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페노프로펜 체내동태 연구를 위한 혈청 중 페노프로펜의 HPLC 정량법 개발 및 검증

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

  • 조혜영 (전남대학교 약학대학 부속 생물학적동등성 및 가교시험연구소, 전남대학교 병원 임상시험센터) ;
  • 강현아 (전남대학교 약학대학 부속 생물학적동등성 및 가교시험연구소, 전남대학교 병원 임상시험센터) ;
  • 김윤균 (단국대학교 의과대학) ;
  • 사홍기 (이화여자대학교 약학대학) ;
  • 이용복 (전남대학교 약학대학 부속 생물학적동등성 및 가교시험연구소, 전남대학교 병원 임상시험센터)
  • 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, Dankook University) ;
  • Sah, Hong-Kee (College of Pharmacy, Ewha Womans University) ;
  • Lee, Yong-Bok (Institute of Bioequivalence and Bridging Study, College of Pharmacy, Chonnam National University, Clinical Trial Center, Chonnam National University Hospital)
  • 발행 : 2005.12.20

초록

A selective and sensitive reversed-phase HPLC method for the determination of fenoprofen in human serum was developed, validated, and applied to the pharmacokinetic study of fenoprofen calcium. Fenoprofen and internal standard, ketoprofen, were extracted from human serum by liquid-liquid extraction with diethyl ether and analyzed on a Luna C18(2) column with the mobile phase of acetonitrile-3 mM potassium dihydrogen phosphate (32:68, v/v, adjusted to pH 6.6 with phosphoric acid). Detection wavelength of 272 nm and flow rate of 0.25 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 fenoprofen concentration $(2\;{\mu}g/mL)$ with respect to its peak area and retention time. And also, 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 $0.05-100\;{\mu}g/mL$ with correlation coefficients greater than 0.999. The lower limit of quantification using 1 mL of serum was $0.05\;{\mu}g/mL$, which was sensitive enough for pharmacokinetic studies. The overall accuracy of the quality control samples ranged from 92.27 to 109.20% for fenoprofen with overall precision (% C.V.) being 5.51-11.71 %. The relative mean recovery of fenoprofen for human serum was 81.7%. Stability (freeze-thaw, short and long-term) studies showed that fenoprofen was not stable during storage. But, extracted serum sample and stock solution were allowed to stand at ambient temperature for 12 hr prior to injection without affecting the quantification. The peak area and retention time of fenoprofen 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 fenoprofen in human serum samples for the pharmacokinetic studies of orally administered Fenopron tablet (600 mg as fenoprofen) at three different laboratories, demonstrating the suitability of the method.

키워드

참고문헌

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