title: Rapid UPLC-MS/MS method for routine analysis of plasma pristanic, phytanic, and very long chain fatty acid markers of peroxisomal disorders.
abstract: Quantification of pristanic acid, phytanic acid, and very long chain fatty acids (i.e., hexacosanoic, tetracosanoic, and docosanoic acids) in plasma is the primary method for investigateing a multitude of peroxisomal disorders (PDs). Typically based on GC-MS, existing methods are time-consuming and laborious. In this paper, we present a rapid and specific liquid chromatography tandem mass spectrometric method based on derivatization with 4-[2-(N,N-dimethylamino)ethylaminosulfonyl]-7-(2-aminoethylamino)-2,1,3-benzoxadiazole (DAABD-AE). Derivatization was undertaken to improve the poor mass spectrometric properties of these fatty acids. Analytes in plasma (20 mul) were hydrolyzed, extracted, and derivatized with DAABD-AE in approximately 2 h. Derivatives were separated on a reverse-phase column and detected by positive-ion electrospray ionization tandem mass spectrometry with a 5 min injection-to-injection time. Calibration plots were linear over ranges that cover physiological and pathological concentrations. Intraday (n = 12) and interday (n = 10) variations at low and high concentrations were less than 9.2%. Reference intervals in normal plasma (n = 250) were established for each compound and were in agreement with the literature. Using specimens from patients with established diagnosis (n = 20), various PDs were reliably detected. In conclusion, this method allows for the detection of at least nine PDs in a 5 min analytical run. Furthermore, this derivatization approach is potentially applicable to other disease markers carrying the carboxylic group.
authors.affiliation: National Laboratory for Newborn Screening, Department of Genetics, King Faisal Specialist Hospital and Research Center, Riyadh, Saudi Arabia. dirbashi@kfshrc.edu.sa
authors.forename: Osama Y
authors.initials: OY
authors.lastname: Al-Dirbashi
authors.forename: Tomofumi
authors.initials: T
authors.lastname: Santa
authors.forename: Mohamed S
authors.initials: MS
authors.lastname: Rashed
authors.forename: Zuhair
authors.initials: Z
authors.lastname: Al-Hassnan
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Shimozawa
authors.forename: Aziza
authors.initials: A
authors.lastname: Chedrawi
authors.forename: Minnie
authors.initials: M
authors.lastname: Jacob
authors.forename: Manhal
authors.initials: M
authors.lastname: Al-Mokhadab
mesh_terms: D015415:Biomarkers / Q000097:blood*; D002853:Chromatography, Liquid / Q000379:methods*; D005227:Fatty Acids / Q000097:blood*; D006801:Humans; D018901:Peroxisomal Disorders / Q000097:blood* / Q000175:diagnosis; D010831:Phytanic Acid / Q000097:blood*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D015415:Biomarkers; D005227:Fatty Acids; D010831:Phytanic Acid; C002844:pristanic acid