Liquid Chromatography

I. Molnar-Perl, L. Eotvos University, LC of Underivatized AAs

Budapest, Hungary

Figure 1 Separation of the phenylthiocarbamyl AAs separated on TSKgel Super-ODS (for details see Table 3). Peaks: 1, ASP; 2, Glu; 3, Ser; 4, Gly; 5, His; 6, Arg; 7, Thr; 8, Ala; 9, Pro; 10, NH/; 11, Tyr; 12, Val; 13, Met; 14, Cys; 15, Ile, 16, Leu; 17, Phe; 18, Lys. (Reproduced with permission from Tosohaas, The Bioseparation Specialist, (1995) Catalogue p. 157, Figure 9/2.)

Figure 1 Separation of the phenylthiocarbamyl AAs separated on TSKgel Super-ODS (for details see Table 3). Peaks: 1, ASP; 2, Glu; 3, Ser; 4, Gly; 5, His; 6, Arg; 7, Thr; 8, Ala; 9, Pro; 10, NH/; 11, Tyr; 12, Val; 13, Met; 14, Cys; 15, Ile, 16, Leu; 17, Phe; 18, Lys. (Reproduced with permission from Tosohaas, The Bioseparation Specialist, (1995) Catalogue p. 157, Figure 9/2.)

gradient mobile phase and pulsed amperometric detection (Figure 2C). For tryptophan and the sulfur-containing AAs (cysteine/cystine, methionine, glutathione, etc.), the fast isocratic elution of the underivatized samples has gained wide acceptance and is a powerful tool in their quantitation.

Tryptophan can be measured directly, within 8 min, in neutralized alkaline hydrolysates of feed and foodstuffs, using an RP column, 5% methanol containing acetate buffer (pH & 4.0) and UV detection at 280 nm. The pulsed amperometric detection of sulfur-containing AAs, at the low pmol level, was carried out with an Au working- and an Ag/AgCl reference electrode, subsequent to their separation on both cation exchange and on RP columns, applying as mobile phase 0.1 mol L"1 HClO4/0.15 mol L"1 NaClO4/5% ACN.

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