Objective: To develop three novel, sensitive, simple validated visible spectrophotometric methods for the quantitative estimation of sulfamethoxazole (SMZ) and trimethoprim (TMP) in bulk form.Methods: Methods were based on coupling the diazotized aromatic primary amino group of the studied drugs with o-phenylenediamine (OPD) in an acidic medium. The first two methods have been proposed for estimation of SMZ and rest for TMP. The resulting products were measured by spectrophotometric (method I, II and III) tools. The methods were validated as per ICH guidelines.Results: In method I, the absorbance was measured at 482 and 457 nm with linearity ranges of 4.0-40.0 and 5.0-45.0 µg/ml for SMZ. On the other hand, method III was devoted to estimate TMP spectrophotometrically at 457 nm with linearity range of 5-30 μg/ml. The r2 value for all methods were found to be 0.99. The percentage recoveries of SMZ and TMP were found to be 97.98%, 97.56% and 97.55% respectively. The developed methods were subjected to detailed validation procedure in their pure forms.Conclusion: The study concludes that visible spectrophotometric validation methods can be very efficient and economically promising technique for the quantitative analysis of SMZ and TMP in bulk form. The statistical analysis of data indicates that the developed methods were reproducible and specific. It was found that there is a good agreement between the obtained results and those obtained by the reported methods; moreover they can be used for the routine estimations of SMZ and TMP in bulk form.

The present work was focused on developing a new RP-HPLC method for the simultaneous estimation of paracetamol and tramadol hydrochloride in bulk and tablet dosage form and to validate it as per ICH and USP guidelines. The method involves use of water and acetonitrile in 9:1 ratio as mobile phase pumped at a rate of 1ml/min. The optimum wavelength selected for monitoring was 268nm. C18 column (4.6mm×250mm) of 5µ particle size was used as stationary phase. The method was finally validated, and parameters were reported. The system suitability parameters passed in which the asymmetric factors for Paracetamol and Tramadol were 1.54 and 1.09 respectively. Linearity ranges were found to be 20 to 100µg/ml with a correlation coefficient of 0.998. Accuracy studies reported a mean recovery of 98.7% for both the drugs. Faster retention times (1.1min and 4.1min) make the method simple and economic. Thus a validated and sensitive RP-HPLC method was developed for simultaneous estimation of Paracetamol and tramadol in bulk and tablet dosage form.

A simple, sensitive and specific UFLC method was developed to estimate Chlorhexidine in bulk drug. Acetonitrile and Water were used in 60:40 v/v ratio as mobile phase. The flow rate of eluent was fixed at 0.8 mL/min. Absorbance was monitored at λmax of 235 nm. A reverse phase column C18, (250mm x 4.6mm i.d., 5µm) was used as stationary phase. The retention time was found to be 2.99 minutes. The linearity range of Chlorhexidine was found to be 1-6 μg/ml at 235nm wavelength.

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