Separation of toxic peptides (microcystins) in capillary electrophoresis, with the aid of organic mobile phase modifiers.
A capillary electrophoretic (CE) method incorporating sodium dodecyl sulphate (SDS)-organic modifier solvents in the CE buffer was developed for the detection of toxic cyclic heptapeptide toxins (microcystins) produced by blue-green algae (cyanobacteria). The applicability of these run buffers for the analysis of microcystins was evaluated and optimum conditions for separation were determined. The migration times, elution order and selectivity of the toxic peptides were influenced by modifying the composition of the electrophoretic buffer with organic solvents [0 to 20% (v/v)]. At maximum addition, the organic solvents with the exception of acetonitrile, increased the viscosity of the buffer solution. In contrast to the migration time of the other microcystins, that of microcystin-RR was not increased by the addition of 2-propanol to the buffer solution. Rather, microcystin-RR eluted more quickly with the increase in 2-propanol, thereby effecting changes in the elution order of the microcystins. In addition, this solvent resulted in comigration of microcystin-LR and microcystin-YR. No significant relationship was found between the elution order and separation and the structure of the toxic peptides studied in micellar electrokinetic capillary chromatography with an organic modifier in the buffer solution; but there is an agreement between the effects of the organic modifiers and their dipole moments. Parameters such as linearity, sensitivity and reproducibility were also evaluated. High-efficiency separations of toxic peptide molecules having equal or nearly equal mass to charge ratios have been achieved using SDS as an additive to the running buffer. The influence of the pH has been examined.
Onyewuenyi
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Australian Water Technologies, West Ryde, NSW, Australia.
Hawkins
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P
D019075:Electrophoresis, Capillary / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D052998:Microcystins; D010456:Peptides, Cyclic / Q000737:chemistry / Q000302:isolation & purification*; D015203:Reproducibility of Results; D012967:Sodium Dodecyl Sulfate
D052998:Microcystins; D010456:Peptides, Cyclic; D012967:Sodium Dodecyl Sulfate; C078588:microcystin