• L. VAIKUNTA RAO Department of Chemistry, Gitam Institute of Science, GITAM University (Deemed to be), Visakhapatnam 530045, India
  • K. TIRUMALA RAO Department of Chemistry, Gitam Institute of Science, GITAM University (Deemed to be), Visakhapatnam 530045, India, Product and Technology Development, Eco Logic Technologies Ltd, Hyderabad 500072, India
  • V. V. KRISHNA MOHAN KANDEPI Product and Technology Development, Eco Logic Technologies Ltd, Hyderabad 500072, India




Trichostatin A, Assay, Related substances, Method Validation, RRLC, UHPLC


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.


Download data is not yet available.


Ahuja S, Dong M. Handbook of pharmaceutical analysis by HPLC. 1st ed. Amsterdam: Elsevier; 2005.

Snyder LR, Kirkland JJ, Glajch JL. Practical HPLC method development. 2nd ed. Hoboken: John Wiley and Sons, Inc.; 1997.

Shivani S, Swapnil G, Kalindi C. A review on analytical method development and validation. Int J Appl Pharm 2018;10:6.

Lutfun N, Alev O, Satyajit DS. A review on the recent advances in HPLC, UHPLC and UPLC analyses of naturally occurring cannabinoids (2010-2019). Phytochem Anal 2019;1-45. Doi:10.1002/pca.2906

Judyta CP, Przemysław Z, Anna J, Piotr G. UHPLC: the greening face of liquid chromatography. Chromatographia 2013;76:1429–37.

Narwate BM, Ghule PJ, Ghule AV, Darandale AS, Wagh JG. Ultra performance liquid chromatography: a new revolution in liquid chromatography. Int J Pharm Drug Anal 2014;2:25-34.

Venu B, Siva RT, Krishna MT, Bhanu TS, Parmita M. Development and validation of stability indicating UPLC–PDA/MS for the determination of Imiquimod and its eight related substances: application to a topical cream. J Chromatographic Sci 2019;57:249–57.

Peterka TR. Forced degradation of tacrolimus and the development of a UHPLC method for impurities determination. Acta Pharm 2019;69:3.

Jayshri RK, Amitkumar JV, Bhupatsinh V, Parth P, Ashok P. DAD Based stability-indicating RP-UPLC method for simultaneous determination of olmesartanmedoxomil and amlodipine besylate. Pharm Chem J 2019;52:959-64.

Mohsin K, Abdulmohsen AA, Badr SA, Musaed A, Fars KA. UHPLC method development for determining sitagliptin and dapagliflozin in lipid-based self-nanoemulsifying systems as combined dose in commercial products and its application to pharmacokinetic study of dapagliflozin in rats. Pharm Chem J 2019;53:79-87.

Vanhaejcke T, Papeleu P, Elaut G, Rogiers V. Trichostatin a-like hydroxamate histone deacetylase inhibitors as therapeutic agents: toxicological point of view. Curr Med Chem 2004;11:1629-43.

You W, Steegborn C. Structural basis of sirtuin 6 inhibition by the hydroxamate trichostatin a: implications for protein deacylase drug development. J Med Chem 2018;61:10922-8.

Drummond DC, Noble CO, Kirpotin DB, Guo Z, Scott GK, Benz CC. Clinical development of histone deacetylase inhibitors as anticancer agents. Annu Rev Pharmacol Toxicol 2005;45:495-528.

Shankar S, Srivastava RK. Histone deacetylase inhibitors: mechanisms and clinical significance in cancer: HDAC inhibitor-induced apoptosis. Adv Exp Med Biol 2008;615:261-98.

Sanderson L, Taylor GW, Aboagye EO, Alao JP, Latigo JR, Coombes RC, et al. Plasma pharmacokinetics and metabolism of the histone deacetylase inhibitor trichostatin an after intraperitoneal administration to mice. Drug Meta Dispos 2004;32:1132-8.

Chan JN, Vuckovic D, Sleno L, Olsen JB, Pogoutse O, Havugimana P, et al. Target identification by chromatographic co-elution: monitoring of drug-protein interactions without immobilization or chemical derivatization. Mol Cell Proteomics 2012;11:M111.016642.

Chen Z, Wen Yuan L, Feng F, Hai Y, Chun Yong W. Determination and stress studies on YK-1101, a potential histone deacetylase, by HPLC-UV and HPLC-TOF/MS methods. J Pharm Anal 2013;3:168-72.

Jeon HG, Yoon CY, Yu JH, Park MJ, Lee JE, Jeong SJ, et al. Induction of caspase-mediated apoptosis and down-regulation of nuclear factor-kB and Akt signaling are involved in the synergistic antitumor effect of gemcitabine and the histone deacetylase inhibitor trichostatin a in human bladder cancer cells. J Urol 2011;186:2084-93.

Li GC, Zhang X, Pan TJ, Chen Z, Ye ZQ. Histone deacetylase inhibitor trichostatin A inhibits the growth of bladder cancer cells through the induction of p21WAF1 and G1 cell cycle arrest. Int J Urol 2006;13:581-6.

Ozawa A, Tanji N, Kikugawa T, Sasaki T, Yanagihara Y, Miura N, et al. Inhibition of bladder tumour growth by histone deacetylase inhibitor. BJU Int 2010;105:1181-6.

ICH: Validation of analytical procedures: text and methodology, International Conference on Harmonization Guidance Documents; 2005;Q2(R1).

ICH: Impurities in new drug substances: international conference on harmonization guidance documents; 2006;Q3A(R1).

FDA. Reviewer guidance validation of chromatographic methods. CDER; 1994.

Shabir GA. Validation of high-performance liquid chromatography methods for pharmaceutical analysis. Understanding the differences and similarities between validation requirements of the US Food and Drug Administration, the US Pharmacopeia and the International Conference on Harmonization. J Chromatogr A 2003;987:57-66.

Gupta V, Ajay DKJ, Gill NS, Kapil G. Development and validation of HPLC method-a review. Int Res J Pharm Appl Sci 2012;2:17-25.

Charde MS, Welankiwar AS, Kumar J. Method development by liquid chromatography with validation. Int J Pharm Chem 2014;4:57-61.

Bhagyasree T, Injeti N, Azhakesan A, Rao UM. A review on analytical method development and validation. Int J Pharm Res Anal 2014;4:444-8.



How to Cite

RAO, L. V., K. T. RAO, and V. V. K. M. KANDEPI. “A NOVEL VALIDATED UHPLC METHOD FOR ESTIMATION OF ASSAY AND ITS RELATED SUBSTANCES OF TRICHOSTATIN-A”. International Journal of Pharmacy and Pharmaceutical Sciences, vol. 12, no. 7, July 2020, pp. 51-58, doi:10.22159/ijpps.2020v12i7.37198.



Original Article(s)