After centrifugation 20 μL of this mixture was injected Selleckchem Cilengitide into the chromatograph. The resulting solution was mixed and filtered through Whatman filter paper and filtrate was appropriately diluted to get approximate concentration and to obtain final concentration of 1000 μg/mL KETO and 400 μg/mL MP, 40 μg/mL respectively. The diluted solution was filtered through 0.20 μ filter. On the TLC plate two bands of standard stock solution D and four bands of sample solution, 5.0 μL each, were applied and the plate was developed and scanned under

the optimum chromatographic condition. After chromatographic development the peak obtained for standard and sample bands was integrated. The amount of KETO, MP and PP

present in applied volume of standard solution was fed to computer. Amount of drug present in applied volume of sample solution was obtained by comparing Rf of sample bands with that of standard bands. Amount of drug estimated in mg/gel and the percent label claim were calculated using the following formula: The content of KETO, MP and PP in sample was calculated using the following formula no. 1. equation(1) Amountofdrugestimated(mg/gel)=Meanamountestimated(μg)inappliedvolumeVolumeofsamplesolutionapplied(μL)×Volumeofstocksolution(mL)Wt.ofgeltaken(mg)×Averagewt.ofgel(mg) C59 wnt price Percent label claim was calculated using above formula no 1. Results of analysis of gel formulation and its statistical evaluation are shown in Table 2 and Table 3 respectively. The proposed method was validated by studying several parameters such as accuracy, precision, linearity, limit of detection (LOD), limit of quantitation (LOQ) and robustness. To as certain found the accuracy of proposed method, recovery studies were carried out by standard addition method, as per ICH guidelines. An accurately weighed quantity of pre-analyzed gel equivalent

to about 1000 mg KETO, 400 mg MP and 40 mg PP was transferred individually in nine different 1000.0 mL volumetric flasks. To each of the flask following quantities of KETO, MP and PP were added: Flask no.1: 800 mg KETO + 320 mg MP + 32 mg PP Then 100 mL methanol was added to each flask and content of the flask was ultrasonicated for 20 min, volume was then made up to the mark with mobile phase. The solution was individually mixed and filtered through Whatman filter paper no. 42. From the filtrate, 1.0 mL solution was diluted to 10.0 mL with mobile phase. The diluted solution was filtered through 0.2 μ membrane filter. On the TLC plate two bands of standard stock solution D and four bands of sample solution, 5.0 μL each, were applied and the plate was developed and scanned under the optimum chromatographic condition. After chromatographic development the peak obtained for standard and sample bands were integrated. The amount of KETO, MP and PP present in applied volume of standard solution was fed to computer.