Analytical Science and Technology (분석과학)

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

LC-MS/MS를 이용한 혈청 중 dutasteride 분석

  • Nam, Hye-Seon (Food Safety Evaluation Department, National Institute of Food and Drug Safety Evaluation, Korea Food and Drug Administration) ;
  • Nam, Kyong-Hee (Food Safety Evaluation Department, National Institute of Food and Drug Safety Evaluation, Korea Food and Drug Administration) ;
  • Jung, Su-Hee (Food Safety Evaluation Department, National Institute of Food and Drug Safety Evaluation, Korea Food and Drug Administration) ;
  • Lee, Jang-Woo (Food Safety Evaluation Department, National Institute of Food and Drug Safety Evaluation, Korea Food and Drug Administration) ;
  • Kang, Jin-Yeong (Food Safety Evaluation Department, National Institute of Food and Drug Safety Evaluation, Korea Food and Drug Administration) ;
  • Hong, Soon-Keun (Food Safety Evaluation Department, National Institute of Food and Drug Safety Evaluation, Korea Food and Drug Administration) ;
  • Kim, Tae-Sung (Food Safety Evaluation Department, National Institute of Food and Drug Safety Evaluation, Korea Food and Drug Administration) ;
  • Jung, Ki-Kyung (Food Safety Evaluation Department, National Institute of Food and Drug Safety Evaluation, Korea Food and Drug Administration) ;
  • Kang, Tae-Seok (Toxicology Evaluation and Research Department, National Institute of Food and Drug Safety Evaluation, Korea Food and Drug Administration) ;
  • Yoon, Hae-Jung (Food Safety Evaluation Department, National Institute of Food and Drug Safety Evaluation, Korea Food and Drug Administration) ;
  • Lee, Kwang-Ho (Food Safety Evaluation Department, National Institute of Food and Drug Safety Evaluation, Korea Food and Drug Administration) ;
  • Rhee, Gyu-Seek (Food Safety Evaluation Department, National Institute of Food and Drug Safety Evaluation, Korea Food and Drug Administration)
  • 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) ;
  • 정기경 (식약청, 식품의약품안전평가원 식품위해평가부) ;
  • 강태석 (식약청, 식품의약품안전평가원 독성평가연구부) ;
  • 윤혜정 (식약청, 식품의약품안전평가원 식품위해평가부) ;
  • 이광호 (식약청, 식품의약품안전평가원 식품위해평가부) ;
  • 이규식 (식약청, 식품의약품안전평가원 식품위해평가부)
  • Received : 2011.10.14
  • Accepted : 2011.12.16
  • Published : 2012.02.25
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Abstract

The determination and confirmation of dutasteride in human serum was performed by a liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI/MS/MS). Beclomethasone as an internal standard (I.S.) was added to the serum and the mixed sample was pretreated by liquid-liquid extraction (LLE) with methyl tert-butyl ether (MTBE). The mass transitions of dutasteride and I.S. monitored in multiple reaction monitoring (MRM) were m/z 529.6${\rightarrow}$461.5 and m/z 409.3${\rightarrow}$391.2, respectively, and the retention times were 6.45 and 5.46 min, respectively. The calibration curve was linear in the concentration range of 0.5~30.0 ng/mL ($R^2$= 0.9999) and the limit of quantitation (LOQ) was found to be 0.5 ng/mL. The recovery of dutasteride was shown to be 66~72%. The intra-day assay precision and accuracy were in the range 3.5~5.5% and 85.7~89.9%, respectively, and the interday assay precision and accuracy were in the range 4.2~5.8% and 90.8~95.8%, respectively.

본 연구는 혈청 중 두타스테라이드의 확인과 정량을 위하여 LC-ESI-MS/MS를 이용한 신뢰성 있는 분석법을 개발하고, 분석방법의 타당도를 검증하고자 수행하였다. 내부표준물질인 베클로메타손을 첨가한 혈청을 methyl tert-butyl ether (MTBE)를 사용하여 액체상추출법(liquid-liquid extraction, LLE)으로 추출하였다. LC-MS/MS의 양이온 모드에서 MRM(multiple reaction monitoring)방법으로 확인한 두타스테라이드와 베클로메타손의 mass transition은 각각 m/z 529.6${\rightarrow}$461.5, m/z 409.3${\rightarrow}$391.2 이었으며, 머무름 시간은 각각 6.45분, 5.46분이었다. 검량선은 0.5~30.0 ng/mL의 농도 범위에서 $R^2$=0.9999의 높은 직선성을 나타내었으며, 정량한계와 회수율은 각각 0.5 ng/mL와 66~72% 이었다. 일내에 대한 정밀도는 3.5~5.5%, 정확도는 85.7~89.9% 범위이었으며, 연속 3일간 수행한 일간 정밀도는 4.2~5.8%, 정확도는 90.8~95.8% 이었다.

Keywords

References

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