Aim: To perform Iron estimation by UV-Visible spectroscopy using an Experimental design approach. Objectives: The robust analytical method was developed for the estimation of iron (III) using 1, 10-phenanthroline reagent. Methods: The analytical method is an exploration of the chemical reaction of iron with 1, 10-phenanthroline reagent to form a colored complex which was measured in the UV-Visible region at 509 nm. To monitor the effect of diverse factors like the concentration of reagent (A), volume of reagent (B), pH (C) and time (D) on the formation of iron 1, 10 -phenanthroline complex the full factorial design (two-level) was used. From the Pareto chart, Normal plot and half normal plot, it was studied that a combination of all factors was initiate to be significant. Then significant variables are optimized by response surface methodology (RSM) via Box-Behnken design. The evaluation of design was performed to study the effect on the selected response by quadratic effects and main interaction effects. The contour plot and surface plot used for the determined response of the selected factors for their optimum value. Results: The prime reaction state, Beer ' s law were obeyed in 2.0-10.0 µg/ml concentration range with a correlation coefficient of 0.998. Conclusion: The method was successfully applied for the estimation of iron in iron sucrose injection. The optimized method was used for the quantitative analysis of iron sucrose injection.
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