title: A routine method for cholesterol and 7-dehydrocholesterol analysis in dried blood spot by GC-FID to diagnose the Smith-Lemli-Opitz syndrome.
abstract: This work was aimed to implement a fast and simple method to quantify cholesterol (CHOL) and 7-dehydrocholesterol (7-DHC) in dried blood spot (DBS) to diagnose the Smith-Lemli-Opitz syndrome (SLOS), an inborn error of CHOL biosynthesis. We developed and validated a GC-FID method for separation and quantification of underivatized CHOL and 7-DHC using a DBS disc of 6mm with a run time of 9 min. Correlation coefficients (r) of calibration curves ranged from 0.998 to 0.999 for CHOL and from 0.997 to 0.998 for 7-DHC. Within-day and between-day imprecision (CV%), accuracy (%), carry-over, and extraction efficacy (%) were also evaluated for validation. CHOL and 7-DHC were analyzed in DBS and plasma samples from 8 SLOS patients and 30 unaffected subjects. In SLOS patients, 7-DHC/CHOL ratios in DBS and plasma samples ranged from 0.035 to 1.448 and from 0.012 to 0.926, respectively. Results from calibration curves, quality controls and patient samples reveal that the method is suitable to analyze DBS to screen patients affected by SLOS.
authors.affiliation: Department of Biochemistry and Medical Biotechnologies, University Federico II of Naples, Naples, Italy.
authors.forename: Monica
authors.initials: M
authors.lastname: Gelzo
authors.forename: Stefano
authors.initials: S
authors.lastname: Clericuzio
authors.forename: Rosalba
authors.initials: R
authors.lastname: Barone
authors.forename: Oceania
authors.initials: O
authors.lastname: D'Apolito
authors.forename: Antonio
authors.initials: A
authors.lastname: Dello Russo
authors.forename: Gaetano
authors.initials: G
authors.lastname: Corso
mesh_terms: D016022:Case-Control Studies; D002784:Cholesterol / Q000097:blood*; D003684:Dehydrocholesterols / Q000097:blood*; D059788:Dried Blood Spot Testing / Q000379:methods*; D005410:Flame Ionization / Q000379:methods*; D006801:Humans; D016014:Linear Models; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D019082:Smith-Lemli-Opitz Syndrome / Q000097:blood* / Q000175:diagnosis
chemical_list: D003684:Dehydrocholesterols; D002784:Cholesterol; C016705:7-dehydrocholesterol