Development and Validation of an HPLC Method for Simultaneous Determination of Rifampicin, Isoniazid, Pyrazinamide, and Ethambutol Hydrochloride in Pharmaceutical Formulations

Abstract

Tuberculosis treatment consists of a fixed dose combination of rifampicin (RIF), isoniazid (INH), pyrazinamide (PYZ), and ethambutol hydrochloride (EMB). The combined treatment using various drugs is necessary for patient curing, without recrudescence, and for prevention of drug-resistant mutants, which may occur during treatment. An HPLC-diode array detector (DAD) method for the simultaneous determination of RIF, INH, PYZ, and EMB in fixed dose combination tablets was developed and validated. Chromatographic experiments were performed on an Agilent 1200 HPLC system, and the separation was carried out on a Purospher STAR RP18e (250×4.6 mm id, 5 μm, Merck) analytical column. Gradient elution was carried out with a mobile phase of 20 mM monobasic sodium phosphate buffer with 0.2% triethylamine (pH 7.0) and acetonitrile at a flow rate of 1.5 mL/min. The total run time was 12 min, and the re-equilibration time was 5 min. EMB detection was performed at 210 nm, and RIF, INH, and PYZ were detected at 238 nm, using a DAD. The method proved to be specific, linear (r2>0.99), precise (RSD<2%), accurate, and robust and may be applied to the QC analysis of pharmaceutical formulations.

Validação De Método Analítico Para Quantificação Do Fipronil Por Cromatografia Líquida De Alta Eficiência

“…. INTRODUÇÃO A busca por metodologias analíticas que sejam confiáveis, comparáveis e reproduzíveis em ampla escala tem sido alvo de pesquisas constantes nos ramos da química qualitativa e quantitativa (CHELLINI et al, 2015). Para que uma metodologia analítica de identificação ou mesmo quantificação seja difundida como parâmetro de determinação para qualidade e certeza de resultados ela deve passar por um sistema de testes e verificações denominado validação (SWARTZ ; IRA, 2018).…”

Development and Validation of RP-HPLC Method for Simultaneous Estimation of Isoniazid &#38; Pyridoxine in Bulk and It Pharmaceutical Formulations

“…Literature survey reveals that few analytical methods have been reported for the estimation of Isoniazid and pyridoxine in pharmaceutical dosage form including UV-Vis spectroscopy [7][8][9], HPTLC [10][11][12] and high performance liquid chromatography HPLC [13][14][15][16][17]. Although reports are available on stability indicating HPLC methods, the information provided is incomplete as well as results are contrast.…”

Development of derivative spectrophotometric method for simultaneous determination of pyrazinamide and rifampicin in cubosome formulation

“…In addition to the pharmacotechnical challenges for the preparation of formulations containing this association, it is necessary to develop new analytical methods for the simultaneous determination of drugs. Several published studies used different analytical techniques for the determination of antituberculosis drugs alone and in combination, including High Performance Liquid Chromatography (HPLC) with UV detector (8,9), High Performance Thin Layer Chromatography (HPTLC) (10), Ultra-high Performance Liquid Chromatography (UHPLC) with UV detector (11,12), Fourier Transform Infrared (FTIR) spectroscopy in combination with multivariate calibration of partial least square (13) and UV spectrophotometry (14). Due to the absence of an analytical method for the simultaneous determination of PYZ and RIF by derivative spectrophotometric method and considering the importance of analytical methods for the simultaneous determination of drugs, this study aims to develop and validate a simple and rapid derivative spectrophotometric method for the determination of PYZ and RIF in cubosome formulation.…”