Determination of sulfur-containing amino acid residues in proteins by pyrolysis-gas chromatography with flame photometric detection

Summary A new method for the determination of sulfur-containing amino acid residues, cysteine (Cys) and methionine (Met) in proteins has been developed with pyrolysis-gas chromatography. Characteristic sulfur compounds among the complicated pyrolysis products formed from protein samples were separated by using a high resolution fused silica capillary column and selectively detected by flame photometric detector (FPD). In order to improve the reproducibility, an internal standard substance, benzothiophene, was pyrolyzed at 600 °C together with the protein samples. Thus the observed pyrograms were interpreted in terms of the compositional analysis. On the basis of the relative peak intensities of CH₃SH and H₂S, the simultaneous determinations of Cys and Met residues in various enzyme proteins were successfully carried out with 10–12% and 5–6% of relative standard deviations, respectively.

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Bestimmung von schwefelhaltigen Aminosäureresten in Proteinen durch Pyrolyse-GC mit flammenphotometrischer Detektion