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Two-dimensional separation by sequential injection chromatography.

Maria Soledad M S F Acevedo1, Mariana R Gama2, Alex D Batista3

  • 1Center of Nuclear Energy in Agriculture, University of São Paulo, Avenida Centenário, 303, 13400-970, Piracicaba, SP, Brazil.

Journal of Chromatography. A
|August 17, 2020
PubMed
Summary
This summary is machine-generated.

This study enhances sequential injection chromatography (SIC) using a two-dimensional approach with distinct columns. This method successfully separates aromatic biogenic amines, overcoming limitations of traditional SIC for complex mixtures.

Keywords:
BioanalysisBiogenic aminesHeart-cuttingLow-pressure chromatographyMultidimensional chromatographySeparation capability

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Area of Science:

  • Analytical Chemistry
  • Chromatographic Techniques

Background:

  • Sequential injection chromatography (SIC) offers fast separations and low solvent consumption but faces limitations in separation capacity due to short columns and gradient elution constraints.
  • Achieving effective separation of aromatic biogenic amines like histamine, tyramine, phenylethylamine, and tryptamine can be challenging with unidimensional chromatographic methods.

Purpose of the Study:

  • To expand the analytical capabilities of SIC by implementing a multidimensional chromatographic approach.
  • To improve selectivity and peak resolution for complex mixtures, specifically aromatic biogenic amines.

Main Methods:

  • A two-dimensional SIC system was developed, utilizing a cyanopropyl monolithic column in the first dimension and a C18 superficially porous particle column in the second dimension (heart-cutting mode).
  • Optimized mobile phases and a sampling loop for heart-cutting were employed to collect and re-inject unresolved fractions.
  • The method was validated for the separation of histamine, tyramine, phenylethylamine, and tryptamine.

Main Results:

  • The multidimensional SIC approach successfully separated aromatic biogenic amines that were unresolved by unidimensional methods.
  • An 8-minute total analysis time was achieved with a resolution of 1.72 between the closest eluting peaks (tyramine and phenylethylamine).
  • The method demonstrated linear responses (r > 0.997) in the 10-50 mg L-1 range, with low detection limits and good precision (CV = 3.0%).

Conclusions:

  • Multidimensional SIC, employing columns with different separation mechanisms, significantly enhances separation capacity and resolution.
  • This approach effectively overcomes the limitations of unidimensional SIC for analyzing complex mixtures of aromatic biogenic amines.
  • The developed method provides a sensitive, rapid, and robust analytical tool for biogenic amine determination.