Analytical Method Validation Report for Essay of Glycopyrrocate and Formoteral Fumerate by RP-HPLC

B. Ashwini, M. Shymala*, JVC Sharma
Department of Pharmaceutical Analysis, Joginapally B.R. Pharmacy College, Yenkapally, Hyderabad, Telangana–500075

A B S T R A C T
A new method was established for simultaneous estimation of Glycopyrrolate and Formoterol fumerate by RP-HPLC method. The chromatographic conditions were successfully developed for the separation of Glycopyrrolate and Formoterol fumerate by using Xterra C18 (4.6 x 150mm, 5.0mm) , flow rate was 1.0ml/min, detection wave length was 220nm. The instrument used was WATERS HPLC Auto Sampler, Separation module 2695, UV Detector, Empower-software version-2. The retention times were found to be 3.1 mins and 4.2 mins. The assay of Glycopyrrolate and Formoterol fumerate was performed with tablets and the % assay was found to be 99.80 and 99.72which shows that the method is useful for routine analysis. The linearity of Glycopyrrolate and Formoterol fumerate was found to be linear with a correlation coefficient of 0.999 and 0.999, which shows that the method is capable of producing good sensitivity. The acceptance criteria of precision is RSD should be not more than 2.0% and the method show  precision 0.3 and 0.6 for Glycopyrrolate and Formoterol fumerate which shows that the method is precise.  The acceptance criteria of intermediate precision is RSD should be not more than 2.0% and the method show  precision 0.3 and 0.4 for Glycopyrrolate and Formoterol fumerate which shows that the method is repeatable when performed in different days also.  The total recovery was found to be 100.01% and 100.34%for Glycopyrrolate and Formoterol fumerate. The validation of developed method shows that the accuracy is well within the limit, which shows that the method is capable of showing good accuracy and reproducibility. The LOD and LOQ for Glycopyrrolate was found to be 3.02 and 3 and LOD and LOQ for Formoterol fumerate l was found to be 10 and 9.98The robustness limit for mobile phase variation and flow rate variation are well within the limit, the % degradation results are in limits. Which shows that the method is having good system suitability and precision under given set of conditions. 
Keywords: Glycopyrrolate, Formoterol fumerate, HPLC