Journal of Pharmaceutical Investigation

The Korean Society of Pharmaceutical Sciences and Technology (한국약제학회)
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Development and Validation of an HPLC Method for the Pharmacokinetic Study of Fexofenadine in Human

테르페나딘 체내동태 연구를 위한 혈청 중 펙소페나딘의 HPLC 정량법 개발 및 검증

  • 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)
  • 조혜영 (전남대학교 약학대학 부속 생물학적동등성 및 가교시험연구소, 전남대학교 병원 임상시험센터) ;
  • 강현아 (전남대학교 약학대학 부속 생물학적동등성 및 가교시험연구소, 전남대학교 병원 임상시험센터) ;
  • 김윤균 (단국대학교 의과대학) ;
  • 최후균 (조선대학교 약학대학) ;
  • 이용복 (전남대학교 약학대학 부속 생물학적동등성 및 가교시험연구소, 전남대학교 병원 임상시험센터)
  • Published : 2005.12.20
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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.

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