Analytical method development and validation of tapentadol hcl by rp hplc
- 1. ANALYTICAL METHOD DEVELOPMENT AND VALIDATION OF TAPENTADOL HYDROCHLORIDE IN TABLET DOSAGE FORM BY RP-HPLC Padmashree Dr. D. Y. Patil College of Pharmacy, Akurdi, Pune-411 044, Maharashtra, India. Prepared by- Shweta Tiwari
- 2. *INTRODUCTION TO HPLC *High Performance Liquid Chromatography/ High Pressure Liquid Chromatography is an analytical chromatographic method to separate ions/molecules from sample using mobile and stationery phases due to difference in adsorption, size, partitioning etc. It consists of 4 basic parts: 1. Pump system 2. The column 3. The injection system 4. The detector system
- 3. Pumps pushes mobile phase from reservoir and pulls to detector. It helps to maintain flow rate. Columns are usually long narrow tubes containing stationery phase. Injection system is a way of introducing sample into mobile phase. Detector in HPLC is used to monitor mobile phase that emerges from column.
- 4. *DRUG PROFILE *Name- Tapentadol Hydrochloride *Solubility- Water soluble *Molecular formula-C14H23NO.HCl *Molecular weight-257.80 *Structure-
- 5. *MATERIALS AND METHODS *Instrument-HPLC Agilent 1120 series with degasser , binary gradient pump and UV detector *Drug- Pure drug of Tapentadol hydrochloride was obtained from Emcure Pharmaceuticals Ltd., Pune. *Stationery phase-Chromasil C18 (4.6 x 250 mm, 5 μm) column maintained at 27°C temperature *Mobile phase- Methanol: water (60:40 v/v) and maintained at flow rate 1.0 ml/min. The mobile phase was kept in ultrasonicator for 30 min. and filtered through a 0.22 μm nylon membrane filter. *Injection volume- 20 μl. *Wavelength- 272.0 nm
- 6. Standard stock solution- 100 μg/ml of Tapentadol hydrochloride. Dilutions made were 5, 10, 20, 30, 40, 50 and 60 μg/ml. Calibration curve- Graph was plotted between peak area and concentration.
- 7. *ANALYSIS OF TABLET FORMULATION 20 tablets of 50mg were weighed and crushed in mortar and pestle 10mg of powder dissolved in 50ml methanol in 100ml volumetric flask and kept in ultrasonicator for 20 mins. Make up the volume with methanol. Filter through 0.2 μ nylon membrane filter. Pipette out 2,3,4,5 ml stock solution in 10ml volumetric flask and make up the volume with mobile phase to get 20,30,40,50 μg/ml of TAP. Inject the solution into HPLC system.
- 8. Fig. 3: Chromatogram of TAP in Tablet Formulation
- 9. Tablet sample Label claim (mg/tabl et) Amount found* (mg/tablet) % Label claim found* Standard deviation Standa rd error TAP 50 49.90 99.80 0.2334 0.0852 Table 2: Analysis of Tablet Formulation *Average of six readings RESULTS AND DISCUSSION The HPLC system was found to best for analysis. The retention time for Tapentadol hydrochloride were found to be 5.185 min.
- 10. Method Validation: Parameter TAP Linearity range* (µg/ml) 5-60 Correlation coefficient* 0.9990 Slope* 10415 Limit of detection (µg/ml) 0.045 Limit of quantitation (µg/ml) 0.76 Retention time* (min) 5.185 Tailing factor* 1.12 Theoretical plates* 9624
- 11. Type of Precision Peak Area % Purity* S.D. % RSD Intraday Precision 8619784 99.51 0.245 0.246 8625869 99.57 8608860 99.46 Interday Precision 8620452 99.53 0.351 0.353 8607565 99.45 8604687 99.42 Table 3: Results of Intraday and Interday Precision
- 12. Level of % recovery % Mean* Standard deviation % RSD Standar d error 80 99.75 0.3620 0.3629 0.1524 100 99.84 0.1365 0.1367 0.0976 120 99.80 0.2586 0.2591 0.1073 Table 4: Results of Accuracy
- 13. CONCLUSION The reversed phase-HPLC method developed was fully validated showing satisfactory data for all the method validation parameters tested. The developed method can be conveniently used by quality control department to determine the assay of pharmaceutical preparations.