Development and validation of analytical spectrophotometric and RP-HPLC methods for the simultaneous estimation of Hydroquinone, Hydrocortisone and Tretinoin ternary mixture in topical formulation
Direct spectrophotometric and RP-HPLC methods were developed, compared and validated for the simultaneous determination of Hydroquinone (HQ), Hydrocortisone (HC) and Tretinoin (TRT) ternary combination in laboratory made mixtures and pharmaceutical preparation. The developed spectrophotometric method characterised by it simplicity and time saving property. The proposed method was able to determine TRT directly from its absorption spectrum at 362 nm, however, HQ and HC from their first derivative spectra at 284 nm and 252 nm, respectively, without any separation step. The RP-HPLC method was developed using a C18 Sunfire© waters column with a mobile phase composed of acetonitrile : phosphate buffer (adjusted to pH 6.1 using ortho-phosphoric acid) in the ratio of 30:70 %, v/v, respectively at a flow rate of 0.8 mL/min. Quantification was based on measuring peak areas at 260 nm, where at this wavelength peaks were well resolved and eluted after 4.0, 8.2 and 19.7 minutes, for Tretinoin (TRT), Hydroquinone (HQ) and Hydrocortisone (HC), respectively.
The signals obtained (A or D1 value or peak areas) plotted against the concentration of each of the three components showed linear response in the concentration ranges of 10-50 µg/mL for HQ, 2-10 µg/mL HC and 0.5-5 µg/mL TRT for the spectrophotometric method and in the concentration range of 2-10 µg/mL for HQ, 0.1-1 µg/mL for HC and 0.05-2 µg/mL for TRT for the RP-HPLC method with excellent linear regression values. Both proposed methods were applied for the determination of the active ingredients in the pharmaceutical formulation and the common excipients present in the formulation did not interfere in the analysis. Both methods were validated in terms of linearity, LOD, LOQ, accuracy, precision and selectivity. The results obtained from the analysis of the co-administered mixture by the proposed spectrophotometric method was statistically compared to those obtained by the applied RP-HPLC method.
7. R, J., P, J. und S, S. (2012) Validated UV spectrophotometric method development for simultaneous estimation of tazarotene and hydroquinone in gel preparation. J Pharm Res, 5, 2273–2275.
8. Kaur, S., Kaur, T., Kaur, G. und Verma, S. (2017) Development and validation of a UV-spectrophotometric method for estimation of hydroquinone in bulk marketed cream and prepared NLC formulation. Int J Appl Pharm, 9, 102–108.
9. García, P.L., Santoro, M.I.R.M., Singh, A.K. und Kedor-Hackmann, E.R.M. (2007) Determination of optimum wavelength and derivative order in spectrophotometry for quantitation of hydroquinone in creams. Brazilian Journal of Pharmaceutical Sciences, 43, 397–404.
10. Htet, A.M., Thin, E.E., Saw, M.M. und Win, S. (206n. Chr.) Chemical analysis of hydroquinone and retinoic acid contents in facial whitening creams. Asian Journal of Pharmaceutical Sciences, 11, 89–90.
11. Lin, Y.-H., Yang, Y.-H. und Wu, S.-M. (2007) Experimental design and capillary electrophoresis for simultaneous analysis of arbutin, kojic acid and hydroquinone in cosmetics. Journal of Pharmaceutical and Biomedical Analysis, 44, 279–282.
12. Abbas, S.S., Elghobashy, a M.R., Bebawyb, a L.I. und Shokry, R.F. (2018) Stability-indicating chromatographic determination of hydroquinone in combination with tretinoin and fluocinolone acetonide in pharmaceutical formulations with a photodegradation kinetic study. Egypt. J. Chem., 61, 143–153.
13. Elghobashy, M.R., Bebawy, L.I., Shokry, R.F. und Abbas, S.S. (2016) Successive ratio subtraction coupled with constant multiplication spectrophotometric method for determination of hydroquinone in complex mixture with its degradation products, tretinoin and methyl paraben. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 157, 16–123.
14. K., S., K., S. und P., K. (2014) Development and validation of analytical method for simultaneous estimation of mometasone furoate, hydroquinone and retinoic acid in the topical formulation by RP-HPLC. Journal of Chemical and Pharmaceutical Research, 6, 934–940.
15. Zayed, M.A. und Abdel-Basset, M.H. Spectrophotometric Microdetermination of Tretinoin, Isotretinoinusing Iodine and Tazarotene Microdetermination Via Reaction with Rose-Bengal Reagent. Chemistry Department, Faculty of Science, Cairo University, 12613 Giza, Egypt.
16. E., R., H., H. und H., S. (2018) Development and Validation Method for Simultaneous Analysis of Retinoic Acid, Hydroquinone and Corticosteroid in Cream Formula by High- Performance Liquid Chromatography. J App Pharm Sci, 8, 87–92.
17. ICH Harmonized Tripartite guideline, Validation of Analytical Procedures text and Methodology Q2 (R1) Currant step 4 version, Parent guideline dated 27 November (Complementary guideline on Methodology dated 6 November 1996 incorporated in November 20 Geneva.
18. J.N. and Mileer, J.C.M. (2005) Statistics and Chemometrics for Analytical Chemistry. 5th Aufl., London, Pearson Prentice Hall.
19. FDA Center for Drug Evaluation Research (CDER), Reviewer Guidance: Validation of Chromatographic Methods, Washington, USA (1994).
20. ICH, International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use, Validation of analytical procedures?: Text and Methodology Q2 (R1), 2005.
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