Development and statistically validated UV spectrophotometric determination of testosterone in gel formulation

Kartik D. Bhagat, Anvesha V. Ganorkar, Atul T. Hemke, Krishna R. Gupta


Objective: A simple, precise and accurate UV-spectrophotometric method is developed and statistically validated for estimation of Testosterone in gel formulation. The proposed method includes using regression equation, area under curve (AUC), first order derivative and second order derivative spectroscopic method.

Methods: based on measurement of absorbance at a selected wavelength using UV-visible spectrophotometer with 1cm matched quartz cell and acetonitrile as a solvent. All developed methods obeyed Beer’s-lambert’s law in the concentration range of 5-25μg/mL, with correlation coefficient value less than 1.

Results: The percent amount of drug estimated was nearly 100%, found to be a good agreement with label claim of marketed gel formulation. The recovery study was carried out at three different levels, the validation study data was found to be statistically significant as all the statistical parameters are within the acceptance range (% RSD <2.0 and S.D. <±2.0).

Conclusions: The results of estimation and validation parameters like accuracy, precision, ruggedness, linearity and range were studied for all the developed methods and were found to be within limits. The results obtained were statistically compared using paired t-test and one way ANOVA analysis. The proposed method can be adopted for routine quality control for estimation of drug in formulation.


Testosterone, UV-Visible spectroscopy, Validation, Derivative, Area under curve

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Bassil N, Alkaade S, Morley JE. The benefits and risks of testosterone replacement therapy: a review. Therapeutics Clinical Risk Management. 2009;5(3):427–48.

Testosterone. American Society of Health-System Pharmacists. December 4, 2015. Accessed on 3 September 2016.

Luetjens CM, Weinbauer GF. Chapter 2: Testosterone: Biosynthesis, transport, metabolism and (non-genomic) actions. In: Nieschlag E, Behre HM, Nieschlag S, eds. Testosterone: Action, Deficiency, Substitution (4th Ed.). Cambridge: Cambridge University Press; 2012: 15–32.

Dabbs M, Dabbs JM. Heroes, rogues, and lovers: testosterone and behavior. New York: McGraw-Hill, 2000.

The monograph for testosterone and drug profile in standard book of Quality, United State Pharmacopoeia-National formulary in issuing year, 2011;3:4374-5.

Orsi DD, Pellegrini M, Marchei E, Nebuloni P, Gallinella B, Scaravelli G, et al. High performance liquid chromatography-diode array and electrospray-mass spectrometry analysis of vardenafil, sildenafil, tadalafil, testosterone and local anesthetics in cosmetic creams sold on the Internet web sites. J Pharma Biomed Analysis. 2009;50:362–9.

Badoud F, Boccard J, Schweizera C, Pralong F, Saugya M. Profiling of steroid metabolites after transdermal and oral administration of testosterone by ultra-high pressure liquid chromatography coupled to quadrupole time-of-flight mass spectrometry Baum. J Steroid Biochem Molecular Biol. 2013;138:222–35.

Blasco M, Carriquiriborde P, Marino D, Ronco AE, Somoza GM. A quantitative HPLC–MS method for the simultaneous determination of testosterone, 11-ketotestosterone and 11-β-hydroxyand-rostenedione in fish serum. Journal of Chromatography B, 2009;877:1509–15.

Petr K, Barbora T, Development of the fast, simple and fully validated high performance liquid chromatographic method with diode array detector for quantification of testosterone esters in an oil-based injectable dosage form. Steroids. 2016;115:34–9.

Youwen Y, Uboh CE, Soma LR, Fuyu G, Li X, Liua Y, et al. Simultaneous separation and determination of 16 testosterone and nandrolone esters in equine plasma using ultra high performance liquid chromatography–tandem mass spectrometry for doping control. J Chromatography A. 2010;1218:3982–93.

Mendham, RC. Denney, JD, Barnes, M. Thomas, B. Sivasankar Vogel textbook of qualitative chemical analysis 6th edition. 2013: 115-119.

Verma RM. Analytical chemistry, Theory and practice third edition. 30-31.



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