Shandong Analysis and Test Center, Shandong Academy of Sciences, Jinan 250014, Shangdong, China.
Molecules. 2013 Aug 15;18(8):9755-69. doi: 10.3390/molecules18089755.
A method combining hydrophilic interaction chromatography (HILIC) and electrospray ionization mass spectrometry (ESI-MS) was developed for the characterization and determination of natural Cordyceps. Separation was achieved on a Waters Xbridge Amide column with gradient elution. Identification of 15 target nucleosides and nucleobases was based on retention time, UV spectra and mass measurements of the protonated molecules ([M+H]⁺) and main fragment ions (ESI-TOF/MS). Eight non-target compounds were tentatively identified by ESI-TOF/MS. The 15 target compounds were quantified by HILIC-ESI-MS/MS using time-programmed selective ion monitoring or multiple reaction monitoring in positive-ion mode under optimized mass conditions. This technique showed good linearity, repeatability and recovery. This approach was also successfully implemented in the analysis of nucleosides and nucleobases in 12 batches of natural Cordyceps samples that were collected from different regions in China. The developed HILIC-ESI-MS method exhibited clear advantages in identifying and determining highly polar bioactive components in Cordyceps, as well as their quality control.
建立了亲水作用色谱（HILIC）和电喷雾电离质谱（ESI-MS）相结合的方法，用于天然虫草的特征化和测定。在 Waters Xbridge Amide 柱上进行梯度洗脱实现分离。基于质子化分子（[M+H]⁺）和主要碎片离子（ESI-TOF/MS）的保留时间、紫外光谱和质量测量，对 15 种目标核苷和碱基进行了鉴定。通过 ESI-TOF/MS 鉴定了 8 种非目标化合物。采用 HILIC-ESI-MS/MS 在正离子模式下，通过时间程序选择离子监测或多反应监测，在优化的质量条件下对 15 种目标化合物进行定量。该技术具有良好的线性、重复性和回收率。该方法还成功地应用于 12 批来自中国不同地区的天然虫草样品中核苷和碱基的分析。所建立的 HILIC-ESI-MS 方法在鉴定和测定虫草中高度极性的生物活性成分及其质量控制方面具有明显的优势。
