Anal Chem. 2010 Dec 1;82(23):9611-6. doi: 10.1021/ac102200q. Epub 2010 Nov 9.
Improving the time resolution in microdialysis coupled to high performance liquid chromatography (HPLC) requires that the volume of the separation system be decreased. A low-volume separation permits smaller microdialysate volumes to be injected without suffering a sensitivity loss from dilution. Thus, improved time resolution can be achieved with offline analysis simply by decreasing the separations system volume. For online (near real-time) analysis, there is a further requirement. The separation speed must be at least as fast as the sampling time. Here, the combined use of high column pressures and temperatures, sub-2-μm stationary phase particles, capillary columns, and sensitive, low dead-volume detection resulted in a retention time for the neurotransmitter serotonin of less than 1 min in a 500 nL dialysate sample volume. Two sensitive detectors, photoluminescence following electron transfer (PFET) and electrochemical, were used for the detection of subnanomolar concentrations of serotonin in brain microdialysate samples. The general principles developed are applicable to a wide range of separations with the additional advantages of increases in sample throughput and decreases in mobile phase usage.
提高与高效液相色谱（HPLC）偶联的微透析的时间分辨率需要减小分离系统的体积。小体积分离允许注入更小的微透析液体积，而不会因稀释而导致灵敏度损失。因此，通过离线分析，仅通过减小分离系统的体积就可以实现更好的时间分辨率。对于在线（接近实时）分析，还有进一步的要求。分离速度必须至少与采样时间一样快。在这里，高压和高温、亚 2-μm 固定相颗粒、毛细管柱的联合使用以及灵敏、低死体积检测，使得神经递质血清素在 500 nL 透析液样品体积中的保留时间小于 1 分钟。两种灵敏的检测器，电子转移后光致发光（PFET）和电化学检测器，用于检测脑微透析液样品中亚纳摩尔浓度的血清素。开发的一般原则适用于广泛的分离，具有增加样品通量和减少流动相使用的额外优点。
