%; 93.34%-102.34%; and 93.33±0.22% (Ahn et al, 2012;Mariutti et al, 2008;Stroher et al, 2012), while Lioe et al (2013) found that the recovery value in HPLC-ELSD was 108%-122.13%. Previously, the validation of cholesterol analytical methods on diverse foodstuffs (sour cream, whole eggs, egg yolks, and some fish species) using HPLC-DAD demonstrated RSD analysis of 0.69%, 0.54%, 1.92%, 3.05%, respectively (Albuquerque et al, 2016;Bavisetty & Narayan, 2015).…”

“…Saponification constitutes initial and crucial step to release cholesterol from the other components since saponification was reported capable of improving cholesterol concentration in egg yolk compared to sample without saponification (John et al, 2015). The use of KOH in saponification enables to hydrolyze disrupting components such as fatty acids and acyl glycerides (Ahn et al, 2012;Park et al, 2013), while its effect may depend on the concentration and heating time (Salimon et al, 2012;Souza et al, 2017). The modification in this procedure was made at the following conditions, including less amount of sample (1 g, from 6 g of standard method), KOH concentration, saponification time, and changing toluene with hexane for extraction.…”

“…Analysis of cholesterol in the food matrices in previous studies could use a spectrophotometer (Santi et al, 2015;Abdurrahman et al, 2016), but in general, the existing methods for assessing cholesterol content in food matrices are based on chromatographic technique. Gas chromatography (GC) was found to be more sensitive for determination of cholesterol in the food matrices (Ahn et al, 2012;Bavisetty & Narayan, 2015;Lioe et al, 2013;Park et al, 2013;Souza et al, 2017;Stroher et al, 2012). Compared to HPLC, GC-based analysis for cholesterol may be hindered by limitations, i.e., time-consuming sample preparation and costly, since derivatization of cholesterol compounds and reliability test using internal standard must be first performed before being used.…”

“…Diode array detector (DAD) or UV seemed to be the most desirable detector for cholesterol quantification (Ahn et al, 2012;Albuquerque et al, 2016;Bavisetty & Narayan, 2015;Lioe et al, 2013;Stroher et al, 2012). MWD (Multi Wavelength Detector) constitutes one of the ultraviolet detectors capable of performing more sensitive and selective detection than fixed wavelength detector of UV-Vis and the other detectors such as ELSD and RI (Mariutti et al, 2008;Wolfender, 2009).…”

“…Studies on the quantification of egg cholesterol using RP-HPLC-MWD have not been reported. Mobile phase-acetonitrile and methanol-is often applied in RP-HPLC at UV detection of 195-210 nm and flow rate of 1-1.5 mL/min (Ahn et al, 2012;Albuquerque et al, 2016;Bavisetty & Narayan, 2015;Stroher et al, 2012). AOAC 994.10:2012 is a standard method for cholesterol analysis in foodstuffs by GC-FID instrument (AOAC, 2012a); however, the technique needs more samples, which in turn requires more chemicals and saponification time.…”