The potential of Agilent Bioanalyzer 2100 for new bioanalytical applications

Within commercially available CE-on-a-chip platforms, the Agilent Bioanalyzer 2100 is well established for applications tailored methods for DNA, RNA and protein analysis. The instrument is equipped with a red diode laser - and optionally a blue LED - for fluorescence detection. The glass chips are polymer encased and contain12 sample reservoirs on one chip enabling high throughput analysis.

This work targeted broadening the applicability of the Bioanalyzer to a wider range of analytes. The glass chips manufactured for DNA analysis were applied for generic CE based on a number of choices of injection and detection parameters. For non-aqueous CE (NACE), the compatibility of the existing chips with solvents such as dimethylsulfoxide (DMSO) was tested and the suitability of this non-volatile, non-hazardous, non-aquous solvent for NACE was verified and demonstrated with NACE separations of basic fluorescent dyes.

The applicability of the instrument is enhanced both for direct and for indirect fluorescence detection by the presence of two excitation light sources: red laser (635 nm) and blue LED (470 nm). Several highly fluorescent dyes including Nile Blue (NB) and fluorescein isothiocyanate (FITC) that can be used as labelling tags were used to determine the sensitivity and LOD values.

The suitability of the chip-based CE for indirect LIF detection using fluorescent dye probes and direct LIF determination of NB-tagged carboxylic acids, FITC-tagged amino acids as well as derivatised carbohydrates, and other biomolecules was investigated and will be illustrated by examples.