Simple and sensitive spectrophotometric method is described based on the coupling reaction of tetracycline hydrochloride(TC. HCl) with diazotized 4-aminopyridine in bulk and pharmaceutical forms. Colored azo dye formed during this reaction is measured at 433 nm as a function of time. Factors affecting the reaction yield were studied and the conditions were optimized. The kinetic study involves initial rate and fixed time (10 minutes) procedures for constructing the calibration graphs to determine the concentration of (TC. HCl). The graphs were linear for both methods in concentration range of 10.0 to 100.0 µg.mL-1. The recommended procedure was applied successfully in the determination of (TC. HCl) in itscommercial formulations.

The measurement of vitamin B1 in pure and pharmaceutical formulations was proposed by using a straightforward and sensitive spectrophotometric approach. Sulfacetamide (SFA) is diazotized, then coupled with vitamin B1 in alkaline media to produce a colored azo dye complex with a stability constant of 5.597 × 105 L/mol. The product is stable, with a maximum absorption wavelength of 489.5 nm, molar absorptivity of 10108 L/mol∙cm, Sandell's sensitivity of 0.0334 μg/cm2, detection limit of 0.0135 μg/mL, and Beer's law being observed over the concentration range of 0.2–20.0 μg/mL. The stability constant and stoichiometry of the produced azo dye were calculated using the continuous variation (Job's) and mole ratio methods. The suggested ap