Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 31105
New Simultaneous High Performance Liquid Chromatographic Method for Determination of NSAIDs and Opioid Analgesics in Advanced Drug Delivery Systems and Human Plasma

Authors: Asad Ullah Madni, Mahmood Ahmad, Naveed Akhtar, Muhammad Usman


A new and cost effective RP-HPLC method was developed and validated for simultaneous analysis of non steroidal anti inflammatory dugs Diclofenac sodium (DFS), Flurbiprofen (FLP) and an opioid analgesic Tramadol (TMD) in advanced drug delivery systems (Liposome and Microcapsules), marketed brands and human plasma. Isocratic system was employed for the flow of mobile phase consisting of 10 mM sodium dihydrogen phosphate buffer and acetonitrile in molar ratio of 67: 33 with adjusted pH of 3.2. The stationary phase was hypersil ODS column (C18, 250×4.6 mm i.d., 5 μm) with controlled temperature of 30 C°. DFS in liposomes, microcapsules and marketed drug products was determined in range of 99.76-99.84%. FLP and TMD in microcapsules and brands formulation were 99.78 - 99.94 % and 99.80 - 99.82 %, respectively. Single step liquid-liquid extraction procedure using combination of acetonitrile and trichloroacetic acid (TCA) as protein precipitating agent was employed. The detection limits (at S/N ratio 3) of quality control solutions and plasma samples were 10, 20, and 20 ng/ml for DFS, FLP and TMD, respectively. The Assay was acceptable in linear dynamic range. All other validation parameters were found in limits of FDA and ICH method validation guidelines. The proposed method is sensitive, accurate and precise and could be applicable for routine analysis in pharmaceutical industry as well as in human plasma samples for bioequivalence and pharmacokinetics studies.

Keywords: Validation, tramadol, Flurbiprofen, diclofenac sodium, HPLCUV detection

Digital Object Identifier (DOI):

