Spectrophotometric determination of trace copper(II) in biological samples with 2,4-bis(4-phenylazophenylaminodiazo)phenol

A highly sensitive and selective chromogenic reagent 2,4-bis(4-phenylazophenylaminodiazo)phenol (BPPAAP) reacted with copper(II) to form a highly stable complex in the ethanolic solution at pH range of 9.0–12.0. The Cu(II)-BPPAAP complex showed maximum absorbance at 540 nm, with molar absorptivity being 1.86 × 10⁵ L/mol cm. Beer’s law was obeyed over the range 0–0.2 μg/100 mL of copper(II) and variation coefficient is found to be 2.4–4.8%. The detection and quantification limit of the method are 2.0 and 6.5 ng/mL, respectively. To eliminate the interference of foreign ions, a convenient and efficient method using a column packed with sulfhydryl dextrose gel as a solid-phase extractant was utilized with satifactory reults. The developed method has been successfully employed for the determination of copper(II) in the biological samples.