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An environmentally begnin second derivative spectrometric approach was developed for the estimation of the dissociation constants pKa(s) of metformin, a common anti-diabetic drug. The ultraviolet spectra of the aqueous solution of metformin were measured at different acidities, then the second derivative of each spectrum was graphed. The overlaid second derivative graphs exhibited two isobestic points at 225.5 nm and 244 nm pointing out to the presence of two dissociation constants for metformin pKa1 and pKa2, respectively. The method was validated by evaluating the reproducibility of the acquired results by comparing the estimated values of the dissociation constants of two different strategies that show excellent matching. As well as, the whole procedure was repeated with a new set of standard solutions and buffers for further verification. The average calculated values of pKa1 and pKa2 were found to be 2.72±0.01 and 11.61±0.08, with correlation coefficients ( R2) of 0.9916 and 0.9614, respectively. The established method was fast, affordable, reproducible, and the mean pKa values obtained were accurate and can be applied for the estimation of the dissociation constants of other active pharmaceutical compounds
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