Use of experimental design to optimize a triple-potential waveform to develop a method for the determination of streptomycin and dihydrostreptomycin in pharmaceutical veterinary dosage forms by HPLC-PAD

• An HPLC-PAD method for the determination of aminoglycosides was developed.
• The triple-potential waveform was optimized using an experimental design.
• The internal standard of glucose improved the method precision.
• No electrode passivation was observed after successive analyses.
• The method allows the simultaneous determination of streptomycin and dihydrostreptomycin.

An HPLC–PAD method using a gold working electrode and a triple-potential waveform was developed for the simultaneous determination of streptomycin and dihydrostreptomycin in veterinary drugs. Glucose was used as the internal standard, and the triple-potential waveform was optimized using a factorial and a central composite design. The optimum potentials were as follows: amperometric detection, E1= −0.15 V; cleaning potential, E2 = +0.85 V; and reactivation of the electrode surface, E3 = −0.65 V. For the separation of the aminoglycosides and the internal standard of glucose, a CarboPac™ PA1 anion exchange column was used together with a mobile phase consisting of a 0.070 mol L−1 sodium hydroxide solution in the isocratic elution mode with a flow rate of 0.8 mL min−1. The method was validated and applied to the determination of streptomycin and dihydrostreptomycin in veterinary formulations (injection, suspension and ointment) without any previous sample pretreatment, except for the ointments, for which a liquid–liquid extraction was required before HPLC-PAD analysis. The method showed adequate selectivity, with an accuracy of 98–107% and a precision of less than 3.9%.

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