High-performance liquid chromatography amino acids

Aguilar, M. I., Hodder, A. N., and Hearn, M. T. W., High-performance liquid chromatography of amino acids, peptides, and proteins. LXV. Studies on the optimisation of the reversed-phase gradient elution of polypeptides. Evaluation of retention relationships with (3-endorphin-related polypeptides, /. Chromatogr., 327, 115, 1985. 

Guo, D, Mant, C. T., Taneja, A. K, Parker, J. M. R., and Hodges, R. S., Prediction of peptide retention times in reversed-phase high-performance liquid chromatography. I. Determination of retention coefficients of amino acid residues of model synthetic peptides, /. Chromatogr., 359, 499, 1986. 

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Heinrikson, R. L. and Meredith, S. C., Amino acid analysis by reverse-phase high-performance liquid chromatography precolumn derivatization with phenylisothiocyanate, Anal. Biochem., 136, 65, 1984. 

Zhao, Q., Sannier, F., Garreau, I., Lecoeur, C., and Piot, J. M., Reversed-phase high-performance liquid chromatography coupled with second-order derivative spectroscopy for the quantitation of aromatic amino acids in peptides application to hemorphins, /. Chromatogr. A, 723, 35, 1996. 

Vasanits, A. and Molnar-Perl, I., Temperature, eluent flow-rate and column effects on the retention and quantitation properties of phenylthiocarbamyl derivatives of amino acids in reversed-phase high-performance liquid chromatography, J. Chromatogr., A, 832,109, 1999. 

Hayakawa, K., Hirano, M., Yoshikawa, K., Katsumata, N, and Tanaka, T., Separation of phenylthiohydantoin-amino acids by temperature-controlled reversed-phase high-performance liquid chromatography, /. Chromatogr. A, 846, 73, 1999. 

Desai,M.J., Armstrong, D.W. (2004). Analysis of native amino acid and peptide enantiomers by high-performance liquid chromatography/atmospheric pressure chemical ionization mass spectrometry. J. Mass. Spec. 39, 177-187. 

Wen-Chen Z, Ling-Jun L, Xian-En Z et al (2008) Application of 2-(l l//-benzo[ i]carbazol-11 -yl) ethyl carbonochloridate as a precolumn derivatization reagent of amino acid by high performance liquid chromatography with fluorescence detection. Chin J Anal Chem 36 1071-1076... 

Watanabe Y, Imai K (1981) High-performance liquid chromatography and sensitive detection of amino acids derivatized with 7-fluoro-4-nitrobenzo-2-oxa-l,3-diazole. Anal Biochem 116 471—472... 

Consider one small molecule, phenylalanine. It is an essential amino acid in our diet and is important in protein synthesis (a component of protein), as well as a precursor to tyrosine and neurotransmitters. Phenylalanine is one of several amino acids that are measured in a variety of clinical methods, which include immunoassay, fluorometry, high performance liquid chromatography (HPLC see Section 4.1.2) and most recently MS/MS (see Chapter 3). Historically, screening labs utilized immunoassays or fluorimetric analysis. Diagnostic metabolic labs used the amino acid analyzer, which was a form of HPLC. Most recently, the tandem mass spectrometer has been used extensively in screening labs to analyze amino acids or in diagnostic labs as a universal detector for GC and LC techniques. Why did MS/MS replace older technological systems The answer to this question lies in the power of mass spectrometer. 

Fig. 1.3 High-performance liquid chromatography of amino acids. Source Own files... 

NL Benoiton, FMF Chen, R Steinauer, M Chouinard. A general procedure for preparing a reference mixture and determining the amount of dimerized contaminant in JV-alkoxycarbony] amino acids by high-performance liquid chromatography. Int J Pept Prot Res 27, 28, 1986. 

S Weinstein, MH Engel, PE Hare. The enantiomeric analysis of a mixture of all common protein amino acids by high-performance liquid chromatography using a new chiral mobile phase. Anal Biochem 121, 370, 1982. 

DW Aswad. Determination of d- and L-aspartate in amino acid mixtures by high-performance liquid chromatography after derivatization with a chiral adduct and o-phthaldialdehyde. Anal Biochem 137, 405, 1984. 

Amino acid formation was monitored over 24 h by chiral high-performance liquid chromatography (CHIROBIOTIC T column) with samples diluted 10-fold (in H2O) to a suitable concentration. 

Ding, GS. etal.. Chiral separation of enantiomers of amino acid derivatives by high-performance liquid chromatography on a norvancomycin-bonded chiral stationary phase, Talanta, 62, 997, 2004. 

Sharp, V.S. et al.. Enantiomeric separation of dansyl amino acids using macrocyclic antibiotics as chiral mobile phase additives by narrow-bore high-performance liquid chromatography. Chirality, 16, 153, 2004. 

Tesafova, E., Bosdkovd, Z., and Zuskova, L, Enantioseparation of selected iV-tert-butyloxycarbonyl amino acids in high-performance liquid chromatography and capillary electrophoresis with a teicoplanin chiral selector. J. Chromatogr. A, 879, 147, 2000. 

Torok, G. et al.. Direct chiral separation of unnatural amino acids by high performance liquid chromatography on a ristocetin A-bonded stationary phase. Chirality, 13, 648, 2001. 

Yarovsky, L, Aguilar, M.I., and Hearn, M.T.W., High-performance liquid-chromatography of amino-acids, peptides and proteins. 125. Molecular-dynamics simulation of n-butyl chains chemically bonded to sihca-based reversed-phase high-performance liquid-chromatography sorbents, 7. Chromatogr. A, 660, 75, 1994. 

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