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Detection of Platinum Species in Plant Material by Preparative Isotachophoresis Using the Model 491 Prep Cell, Rev A

l pollution.1 The emission of platinum as small particles (sub- m range) increases the bio-availability of this element2,3 and has created interest in biological studies of platinum. For the separation and characterization of platinum species in grass cultivations, preparative isotachophoresis with the Model 491 Prep Cell leads to excellent separations. Sharply isolated platinum- containing fractions were obtained, which were further characterized biochemically.


Methods

The combined procedure for the separation, detection, and characterization of platinum-containing species in plant materials is described in references 4 and 5.

Briefly, extracts of untreated and platinum-treated grass cultivations were separated by combined use of two different preparative methods with different separation principles: gel permeation chromatography followed by isotachophoresis in the Model 491 Prep Cell. This combined procedure leads to an excellent recovery of purified platinum binding proteins. The detection of the platinum species was carried out sequentially by absorptive voltammetric platinum determination in individual fractions. Native electrophoresis (PAGE) was used for further characterization of selected species.

Isotachophoresis
Following chromatography4,5, a second preparative separation step of isotachophoresis gave best results. In isotachophoresis, the upper buffer is called the terminating electrolyte and the lower buffer is the leading electrolyte. The non-restrictive polyacrylamide gel is polymerized containing the leading electrolyte. During electrophoresis, charged molecules electrophorese through
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