• Manasi B. Kulkarni Department of Pharmaceutical Chemistry, Niramay, Shree Chanakya Education Society’s Indira College of Pharmacy, New Pune-Mumbai Highway, Tathwade, Pune 411033
  • Anagha M. Joshi Principal, SCES's Indira College of Pharmacy, Tathawade, Pune.
  • Rohini V. Patil Alard College of Pharmacy, Hinjewadi, Pune


Objective: HPTLC Method for Simultaneous quantification of co-enzyme Q10 and α-tocopherol in bulk and capsule dosage form was developed and validated as per International Conference on Harmonization [(ICH) Q2 (R1)] guideline.

Methods: The chromatograms were developed using a mobile phase of Toluene: ethyl acetate: chloroform (10:1:2 v/v/v) on Pre-coated silica 60F 254 plates and quantified by densitometric absorbance mode at 280 nm.

Results: The Rf values were 0.77 and 0.87 for co-enzyme Q10 and α-tocopherol, respectively. The linearity of the method was found to be in the concentration range of 0.6µg-1.8 µg/band for α-tocopherol and 2 µg-6 µg/band for co-enzyme Q10. The limits of detection and quantification were 0.3154 and 0.9559 µg/band for α-tocopherol and 3.441 and 10.42 µg/band for co-enzyme Q10.

Conclusion: Developed densitometric method was found to be robust, precise, accurate, and rapid and can be used to analyse fixed-dose capsule samples of co-enzyme Q10 and α-tocopherol.

Keywords: co-enzyme Q10, α-tocopherol, HPTLC, Validation, ICH Q2 (R1)


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1. Sethi PD. Quantitative analysis of pharmaceutical formulations, high performance thin layer chromatography. 1st edition. CBS publisher and distributor; 2013.
2. Steven Lunetta, Mark Roman. Determination of Co-enzyme Q10 content in raw materials and dietary supplements by high-performance liquid chromatography-UV: collaborative study. J AOAC Int 2008;4:702–8.
3. Crane FL, Hatefi Y, Lester RI, Widmer C. Isolation of a quinone from beef heart mitochondria. Biochim Biophys Acta 1957;25:220–1.
4. Astridani Rizky Putranti, Riesta Primaharinastiti, Esti Hendradi. effectivity and physicochemical stability of nanostructured lipid carrier coenzyme Q10 in different ratio of lipid cetylpalmitate and alpha tocopheryl acetate as carrier. Asian J Pharm Clin Res 2017;10:146-52.
5. Anterpreet Chahal, Adesh K Saini, Anil Kumar Chhillar, Reena V Saini. Natural antioxidants as defense system against cancer. Asian J Pharm Clin Res 2018;11:38-44.
6. Raitakari OT, McCredie RJ, Witting P, Griffiths KA, Letter J, Sullivan D, et al. Coenzyme Q improves LDL resistance to ex vivo oxidation but does not enhance endothelial function in hypercholesterolemic young adults. Free Radical Biol Med 2000;28:1100–5.
7. Langsjoen P, Langsjoen A, Willis R, Folkers K. Treatment of essential hypertension with coenzyme Q10. Mol Aspects Med 1994;Suppl 15:S265-72.
8. Kendler BS. Recent nutritional approaches to prevention and therapy of cardiovascular disease. Prog Cardiovasc Nurs 1997;12:3–23.
9. Fernholz E. On the constitution of alpha-tocopherol. J Am Chem Soc 1938;3:700-5.
10. Flohe RB, Traber MG. α-tocopherol: function and metabolism. FASEB J 1999;13:1145–55.
11. Niki E, Saito T, Kawakami A, Kamiya A. Inhibition of oxidation of methyl linoleate in solution by α-tocopherol and vitamin C. J Biol Chem 1984;7:4177–82.
12. Niki E, Yamamoto Y, Komuro E, Sato K. Membrane damage due to lipid oxidation. Am J Clin Nutr 1991;1:201–5.
13. Dugan RE, Frigerio NA, Sieber JM. Colorimetric determination of vitamin A and its derivatives with trifluoroacetic acid. Anal Chem 1964;1:114–7.
14. Moreno P, Salvado V. Determination of eight water and fat soluble vitamins in multivitamin pharmaceutical formulation by high performance liquid chromatography. Food Chem 2000;4:535-43.
15. Helena M, Cabral M, Jose AG. An isocratic LC method for the simultaneous determination of vitamins A, C, E and β-carotene. J Pharm Biomed Anal 1999;2:399-400.
16. Mieko K, Yoshinori I. Simultaneous determination of retinol and tocopherols by high-performance liquid chromatography. J Chromatography 1992;2:372-6.
17. Kareti Srinivasa Rao. A simple and rapid HPLC method for the simultaneous determination of vitamin-a and vitamin-E in tablet dosage forms. Res J Chem Environ 2013;1:2-6.
18. Tang PH, Miles MV, DeGrauw A, Hershey A, Pesce A. HPLC analysis of reduced and oxidized coenzyme Q in human plasma. Clin Chem 2001;2:256-65.
19. Racz Anita, Vass Andrea, Heberger Karoly, Fodor Marietta. Quantitative determination of coenzyme Q10 from dietary supplements by FT-NIR spectroscopy and statistical analysis. Anal Bioanal Chem 2015;10:2887-98.
20. Johanna K Lang, Lester Packer. Quantitative determination of vitamin E and oxidized and reduced coenzyme Q by high-performance liquid chromatography with in-line ultraviolet and electrochemical detection. J Chromatogr 1987;385:109-17.
21. Claude Leray, Margaret D Andriamampandry, Monique Freund, Christian Gachet, Jean-Pierre Cazenave. Simultaneous determination of homologues of vitamin E and coenzyme Q and products of α-tocopherol oxidation. J Lipid Res 1998;39:2099-105.
22. ICH Q2 (R1). Validation of Analytical Procedure, Text and Methodology, ICH Harmonized Tripartite Guidelines; 2005.
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How to Cite
Kulkarni, M. B., A. M. Joshi, and R. V. Patil. “A NOVEL HPTLC METHOD FOR SIMULTANEOUS DETERMINATION OF CO-ENZYME Q10 AND α-TOCOPHEROL IN BULK AND PHARMACEUTICAL FORMULATION”. International Journal of Pharmacy and Pharmaceutical Sciences, Vol. 10, no. 10, Oct. 2018, pp. 134-41, doi:10.22159/ijpps.2018v10i10.28828.
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