Microchip HPLC separations monitored simultaneously by coherent anti-Stokes Raman scattering and fluorescence detection
Microchip HPLC separations monitored simultaneously by coherent anti-Stokes Raman scattering and fluorescence detection
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DOI：
10.1007/s00604-016-2012-3
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发表时间：
2016
影响因子：
5.7
通讯作者：
D. Geissler;Josef J. Heiland;Carsten Lotter;D. Belder
中科院分区：
文献类型：
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作者：
D. Geissler;Josef J. Heiland;Carsten Lotter;D. Belder
对相干抗斯托克斯拉曼散射（CARS）进行了评价，并将其应用于基于芯片的高效液相色谱（HPLC）过程监测中。通过在1064 nm和858.3 nm激光激发乙腈的C≡n振动，可以实时监测溶剂梯度。该方法也可应用于间接检测，例如用含乙腈的洗脱液分离短链醇。通过集成365 nm LED作为第三激发源，CARS也与外延荧光检测相结合。这使得荧光分析物可以在用CARS追踪洗脱液的乙腈部分的同时进行检测。采用梯度洗脱法在定制的HPLC玻璃芯片上分离120 s内的多环芳烃。在这个概念验证研究中，首次介绍了CARS在色谱中的应用，可以追踪洗脱液中乙腈的百分比或相当高的酒精含量。与荧光结合，在1 μg·mL−1范围内检测到多环芳烃。摘要实现了芯片液相色谱（chip-HPLC）分析多环芳烃的平行相干反斯托克斯拉曼散射（CARS）和外延荧光检测。通过刺激乙腈C≡N振动，可以实时监测溶剂梯度和死区时间（t0）。
Coherent anti-Stokes Raman scattering (CARS) was evaluated and applied to process monitoring in chip-based high-performance liquid chromatography (HPLC). By stimulating the C≡N-vibration of acetonitrile via laser excitation at 1064 nm and 858.3 nm, solvent gradients can be monitored in real-time. The approach can also be applied to indirect detection, which is exemplarily shown for the separation of short chain alcohols with an eluent containing acetonitrile. CARS was also combined with epi-fluorescence detection by integrating a 365 nm LED as a third excitation source. This allows fluorescent analytes to be detected in parallel to tracing the acetonitrile fraction of the eluent by CARS. This was applied to separate polycyclic aromatic hydrocarbons within 120 s on custom HPLC glass chips using gradient elution. In this proof of concept study, presenting the first application of CARS in chromatography, the percentage of acetonitrile in the eluent or rather high amounts of alcohols can be traced. If combined with fluorescence, polycyclic aromatic hydrocarbons were detected at the 1 μg·mL−1range.Graphical abstractParallel coherent anti-Stokes Raman scattering (CARS) and epi-fluorescence detection is realized for microchip liquid chromatography (chip-HPLC) for the analysis of polycyclic aromatic hydrocarbons. Solvent gradients and dead time (t0) can be monitored in real-time by stimulating the acetonitrile C≡N vibration.