November-December 2021 Indian Journal of Pharmaceutical Sciences 1208 Research Paper Development and Validation by Reverse Phase High Performance Liquid Chromatography Method for the Estimation of Piperine and Coenzyme Q10 in Bulk and Pharmaceutical Dosage Form M. SARKAR, PUJA BAG, B. KUMAR AND POOJA A.CHAWLA* Department of Pharmaceutical Analysis, ISF College of Pharmacy, Moga, Punjab 142001, India Sarkar et al.: Reverse Phase High Performance Liquid Chromatography Method for Estimation of Piperine and Coenzyme Q10 Today people are focused on their health, as good health is a key to the happiness of our life. Medicine takes a big responsibility to take care of our health. Various supplements are available to meet the nutritional requirement. One such supplement is a combination of piperine and coenzyme Q10. The medicines have to be analyzed properly before introducing them to the market. As no analytical method is available for simultaneous estimation of the combination, hence an attempt was made to develop a validated method for the estimation of piperine and coenzyme Q10 simultaneously in bulk and its dosage form. A simple, precise, accurate and validated reversed phase high performance liquid chromatography technique was developed for the estimation of piperine and coenzyme Q10 in bulk and its tablet formulation. In this developed method, Waters X Bridge C 18 column (250 mm×4.6 mm, 5 µm) was used as a stationary phase and acetonitrile, tetrahydrofuran and water was used in 65:32:3 (v/v) ratio as mobile phase with 1 ml/min fow rate. This isocratic separation was accomplished using Waters 2707 Autosampler high performance liquid chromatography system, Waters 515 solvent delivery system, with photodiode array detector detection at 275 nm. Chromatographic data was processed by Empower 2 software. The retention times of coenzyme Q10 and piperine were 4.56 and 8.19 min respectively. The linearity ranges have lied between 4-6 µg/ml, 240-360 µg/ml for piperine and coenzyme Q10 respectively with 0.997 as correlation coeffcient for both. Key words: Piperine, coenzyme Q10, reversed phase high performance liquid chromatography, method validation, limit of detection, limit of quantitation Coenzyme Q10 (CoQ10) is a vitamin like nutraceutical which was found out by Frederick Crane and his colleagues at the University of Wisconsin in 1957 [1] . Chemically, it is a 2,3 dimethoxy-5 methyl-6 decaprenyl benzoquinone, having 10 isoprene units associated in the side chain and also known as ubiquinone or ubidecarenone [2] (fg. 1). It is chemically produced in the body and found in fsh, meat, nut and various oils [3] . CoQ10 is very popular for its potent antioxidant property, has a vital role in Adenosine Triphosphate (ATP) productions through the electron transport chain in mitochondria, cardioprotective, nephroprotective and membrane stabilizer [4] . CoQ10 is a higher molecular weight (863.34 g/mol) highly lipophilic compounds which come under Biopharmaceutical Classifcation System (BCS) class II [5] (fg. 1), practically insoluble in water [6] , dietary absorption of Q10 is very mild and confned [7] . CoQ10 has an outstanding safety profle, as proved by a large number of preclinical and clinical studies, in both adults and children [8-10] . For adults and pediatrics doses were up to 1200 mg/kg/d and 8 mg/ kg/d respectively. Several clinical trials have identifed the use of CoQ10 as adjuvant care in cardiovascular, neurodegenerative disorders and myopathies of the mitochondria [11-13] . According to the National Cancer *Address for correspondence E-mail: pvchawla@gmail.com Accepted 01 November 2021 Revised 06 April 2021 Received 29 November 2020 Indian J Pharm Sci 2021;83(6):1208-1214 This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike 3.0 License, which allows others to remix, tweak, and build upon the work non-commercially, as long as the author is credited and the new creations are licensed under the identical terms