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A Robust LC/MS/MS Method for the Quantitation of Very Long Chain Fatty Acids for Research into the Characterization of Peroxisomal Disorders

ollision cell for multi-component analysis at reduced MRM dwell times, maintaining sensitivity and preventing cross talk. This reduces significantly the occurrence of false positive results.

Rugged and reliable triple quadrupole tandem MS system for maximum uptime

Compatibility with the broadest range of ionization sources for the analysis of a wide range of compounds.

Experimental Conditions

Samples were prepared according to the procedure described by Johnson(1) with some modifications. The hardware configuration included an API 3000TM LC/MS/MS System Triple Quadrupole Mass Spectrometer equipped with a TurboIonSpray source. The source operates in positive ion mode at a voltage of +5500 volts and with a turbo gas flow of 8 L/min of air heated at 300 degrees C (nominal heating-gun temperature).

MRM measurements were made using declustering potential (DP) and collision energy (CE) values as optimized automatically by the software for each of the analytes (DP value at 40 V; CE between 30 and 45 eV for the singly charged species measured here).

Chromatography was performed through an Agilent 1100 LC Binary Pump, and separation was accomplished via a Kromasil C8 5μm, 2 x 50 mm column housed in an Agilent 1100 Column Oven at 60C. 3 μL of the reconstituted sample extract were injected into the column using an Agilent 1100 Wellplate Autosampler. All hardware set-up details, solution and reagents requirements, and operating parameters were stored on CD for quick transfer of the proposed methodology.

Results and Discussion

The results of this research study are summarized in the Figures below.

As shown in Figure 1, chromatographic separation and identification of the derivatized pristanic, phytanic, docosanoic, tetracosanoic, and hexacosanoic VLCFAs, and the derivatized intern
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