以即時直接分析游離法（DART）質譜儀搭配紙片取樣快速偵測血清中膽固醇含量

Quantification of Endogenous Cholesterol in Human Serum on Paper Using Direct Analysis in Real Time Mass Spectrometry

10.6134/tjfm.2017.0901.04

黃映晨(Ying-Chen Huang)；謝樺儀(Hua-Yi Hsieh)；鄒欣蓓(Hsin-Bai Zou)；李麗花(Li-Hua Li)；許仁裕(Ren-Yu Hsu)；徐丞志(Cheng-Chih Hsu)

即時直接分析 ； 質譜分析裝置 ； 臨床檢測 ； 膽固醇 ； pDART ； Mass Spectrometry ； Clinical Detection ； Cholesterol

台灣法醫學誌

9卷1期（2017 / 06 / 01）

25 - 35

繁體中文

血液測試為目前臨床最常用來檢測病患健康狀態的一種常規檢測方式。本研究以即時直接分析法（direct analysis in real time, DART）游離源結合離子阱質量分析器再搭配紙片取樣，建立一快速偵測血清中游離膽固醇含量的平台，我們稱此檢測技術為paper-loaded DART mass spectrometry （pDART-MS）。血液中膽固醇含量的變化與許多疾病相關，包含動脈粥狀硬化、糖尿病、腦部病變相關疾病等等，除此之外，膽固醇含量多寡，廣泛的被應用在評估罹患心血管疾病風險的高低，所以臨床上將血液中膽固醇含量視為一重要指標，並且透過這個指標可以做疾病的判斷與預防。此技術中以同位素氘標定的膽固醇做為內標準品，對血清中內源性游離膽固醇進行定量，並且將其定量結果與傳統酵素螢光分析法以及高效能液相層析質譜法進行比較，統計分析結果顯示3種方式所得結果沒有顯著差異，由此得知此檢測方式具有相當的準確性，同時利用此技術偵測21個超級馬拉松跑者分別在3個不同時間點收集之血清檢體內膽固醇含量，並與酵素螢光分析法做比較。此檢測方式幾乎不需要樣本前處理就能快速偵測到樣品中的膽固醇濃度，滿足了現代社會對高通量樣品快速分析的需求，這樣的分析技術有潛力成為臨床上檢測膽固醇的新方法，且有機會可被應用在重點照護檢驗。

Blood testing for endogenous small metabolites to determine physiological and biochemical states is routine for laboratory analysis. Here we demonstrate that by combining the commercial direct analysis in real time (DART) ion source with an ion trap mass spectrometer, native cholesterol in its free alcohol form is readily detected from a few hundred nanoliters of human serum loaded onto chromatography paper. Deuterium labeled cholesterol was used as the internal standard to obtain the absolute quantity of the endogenous cholesterol. The amount of the cholesterol measured by this paper-loaded DART mass spectrometry (pDART-MS) is statistically comparable with that obtained by using commercially available fluorometric-enzymatic assay and liquid chromatography/mass spectrometry. Furthermore, sera from 21 participants at three different time points in an ultramarathon were collected to obtain their cholesterol levels. The test requires only very minimal sample preparation, and the concentrations of cholesterol in each sample were acquired within a minute.

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