Browse > Article
http://dx.doi.org/10.4333/KPS.2005.35.6.437

Development and Validation of an HPLC Method for the Pharmacokinetic Study of Fexofenadine in Human  

Cho, Hye-Young (Institute of Bioequivalence and Bridging Study, College of Pharmacy, Chonnam National University, Clinical Trial Center, Chonnam National University Hospital)
Kang, Hyun-Ah (Institute of Bioequivalence and Bridging Study, College of Pharmacy, Chonnam National University, Clinical Trial Center, Chonnam National University Hospital)
Kim, Yoon-Gyoon (Medical School, Dankok University)
Choi, Hoo-Kyun (College of Pharmacy, Chosun University)
Lee, Yong-Bok (Institute of Bioequivalence and Bridging Study, College of Pharmacy, Chonnam National University, Clinical Trial Center, Chonnam National University Hospital)
Publication Information
Journal of Pharmaceutical Investigation / v.35, no.6, 2005 , pp. 437-443 More about this Journal
Abstract
A rapid, selective and sensitive reversed-phase HPLC method for the determination of a major metabolite of terfenadine, fexofenadine, in human serum was developed, validated, and applied to the pharmacokinetic study of terfenadine. Fexofenadine and internal standard, haloperidol were extracted from human serum by liquid-liquid extraction with acetonitrile and analyzed on a $Symmetry^{TM}$ C8 column with the mobile phase of 1% triethylamine phosphate (pH 3.7)-acetonitrile (67:33, v/v, adjusted to pH 5.6 with triethylamine). Detection wavelength of 230 nm for excitation, 280 nm for emission and flow rate of 1.0 mL/min were fixed for the study. The assay robustness for the changes of mobile phase pH, organic solvent content, and flow rate was confirmed by $3^{3}$ factorial design using a fixed fexofenadine concentration (50 ng/mL) with respect to its peak area and retention time. In addition, the ruggedness of this method was investigated at three different laboratories using same quality control (QC) samples. This method showed linear response over the concentration range of 10-500 ng/mL with correlation coefficients greater than 0.999. The lower limit of quantification using 0.5 mL of serum was 10 ng/mL, which was sensitive enough for the pharmacokinetic studies of terfenadine. The overall accuracy of the quality control samples ranged from 95.70 to 114.58% for fexofenadine with overall precision (% C.V.) being 3.53-14.39%. The relative mean recovery of fexofenadine for human serum was 90.17%. Stability studies (freeze-thaw, short-term, extracted serum sample and stock solution) showed that fexofenadine was stable during storage, or during the assay procedure in human serum. However, the storage at $-70^{\circ}C$ for 4 weeks showed that fexofenadine was not stable. The peak area and retention time of fexofenadine were not significantly affected by the changes of mobile phase pH, organic solvent content, and flow rate under the conditions studied. This method showed good ruggedness (within 15% C.V.) and was successfully used for the analysis of fexofenadine in human serum samples for the pharmacokinetic studies of orally administered Tafedine tablet (60 mg as terfenadine) at three different laboratories, demonstrating the suitability of the method.
Keywords
Terfenadine; Fexofenadine; Human serum; Validation; Pharmacokinetics; HPLC;
Citations & Related Records
연도 인용수 순위
  • Reference
1 Martindale, The complete drug reference, 32 editions 1999, p. 418 (1999)
2 J.E. Countant, P.A. Westmark, P.A. Nardella, S.M. Walter and R.A.Okerholm, Determination of terfenadine and terfenadine acid metabolite in plasma using solid-phase extraction and high-performance liquid chromatography with fluorescence detection, J. Chromatogr., 570, 139-148 (1991)   DOI   ScienceOn
3 K.Y. Chan, R.C. George, T.M. Chen and R.A. Okerholm, Direct enantiomeric separation of terfenadine and its major acid metabolite by high-performance liquid chromatography, and the lack of stereoselective terfenadine enantiomer biotransformation in man, J. Chromatogr., 571, 291-297 (1991)   DOI   ScienceOn
4 식품의약품안전청 고시 제 2002-60호, 생물학적동등성시험 기준 (2002. 11. 22)
5 $WinNonlin^{TM}$ Users Guide Ver. 3.0, Pharsight Corp. Mountain View, CA, USA (1998-1999)
6 T. Uno, N. Yasui-Furukori, T. Takahata, K. Sugawara and T. Tateishi, Liquid chromatographic determination of fexofenadine in human plasma with fluorescence detection, J. Pharm. Biomed. Anal., 35(4), 937-942 (2004)   DOI   ScienceOn
7 Martindale, The complete drug reference, 32 editions 1999, p. 410 (1999)
8 D.K. Robbins, M.A. Castles, D.J. Pack, V.O. Bhargava and S.J. Weir, Dose proportionlity and comparison of single and multiple dose pharmacokinetics of fexofenadine (MDL 16455) and its enantiomers in healthy male volunteers, Biopharm. Drug Dispos., 19, 455-463 (1998)   DOI   ScienceOn
9 U. Hofmann, M. Seiler, S. Drescher and M.E. Fromm, Determination of fexofendine in human plasma and urine by liquid chromatography-mass spectrometry, J. Chromatogr. B, 776, 227-233 (2002)