Analysis of dansyl amino acids in feedstuffs and skin by micellar electrokinetic capillary chromatography.
Micellar electrokinetic capillary chromatography (MECC) using sodium dodecyl sulphate (SDS) and sodium cholate have been used for analyses of 30 dansylated (Dns) amino acids. The influences of sample preparation, Dns/amino acid ratio, sample solvent composition, and separation conditions including voltage, temperature, pH and buffer composition were investigated. Complete separations of acidic and neutral amino acids were obtained within 45 min in the SDS system. The efficiency expressed as number of theoretical plates for the applied capillary 0.52 m long were between 210,000 and 343,000, and the repeatability was very good with relative standard deviations on relative migration times between 0.09 and 0.70% and on relative normalised peak areas (RNPAs) between 0.85 and 3.41%. The linearity studies gave correlation coefficients between 0.9957 and 0.9993 for RNPAs against concentration. Detection limits were between 3 and 6 fmol or approximately 2 pg of each amino acid. Basic amino acids were separated in a MECC system using sodium cholate. Procedures and problems using Dns derivatisation for amino acids analysed by the MECC methods are described. Finally, examples of analyses of hydrolysates of real complex samples show, that this method can be applied to determine the amino acid composition of proteins in feedstuffs and skin.
Department of Research in Small Animals, National Institute of Animal Science, Research Centre Foulum, Tjele, Denmark.
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S
Michaelsen
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P
Møller
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H
Sørensen
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis*; D000821:Animal Feed / Q000032:analysis*; D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods*; D003619:Dansyl Compounds / Q000032:analysis* / Q000737:chemistry; D004563:Electrochemistry; D008823:Micelles; D008907:Mink; D008969:Molecular Sequence Data; D015203:Reproducibility of Results; D012867:Skin / Q000737:chemistry*; D013056:Spectrophotometry, Ultraviolet
D000596:Amino Acids; D003619:Dansyl Compounds; D008823:Micelles