New Simultaneous High Performance Liquid Chromatographic Method for Determination of NSAIDs and Opioid Analgesics in Advanced Drug Delivery Systems and Human Plasma
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.
[1] N. R. Srinivasa, A. H. Jennifer. (2005, March). Combination Therapy for
Neuropathic Pain. New Eng. J. Med. 352 (11):1373-1375. Available:
http://content.nejm.org/cgi/content/ 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: http://www.ncbi.nlm.nih.gov/pubmed
/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: http://www.ncbi.nlm.nih.gov/pubmed/ 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:
http://www.sciencedirect.com
[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: http://www.sciencedirect.com
[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:
http://www.sciencedirect.com
[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: http://www.sciencedirect.com
[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: http://www.sciencedirect.com
[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:
http://www.sciencedirect.com
[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:
http://www.sciencedirect.com
[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: http://www.sciencedirect.com
[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: http://www.sciencedirect.com
[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: http://www.sciencedirect.com
[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:
http://www.sciencedirect.com
[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: http://www.sciencedirect.com
[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:
http://www.sciencedirect.com
[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: http://www.sciencedirect.com
[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: http://www.sciencedirect.com
[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:http://www.sciencedirect.com
[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: http://www.sciencedirect.com
[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: http://www.sciencedirect.com
[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: http://www.sciencedirect.com
[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:
http://www.sciencedirect.com
[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:
http://www.sciencedirect.com
[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: http://www.sciencedirect.com
[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: http://www.sciencedirect.com
[1] N. R. Srinivasa, A. H. Jennifer. (2005, March). Combination Therapy for
Neuropathic Pain. New Eng. J. Med. 352 (11):1373-1375. Available:
http://content.nejm.org/cgi/content/ 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: http://www.ncbi.nlm.nih.gov/pubmed
/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: http://www.ncbi.nlm.nih.gov/pubmed/ 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:
http://www.sciencedirect.com
[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: http://www.sciencedirect.com
[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:
http://www.sciencedirect.com
[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: http://www.sciencedirect.com
[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: http://www.sciencedirect.com
[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:
http://www.sciencedirect.com
[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:
http://www.sciencedirect.com
[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: http://www.sciencedirect.com
[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: http://www.sciencedirect.com
[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: http://www.sciencedirect.com
[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:
http://www.sciencedirect.com
[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: http://www.sciencedirect.com
[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:
http://www.sciencedirect.com
[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: http://www.sciencedirect.com
[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: http://www.sciencedirect.com
[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:http://www.sciencedirect.com
[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: http://www.sciencedirect.com
[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: http://www.sciencedirect.com
[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: http://www.sciencedirect.com
[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:
http://www.sciencedirect.com
[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:
http://www.sciencedirect.com
[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: http://www.sciencedirect.com
[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: http://www.sciencedirect.com
@article{"International Journal of Medical, Medicine and Health Sciences:64698", author = "Asad Ullah Madni and Mahmood Ahmad and Naveed Akhtar and Muhammad Usman", title = "New Simultaneous High Performance Liquid Chromatographic Method for Determination of NSAIDs and Opioid Analgesics in Advanced Drug Delivery Systems and Human Plasma", abstract = "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 = "Diclofenac Sodium, Flurbiprofen, Tramadol, HPLCUV
detection, Validation.", volume = "3", number = "7", pages = "174-6", }
