Defined because the lowest concentration of an analyte that can reliably be differentiated from background levels. Limit ofNovember – DecemberMATERIALS AND METHODSAnalytically pure DIC and MEF had been obtained as gift samples from Balaji Laboratory limited, Mumbai, India and PCM was obtained as gift sample from Zydus Cadila Ltd., Ahmedabad, India, respectively. HPLC grade acetonitrile and water were obtained from SRL Ltd., Mumbai, India. Potassium dihydrogen phosphate and orthophoshoric acid have 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 quantity of 20 mg of diclyclomine, 250 mg of mefenamic acid and 500 mg of paracetamol were procured in the industry. The liquid chromatographic system consist of PerkinElmer series 200 LC (Shelton, USA) equipped having a series 200 UV detector, series 200 quaternary gradient pump and manual injector rheodyne valve with 20 fixed loop. The analytes have been monitored at 220 nm. Chromatographic analysis was performed on a Brownlee C18 column obtaining 250?.6 mm i.d. and 5 particle size. All of the drugs and chemical compounds have been weighed on Shimadzu electronic balance (AX200, Shimadzu Corp., Japan). The mobile phase was degassed by ultrasonic vibrations prior to use. All determinations were performed at ambient temperature. Chromatographic conditions: The Brownlee C18 column was equilibrated together with the mobile phase, acetonitrile:20 mM potassium dihydrogen phosphate 70:30 (v/v); pH 4. The flow rate was maintained at 1 ml/min. Eluent have been monitored with UV detector at 220 nm, and also the injection volume was 20 . Total run time was kept 12 min.Indian Journal of IRAK4 Inhibitor custom synthesis Pharmaceutical Sciencesijpsonlinequantification (LOQ) of an individual analytical process would be the lowest level of analyte that may be quantitatively determined with appropriate precision and accuracy. LOD and LOQ have been calculated making use of following Eqns. as per ICH guidelines, LOD=3.3?S and LOQ=10?S, where will be the regular deviation of yintercepts of regression lines and S would be the slope with the calibration curve. Robustness was studied by evaluating the impact of modest but deliberate variations in the chromatographic circumstances. The conditions studied have been flow price (altered by ?.two ml/min) and percentage of organic phase. Stability of sample options had been studied at 25??for 24 h. Program suitability test was an integral aspect of the approach improvement to verify that the system is sufficient for the analysis of DIC, MEF and PCM to be performed. Program suitability test in the chromatography program was performed prior to validation in the process. Five replicate injections of very same concentration (50 /ml of DIC, 1 /ml of MEF, two /ml of PCM) of technique suitability requirements and 1 injection of a verify regular were created. Area, retention time (RT), HIV-1 Activator custom synthesis asymmetry element, and theoretical plates for the 5 suitability injections were determined. Analysis of marketed formulation: Twenty tablets were weighed accurately and finely powdered. Tablet powder equivalent to 20 mg DIC (250 mg of MEF and 500 mg of PCM) was taken in one hundred ml volumetric flask. Methanol (50 ml) was added towards the above flask and the flask was sonicated for 15 min. The solution was filtered usingWhatman filter paper No. 41 and volume was created up to the mark using the mobile phase. Suitable volume from the aliquot was transferred to a ten ml volumetric flask plus the volume.