Flow Injection Tutorial

Determination of Fe(II) by reaction with ferrozine was automated by programmable Flow Injection method, by using the instrumental setup (A), reagents and standards described in (B) and in the assay protocol (C).

The apparatus and protocol are version of the system designed for SE (1.3.14.B.), modified by adding reagent container and “reagent” section in the protocol. This section is designed to mix Fe(II) sample with ferrozine reagent in ratio 5+1, within the volume of 1400 mcrL. The reaction mixture was heated by thermostating both holding coils at 35 C. Because on column adsorption, carried out at 10mcrL/sec, is time consuming, only 1500mcrL of was delivered on the column, leaving some of the tail section of reaction mixture in HC2. Elution was carried out at flow rate of 10mcrL/sec into 20 cm long flow cell, to allow reliable capture of peak maximum within the window WIN (D). The results and the calibration graph (E) obtained by means of protocol (C) are discussed in the next section.