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 1583


[1] N. R. Srinivasa, A. H. Jennifer. (2005, March). Combination Therapy for Neuropathic Pain. New Eng. J. Med. 352 (11):1373-1375. Available: full/352/13/1373
[2] R. B. Raffa. (2001, August). Pharmacology of oral combination analgesics: rational therapy for pain. J. Clin. Pharm. Ther. 26 (4), 257 - 264. Available: /11493367?dopt=Abstract
[3] N. Singla, A. Pong, K. Newman. (2005, January). Combination oxycodone 5 mg/ibuprofen 400 mg for the treatment of pain after abdominal or pelvic surgery in women. Clin. Ther. 27(1) 45-57. Available: 15763605
[4] S. Perrot, D. Krause, P. Crozes, N. Naim. (2006, October). Efficacy and tolerability of paracetamol/tramadol (325 mg/ 37.5 mg) combination treatment compared with tramadol (50 mg) monotherapy in patients with subacute low back pain. Clin. Ther. 28(10). 1592-1606. Available:
[5] E. Mohammed, A. Hamid, L. Novotny, H. Hamza. (2001, February). Determination of diclofenac sodium, flufenamic acid, indomethacin and ketoprofen by LC-APCI-MS. J Pharm. Biomed. Anal. 24. 587-594. Available:
[6] H. S. Lee, C. K. Jeong, S. J. Choi, S. B. Kim, M. H. Lee, G. Ko, D. H. Sohn. (2000, October). Simultaneous determination of aceclofenac and diclofenac in human plasma by narrowbore HPLC using columnswitching. J Pharm. Biomed. Anal. 23: 775-781. Available:
[7] O. Kuhlmann, G. Stoldt, H. Gert, G. J. Krauss. (1998, September). Simultaneous determination of diclofenac and oxybuprocaine in human aqueous humor with HPLC and electrochemical detection. J Pharm. Biomed. Anal. 17. 1351-1356. Available:
[8] T. Hirai, S. Matsomoto, I. Kishi. (1997, May). Simultaneous analysis of several non-steroidal anti inflammatory drugs in human urine by high performance liquid chromatography with normal solid phase extraction. J Chrom. B. 692. 375-388. Available:
[9] C. Arcelloni, R. Lanzi, S. Pedercini, G. Moltani, I. Fermo. A. Pontiroli, R. Paroni. (2001, September). High performance liquid chromatographic determination of diclofenac in human plasma after solid phase extraction. J Chrom. B. 763. 195-200. Available:
[10] A. Panusa, A. Multari, G. Incarnato, L. Gangliardi. (2007, March). High performance liquid chromatography analysis od anti inflammatory pharmaceuticals with UV Detection and electron spray-mass spectrometry detection in suspected counterfeit Homeopathic medicinal products. J Pharm. Biomed. Anal. 43. 1221-1227. Available:
[11] S A. M. Taburet, E. T. Singlas, R. C. Glass, F. M. Thomas, E.M. Leutenegger. (1995, April). Pharmacokinetic comparison of oral and local action transcutaneous Flurbiprofen in healthy volunteers. J Clin Pharm Ther; 20 (2). 101-7. Available:
[12] S.H. Gun, R. Ismail. (2001, August). Validation of a high performance liquid chromatographic method for Tramadol and 0-desmethyl Tramadol in human plasma using solid phase extraction. J Chrom. B. 759. 325- 335. Available:
[13] M. G. Soledad, M. A. Isabel, N. Concepcio, S. Pedren, J. Molina. (1998, June). Flow-injection spectrophotometric determination of diclofenac sodium in pharmaceuticals and urine samples. J.Pharm.Biomed. Anal. 17. 267-273. Available:
[14] A. Kugtrin, L. Zivanovio, M. Zeoevio, D. Radulovio. (1997, September). Spectrophoto-meteric study of diclofenac-Fe (III) complex S. J.Pharm.Biomed. Anal. 16. 147-53. Available:
[15] B. X. Mayer, K. Namiranian, P. dehghanyar, R. Stroh, H. Mascher, M. Muller. (2003). Comparison of UV and tandem mass spectrometric detection for the high-performance liquid chromatographic determination of diclofenac in microdialysis samples. J. Pharm. Biomed. Anal. 33. 745-754. Available:
[16] P. C. Damiani, M. Bearzoti, M. A. Cabezon. (1999, July). Spectrofluorometric determination of diclofenac sodium in tablets and ointments. J. Pharm. Biomed. Anal. 20. 587-90. Available:
[17] O. Kuhlmann, G. J. Krauss. (1997, December). Crocheted ETFE-reactor for on-line post-column photoderivatization of diclofenac in highperformance liquid chromatography. J. Pharm. Biomed. Anal. 16. 553- 559. Available:
[18] J.S. Millership, L.G. Hare, M. Farry, P.S. Collier, J.C. McElnay, M.D. Shields, D.J. Carson. (2001, July). The use of hydrophilic lipophilic balanced (HLB) copolymer SPE cartridges for the extraction of diclofenac from small volume paediatric plasma samples. J. Pharm. Biomed. Anal. 25. 871-879. Available:
[19] A. Jankowski, W. Stefanik, E. Dec, Nidal Laraki. (2006).The simultaneous determination of carbamazepine and diclofenac in biological samples. Ann. Acad. Med. Siles. 60 (2). 130-33. Available:
[20] Y. Sun, K. Takaba, H. Kido, M. N. Nakashima, K. NakaShima. (2003, January). Simultaneous determination of arylpropionic acid nonsteroidal anti-inflamatory drugs in Pharmaceutical formulations and human plasma by HPLC with UV detection. J. Pharm. Biomed. Anal. 30. 1611-1619. Available:
[21] L.G. Lala, P.M. Mello, S.R. Naik. (2002,July). HP-TLC determination of diclofenac sodium from serum. J. Pharm. Biomed. Anal. 29. 539-544. Available:
[22] M. Shamsipur, F. Jalali, S. Ershad. (2005,April). Preparation of a diclofenac potentiometric sensor and its application to pharmaceutical analysis and to drug recovery from biological fluids. J. Pharm. Biomed. Anal. 37. 943-47. Available:
[23] A. Sipos, S. Maria, S. Andras Szab, E. Istvan, S. Piroska. (2008, January). An assessment of the interactions between diclofenac sodium and ammonio methacrylate copolymer using thermal analysis and Raman spectroscopy. J. Pharm. Biomed. Anal. 46. 288-294. Available:
[24] S. Mazurek, R. Szostak. (2006, March). Quantitative determination of diclofenac sodium and aminophylline ininjection solutions by FT-Raman spectroscopy. J. Pharm. Biomed. Anal. 40. 1235-1242. Available:
[25] T. Nii, F. Ishii. (2005, July). Encapsulation efficiency of water soluble and insoluble drug in liposomes prepared by MCV method. Int. J. Pharm. 298.198-205. Available:
[26] C. Sajeev, G. Vinay, R. Archna, R. N. Saha. (2002, November). Oral controlled release formulation of diclofenac sodium by microencapsulation with ethyl cellulose. J Microencap. 19 (6). 753-760. Available: