Original Article A NOVEL VALIDATED UHPLC METHOD FOR ESTIMATION OF ASSAY AND ITS RELATED SUBSTANCES OF TRICHOSTATIN-A L. VAIKUNTA RAO 1* , K. TIRUMALA RAO 1,2 , V. V. KRISHNA MOHAN KANDEPI 2 1 Department of Chemistry, Gitam Institute of Science, GITAM University (Deemed to be), Visakhapatnam 530045, India, 2 Product and Technology Development, Eco Logic Technologies Ltd, Hyderabad 500072, India Email: tirumalwithu@gmail.com Received: 19 Feb 2020, Revised and Accepted: 21 May 2020 ABSTRACT Objective: The main objective of the research work is to develop and validate a rapid UHPLC method for the estimation of assay and its related substances of Trichostatin A (TSA) in pharmaceutical samples. Methods: The UHPLC method developed for chromatographic separation between TSA and its related compounds on Poroshell 120 SB C18(50×4.6) mm; 2.7 µm RRLC column using Agilent RRLC (UHPLC) system with linear gradient elution. Results: The developed UHPLC method has shown excellent chromatographic separation between TSA and its related compounds within 12 min run time, during validation experiments, specificity study revealed that the peak threshold was more than the peak purity and no purity flag was observed. Repeatability, intra, and inter-day precision results were well within the tolerable limits. Limits of detection concentrations were found between 0.075 to 0.077 ppm and the limit of quantitation is between 0.252 to 0.258 ppm for related compounds and TSA. The related substances method recoveries were found between 80 and 120 % and assay method recovery was found between 98.0 to 102.0%. Conclusion: The developed method capability was proven for the assay of TSA and its related compounds in pharmaceutical samples and the method shows eco-friendlier than routine, conventional HPLC methods in terms of analysis time, cost and HPLC effluent waste. Keywords: Trichostatin A, Assay, Related substances, Method Validation, RRLC, UHPLC © 2020 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/) DOI: http://dx.doi.org/10.22159/ijpps.2020v12i7.37198. Journal homepage: https://innovareacademics.in/journals/index.php/ijpps INTRODUCTION The HPLC method is an analytical method in which proving the separation-related impurities and its drug compound, and it helps to estimate the content of impurities and drugs to prove the quality of the drug substances [1-3]. In this study used UHPLC system and short column during development and the application of the UHPLC method is chromatographic method gained increasing importance because of their high selectivity, sensitivity and quick chromatographic separation method. In order to prove the quality of the drug substance being analyzed in sample solutions, it is necessary to measure the amount of impurities and drugs. By applying UHPLC concepts, it can reduce the run times of chromatographic methods and effluents and decrease the turnaround times [4-6]. Various UHPLC methods have been developed for the determination of related compounds and potency of the drug in pharmaceutical drug substances and formulations and many bioanalytical methods were developed to estimate the drug content in the pharmacy kinetic study and dietary samples [7-10]. Trichostatin A (TSA) is chemically known as, [R-(E,E)]-7-[4- (Dimethylamino)phenyl]-N-hydroxy-4,6-dimethyl-7-Oxo-2,4-hepta- dienamide (fig. 1). Trichostatin A is a hydroxamic acid. It has a role as a bacterial metabolite. It derives from a (R)-Trichostatic acid. TSA serves as an antifungal antibiotic and selectively inhibits the class I and II mammalian histone deacetylase (HDAC) families of enzymes, but not class III HDACs (i.e., sirtuins) [11, 12]. It is a member of a larger class of histone deacetylase inhibitors (HDIs or HDACIs) that have a broad spectrum of epigenetic activities. Thus, TSA has some potential as an anti-cancer drug [13, 14]. Fig. 1: Chemical structure of trichostatin A (TSA) TSA concentration was determined by high-performance liquid chromatography (HPLC)-UV assay (high dose) or by HPLC-multiple reaction monitoring assay (low dose) in plasma sample [15] and identification of the TSA by IEX-HPLC [16] and reported on stress studies of HDAC family compound YK-1101by HPLC–UV and HPLC– TOF/MS methods [17] and also the majority of the reports were published on drug activity and metabolism [18-20]. Screening the literature, no related substances and assay method has been reported for the determination of TSA in pharmaceutical International Journal of Pharmacy and Pharmaceutical Sciences Print ISSN: 2656-0097 | Online ISSN: 0975-1491 Vol 12, Issue 7, 2020