ISSN: 2320 – 3471(Online) Krupa Rani Gorthi and Aravind G Indian Journal of Research in Pharmacy and Biotechnology Volume 1 Issue1 January – February 2013 Page 1 QUANTITATIVE ANALYSIS OF ACETAMINOPHEN IN NUROMOL TABLETS BY HIGH PRESSURE LIQUID CHROMATOGRAPHY Krupa Rani Gorthi 1 * Aravind G 2 1. Sheffield Hallam University, Sheffield, United Kingdom 2. Nimra College of Pharmacy, Ibrahimpatnam, Vijayawada *Corresponding author: Email:krurani@gmail.com ABSTRACT In this study quantitative analysis of Acetaminophen in Nuromol tablets by High pressure liquid chromatography were reported and discussed. The mobile phase as a mixture of the acetonitrile and water 25:75 v/v, the pH was adjusted to 2.5 with the help of phosphoric acid and the flow rate was set to 1.4ml/min; the UV detection was set to 207nm. The column used for the separation of the compounds was the C18 column. The results showed that the calibration curve was linear when plotted and was tested by running the standards from 1 to 10 micrograms/ml and best fit line was obtained. Accuracy achieved is 86.21% and different parameters for validation of a method like precision, limit of detection, limit of quantification, linearity, robustness, repeatability and reproducibility were calculated and the results were within the limits. It is therefore reasonable to conclude that the method can be used for quantification of the acetaminophen in Nuromol tablets by high pressure liquid chromatography. KEYWORDS: Acetaminophen, quantitative analysis, high pressure liquid chromatography 1. INTRODUCTION Nuromol is a painkiller contains combinations of 200mg of ibuprofen and 500mg of acetaminophen. It is being used for the temporary relief of backache, migraine, headache rheumatic, dental and non dangerous arthritis, cold and fever. Acetaminophen is one among the most used non-narcotic analgesic - antipyretic agents. Ibuprofen is also called as an anti inflammatory and non steroidal drug. Pharmacologically it inhibits action of cyclo oxygenase which produces prostaglandins. Prostaglandins are produced in body when an injury occurs and cause inflammation, pain and swelling, therefore the ibuprofen reduces the pain and inflammation. By minimising the prostaglandins ibuprofen reduces the fever which raises the temperature in the body (Rang and Dales, 2007). The objective of the present scientific method is to develop and validate a simple, reliable, and accurate HPLC method for acetaminophen and ibuprofen. 2. MATERIALS AND METHODS Chemicals that were used for the experiment were Triethylamine, which is ion pair reagent, purity of 99.6% (from VMR international ltd), Acetonitrile, is HPLC grade which is stored at <30 degree centigrade (from Fisher scientific UK ltd), Methanol, which is HPLC grade (code: M/4056/17), orthophosphoric acid is from sigma Aldrich. The HPLC used was Perkin Elmer and series 200 pump and series is 200 UV/visible detector. The column used was C18 and the drugs that were used in the experiment were acetaminophen and ibuprofen (Nuromol) tablets. Chromatographic conditions that were used for the analysis is the mobile phase as a mixture of the acetonitrile and water 25:75 v/v, the pH was adjusted to 2.5 with the help of phosphoric acid and the flow rate was set to 1.4ml/min; the UV detection was set to 207nm. 2.1. Preparation of the standard solutions: Different standards ranging from 1, 2, 3, 4 and 5 microgram/ml from a stock solution of 100 microgram/ml concentration. All the standards were run for several times the mean of the peak area was calculated. The calibration curve was plotted and it was a good best fit line. Again the standards were prepared ranging from the 1 to 10 microgram/ml to see the linearity, the standards were run through the HPLC in triplicate and the mean of the peak area was calculated and plotted the calibration curve, by taking the peak area on Y-axis and the concentration on X-axis it was a good linear curve. Acetaminophen y = 110288x R 2 = 0.9987 0 100000 200000 300000 400000 500000 600000 0 1 2 3 4 5 6 Concentration Peak area Series1 Series2 Linear (Series1) Figure: 1 Chromatogram of Acetaminophen Figure: 2 Calibration curve for Acetaminophen