+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Micellar liquid chromatographic separation of amino acids using pre- and post-column o-phthalaldehyde/N-acetylcysteine derivatization

Micellar liquid chromatographic separation of amino acids using pre- and post-column o-phthalaldehyde/N-acetylcysteine derivatization

Analytica Chimica Acta 418(2): 153-165

The isocratic analysis of primary proteic amino acids using a C18 column, micellar mobile phases of sodium dodecyl sulphate (SDS) and 1-propanol at varying pH, and pre- and post-column formation of the o-phthalaldehyde (OPA)/N-acetyl-L-cysteine (NAC) isoindoles, is examined. An interpretive optimization strategy was applied to find the best separation conditions. For this purpose, empirical polynomial models were used to describe the elution behavior of the isoindoles and free amino acids, using as separation factors the concentrations of surfactant and modifier (two-factor problem), or these two factors and the pH of the mobile phase (three-factor problem). The separation of the OPA/NAC amino acid isoindoles was very poor: only eight compounds could be separated satisfactorily. In contrast, 14 free amino acids (ordered according to their elution times: aspartic acid, glutamine, threonine, cysteine, alanine, tyrosine, valine, methionine, phenylalanine, leucine, lysine, histidine, tryptophan and arginine) could be well resolved with 0.05 M SDS-1.2% propanol at pH 3 in 42 min, partially due to the higher efficiencies frequently five- to 10-fold those achieved for the isoindoles at this pH. A mobile phase of 0.05 M SDS without alcohol allowed the resolution of a mixture of aspartic acid, glutamine, asparagine, glutamic acid, threonine, cysteine and alanine, which eluted at the lower retention times. In these conditions, only serine and glycine overlapped with glutamine and threonine, respectively.

Please choose payment method:

(PDF emailed within 0-6 h: $19.90)

Accession: 010981608

Download citation: RISBibTeXText

DOI: 10.1016/s0003-2670(00)00965-x

Related references

Liquid chromatographic analysis of amino and imino acids in protein hydrolysates by post-column derivatization with o-phthalaldehyde and 3-mercaptopropionic acid. Journal of Chromatography 476: 83-92, 1989

Determination of amino acids by micellar high-performance liquid chromatography and pre-column derivatization with o-phthalaldehyde and N-acetyl-L-cysteine. Journal of Liquid Chromatography 18(14): 2827-2841, 1995

Pre column o phthalaldehyde derivatization of amino acids and their separation using reversed phase high performance liquid chromatography 2. simultaneous determination of amino acids and imino acids in protein hydrolysates. Journal of Chromatography 285(3): 490-494, 1984

Pre column o phthalaldehyde derivatization of amino acids and their separation using reversed phase high performance liquid chromatography 1. detection of the imino acids hydroxy proline and proline. Journal of Chromatography 285(3): 484-489, 1984

Pre-column derivatization of amino acids with N,N-diethyl-2,4-dinitro-5-fluoroaniline and reversed-phase liquid chromatographic separation. Journal of Chromatography 433: 53-62, 1988

Ion pair liquid chromatographic analysis of phenylpropanolamine in plasma and urine by post column derivatization with o phthalaldehyde. Journal of Liquid Chromatography 8(8): 1489-1500, 1985

High-performance liquid chromatographic method for post-column, in-line derivatization with o-phthalaldehyde and fluorometric detection of phenylpropanolamine in human urine. Journal of Chromatography 284(2): 457-461, 1984

Automated pre-column derivatization with o-phthalaldehyde for the determination of neurotransmitter amino acids using reversed-phase liquid chromatography. Journal of Chromatography 417(2): 385-390, 1987

Liquid chromatographic enantiomeric resolution of amino acids with beta cyclodextrin bonded phases and derivatization with o phthalaldehyde. Analytica Chimica Acta 234(1): 127-132, 1990

Whole blood analysis of gluconeogenic amino acids for estimation of de novo gluconeogenesis using pre-column o-phthalaldehyde derivatization and high-performance liquid chromatography. Journal of Chromatography B Biomedical Applications 676(1): 1-6, 1996

The separation of pre column o phthalaldehyde derivatized amino acids by high performance liquid chromatography. Journal of Liquid Chromatography 2(7): 1047-1059, 1979

Enantiomeric separation of amino acids using micellar electrokinetic chromatography after pre-column derivatization with the chiral reagent 1-(9-fluorenyl)-ethyl chloroformate. Electrophoresis 16(4): 504-509, 1995

On-line solid-phase extraction with surfactant accelerated on-column derivatization and micellar liquid chromatographic separation as a tool for the determination of biogenic amines in various food substrates. Analytical Chemistry 76(5): 1289-1294, 2004

Determination of the renin inhibitor Ro 42-5892 in human plasma by automated pre-column derivatization, reversed-phase high-performance liquid chromatographic separation and electrochemical detection after post-column irradiation. Journal Of Chromatography B Biomedical Applications. 665(2): 373-381, 1995

Derivatization methods for liquid chromatographic separation of amino acids. Nollet, L M L (Ed ) Food Science And Technology (New York), Vol 52 Food Analysis By Hplc Ix+759p Marcel Dekker, Inc : New York, New York, Usa; Basel, Switzerland Illus 53-74, 1992