Quantitative Determination of Clavulanic Acid in Human Plasma by Liquid Chromatography–Tandem Mass Spectrometry and its Application to Pharmacokinetics Study

Document Type: Original research article

Authors

1 Pharmazone Research Partner, S.G. Highway, Ahmedabad, India

2 K. B. Institute of Pharmaceutical Education and Research, Kadi Sarva Vishwavidyalaya, Gandhinagar, India

3 K. B. Institute of Pharmaceutical Education and Research, Kadi Sarva Vishwavidyalaya, Gandhinagar, India

Abstract

 
A rapid, selective and sensitive liquid chromatography–tandem mass spectrometry (LC–MS/MS) assay has been proposed for the determination of Clavulanic acid (CA) in human plasma using Diclofenac sodium as internal standard (IS). The analyte and IS were extracted from human plasma via solid phase extraction and the chromatographic separation was achieved on Inertsil ODS-3, 50 x 4.6 mm, 5µ column under isocratic conditions. Detection of CA and IS was done by tandem mass spectrometry, operating in positive ionization and multiple reaction monitoring (MRM) acquisition mode. The protonated precursor to product ion transitions monitored for CA and IS were m/z 365.2→240.2 and 409.2→228.2, respectively. The method was fully validated as per the US FDA guidelines. The linear dynamic range of CA was 7.564 - 897.893 ng/mL. The intra-batch and inter-batch precision (%CV) was ≤ 14.1% while the mean extraction recovery was 84.48 % across quality control levels. It was successfully applied to a bioequivalence study of Cefdinir/CA (125 mg/62.5 mg) suspension formulation in 32 healthy Indian male subjects under fasting condition.

Keywords


 

[1]     H.C. Neu, The crisis in antibiotic resistance, Science 257 (1992) 1064-1073.

[2]     P. Ball, A. Geddes and G. Rolinson, Amoxycillin clavulanate: an assessment after 15 years of clinical application, J. Chemother. 9 (1997) 167-198.

[3]     S.C. Sweetman, Martindale, The Complete Drug Reference, 34th ed., The Pharmaceutical Press, London, 2005.

[4]     P.A. Todd and P. Benfield, Amoxicillin/clavulanic acid. An update of its antibacterial activity, pharmacokinetic properties and therapeutic use, Drugs 39 (1990) 264-307.

[5]     J. Easton, S. Noble and C.M. Perry, Amoxicillin/clavulanic acid: a review of its use in the management of paediatric patients with acute otitis media, Drugs 63 (2003) 311-340.

[6]     G. Hoizey, D. Lamiable, C. Frances, T. Trenque, M. Kaltenbach, J. Denis and H. Millart, Simultaneous determination of amoxicillin and clavulanic acid in human plasma by HPLC with UV detection, J. Pharm. Biomed. Anal. 30 (2002) 661-666.

[7]     A.P. Ball, A.M. Geddes, P.G. Davey, I.D. Farrell and G.R. Brookes, Clavulanic acid and amoxycillin: a clinical, bacteriological, and pharmacological study, Lancet 1 (1980) 620-623.

[8]     W. Cullmann and W. Dick, A simple enzyme assay for the simultaneous determination of penicillin derivatives and clavulanic acid in biological fluids, Immun. Infekt. 14 (1986) 188-190.

[9]     A.E. Bird, J.M. Bellis and B.C. Gasson, Spectrophotometric assay of  clavulanic acid by reaction with imidazole, Analyst 107 (1982) 1241-1245.

[10] G. Pajchel, K. Pawlowski and S. Tyski, CE versus LC for simultaneous determination of amoxicillin/clavulanic acid and ampicillin/sulbactam in pharmaceutical formulations for injections, J. Pharm. Biomed. Anal. 29 (2002) 75-81.

[11] M.A. Abounassif, E.M. Abdel-Moety, M.E. Mohamed and R.A. Gad-Kariem, Liquid chromatographic determination of amoxycillin and clavulanic acid in pharmaceutical preparations, J. Pharm. Biomed. Anal. 9 (1991) 731-735.

[12] T.L. Tsou, J.R. Wu, C.D. Young and T.M. Wang, Simultaneous determination of amoxycillin and clavulanic acid in pharmaceutical products by HPLC with beta-cyclodextrin stationary phase, J. Pharm. Biomed. Anal. 15 (1997) 1197-1205.

[13] H. Ellis, H.K. Hundt, K.J. Swart, A.F. Hundt, A.L. Joubert, G.H. van Essen and J.B. Du Plessis, A fast and simple method for the determination of clavulanic acid in human plasma using derivatisation reaction kinetics, J. Pharm. Biomed. Anal. 22 (2000) 933-937.

[14] M. Foulstone and C. Reading, Assay of amoxicillin and clavulanic acid, the components of Augmentin, in biological fluids with high-performance liquid chromatography, Antimicrob. Agents. Chemother. 22 (1982) 753-762.

[15] C. Martin, M.N. Mallet, B. Sastre, X. Viviand, A. Martin, P. De Micco and F. Gouin, Comparison of concentrations of two doses of clavulanic acid (200 and 400 milligrams) administered with amoxicillin (2,000 milligrams) in tissues of patients undergoing colorectal surgery, Antimicrob. Agents. Chemother. 39 (1995) 94-98.

[16] S. Chulavatnatol and B.G. Charles, High-performance liquid chromatographic determination of amoxicillin in urine using solid-phase, ion-pair extraction and ultraviolet detection, J. Chromatogr. 615 (1993) 91-96.

[17] Q.B. Cass, R.F. Gomes, S.A. Calafatti and J. Pedrazolli Junior, Determination of amoxycillin in human plasma by direct injection and coupled-column high-performance liquid chromatography, J. Chromatogr. A 987 (2003) 235-241.

[18] A. Aghazadeh and G. Kazemifard, Simultaneous determination of amoxycillin and clavulanic acid in pharmaceutical dosage forms by LC with amperometric detection, J. Pharm. Biomed. Anal. 25 (2001) 325-329.

[19] H. Sourgens, M.A. Bertola, J.S. Verschoor, M. Kuipers and B. Rayer, Amoxicillin/clavulanic acid (875/125): bioequivalence of a novel Solutab tablet and rationale for a twice-daily dosing regimen, Int. J. Clin. Pharmacol. Ther. 42 (2004) 165-173.

[20] H. Sourgens, H. Steinbrede, J.S. Verschoor, M.A. Bertola and B. Rayer, Bioequivalence study of a novel Solutab tablet formulation of amoxicillin/clavulanic acid versus the originator film-coated tablet, Int. J. Clin. Pharmacol. Ther. 39 (2001) 75-82.

[21] C. Li, Q. Geng, D.P. Nicolau and C.H. Nightingale, Simultaneous determination of ticarcillin and clavulanate in rabbit serum and tissue cage fluid by liquid chromatography, J. Chromatogr. B Analyt. Technol. Biomed. Life Sci. 794 (2003) 227-236.

[22] D. Passali, T. Mazzei, A. Novelli, S. Fallani, P. Dentico, C. Mezzedimi, G.C. Passali and L. Bellussi, Amoxicillin/clavulanate in chronic rhinosinusitis: tissue and serum distribution, Acta Otorhinolaryngol. Belg. 55 (2001) 259-264.

[23] USFDA, Center for Drug Evaluation and Research, in, 1996.

[24] USFDA, Guidance for Industry, Bioanalytical Method Validation, Center for Drug Evaluation and Research, Rockville, USA, in, 2001.