title: HPLC method for quantification and characterization of cholesterol and its oxidation products in eggs.
abstract: A new method was developed for the simultaneous determination of cholesterol and its oxidation products in eggs, using HPLC with UV and refractive index (RI) detectors, and HPLC interfaced with atmospheric pressure chemical ionization coupled to MS (HPLC-APCI-MS). The best conditions for direct saponification of the sample and extraction of the non-saponifiable material were defined using complete factorial designs with central points. The method showed accuracy and precision with a detection limit between 0.002 and 0.079 microg/g. The oxides cholest-5-ene-3beta,20alpha-diol and cholest-5-ene-3beta,25-diol identified by HPLC-UV-RI were not confirmed by HPLC-APCI-MS.
authors.affiliation: Department of Food Science, Faculty of Food Engineering, State University of Campinas, 13083-862 Campinas, SP, Brazil.
authors.forename: Mônica R
authors.initials: MR
authors.lastname: Mazalli
authors.forename: Alexandra C H F
authors.initials: AC
authors.lastname: Sawaya
authors.forename: Marcos N
authors.initials: MN
authors.lastname: Eberlin
authors.forename: Neura
authors.initials: N
authors.lastname: Bragagnolo
mesh_terms: D000390:Air Ionization; D001274:Atmospheric Pressure; D002784:Cholesterol / Q000031:analogs & derivatives* / Q000032:analysis / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D002853:Chromatography, Liquid; D003897:Desmosterol / Q000032:analysis / Q000302:isolation & purification; D004531:Eggs / Q000032:analysis*; D005504:Food Analysis; D006888:Hydroxycholesterols / Q000032:analysis / Q000302:isolation & purification; D007653:Ketocholesterols / Q000032:analysis / Q000302:isolation & purification; D013058:Mass Spectrometry / Q000379:methods; D010084:Oxidation-Reduction; D012680:Sensitivity and Specificity; D013056:Spectrophotometry, Ultraviolet; D014466:Ultraviolet Rays
chemical_list: D006888:Hydroxycholesterols; D007653:Ketocholesterols; C003585:22-hydroxycholesterol; C038027:20-hydroperoxycholesterol; D003897:Desmosterol; D002784:Cholesterol