70-min Amino Acid Analysis of Physiological Samples

Pickering Laboratories specializes in the manufacturing of cation-exchange columns and eluants for amino acid analysis. Post-column derivatization with Ninhydrin offers unmatched selectivity and reproducibility of the analysis for most challenging matrixes.

Table I: HPLC program

Pinnacle PCX post-column derivatization system allows you to combine eluant gradient capabilities of modern HPLC instruments with column temperature gradients. We introduce accelerated amino analysis method for physiological samples that utilizes temperature and eluant gradients.

Figure 1: List of amino acids: 1) Phosphoserine, 2) Taurine, 3) Phosphoethanolamine, 4) Urea, 5) Aspartic acid, 6) Hydroxyproline, 7) Threonine, 8) Serine, 9) Asparagine, 10) Glutamic acid, 11)Glutamine, 12) Sarcosine, 13) α-Aminoadipic acid, 14) Proline, 15) Glycine, 16) Alanine, 17) Citrulline, 18) α-Amino-n-butyric acid, 19) Valine, 20) Cystine, 21) Methionine, 22) Cystathionine, 23) Isoleucine, 24) Leucine, 25) Tyrosine, 26) Phenylalanine, 27) β-Alanine 28) β-Amino-i-butyric acid, 29) Homocystine, 30) γ-Aminobutyric acid, 31) Tryptophan, 32) Ethanolamine, 33) Ammonia, 34) Hydroxylysines, 35) Ornitine, 36) Lysine, 37) 1-Methylhistidine, 38) Histidine, 39) 3-Methylhistidine, 40) Anserine, 41) Carnosine, 42) Arginine.

Method

Analytical Conditions

Column: High-efficiency Lithium cation-exchange column (4.6 × 75 mm), 5 µm Catalog No. 0354675T

Flow Rate: 0.55 mL/min

Mobile Phase: 1700–1125, Li365, Li375, RG003

Table II: Column oven program

Post-column conditions

Post-column system: Pinnacle PCX

Reactor Volume: 0.5 mL

Temperature: 130 °C

Reagent: Trione

Flow Rate: 0.5 mL/min

Detection: UV-vis 570 nm for primary amino acids 440 nm for secondary amino acids

Run Time: 72 min

Equilibration Time: 15 min

Pickering Laboratories, Inc

1280 Space Park Way, Mountain View, CA 94043

tel. (800) 654-3330, (408) 694-6700

Website: www.pickeringlabs.com