Validation of a gas chromatography-mass spectrometry method for the analysis of sterol oxidation products in serum.
A validated gas chromatography-mass spectrometry (GC-MS) detection method for the quantitative analysis of sterol oxidation products (SOPs) in serum is described. After a lipid extraction procedure with chloroform-methanol, a cold saponification and purification by solid phase extraction, oxysterols were derivatized to form trimethyl-sylil-ethers which were subjected to GC-MS analysis. Calibration curves for cholesterol oxidation products showed determination coefficient (R(2)) of 1.0, with low values for the coefficient of variation of the response factors (< 1%). Detection and quantification limits were below 5 ng/mL and 10 ng/mL, respectively. Recovery data were between 77.65% and 110.29% (CV < 10% for all compounds). Good results were obtained for within- and between-day repeatability, with values below 10%. In conclusion, the method performed is suitable for the determination and quantification of SOPs in serum.
Menéndez-Carreño
María
M
Departamento de Ciencias de la Alimentación, Fisiología y Toxicología, Faculty of Pharmacy, University of Navarra, C/Irunlarrea s/n 31080-Pamplona, Spain.
García-Herreros
Cecilia
C
Astiasarán
Iciar
I
Ansorena
Diana
D
D002784:Cholesterol / Q000097:blood; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D006801:Humans; D006888:Hydroxycholesterols / Q000097:blood; D010084:Oxidation-Reduction; D010840:Phytosterols / Q000097:blood; D011786:Quality Control; D015203:Reproducibility of Results; D012503:Saponins / Q000737:chemistry; D012680:Sensitivity and Specificity; D013261:Sterols / Q000097:blood*
D006888:Hydroxycholesterols; D010840:Phytosterols; D012503:Saponins; D013261:Sterols; D002784:Cholesterol