Defined as the lowest concentration of an analyte that could reliably be differentiated from background levels. Limit ofNovember – DecemberMATERIALS AND METHODSAnalytically pure DIC and MEF had been obtained as gift samples from Balaji Laboratory restricted, Mumbai, India and PCM was obtained as present sample from Zydus Cadila Ltd., Ahmedabad, India, respectively. HPLC grade acetonitrile and water had been obtained from SRL Ltd., Mumbai, India. Potassium dihydrogen phosphate and orthophoshoric acid had been of analytical reagent grade obtained from S. D. Fine Chem Ltd., Mumbai. Marketed tablet formulation A (Cyclopam plus, Indoco Remedies, India) and B (Trigan MF, Cadila Pharmaceuticals Ltd., India) containing labeled amount of 20 mg of diclyclomine, 250 mg of mefenamic acid and 500 mg of paracetamol had been procured in the market. The liquid Chromatographic program consist of PerkinElmer series 200 LC (Shelton, USA) equipped using a series 200 UV detector, series 200 quaternary gradient pump and manual injector rheodyne valve with 20 fixed loop. The analytes had been monitored at 220 nm. Chromatographic analysis was performed on a Brownlee C18 column getting 250?.six mm i.d. and 5 particle size. All of the drugs and chemicals had been weighed on Shimadzu electronic balance (AX200, Shimadzu Corp., Japan). The mobile phase was degassed by ultrasonic vibrations before use. All determinations have been performed at ambient temperature. Chromatographic circumstances: The Brownlee C18 column was equilibrated together with the mobile phase, acetonitrile:20 mM potassium dihydrogen phosphate 70:30 (v/v); pH four. The flow price was maintained at 1 ml/min. Eluent were monitored with UV detector at 220 nm, as well as the injection volume was 20 . Total run time was kept 12 min.Indian Journal of Pharmaceutical Sciencesijpsonlinequantification (LOQ) of an individual analytical process is definitely the lowest amount of analyte that may be quantitatively determined with suitable precision and accuracy. LOD and LOQ have been calculated using following Eqns. as per ICH recommendations, LOD=3.three?S and LOQ=10?S, where will be the standard deviation of yintercepts of regression lines and S could be the slope on the calibration curve. Robustness was studied by evaluating the impact of modest but deliberate variations inside the chromatographic circumstances. The conditions studied had been flow price (altered by ?.two ml/min) and percentage of organic phase. Stability of sample solutions had been studied at 25??for 24 h. Program H-Ras Inhibitor custom synthesis suitability test was an integral portion on the process improvement to verify that the technique is sufficient for the evaluation of DIC, MEF and PCM to become performed. Program suitability test on the chromatography technique was performed before validation with the system. Five replicate injections of same concentration (50 /ml of DIC, 1 /ml of MEF, two /ml of PCM) of system suitability standards and a single injection of a check common had been produced. Location, retention time (RT), asymmetry element, and theoretical plates for the 5 suitability injections had been determined. Evaluation of marketed formulation: Twenty tablets have been weighed accurately and finely powdered. Tablet powder equivalent to 20 mg DIC (250 mg of MEF and 500 mg of PCM) was taken in 100 ml volumetric flask. Methanol (50 ml) was added for the above flask and the flask was sonicated for 15 min. The Dopamine Receptor Antagonist MedChemExpress answer was filtered usingWhatman filter paper No. 41 and volume was made up to the mark using the mobile phase. Acceptable volume from the aliquot was transferred to a 10 ml volumetric flask and also the volume.