European Journal of Biomedical and Pharmaceutical Sciences

SYNTHESIS OF 2, 6-DIHYDROXY-3-PHENYLDIAZENYLPYRIDINE AND 6-AMINO-2-HYDROXY-3-PHENYLDIAZENYLPYRIDINE: AN IMPURITY IN THE PROCESS FOR PHENAZOPYRIDINE HYDROCHLORIDE A GENITO-URINARY ANTISEPTIC DRUG

Dr. Ajit Anerao*, Dr. Vihar Telange, Nitin Bondre, Vikram Dighe, K. Swamy Reddy, A. V. V. Somasekhar, Dr. Nitin Pradhan

ABSTRACT

2,6-Dihydroxy-3-phenyldiazenylpyridine and 6-Amino-2-hydroxy-3-phenyldiazenylpyridine an impurity formed in the process for the genitourinary antiseptic drug Phenazopyridine hydrochloride. Impurities were synthesized; characterized and reverse phase high-pressure liquid chromatography (HPLC) method has been developed and validated for estimation of related substances of Phenazopyridine hydrochloride active pharmaceutical ingredient (API). Efficient chromatographic separation was achieved on a Cosmosil 5μm particles size, 250mm × 4.6mm column. The method was validated for accuracy, precision, specificity, robustness, and detection and quantification limits, in accordance with ICH guidelines. Statistical analysis proved the method was precise, reproducible, selective, specific, and accurate for analysis of Phenazopyridine hydrochloride and its related impurities. HIGHLIGHTS • Related substances method is developed for quantification of specified and unspecified impurities of Phenazopyridine hydrochloride. • Reliability and reproducibility of results is confirmed by performing analytical method validation as per ICH guideline. • Force degradation study is carried out by exposing the API to stress conditions and found all degradants are separating from the main component and can be quantified accurately. • In acidic degradation two degrading impurities generated are synthesized and identified.

Keywords: Phenzopyridine; Synthesis; characterization; impurity; NMR; validation.