以電灑法串聯式液相層析質譜儀定量分析含heparin 之人體血漿中的fentanyl

Quantitative Analysis of Fentanyl in Human Heparin Plasma by LC/ ESI-MS/MS

張慧兒(Hei-Erh Chang)

指導教授 ： 林銘澤

大同大學/工程學院/生物工程學系所/碩士(2010年)

摘要

Fentanyl 為一種強效鎮痛貼片，傳統上使用於靜脈內在外科或診斷操作。本研究發展一簡單、靈敏、低樣品量且不需衍生反應之萃取前處理方法，並結合高效能液相層析儀及電灑法串聯式液相質譜儀，以定量分析含heparin之人體血漿中的fentanyl 濃度。測定的線性範圍是20~2000 pg/mL，而最低定量極限 (lower limit of quantitation, LLOQ) 為20 pg/mL，其訊號與背景值所產生的干擾比，S/N (signal to noise ratio)，設定為大於五倍。
應用乙醚萃取前處理方法於fentanyl定量的確效評估項目，包括有選擇性/專一性測試、最低偵測極限、線性測試、變異係數(代表精確度)及相對誤差(代表精準度)、樣品製備的回收率、安定性及系統的穩定性等。選用六個不同來源的空白人體血漿測試中，皆無明顯的干擾及基質干擾的現象。在批次內分析 (within-run) 中，品管檢體代表精確度的變異係數及代表準確度的相對誤差分別為1.2~6.0%與-4.5~7.5%；而批次間分析 (between-run) 品管檢體的變異係數及相對誤差為3.2~5.7% 與-1.5~ 2.3%。批次內分析與批次間分析的確效分析，獲得LLOQ 變異係數及相對誤差介於4.9%與6.8%及-10.5%與17.0%；LLOQ相對誤差及相對誤差則為10.8%與6.5%，所以此分析方法符合生物樣品分析方法可接受的標準。

關鍵字

Fentanyl ； 高效液相色層分析 ； 血漿 ； 串聯式質譜法 ； 電灑法 ； 定量 ； 驗證

並列摘要

Fentanyl, a potent analgesic drug, has traditionally been used intravenously in surgical or diagnostic operation. In this study, a simple, sensitive, low sample consumption, and non-derivative method was developed for the quantitative determination of fentanyl in human heparin plasma by liquid chromatography/tandem mass spectrometry (LC/MS/MS). Quantification limit ranging from 20 to 2000 pg/mL was obtained. The analyte response at the lower limit of quantification (LLOQ: 20 pg/mL) was defined as at least 5 times the response compared to blank.
Validation parameters such as specificity/selectivity, limit of quantification (LOQ), linearity, precision, accuracy, recovery, stability and system suitability were evaluated for this diethylether extraction method. No significant interference or matrix effect caused from endogenous compounds was observed at the blank plasma from 6 different sources. The precision (%CV) and accuracy (%RE) of within-run for quality control (QC) of samples ranged from 1.2% to 6.0% and -7.5% to 7.5%, respectively. The precision (%CV) and accuracy (%RE) of between-run for QC of samples ranged from 3.2% to 5.7% and -1.5% to 2.3%, respectively. The precision (%CV) and accuracy (%RE) of LLOQ were 4.9% to 6.8% and -10.5% to 17.0% in within-run, and they were 10.8% and 6.5% in between-run. The method described here is thus eligible for the analysis of biological samples.

並列關鍵字

tandem mass spectrometry ； Eelectrospray ； Fentanyl ； HPLC ； plasma ； quantitation ； validation.

參考文獻

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