method development

The aim of this work is to establish simple economical, and rapid spectrophotometric and chromatographic identification method for qualification of captopril in Active Pharmaceutical Ingredient. The work was carried out to for estimation of captopril in bulk pharmaceutical from by utilizing area under curve (AUC)method using UV –Visible Spectrophotometry .For this purpose the wavelength range 200-400 nm was selected. Thin-Layer chromatography (TLC) is a chromatography used to separate non-volatile mixtures. TLC can be used for monitoring the progress of a reaction ,identification compounds present in a given mixture, and determination of active pharmaceutical ingredient .TLC analysis of captopril in pharmaceutical and for using this method of analysis in future for the development of bio analytical method .for TLC the mobile phase with different concentration was used .we have established that the most perfect RF observed using mobile phase .chloroform –methanol are used (9:1)and (8:2) ratio .the development method was found to be simple ,linear ,precise ,accurate and sensitive which can be used for routine analysis for spectrophotometric and Thin-layer chromatographic method estimation of active pharmaceutical Ingredient.

A stable, simple, accurate, precise, robust and highly selective Reverse Phase High Performance Liquid Chromatographic (RP-HPLC) method was developed and validated using ritonavir. Chromatographic separation was achieved using cyber labs, LC 100 separation module, Agilent C18 column at temperature 30°C. Flow rate selected was 1ml/min. Both drugs are identified with UV detector at 256nm. Mobile phase employed was Methanol: Water (50:50), which resulted best   sensitivity. Developed method was validated in terms of linearity, range (25-150µg/ml), precession (correlation coefficient is less than 0.999), robustness, accuracy(recovery was 101.96%) and  under stress conditions drug degradation was less than 10%.The validation of proposed stability indicating method was verified by recovery studies and can be applicable in routine pharmaceutical analysis.

This research paper describes a procedure for determination of valproic acid (VA) in human serum using salicylic acid (SA) as internal standard (I.S.). The two drugs were separated by using 60% acetonitrile and 40% acidified water with phosphoric acid adjusted pH to 3 ±0.1 as mobile phase.  The column employed was C18 150×4.65µm at ambient temperature. Detection wave length, 220nm, flow rate 1ml/min. injection volume,20µL The method was linear over the range of 10-100ug/ml valproic acid. The validated limit of quantification and the limit of detection for valproic acid were11,1 and 1.11ug/ml, respectively. The precision of the method was evaluated and the intra and the interday precision were found to be 0.2 and 2.0, respectively.  The method is simple rapid, sensitive, robust and accurate and can be used for therapeutic drug monitoring (TDM).

A simple, accurate and precise high performance thin layer chromatographic (HPTLC) method has been developed for the estimation of Acenocoumerol in bulk drug and dosage form. The method employed silica gel 60 F254 precoated plates as stationary phase and mixture of toluene: isopropyl alcohol:formic acid (7.5:2:0.5) as mobile phase. Densitometric scanning was performed at 290 nm using Camag TLC scanner 3 with WINCAT software of version 1.4.3 Camag. Beer’s law was obeyed in the concentration range of 30ng/spot-90ng/spot. The Retention factor for Acenocoumerol was found to be 0.68. The limit of detection and limit of quantitation were found to be 5 ng/spot & 15 ng/spot respectively. The % RSD of intra-day variation and inter day variation were found to be 1.10 and 1.12 respectively. As per ICH guidelines the results of the analysis were validated in terms of linearity, precision, accuracy, limit of detection and limit of quantification, and were found to be satisfactory. The proposed method can also be used for routine quality control to accurately determine Acenocoumerol in bulk and dosage form.