Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry-based amino acid analysis.
Amino acid analysis has been an integral part of analytical biochemistry for more than 50 years. However, its experimental design, which includes derivatization of amino acids followed by some kind of chromatographic separation, has not changed over the years. We have developed a matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF)-based method for the quantitative analysis of amino acids. This method does not require any amino acid modification, derivatization, or chromatographic separation. The data acquisition time is decreased to several seconds for a single sample. No significant ion suppression effects were observed with the developed sample deposition technique, and the method was found to be reproducible. Linear responses between the amino acid concentration and the peak intensities ratio of corresponding amino acid to internal standard were observed for all amino acids analyzed in the range of concentrations from 20 to 300 microM, and correlation coefficients were between 0.983 (for arginine) and 0.999 (for phenylalanine). Limits of quantitation were between 0.03 microM (for arginine) and 3.7 microM (for histidine and homocysteine). This method was applicable to the mixtures of free amino acids as well as to HCl hydrolysates of proteins. Furthermore, we have shown that this method can be applied to other biologically important low-molecular weight compounds such as glucose.
Alterman
Michail A
MA
Biochemical Research Service Laboratory and Analytical Proteomics Laboratory, University of Kansas, Lawrence, KS 66045, USA. malterman@ku.edu
Gogichayeva
Natalya V
NV
Kornilayev
Boris A
BA
D000596:Amino Acids / Q000032:analysis*; D005947:Glucose / Q000032:analysis; D015203:Reproducibility of Results; D012710:Serum Albumin, Bovine / Q000032:analysis* / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D013997:Time Factors
D000596:Amino Acids; D012710:Serum Albumin, Bovine; D005947:Glucose