Analytical Science and Technology (분석과학)

The Korean Society of Analytical Sciences (한국분석과학회)
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Determination of finasteride in human serum by LC-MS/MS

LC-MS/MS를 이용한 혈청 중 finasteride 분석

  • Nam, Hye-Seon (Food Safety Evaluation Department, National Institute of Food and Drug Safety Evaluation, Korea Food and Drug Administration Osong Health Technology Administration Complex) ;
  • Nam, Kyong-Hee (Food Safety Evaluation Department, National Institute of Food and Drug Safety Evaluation, Korea Food and Drug Administration Osong Health Technology Administration Complex) ;
  • Jung, Su-Hee (Food Safety Evaluation Department, National Institute of Food and Drug Safety Evaluation, Korea Food and Drug Administration Osong Health Technology Administration Complex) ;
  • Lee, Jang-Woo (Food Safety Evaluation Department, National Institute of Food and Drug Safety Evaluation, Korea Food and Drug Administration Osong Health Technology Administration Complex) ;
  • Kang, Jin-Yeong (Food Safety Evaluation Department, National Institute of Food and Drug Safety Evaluation, Korea Food and Drug Administration Osong Health Technology Administration Complex) ;
  • Hong, Soon-Keun (Food Safety Evaluation Department, National Institute of Food and Drug Safety Evaluation, Korea Food and Drug Administration Osong Health Technology Administration Complex) ;
  • Kim, Tae-Sung (Food Safety Evaluation Department, National Institute of Food and Drug Safety Evaluation, Korea Food and Drug Administration Osong Health Technology Administration Complex) ;
  • Kang, Tae-Seok (Toxicology Evaluation and Research Department, National Institute of Food and Drug Safety Evaluation, Korea Food and Drug Administration Osong Health Technology Administration Complex) ;
  • Yoon, Hae-Jung (Food Safety Evaluation Department, National Institute of Food and Drug Safety Evaluation, Korea Food and Drug Administration Osong Health Technology Administration Complex) ;
  • Lee, Kwang-Ho (Food Safety Evaluation Department, National Institute of Food and Drug Safety Evaluation, Korea Food and Drug Administration Osong Health Technology Administration Complex) ;
  • Rhee, Gyu-Seek (Food Safety Evaluation Department, National Institute of Food and Drug Safety Evaluation, Korea Food and Drug Administration Osong Health Technology Administration Complex)
  • 남혜선 (식약청, 식품의약품안전평가원 식품위해평가부) ;
  • 남경희 (식약청, 식품의약품안전평가원 식품위해평가부) ;
  • 정수희 (식약청, 식품의약품안전평가원 식품위해평가부) ;
  • 이장우 (식약청, 식품의약품안전평가원 식품위해평가부) ;
  • 강진영 (식약청, 식품의약품안전평가원 식품위해평가부) ;
  • 홍순근 (식약청, 식품의약품안전평가원 식품위해평가부) ;
  • 김태성 (식약청, 식품의약품안전평가원 식품위해평가부) ;
  • 강태석 (식약청, 식품의약품안전평가원 독성평가연구부) ;
  • 윤혜정 (식약청, 식품의약품안전평가원 식품위해평가부) ;
  • 이광호 (식약청, 식품의약품안전평가원 식품위해평가부) ;
  • 이규식 (식약청, 식품의약품안전평가원 식품위해평가부)
  • Received : 2011.08.03
  • Accepted : 2011.09.01
  • Published : 2011.10.25
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Abstract

A liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI/MS/MS) method was developed and validated for the determination of finasteride in human serum. Beclomethasone was used as internal standard (IS) and liquid-liquid extraction (LLE) using methyl tert-butyl ether (MTBE) was carried out to isolate analyte. The mass transitions monitored in multiple reaction monitoring (MRM) in positive ion mode were m/z 373.2${\rightarrow}$305.2 for finasteride and m/z 409.3${\rightarrow}$391.2 for IS. Retention times of finasteride and IS were 5.81 and 5.46 min, respectively. The limit of quantitation (LOQ) was 0.1 ng/mL and the calibration curve showed good linearity in the range of 0.1~20.0 ng/mL ($R^2$=0.9997). The intra-day assay precision and accuracy were in the range 6.3~10.6% and 97.3~103.6%, respectively, and the inter-day assay precision and accuracy were in the range 0.8~5.2% and 99.8~102.5%, respectively. The sample extract recovery of the method was 80~83%.

본 연구는 LC-ESI-MS/MS를 이용하여 혈청 중의 피나스테라이드의 분석법을 확립하고, 밸리데이션을 통하여 분석방법의 타당도를 검증하였다. 혈청에 내부표준물질로 베클로메타손을 첨가한 후 methyl tert-butyl ether (MTBE)을 사용하여 액체상추출법(liquid-liquid extraction, LLE)으로 전처리하였다. LC-MS/MS의 양이온 모드에서 피나스테라이드와 베클로메타손의 MRM (multiple reaction monitoring) mass transition은 각각 m/z 373.2${\rightarrow}$305.2, m/z 409.3${\rightarrow}$391.2 이었으며, 머무름 시간은 각각 5.81분, 5.46분이었다. 피나스테라이드의 정량한계는 0.1 ng/mL로 나타났으며, 0.1~20.0 ng/mL의 농도 범위에서 검량선은 우수한 직선성($R^2$=0.9997)을 확인할 수 있었으며, 회수율은 80~83% 범위이었다. 피나스테라이드의 일내에 대한 정밀도는 6.3~10.6%, 정확도는 97.3~103.6% 범위이었으며, 연속 3일간 수행한 일간 정밀도는 0.8~5.2%, 정확도는 99.8~102.5%로 나타났다.

Keywords

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