The Pharmaceutical and Chemical Journal, 2015, 2(2):52-59 The Pharmaceutical and Chemical Journal 52 Available online www.tpcj.org Research Article ISSN: 2349-7092 CODEN(USA): PCJHBA A New Validated UV Spectrophotometric Method for Quantitative Determination of Pefloxacin Mesylate in Bulk Form and Developed In-Situ Gelling Ocular Formulations Ashutosh Pareek 1 , Vivek Jain 1 , Yashumati Ratan 1 , Mahendra Singh Ashawat 1, 2, 3* 1 Department of Pharmacy, Banasthali University, Banasthali, (Rajasthan) 2 Rungta College of Pharmaceutical Science & Research, Bhilai, (Chhattisgarh) 3 Laureate Institute of Pharmacy, Kathog, Distt. Kangra (Himachal Pradesh) *Corresponding author Email: msaresearch1@gmail.com, msaresearch@rediffmail.com Abstract The present study was conducted to develop a new simple, rapid, precise, sensitive, eco-friendly UV- spectroscopic method for the quantitative determination of Pefloxacin Mesylate in bulk form and prepared in-situ gelling ocular formulations. Method was successfully developed in simulated tear fluid pH 7.4 and further validated in accordance with International Conference on Harmonization (ICH) Q2B guidelines. In line to this, it was tested for linearity, accuracy, precision, detection limit, quantification limit, stability testing, Sandell’s sensitivity and molar absorptivity. Finally developed method was applied to conduct assay of Pefloxacin Mesylate and recovery study in developed in-situ gelling ocular formulations. The absorption maximum of the drug was found to be 272 nm and linearity was observed from 0.5-20 μg/ml with a regression coefficient of 0.999. Validation and statistical results strongly suggested that the developed UV spectroscopic method was accurate, precise, sensitive, versatile and stable. It could be a feasible eco-friendly alternate for the rapid analysis of Pefloxacin Mesylate in bulk as well as in- situ gelling ocular formulations. Keywords Pefloxacin Mesylate, In-Situ Gelling, UV Spectrophotometric Method, Ocular. INTRODUCTION Pefloxacin is a synthetic broad-spectrum third generation fluoroquinolone antibiotic. Chemically pefloxacin is 1- ethyl-6-fluoro-7- (4-methyl- 1-piperazinyl)-4-oxo- 1, 4- dihydro-3-quinolone carboxylic acid and structurally it is analog of norfloxacin [1]. Dihydrate mesylate salt of pefloxacin has been frequently used worldwide in the form of oral tablet, parentral infusion and topical eye drops [2]. It is effective against most gram-negative (Enterobectar, E. coli, klebsiella [3], Pseudomonas aeruginosa [4], Acinetobacter spp., Alcaligenes [1], Pseudomonas spp. including Xanthomonas maltophilia [1], H. influenza [1], N. gonorrhoeae [5] and G. vaginalis [6]) and gram-positive bacteria (S. aureus [7], S. epidermidis [7], S. pneumonia [6] and many other mycobecteria [8]). It inhibits the bacterial enzymes DNA gyrase and topoisomerase IV, which are responsible for transcription and replication of bacterial DNA. DNA gyrase is considered as primary quinolone target for gram-negative bacteria while topoisomerase IV for gram-positive organisms. Systemically PM is prescribed in uncomplicated cystitis in women, uncomplicated gonococcal urethritis in males and for gram-negative bacterial infections in gastrointestinal