超高效能液相層析電灑質譜法分析乙型受體素之研究與探討

Determination of β-agonists Using Ultra High Performance Liquid Chromatography Mass Spectrometry

張璇璽

超高效液相層析法 ； 電灑法質譜 ； 乙型受體素類 ； 管柱後分流 ； UPLC ； ESI/MS ； β-agonists ； post-column splitting

臺灣大學化學研究所學位論文

2014年

碩士

何國榮

繁體中文

超高效能液相層析法使用填充顆粒為1.7 μm之層析管柱，根據Van Deemter equation，其最佳線性流速為0.6 mL/min，在此流速下可提供較高的管柱效能，擁有較好的解析度，並且可以有更快的分析速度。由於液相層析的移動相流速會影響液相層析電灑質譜法之靈敏度，當流速較高時需要揮發的溶劑較多，無法有效的揮發，使游離化效率較差而造成靈敏度下降。因此目前液相層析質譜法的文獻中，使用1.7 μm顆粒填充的層析管柱時多是操作在較低之流速，未能達到其管柱之效能及快速分析之優點。 本研究目的是希望利用超高效能液相層析電灑質譜法操作在最佳線性流速下，並結合管柱後分流的方式，使進入電灑質譜法的移動相流速為最佳噴灑的流速，以達到低流速的感度及高流速的解析度與快速分析的訴求。本實驗以七種乙型受體素進行分析，首先比較各個流速在最佳游離條件下的感度，實驗結果得到在沖提流速為0.2 mL/min時的感度最好。因此本實驗將以0.6 mL/min的移動相流速進行沖提，在移動相流出層析管柱後將其分流(分流比1：3)，使移動相以0.2 mL/min的流速進入游離源進行氣化及離子化。 以超高效能液相層析電灑質譜結合管柱後分流的方式分析乙型受體素類，其感度較沖提流速為0.6 mL/min未分流直接進入電灑質譜法偵測所得的感度佳，但未能達到以沖提流速為0.2 mL/min未分流直接進入電灑質譜法所偵測之感度，推測是由於質譜偵測器於高流速偵測下，離子由離子源傳送入質譜儀速度之限制，導致分流方法達不到預期感度。

The ultra performance liquid chromatography (UPLC) is based on the principle of using stationary phase with particles less than 2 μm. According to the Van Deemter equation, as the particle size decreases to less than 2 μm, not only increased efficiency, but also the ability to work at higher linear velocity without a loss of efficiency, providing both resolution and speed. The flow rate of mobile phase is an important parameter affecting the sensitivity of LC-ESI-MS. High flow rate can reduce the ionization efficiency. Therefore, low flow rate was often used. However, low flow rate failed to achieve column efficiency and rapid analysis. In this study, post-column splitting was used to reduce the flow into the mass spectrometer. This would enhance the ionization efficiency under ESI. First part of this work was to compare the sensitivity at different flow rates. The results showed that flow rate at 0.2 mL/min provided the best sensitivity. Therefore, the flow rate of mobile phase was at to 0.6 mL/min. Using post-column splitting (split ratio1:3), a flow rate of 0.2 mL/min was introduced into the ESI-MS system for detection. UPLC-ESI-MS/MS with post-column splitting (0.2 mL/min) provided better sensitivity than elution flow rate of 0.6 mL/min without splitting. However, post-column splitting did not achieve the sensitivity under 0.2 mL/min without splitting. One possible reason is the limitation in the rate of transferring the ions from ESI source to the mass spectrometer.

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1. 游孟儒(2015)。動物用藥於液相層析電灑質譜之訊號改善研究。臺灣大學化學研究所學位論文。2015。1-116。