Liquid chromatographic determination of D- and L-amino acids by derivatization with o-phthaldialdehyde and N-isobutyryl-L-cysteine. Applications with reference to the analysis of peptidic antibiotics, toxins, drugs and pharmaceutically used amino acids.
In order to evaluate and extend the applicability of an analytical method that enables the quantitative and simultaneous high-performance liquid chromatographic determination of D- and L-amino acids (DL-AAs) by automated precolumn derivatization with o-phthaldialdehyde together with the chiral thiol N-isobutyryl-L-cysteine [J. Chromatogr., 666 (1994) 259] selected natural and synthetic bioactive peptides, as well as pharmaceutically used formulations of AA, were investigated and the amounts of D- and L-AA determined by fluorescence detection. Peptides containing cys(e)ine were oxidized with performic acid prior to hydrolysis with 6 M HCl, and those containing Trp were hydrolyzed with 4 M methanesulfonic acid (24 h at 110 degrees C in both cases). Peptides analyzed were the peptide antibiotics bacitracin, gramicidins A and S, polymyxin B, metanicin C, the peptide toxin malformin A and the peptide drugs D-Arg-[Hyp3,Thi5,8,D-Phe7]-bradykinin, beta-casomorphin and alpha s1-exorphin. Further, the enantiomeric ratios of pharmaceutically used AA formulations containing racemic DL-Ser, DL-Asp and DL-Met were determined, and the AA drugs L-Asp and L-Trp were tested negatively for the presence of the respective D-enantiomers. In two aqueous formulations of L-AA used for parenteral nutrition, low amounts of D-AA (0.1-0.9% with respect to certain L-AA enantiomers and of totally 128 mg and 149 mg D-AAs per liter infusion solution) were determined.
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis* / Q000737:chemistry; D000900:Anti-Bacterial Agents / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid; D003545:Cysteine / Q000031:analogs & derivatives* / Q000737:chemistry; D004345:Drug Industry; D008969:Molecular Sequence Data; D010455:Peptides; D004364:Pharmaceutical Preparations / Q000737:chemistry*; D013050:Spectrometry, Fluorescence; D013237:Stereoisomerism; D014118:Toxins, Biological / Q000737:chemistry*; D009764:o-Phthalaldehyde / Q000032:analysis* / Q000737:chemistry
D000596:Amino Acids; D000900:Anti-Bacterial Agents; D010455:Peptides; D004364:Pharmaceutical Preparations; D014118:Toxins, Biological; C096972:N-isobutyrylcysteine; D009764:o-Phthalaldehyde; D003545:Cysteine