LC/MS/MS를 이용한 원숭이 및 비글견의 간 및 장관 조직에서의 Doxifluridine과 대사체 5-FU 동시분석법 개발

• 영문명: Simultaneous Determination of Doxifluridine and 5-FU in Liver and Intestine Tissue Using LC/MS/MS
• 발행기관: 대한약학회
• 저자명: 우영아(Young Ah Woo) 김기환(Ghee Hwan Kim) 정은주(Eun Ju Jeong) 김충용(Choong Yong Kim)
• 간행물 정보: 『약학회지』제52권 제2호 (2008년), 93~100쪽, 전체 8쪽
• 주제분류: 의약학 > 기타의약학
• 파일형태: PDF
• 발행일자: 2008.04.30

영문 초록

A liquid chromatographic method with tandom spectrometric detection (LC/MS/MS) for the simultaneous determination of doxifluridine and its active metabolite, 5-fluorouracil (5-FU) was developed over the concentration range of 5~2000ng/㎖, respectively. Doxifluridine, 5-FU and internal standard, 5-chlorouracil (5-CU), were extracted from liver and intestine tissue via protein precipataion. Acetonitrile was used as the extraction solvent and the supernatant was evaporated and reconstructed in mobile phase. Optimum chromatographic separation was achieved on a Agilent Zorbax C18 (100mm×2.1mm, 3.5㎛) column with mobile phase run in isocratic with methanol : water (20 : 80, v/v). The flow rate was 0.2㎖/min with total cycle time of 5 min. The lower limit of quantification was validated at 5.0 ng/㎖ of liver and intestine tissue, for both doxifluridine and 5-FU, respectively. The intra-day and inter-day precision and accuracy of quality control (QC) samples were <11% coefficient of variation and <7% relative error from theoretical concentration for both analytes. In addition, the special designed stability study was performed, because the metabolism of doxifluridine occurs spontaneously even in ice bath for monkey liver. The stability of doxifluridine in liver and intestine of monkey and beagle dog was compared. It was found that bioanalytical validation could not be performed for the monkey liver; however, beagle dog's liver has relatively low speed of metabolism compared to monkey liver and instead of monkey liver, beagle dog's liver could be used for the validation. Bioanalytical validation could be performed in monkey intestine. Eventually, this developed method for liver and intestine will be useful in support of the toxicokinetic and pharmacokinetic studies of doxifluridine and 5-FU.

키워드

doxifluridine 5-FU LC/MS/MS monkey beagle dog liver intestine simultaneous determination stability