Analytical method development and validation for the simultaneous estimation of sulfadiazine and pyrimethamine by rp-hplc method
Keywords:Zodiac silica C18 column, Sulfadiazine and Pyrimethamine, RP-HPLC
Estimation of Sulfadiazine and Pyrimethamine is a combination drug of choice used to treat malaria who are living in or will be travelling to an area where there is chance of getting malaria.
The regimen above is in short supply and to evaluate efficacy of twice weekly maintenance therapy to prevent recurrent toxoplasmic encephalitis in patients with Acquired Immnuno deficiency syndrome.
Materials and methods
A HPLC (Alliance,Water2695) with UV/VIS Detector/PDA detector, UV (lab India,UV 3000 series) and Agilent C18 250mm × 4.6mm × 5µm column was used. A new method was established for simultaneous estimation of Sulfadiazine and Pyrimethamine by RP-HPLC method. The chromatographic conditions were successfully developed for the separation of Sulfadiazine and Pyrimethamine by using Zodiac sil C18 column
(4.6×150mm)5µ, flow rate was
1ml/min, mobile phase ratio was (70:30 v/v) methanol: phosphate
buffer(KH2PO4 and K2HPO4) phosphate pH 3 (pH was adjusted with orthophosphoricacid),detection wavelength was 240nm.
The results were in good agreement with those obtained with official HPLC with absorption maximum of 240 nm by preparing mobile phase 70:30 methanol : phosphate buffer with flow rate 1 ml/min and it run for 10 minutes by selecting column Zodiac silca RP C18 4.5×100 mm 3.0 µm of ambient temperature .All the results obtained with good precise ,accurate and robustness as per ICH guidelines.
It can be concluded that the proposed RPHPLC method is accurate, precise, sensitive, specific, robust and reproducible for the simultaneous analysis of Sulfadiazine and Pyrimethamine with less tailing factor and is also economical. Zodiac sil C18 column (4.6×150mm)5µ, flow rate was 1ml/min, mobile phase ratio was (70:30 v/v) methanol: phosphate buffer(KH2PO4and K2HPO4) phosphate pH 3 (pH was adjusted with orthophosphoricacid),detection wavelength was 240nm.