Published:Journal of Chromatographic Science,
ISSN 0021-9665Volume
37, Number 7, July 1999, pp. 229-236
Osmolarity Effects on Red
Blood Cell Elution in Sedimentation Field-Flow FractionationN. Emmanuel Assidjo, Thierry Chianéa,
Igor Clarot, Marie F. Dreyfuss, and Philippe J.P. Cardot
Field-flow fractionation (FFF) is an analytical technique particularly suitable
for the separation, isolation, and characterization of macromolecules and micrometer-
or submicrometer-sized particles. This chromatographic-like methodology can
modulate the retention of micron-sized species according to an elution mode
described to date as “steric hyperlayer”. In such a model, differences
in sample species size, density, or other physical parameters make particle
selective elution possible depending on the configuration and the operating
conditions of the FFF system. Elution characteristics of micron-sized particles
of biological origin, such as cells, can be modified using media and carrier
phases of different osmolarities. In these media, a cells average size, density,
and shape are modified. Therefore, systematic studies of a single reference
cell population, red blood cells (RBCs), are performed with 2 sedimentation
FFF systems using either gravity (GrFFF) or a centrifugational field (SdFFF).
However, in all cases, normal erythrocyte in isotonic suspension elutes as a
single peak when fractionated in these systems. With carrier phases of different
osmolarities, FFF elution characteristics of RBCs are modified. Retention modifications
are qualitatively consistent with the “steric-hyperlayer” model. Such
systematic studies confirm the key role of size, density, and shape in the elution
mode of RBCs in sedimentation FFF for living, micron-sized biological species.
Using polymers as an analogy, the RBC population is described as highly “polydisperse”.
However, this definition must be reconsidered depending on the parameters under
concern, leading to a matricial concept: multipolydispersity. It is observed
that multipolydispersity modifications of a given RBC population are qualitatively
correlated to the eluted sample band width.
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