Measurement of amino acid isotope enrichment by liquid chromatography mass spectroscopy after derivatization with 9-fluorenylmethylchloroformate.
An automated method is described to measure tracer-tracee-ratios (TTR) of plasma amino acids after their separation as 9-fluorenylmethylchloroformate derivatives. In a 45 min cycle, 5 microl plasma aliquots were derivatized, HPLC separated and subjected to electrospray ionization. By applying source collision induced dissociation, derivatives were dissociated at the peptide bond releasing the originating amino acids into the mass spectrometer. This approach enabled the determination of plasma amino acid TTRs with a standard deviation between 0.15 and 0.36%, which is sufficient to study the fate of infused tracers and their conversion products in an in vivo experiment in humans.
van Eijk
Hans M H
HM
Department of Surgery, Maastricht University, Nutrition and Toxicology Research Institute Maastricht (NUTRIM), PO Box 616, NL-6200 MD, Maastricht, The Netherlands. HMH.vaneijk@ah.unimaas.nl
Suylen
Dennis P L
DP
Dejong
Cornelis H C
CH
Luiking
Yvette C
YC
Deutz
Nicolaas E P
NE
D000368:Aged; D000596:Amino Acids / Q000097:blood*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005449:Fluorenes / Q000737:chemistry*; D006801:Humans; D007202:Indicators and Reagents / Q000737:chemistry*; D008297:Male; D008875:Middle Aged; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
C496789:9-fluorenylmethoxycarbonyl; D000596:Amino Acids; D005449:Fluorenes; D007202:Indicators and Reagents