Simultaneous Estimation of Clonazepam and Metronidazole in Pharmaceutical Tablets by Reversed-Phase High-Performance Liquid Chromatography Mode With Uv Detection

Abstract

Clonazepam (CLO) plays a significant role in treating seizures, myoclonic seizures, and other clinical infections, while metronidazole (MTZ) is an antiprotozoal, antibacterial drug. The reversed-phase high performance liquid chromatographic (RP-HPLC) technique has provided greater precision and sensitivity over other methods, especially the colorimetric and spectrophotometric. This study aimed to implement a simple method in pure and pharmaceutical tablets for the simultaneous determination of CLO and MTZ. The RPHPLC method's development and optimization for validating a separation method for the simultaneous estimation of both drugs in pharmaceutical formulation involved studying the optimum mobile phase, buffer concentration, and the pH value. Under the chromatographic conditions, RP-HPLC system achieved excellent separation on a Phenomenex HyperClone BDS (250 x 4.60 mm, 130A, and 5μ) at a temperature of 45°C and the following conditions: 65:35% acetonitrile:acetic acid and sodium acetate buffer solution (NaOAc/HAc) as mobile phase (pH 3.5); 20 μL injection volume at a flow rate of 0.75 mL/min and detection wavelength of 310 nm. The proposed RP-HPLC procedure demonstrated high precision (RSD% 1%), as well as the good linear relationship of the calibration graph at concentration ranges of 50-160, 35-100 ppb with a coefficient of determination (r2) of the regression line of 0.9996 and 0.9997 for metronidazole and clonazepam respectively. The proposed method offered excellent validated values for both LOD (4.24 and 3.06 ppb) and LOQ (14.15– 10.21 ppb) for both metronidazole and clonazepam drugs, respectively. The RP-HPLC method has been successfully applied to estimate metronidazole and clonazepam in the well-known commercial pharmaceutical tablets and gave an excellent recovery> 98% for both drugs. The results of the method was compared with the standard pharmacopeial method for both drugs using statistical tests, which indicated no difference in accuracy between the methods

Interpol Review of Drug Analysis 2019-2022

“…2019 electrochemical sensor for the determination of clonazepam [ 500 ]; electrochemical sensor for the determination of five benzodiazepines (clonazepam, diazepam, alprazolam, chiordiazepoxide, oxazepam) with poly dopamine-poly folic acid (P(DA-FA)) nanocomposite modified glassy carbon electrode (P(DA-FA)-GCE) [ 501 ]; HPLC method for the simultaneous determination of three benzodiazepines [ 502 ]; Air-assisted liquid-liquid microextraction (AALLME) as an extraction method for three benzodiazepines (chlordiazepoxide, alprazolam, and lorazepam) followed by HPLC-UV for separation and determination [ 503 ]; simultaneous detection of a ternary mixture of the benzodiazepines diazepam, lorazepam, and flunitrazepam using an array of voltammetric sensors [ 504 ]; review of electrochemical-based approaches for the determination of the benzodiazepine class of drugs [ 505 ]; synthesis of flubromazepam positional isomers for forensic analysis [ 506 ]; analytical properties (LC-QTOF-MS, GC-MS and NMR) of the designer benzodiazepine 8-chloro-6-(2-fluorophenyl)-1-methyl-4H- [1,2,4]triazolo[4,3-a] [1,4]benzodiazepine (flualprazolam) seized in an anesthesia robbery case [ 507 ]; review provides a summary of sample preparation techniques (solid-phase extraction and Liquid-liquid phase extraction) and the methods for the detection and quantification of BZDs molecules [ 508 ]; liquid-chromatography high-resolution mass spectrometry method for the detection of 15 non-FDA approved BZDs [ 509 ]; 2020 spectrophotometric-reverse flow injection analysis (rFIA) method for the determination of Nitrazepam in pure and pharmaceutical preparations [ 510 ]; RP-HPLC method for the simultaneous determination of Clonazepam and metronidazole in pharmaceutical tablets [ 511 ]; microsolid-phase extraction for the preconcentration of Nitrazepam and Oxazepam before HPLC-DAD analysis [ 512 ]; FTIR and Raman spectroscopy method supported by Differential Scanning Calorimetry (DSC) for the screening and detection of benzodiazepine [ 513 ]; TD-DART-MS for the rapid and sensitive detection of a suite of 19 benzodiazepines [ 514 ]; sensor for detecting nitro-containing BZDs in beverages at DFSA crime scenes [ 515 ]; batch and cloud point extraction kinetic spectrophotometric method for determination of Nitrazepam and Clonazepam in pharmaceutical preparations [ 516 ]; MALDI-TOF-MS methodology for the quick and qualitative detection of benzodiazepines [ 517 ]; supercritical fluid chromatography coupled with tandem mass spectrometry method for the analysis of 140 chiral and non-chiral chemicals of emerging concern in environmental samples including benzodiazepines [ 518 ]; pipette-tip micro-solid phase extraction (PT-mu SPE) with corona discharge ionization-IMS (CD-IMS) for on-site fast detection of benzodiazepines in dietary supplements [ 519 ]; electrochemical sensor for measuring lorazepam in various matrices [ ...…”