Pharmaceutical Sciences Division, University of Wisconsin-Madison, 777 Highland Avenue, Madison, Wisconsin 53705, USA.
J Nat Prod. 2012 Apr 27;75(4):802-6. doi: 10.1021/np300034c. Epub 2012 Apr 3.
A lack of good methods for absolute quantification of natural products has limited the accuracy of high-throughput screening. Many currently used methods for quantification are either too slow or not amenable to the structural diversity of natural products. Recent developments in low-temperature evaporative light scattering detectors (ELSD-LT) have overcome several historical limitations of ELSDs, including analyte decomposition and low sensitivity. Primarily, ELSDs have been used for relative quantification and detection of compounds that lack a UV chromophore. In this study, we employ an ELSD-LT for absolute quantification of natural products. Calibration curves were constructed using a weighted least-squares analysis for a diverse set of natural products and other compounds. An average calibration curve was evaluated for the "universal" quantification of natural products. Optimization of ELSD-LT hardware and parameters improved sensitivity and throughput and established the utility of ELSD-LT for quantification of large natural product libraries.
缺乏对天然产物进行绝对定量的好方法，限制了高通量筛选的准确性。目前用于定量的许多方法要么太慢，要么不适合天然产物的结构多样性。低温蒸发光散射检测器（ELSD-LT）的最新发展克服了 ELSD 的几个历史局限性，包括分析物分解和低灵敏度。主要地，ELSD 用于相对定量和缺乏 UV 发色团的化合物的检测。在这项研究中，我们使用 ELSD-LT 对天然产物进行绝对定量。使用加权最小二乘法分析为各种天然产物和其他化合物构建了校准曲线。评估了平均校准曲线，以实现天然产物的“通用”定量。ELSD-LT 硬件和参数的优化提高了灵敏度和通量，并确立了 ELSD-LT 用于大量天然产物文库定量的实用性。
