Citation: Palled PJ, Dushyanth Reddy V, Mannor VS, et al. Validated Isocratic/Gradient RP-HPLC for Simultaneous Estimation of Paracetamol Ibuprofen and Caffeine in Marketed Formulations Using Diclofenac as Internal Standard. Anal Chem Ind J. 2017;17(1):116. © 2017 Trade Science Inc. 1 Validated Isocratic/Gradient RP-HPLC for Simultaneous Estimation of Paracetamol Ibuprofen and Caffeine in Marketed Formulations Using Diclofenac as Internal Standard Palled PJ, Dushyanth Reddy V * , Mannor VS and Bharat Chowdary Department of Pharmaceutical Chemistry, KLE's University College of Pharmacy, KLE University Belagavi 590001, India * Corresponding author: Dushyanth Reddy V, Department of Pharmaceutical Chemistry, KLE's University College of Pharmacy, KLE University, Belagavi 590001, India, Tel: 08312471399; E-mail: dushyanthreddy233@gmail.com Received: October 16, 2016; Accepted: January 16, 2017; Published: January 22, 2017 Introduction Paracetamol (Acetaminophen) is chemically N-(4-hydroxyphenyl) acetamide is a crystalline solid is a sparingly soluble compound which is classified under antipyretic analgesics. Drugs Classified under this class possess analgesic and antipyretic activity but lacks anti-inflammatory effects indicated for use in patients who are sensitive to aspirin with usual adult dosage 325 mcg to 650 mcg dose greater than 2.6 g/day are not advisable for prolonged treatment owing to its hepatotoxicity. Antipyretic effect of acetaminophen affords to the inhibition of endogenous leukocytic pyrogens released from cells upon external stimuli or upon activation with exogenous pyrogen. acetaminophen possesses analgesic activity in arthritis and musculoskeletal disorders. Abstract Objective: To develop a validated RP-HPLC method for simultaneous estimation of Paracetamol Ibuprofen and Caffeine. Methods: The HPLC instrument used was Shimadzu LC-20AD with reverse phase ODS-Hypersil-C18 (250 mm × 4.6 mm, 5 μm) column using Acetonitrile:water (90:10) pH adjusted to 2.8 as mobile phase. The flow rate was maintained at 0.3 ml/min and UV detection was carried at 203 nm. Results: The method was validated for linearity, accuracy, precision, specificity, robustness and ruggedness according to ICH guidelines. The retention time for Paracetamol, Ibuprofen and Caffeine was found to be 9.7, 12.66 and 10.48 respectively. The regression analysis showed good linearity over the concentration range of 1.25 μg/ml to 20 μg/ml for paracetamol, 0.625 μg/ml to 10 μg/ml for Ibuprofen and 0.625 μg/ml to 10 μg/ml for Caffeine. The recovery studies of the method gave good results in the range of 99.89% to 100.48% with less than 2% of RSD. Conclusion: The method was found to be suitable for the analysis of marketed formulation in presence of other excipients. Keywords: Paracetamol; HPLC; Chromatography; Ibuprofen; Anti-inflammatory