Seger Christoph, Godejohann Markus, Spraul Manfred, Stuppner Hermann, Hadacek Franz
Institute of Pharmacy, Center of Molecular Biosciences, Leopold Franzens University Innsbruck, Innrain 52, A-6020 Innsbruck, Austria.
J Chromatogr A. 2006 Dec 8;1136(1):82-8. doi: 10.1016/j.chroma.2006.09.058. Epub 2006 Oct 5.
The absolute configuration of secondary hydroxy functions of seven natural occurring polyyne derivatives has been elucidated by the application of Mosher method of diastereomeric methoxy-2-trifluoromethyl-phenylacetyl (MTPA) ester formation. High-performance liquid chromatography with diode array detection (HPLC-DAD) of the reaction mixture using a water/acetonitrile gradient allowed monitoring the reaction progress. Coupling of high-performance liquid chromatography to solid-phase extraction combined with nuclear magnetic resonance (HPLC-SPE-NMR) was utilized to generate highly reproducible (1)H and (19)F NMR data needed as input for the absolute configuration determination based on the analysis of relative shift differences. Chromatographic peaks of reaction substrates and reaction products bearing less 10mug analyte were trapped on SPE cartridges with the aid of water as makeup solvent. Deuterated chloroform was used to elute and transfer the peak content from the SPE to the 60mul flow cell of a 500MHz NMR spectrometer. For each analyte (1)H NMR spectra were obtained within 15min. Additionally (19)F NMR spectra were recorded for selected analytes in the same timeframe. Based on the obtained NMR data, the absolute configuration of all polyynes under investigation was successfully designated.
通过应用非对映体甲氧基-2-三氟甲基苯乙酰（MTPA）酯形成的莫舍尔方法，阐明了七种天然存在的多炔衍生物仲羟基官能团的绝对构型。使用水/乙腈梯度对反应混合物进行二极管阵列检测的高效液相色谱法（HPLC-DAD），可以监测反应进程。利用高效液相色谱与固相萃取相结合并与核磁共振联用（HPLC-SPE-NMR），来生成高度可重现的¹H和¹⁹F NMR数据，这些数据是基于相对位移差异分析进行绝对构型测定所需的输入数据。含有少于10μg分析物的反应底物和反应产物的色谱峰，借助水作为补充溶剂被捕集在固相萃取柱上。用氘代氯仿洗脱并将峰内容物从固相萃取柱转移到500MHz核磁共振光谱仪的60μl流通池中。对于每种分析物，在15分钟内获得¹H NMR光谱。此外，在相同的时间范围内，对选定的分析物记录¹⁹F NMR光谱。基于获得的NMR数据，成功确定了所有被研究多炔的绝对构型。
