Stoll Dwight R, Paek Changyub, Carr Peter W
University of Minnesota, Department of Chemistry, Smith and Kolthoff Halls, 207 Pleasant Street SE, Minneapolis, MN 55455, USA.
J Chromatogr A. 2006 Dec 29;1137(2):153-62. doi: 10.1016/j.chroma.2006.10.017. Epub 2006 Oct 31.
A new approach for the high-throughput screening of biological samples to detect the presence of regulated intoxicants has been developed by modifying a conventional gradient elution high-performance liquid chromatograph (HPLC). The goal of this work was to improve the speed of gradient elution screening methods over current approaches by optimizing the operational parameters of both the column and the instrument without compromising the reproducibility of the retention times, which is the basis for the identification of intoxicant compounds. Most importantly, the novel instrument configuration substantially reduces the time needed to re-equilibrate the column between consecutive gradient runs, thereby reducing the total time for each analysis. The total analysis time for each gradient elution run is only 2.80 min, including 0.30 min for column re-equilibration between analyses. Retention times of standard calibration solutes are reproducible to better than 0.002 min in consecutive runs. A corrected retention index was adopted to account for day-to-day and column-to-column variations in retention time. For a set of forty-seven target compounds, the discriminating power and mean list length were found to be 0.95 and 3.26, respectively. In comparison to previous work with similar numbers of target compounds, the current approach provides an order of magnitude improvement in analysis time, and a four-fold decrease in mean list length.
通过对传统梯度洗脱高效液相色谱仪（HPLC）进行改进，开发出了一种用于高通量筛选生物样品以检测管制毒物存在的新方法。这项工作的目标是通过优化色谱柱和仪器的操作参数，在不影响保留时间重现性的前提下，提高梯度洗脱筛选方法相对于现有方法的速度，而保留时间的重现性是鉴定毒物化合物的基础。最重要的是，这种新颖的仪器配置大幅减少了连续梯度运行之间色谱柱重新平衡所需的时间，从而减少了每次分析的总时间。每次梯度洗脱运行的总分析时间仅为2.80分钟，包括分析之间0.30分钟的色谱柱重新平衡时间。标准校准溶质的保留时间在连续运行中的重现性优于0.002分钟。采用校正保留指数来考虑保留时间的日常变化和柱间变化。对于一组47种目标化合物，发现鉴别能力和平均列表长度分别为0.95和3.26。与之前针对类似数量目标化合物的工作相比，当前方法在分析时间上提高了一个数量级，平均列表长度减少了四倍。
