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A more recent version of this article appeared on August 1, 2008

Papers In Press, published online ahead of print April 25, 2008
J. Lipid Res., doi:10.1194/jlr.D800019-JLR200
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Submitted on April 2, 2008
Revised on April 17, 2008
Accepted on April 25, 2008

Rapid UPLC-MS/MS method for routine analysis of plasma pristanic, phytanic and very-long chain fatty acid markers of peroxisomal disorders

Osama Y. Al-Dirbashi, Tomofumi Santa, Mohamed S. Rashed, Zuhair Al-Hassnan, Nobuyuki Shimozawa, Aziza Chedrawi, Minnie Jacob, and Manhal Al-Mokhadab

Genetics, King Faisal Specialist Hospital and Research Center, Riyadh 11211

Corresponding Author: dirbashi{at}kfshrc.edu.sa

Quantification of pristanic, phytanic and very long-chain fatty acids (i.e. hexacosanoic, tetracosanoic and docosanoic acids) in plasma is the primary test to investigate for 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 (LC-MS/MS) method based on derivatization with 4-[2-(N,N-dimethylamino)ethylaminosulfonyl]-7-(2-aminoethylamino)-2,1,3-benzoxadiazole (DAABD-AE). Derivatization was taken to improve the poor mass spectrometric properties of these fatty acids. Analytes in plasma (20 mu l) were hydrolyzed, extracted and derivatized with DAABD-AE in about 2 hours. Derivatives were separated on a reversed-phase column and detected by positive-ion electrospray ionization (ESI)-MS/MS 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 carboxylic group


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