Shown below, sample chromatograms of the routine physiological analysis program with Arginine eluting at 103 minutes and the physiological extended program separating additional amino acids with Arginine eluting at 157 minutes. Figure 8 and 9 illustrate short programs for some specific amino acids and Glutathione. The graphs are 700×400 pixels. Zooming in could reveal more detail on your monitor.
Fig.1. Physiological standard. Sigma Acidics/Neutrals/Basics. Routine program with Arginine eluting at 103 minutes. Injection to injection time 128 minutes.
Fig.2. Same as Fig.1. showing p-Ser, Tau, p-ethanolamine, Asp, OH-Pro, Thr, Ser, Asn, Glu, Sar, alpha-Aminoadipic acid, Pro.
Fig.3. Same as Fig.1. showing Gly, Ala, Cit, alpha-Aminobutyric acid, Val, Cys, Cysth., Ile, Leu, Nle, Tyr, Phe, Hcys, ß-Ala, ß-AIBA.
Fig. 4. Same as Fig.1. showing gaba, His, 1M-His, 3M-His, Trp, Car, Ans, OH-Lys, Orn, Lys, Ammonia, Eth-Amine, Arg (RT-103min.)
Fig. 5. Extended Physiological – extended – program with separation of additional amino acids. Internal standards: d-Glucosaminic acid, Norleucine, Aminoethylcystein. Additional amino acids: Pipecolic acid, Sacharopine, Homocitrulline, allo-Isoleucine, Arginine succinic acid, Cystine-homocysteine-disulfide, d-Amino-levolinic acid, 3-hydroxy-kynurenine, Kynurenine, Homocarnosine. Retention time to Arginine 158 minutes. Injection to injection time 184 minutes.
Fig. 6. Same as Fig.5. showing middle part of chromatogram. alfa-Amino-adipic-acid, Pro, Gly, Ala, Cit, alpha-Aminobutyric acid, Val, Pipecolic acid, Cystine, Sacharopine, Homocitrulline, Met, allo-Isoleucine, Isoleucine, Arginine-succinic acid, Leucine, Cystine-homocystine-disulfide, Nle, Tyr, ß-Ala, Phe. Cystathionine (not present) elutes between allo-Isoleucine and Isoleucine.
Fig. 7. Same as Fig. 5, showing end part of chromatogram. Phe, ß-AIBA, Homocystine, d-Amino-levolinic acid, GABA, Hydroxykynurenine, Kynurenine, His, 1M-His, 3M-His, Trp, Car, Homocarnosine, Ans, Aminoethylcysteine (ISTD), Hydroxylysine, Orn, Lys, NH3, Ethanolamine, Arginine. Note: Tryptophan elutes after the histidines.
Fig. 8. Example of a short program containing Argininosuccinic acid (RT 8.0 min), Homocysteine, Methionine, allo-Isoleucine (RT 14.4 min.), Leucine, Tyrosine, Phenylalanine and Homocystine. Norleucine (ISTD, not present) elutes between Leucine and Tyrosine. Time from injection to injection 44 minutes.
Fig. 9. Example of a short program for the analysis of Reduced Glutathione (GSH) eluting at 12 minutes and Oxidized Glutathione (GSSH) eluting at 26 minutes.
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