Profiling of human body fluids in healthy and diseased states using gas chromatography and mass spectrometry, with special reference to organic acids.

A very sensitive and specific method of 2-ketoacid determination in various biological fluids using gas chromatography chemical ionization mass spectrometry of O-trimethylsilyl-quinoxalinol derivatives is described. After derivatization with o-phenylenediamine in acidic medium and extraction, the 2-ketoacids, and the 2-ketovaleric acid used as internal standard, are silylated and resolved on a capillary column and their concentrations automatically determined by monitoring the m/z values corresponding to their respective protonated molecular ions, using ammonia as reactant gas. The detection limit is below 50 ng ml-1 of each 2-ketoacid. The recoveries of 2-ketoacids from urine and plasma were between 97 and 104%. Problems encountered with interfering substances were tested for and discussed. This method has been applied successfully to various metabolic disorders characterized by an accumulation of aliphatic 2-ketoacids. The metabolic interrelationship of branched chain 2-ketoacids and other compounds, especially pyruvate, is discussed.

Quantitation of 2‐ketoacids in biological fluids by gas chromatography chemical ionization mass spectrometry of O‐trimethylsily‐quinoxalinol derivatives

We assessed the quantitative performances of a classical method for profiling urinary organic acids: ethyl acetate extraction/oxime-trimethylsilyl derivatization/GC-MS. Twenty-seven acids were quantified on the basis of specific ions in both scan and selected-ion monitoring modes. We found that the tuning of the mass detector severely affects the calibration factors, being critical to achieve quantitative results, and we propose a practical procedure for reproducible tuning. Of seven compounds tested, tropic acid was retained as the internal standard suitable for most of the acids of clinical interest; a second internal standard, 2-ketocaproic acid, was used in quantifying keto-acids. The within-day and total relative standard deviations (CVs), estimated from scan-mode analyses of urine, ranged from 2.6% to 12.7% and from 4.2% to 11.8%, respectively. Curvilinear relationships between analytical response and concentration were observed for most of the acids investigated.

/pdf/gc-ms-profiling-of-urinary-organic-acids-evaluated-as-a-4tq7s6avzw.pdf

GC-MS profiling of urinary organic acids evaluated as a quantitative method.

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Quantitative gas chromatography and single-ion detection of aliphatic α-keto acids from urine as their o-trimethylsilylquinoxalinol derivatives

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Quantitative gas chromatography and single-ion detection of aliphatic α-keto acids from urine as their o-trimethylsilylquinoxalinol derivatives