Development and Validation of an Ultra Performance Liquid Chromatography Photodiode Array Method for Quantification of Low Levels of Triamcinolone Acetonide in Skin Permeation Studies

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Alisa Wongman
Paiboon Tummarintra
Premrutai Thitilertdecha
Rasda Boonprasert


An analytical method using ultra-performance liquid chromatography coupled with photodiode array (UPLC-PDA) was developed and validated to quantify triamcinolone acetonide (TA) at a low level in samples from skin permeation studies which were based in phosphate buffer solution. Dexamethasone was used as an internal standard (IS) for evaluation. The separation of analytes and IS was performed on an ACQUITY UPLCTM BEH Shield RP18 column (1.7 μm, 100 mm x 2.1 mm I.D.) with a gradient system using 0.05% trifluoroacetic acid and acetonitrile as mobile phases with 5 min total run time and a detectable wavelength of 242 nm for a maximum absorbance of TA. Calibration curve linearity was linear over concentrations ranging from 50 ng/mL to 1,000 ng/mL with a correlation coefficient (r2) over 0.998. The lower limit of quantification (LLOQ) for TA was also as low as 50 ng/mL. Accuracy, precision, stability, and dilution integrity were evaluated to be within the acceptance criteria. Therefore, this newly developed UPLC-PDA method of high sensitivity and accuracy is promising for TA determination in samples from in vitro permeation experiments for dermatopharmacokinetic studies.


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Frances JS. Use and abuse of systemic corticosteroid therapy. J Am Acad Dermatol. 1979;1(2):95-105.

Ahluwalia A. Topical glucocorticoids and the skin--mechanisms of action: an update. Mediators Inflamm. 1998; 7(3):183-193.

van Heugten AJP, de Boer W, de Vries WS, Markesteijn CMA, Vromans H. Development and validation of a stability-indicating HPLC-UV method for the determination of triamcinolone acetonide and its degradation products in an ointment formulation. J Pharm Biomed Anal. 2018;149:265-270.

Abbas SS, Hegazy MA, Hendawy HAM, Weshahy SA, Abdelwahab MH. Resolution and Quantitation of Triam-cinolone Acetonide and Its Coformulated Drug in the Presence of Its Impurities and Degradation Products by HPTLC and HPLC. J AOAC Int. 2018;101(4): 981-991.

Cesar IC, Byrro RM, de Santana e Silva Cardoso FF, Mundim IM, de Souza Teixeira L, de Sousa WC, et al. Determination of triamcinolone in human plasma by a sensitive HPLC-ESI-MS/MS method: application for a pharmacokinetic study using nasal spray formulation. J Mass Spectrom. 2011;46(3):320-326.

Korodi T, Lachmann B, Kopelent-Frank H. Evaluation of different preparation methods for a preservative free triam-cinolone acetonide preparation for intravitreal administration: a validated stability indicating HPLC-method. Pharmazie. 2010;65(12):860-866.

Vieira Mde L, Singh RP, Derendorf H. Simultaneous HPLC analysis of triam-cinolone acetonide and budesonide in microdialysate and rat plasma: application to a pharmacokinetic study. J Chromatogr B Analyt Technol Biomed Life Sci. 2010;878(29):2967-2973.

Glowka FK, Karazniewicz M, Lipnicka E. RP-HPLC method with fluorescence detection for determination of small quantities of triamcinolone in plasma in presence of endogenous steroids after derivatization with 9-anthroyl nitrile; pharmacokinetic studies. J Chromatogr B Analyt Technol Biomed Life Sci. 2006;839(1-2):54-61.

Sudsakorn S, Kaplan L, Williams DA. Simultaneous determination of triam-cinolone acetonide and oxymetazoline hydrochloride in nasal spray formulations by HPLC. J Pharm Biomed Anal. 2006; 40(5):1273-1280.

Matysova L, Hajkova R, Sicha J, Solich P. Determination of methylparaben, propyl-paraben, triamcinolone acetonide and its degradation product in a topical cream by RP-HPLC. Anal Bioanal Chem. 2003; 376(4):440-443.

Liu H, Yang M, Wu P, Guan J, Men L, Lin H, et al. Simultaneous determination of triamcinolone acetonide palmitate and triamcinolone acetonide in beagle dog plasma by UPLC-MS/MS and its application to a long-term pharmaco-kinetic study of triamcinolone acetonide palmitate lipid emulsion injection. J Pharm Biomed Anal. 2015;104:105-111.

Sun W, Ho S, Fang XR, O'Shea T, Liu H. Simultaneous determination of triam-cinolone hexacetonide and triamcinolone acetonide in rabbit plasma using a highly sensitive and selective UPLC-MS/MS method. J Pharm Biomed Anal. 2018;153:267-273.

El-Saharty YS, Hassan NY, Metwally FH. Simultaneous determination of terbinafine HCL and triamcinolone acetonide by UV derivative spectro-photometry and spectrodensitometry. J Pharm Biomed Anal. 2002;28(3-4): 569-580.

U.S. Department of Health and Human Services, Food and Drug Administration. Center for Drug Evaluation and Research (CDER), Center for Veterinary Medicine (CVM), Guidance for Industry Bioana-lytical Method Validation, Rockville, Maryland. May 2018 BP.