P. Avino et al., High-performance liquid chromatography intercomparative study for amino acid analysis in two tissues by PITC- and OPA-derivatizations, ANAL LETTER, 34(6), 2001, pp. 867-882
In this paper, we have extensively investigated the pre-column derivatizati
on method with o-phthaldialdehyde (OPA) to analyze cysteine, cystine, among
with other 18 amino acids, using reversed-phase high-performance liquid ch
romatography analysis (RP-HPLC), and after we have compared it with an othe
r method involving phenylisothiocyanate (PITC) in order to have a two sensi
tive methods to use accordingly the matrix. We have studied the analytical
conditions for the OPA-reaction together with precision, linearity range, c
orrelation coefficient (>0.998) and limit of detection (between 14 fmol and
1 pmol) for each amino acid. Then, we have applied the two methods to the
analysis of larynx and kidney tissues: between the two methods, the amino a
cid recoveries (between 80% and 100%) and the correspondence of the OPA and
PTC results are satisfactory considering the different involved derivatiza
tion procedures. PTC- and OPA-chromatograms obtained in the analysis of act
ual samples are reported together with the amino acid level, with particula
r regard to the cysteine/cystine determination.