Comparison Between Solvent Extraction Using Gas Chromatography Mass Spectrometry Triple Quadrupole (gcms-qqq) Of Cholesterol And Cholesterol Oxidation Derivatives

Abstract: Cholesterol Oxidation Products (COPS) are well known for their negative biological effects. Till now,there is no study reported on solvent extraction used for COPs. In this study, two solvent extractiontechniques, namely chloroform/methanol (2:1, v/v) or Folch method and n-hexane/2-propanol (3:2, v/v) orHara-Radin method of cholesterol and COPs in beef tallow and lard were compared. Cholesterol and COPs(5-cholestane, 7-ketocholesterol and 25-hydroxycholesterol) contents in both fats also analyzed. The analysispassed through four major steps; extraction of lipids, saponification, enrichment of COPs and quantificationby gas chromatography mass spectrometry triple quadrupole. All standards showed good linearity withcorrelation coefficient (r ) of 0.9998 (5-cholestane), 0.9999 (cholesterol), 0.9873 (25-hydroxycholesterol) and20.9693 (7-ketocholesterol). Data obtained by this method was analyzed based on precision and recovery criteria.Precision measured as standard deviation (SD) was between 0.0040 and 2.2460; and no significant different(P>0.05) for the recovery testing using 5 -cholestane in both method. Recovery by Hara Radin method in lard(27.23%) and beef tallow (8.73%) was higher than the Folch method, 14.25% and 2.97% respectively. This studyimplies that Hara-Radin method can be an alternative method to avoid the use of harmful solvent such aschloroform in Folch method.
Key words:Cholesterol Oxidation Products Gas Chromatography Mass Spectrometer Triple Quadrupole Fat Solvent Extraction