Development and Validation of HPLC method for simultaneous estimation of Rifampicin and Ofloxacin using experimental design

Purvi Shah,Tosha Pandya,Mukesh Gohel,Vaishali Thakkar

doi:10.1080/16583655.2018.1548748

Abstract

ABSTRACTA accurate, precise and robust isocratic HPLC method has been developed and validated for simultaneous determination of Rifampicin and Ofloxacin. The chromatographic separation was carried out on Kinetex C18, 100 A Phenomenex column with a mixture of 0.03M Potassium dihydrogen phosphate buffer pH 3.0: Acetonitrile (55:45) as mobile phase at 230 nm. The retention times were 2.91 and 4.87 min for Ofloxacin and Rifampicin, respectively. Calibration plots were linear over the concentration range 1–5 and 2–10 µg/ml for Rifampicin and Ofloxacin, respectively. The method was validated for linearity, sensitivity accuracy, precision, and robustness. Percent recoveries were found to be close to 100% with low variability. Fractional factorial design with four factors was chosen for robustness testing. The volume of acetonitrile and flow rate showed significant effect on retention factor of both the drugs and asymmetry factor of ofloxacin. The method may be adopted for routine analysis at industry.

Highlights

Tuberculosis (TB) remains a major global threat, the control of which has become more complicated since the emergence of multidrug-resistant (MDR) and extensively drug-resistant strains of Mycobacterium tuberculosis
Rifampicin (RIF) (Figure 1(a)) is a complex semisynthetic macrocyclic rifamycin class of antibiotic derived from Streptomyces mediterranei [3], used for the treatment of tuberculosis and other infectious diseases [4,5,6,7]
The optimization of chromatographic conditions were done with a view to develop HPLC method for the simultaneous determination of RIF and OFX in bulk and in pharmaceutical dosage form

Summary

Introduction

Tuberculosis (TB) remains a major global threat, the control of which has become more complicated since the emergence of multidrug-resistant (MDR) and extensively drug-resistant strains of Mycobacterium tuberculosis. In 2015, there were an estimated 10.4 million new (incident) TB cases worldwide and 480,000 new cases of multidrug-resistant TB (MDR-TB). Rifampicin (RIF) (Figure 1(a)) is a complex semisynthetic macrocyclic rifamycin class of antibiotic derived from Streptomyces mediterranei [3], used for the treatment of tuberculosis and other infectious diseases [4,5,6,7]. It is categorized one of the first line antituberculous agents. OFX (Figure 1(b)) is a synthetic fluoroquinolone antibacterial agent that inhibits the supercoiling activity of bacterial DNA gyrase, halting DNA replication [11]

Objectives

Methods

Results