A routine experimental protocol for qHNMR illustrated with taxol
A routine experimental protocol for qHNMR illustrated with taxol
复制标题
DOI：
10.1021/np060535r
复制
发表时间：
2007-04-01
影响因子：
5.1
通讯作者：
Lankin, David C.
中科院分区：
文献类型：
--
作者：
Pauli, Guido F.;Jaki, Birgit U.;Lankin, David C.
定量 H-1 NMR (qHNMR) 为结构分析中涉及的标准光谱数据集提供了增值维度，特别是在分析生物活性分子和阐明新天然产物时。 qHNMR 方法可以集成到任何常规定性工作流程中，只需为标准溶液 H-1 NMR 实验建立定量条件，无需太多额外工作。此外，对不同化学批次的紫杉醇（紫杉醇）和短叶红豆杉提取物的检查作为工作示例，得出了进行常规实践的 C-13 解耦 qHNMR 实验并认识其潜力和主要局限性的通用方法的蓝图。拟议的协议基于新组装的 C-13 GARP 宽带解耦质子采集序列，该序列通过去除碳卫星来降低光谱复杂性。该方法能够同时提供定性和定量 NMR 数据，涵盖从纯化合物到代谢组等复杂混合物的各种分析物。由于常规可实现 300:1 (0.3%) 或更好的动态范围，qHNMR 适合从参考标准品到生物活性化合物再到代谢组分析的各种应用。提供了 qHNMR 的“食谱”方法，描述了可适用于所有主要制造商的当代 NMR 波谱仪的采集条件。
Quantitative H-1 NMR (qHNMR) provides a value-added dimension to the standard spectroscopic data set involved in structure analysis, especially when analyzing bioactive molecules and elucidating new natural products. The qHNMR method can be integrated into any routine qualitative workflow without much additional effort by simply establishing quantitative conditions for the standard solution H-1 NMR experiments. Moreover, examination of different chemical lots of taxol (paclitaxel) and a Taxus brevifolia extract as working examples led to a blueprint for a generic approach to performing a routinely practiced C-13-decoupled qHNMR experiment and for recognizing its potential and main limitations. The proposed protocol is based on a newly assembled C-13 GARP broadband decoupled proton acquisition sequence that reduces spectroscopic complexity by removal of carbon satellites. The method is capable of providing qualitative and quantitative NMR data simultaneously and covers various analytes from pure compounds to complex mixtures such as metabolomes. Due to a routinely achievable dynamic range of 300:1 (0.3%) or better, qHNMR qualifies for applications ranging from reference standards to biologically active compounds to metabolome analysis. Providing a "cookbook" approach to qHNMR, acquisition conditions are described that can be adapted for contemporary NMR spectrometers of all major manufacturers.