title: Retention Mechanism Studies of Selected Amino Acids and Vitamin B6 on HILIC Columns with Evaporative Light Scattering Detection.
abstract: The goal of the study was to investigate separation mechanism of selected "essential" amino acids (leucine, isoleucine, threonine, tryptophan, proline, and glycine) and vitamin B6 in hydrophilic interaction liquid chromatography (HILIC) with the evaporative light scattering detection. Chromatographic measurements were made on three different HILIC columns: amide-silica (TSK-gel Amide-80), amino-silica (TSK-gel NH2-100), and cross-linked diol (Luna HILIC). The retention behaviour of the analytes was investigated as a function of different binary hydro-organic mobile phases containing 10-90 % (v/v) acetonitrile. The compounds studied were separated under isocratic and gradient conditions. The best results of tested biologically active compounds separation were obtained on the TSK-gel NH2-100 column. TSK-gel NH2column showed mixed HILIC-ion-exchange mechanism, the highest separation efficiency and better selectivity and resolution for tested analytes than the other studied column, especially at concentration of water in mobile phase lower than 30 % (v/v). Special attention was dedicated to the study of interactions among the stationary phase, mobile phase and the analytes.
authors.affiliation: Chair of Environmental Chemistry & Bioanalytics, Faculty of Chemistry, Nicolaus Copernicus University, 7 Gagarin St., 87-100 Toruń, Poland.
authors.forename: Sylwia
authors.initials: S
authors.lastname: Noga
authors.forename: Pavel
authors.initials: P
authors.lastname: Jandera
authors.forename: Bogusław
authors.initials: B
authors.lastname: Buszewski
keywords: Amino acids; Evaporative light scattering detection; Hydrophilic interaction liquid chromatography; Retention mechanism; Vitamin B6
references.citation: Int J Pharm. 2000 Jul 20;202(1-2):113-20
references.pmid: 10915933
references.citation: J Chromatogr B Biomed Appl. 1996 Jun 28;682(1):3-22
references.pmid: 8832421
references.citation: J Chromatogr A. 1999 Sep 3;855(1):191-202
references.pmid: 10514984
references.citation: J Sep Sci. 2006 Aug;29(12):1784-821
references.pmid: 16970185
references.citation: J Chromatogr A. 2008 Mar 14;1184(1-2):474-503
references.pmid: 18294645
references.citation: Anal Bioanal Chem. 2012 Jan;402(1):231-47
references.pmid: 21879300
references.citation: J Chromatogr A. 2000 Feb 18;870(1-2):245-54
references.pmid: 10722083
references.citation: J Chromatogr. 1990 Jan 19;499:177-96
references.pmid: 2324207
references.citation: J Chromatogr. 1993 May 19;615(1):1-22
references.pmid: 8340448
references.citation: J Nutr. 2004 Jun;134(6 Suppl):1558S-1565S
references.pmid: 15173430
references.citation: J Chromatogr B Analyt Technol Biomed Life Sci. 2009 Dec 15;877(32):4133-9
references.pmid: 19926350
references.citation: J Nutr. 1994 Aug;124(8 Suppl):1517S-1523S
references.pmid: 8064412
references.citation: Anal Chem. 2000 Apr 15;72(8):1736-9
references.pmid: 10784135
references.citation: J Chromatogr A. 2000 Sep 1;891(1):1-32
references.pmid: 10999622
references.citation: J Chromatogr Sci. 2004 Oct;42(9):484-90
references.pmid: 15693189
references.citation: J Chromatogr B Biomed Sci Appl. 2000 Sep 29;747(1-2):1-19
references.pmid: 11103896
references.citation: Electrophoresis. 1997 Nov;18(12-13):2377-92
references.pmid: 9456052
references.citation: J Chromatogr A. 2010 May 14;1217(20):3408-17
references.pmid: 20362994