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ANALIS

PhoEBuS software for electrolyte prep in CE, HPLC, IC and titration

The PHoEBuS application program is designed to assist the scientist in creating and preparing the electrolyte to be used in a separation via capillary electrophoresis. It provides the following information for electrolytic solutions at 25°C.
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lifescience@analis.be ++(0)81 25 50 50 ++(0)9 243 77 10
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DESCRIPTION

PHoEBuS software overview

1. General Overview

The PHoEBuS application program is designed to assist the scientist in creating and preparing the electrolyte to be used in a separation via capillary electrophoresis. It provides the following information for electrolytic solutions at 25°C:

" chemical characteristics (pH, ionic composition, buffer capacity)

" electrical characteristics (conductivity, estimation of the current, electrical power)

" electrophoretic characteristics (mean electrophoretic mobilities of the co-ion and the counter ion)

In addition, the application program is also an useful tool to determine the effect of changes in the pH, the ionic strength or the concentration of one of the components when such solutions are developed for other analytical methods such as liquid chromatography, ion chromatography and titrimetry.

There are two general modes of operation in Capillary Electrophoresis:

" Detection by Direct Spectrometry

" Detection by Indirect Spectrometry

2. Detection by Direct Spectrometry

In many cases, the electrophoretic separation is performed using an electrolyte that does not absorb at the wavelength that is used to detect the compounds of interest. When this mode is used, a direct UV spectrometric detection is possible, with the wavelength set at the absorbance maximum of the analyte(s).

The optimization of the electrophoretic separation depends on two physico-chemical parameters; the ionic strength and the pH of the electrolyte. PHoEBuS can be used to describe how to make an electrolyte with a pH and the ionic strength that is selected by the user.

3. Detection by Indirect Spectrometry

If the analyte does not absorb in the UV-visible region of the spectrum, an indirect spectrometric detection is used. In this approach, the electrolyte contains an ion that includes a chromophoric group and the spectrometric detector is set at the maximum absorbance wavelength of the ion. This mode of detection is commonly used for the analysis of inorganic cations and anions in capillary electrophoresis.

The sensitivity of detection via an indirect spectrometric method can be optimized by varying the pH and the concentration of the co-ion of the electrolyte. The application program can determine the composition that will provide the desired pH and the concentration of the co-ion of the electrolyte which are imposed by the user. In addition, the program allows for selection of the chemical form under which the co-ion will be mainly present in the electrolyte so as to optimize the transfer ratio.

In summary, the application program allows the user to select the pH and the concentration of an ion of the electrolyte (in general the co-ion). As an alternative, the user can select the pH and the concentration of one of the chemical forms of this ion (if imposed values have a chemical reality).

4. Comparing Calculated Results with Buffer

Properties

PHoEBuS employs a number of theoretical models which simulate inter-ionic interactions to assist the analyst in the preparation of an electrolyte for capillary electrophoresis. It is strongly recommended that the analyst checks the properties of the buffer generated by the application program before the buffer is used since the buffer properties may differ slightly from calculations. To ensure that the buffer meets the requirements of the separation, the following points should be kept in mind:

" Calculations are based on physico-chemical parameters at 25°C (pKa, ionic product of the water, electrophoretic mobility).

" Measure the pH at 25°C using a calibrated pH meter.

" Make certain that the concentrations of stock solutions are accurate. If the stability of the stock solution is questionable, prepare a fresh solution.

" Verify that the correct value of pKa (the acidity constant) and the electrophoretic mobilities are used. This latter physico-chemical parameter is frequently poorly defined.

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