Zong Shi-Yu, Han Han, Wang Bing, Li Ning, Dong Tina Ting-Xia, Zhang Tong, Tsim Karl W K
School of Pharmacy, Shanghai University of Traditional Chinese Medicine, No. 1200 Cailun Road, Pudong New District, Shanghai 201203, China.
Experimental Center of Teaching and Learning, Shanghai University of Traditional Chinese Medicine, No. 1200 Cailun Road, Pudong New District, Shanghai 201203, China.
Molecules. 2015 Dec 4;20(12):21816-25. doi: 10.3390/molecules201219807.
A reliable ultra-high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry (UHPLC-ESI-MS/MS) method for the fast simultaneous determination of 13 nucleosides and nucleobases in Cordyceps sinensis (C. sinensis) with 2-chloroadenosine as internal standard was developed and validated. Samples were ultrasonically extracted in an ice bath thrice, and the optimum analyte separation was performed on an ACQUITY UPLC(TM) HSS C18 column (100 mm × 2.1 mm, 1.8 μm) with gradient elution. All targeted analytes were separated in 5.5 min. Furthermore, all calibration curves showed good linear regression (r > 0.9970) within the test ranges, and the limits of quantitation and detection of the 13 analytes were less than 150 and 75 ng/mL, respectively. The relative standard deviations (RSDs) of intra- and inter-day precisions were <6.23%. Recoveries of the quantified analytes ranged within 85.3%-117.3%, with RSD < 6.18%. The developed UHPLC-ESI-MS/MS method was successfully applied to determine nucleosides and nucleobases in 11 batches of C. sinensis samples from different regions in China. The range for the total content in the analyzed samples was 1329-2057 µg/g.
建立并验证了一种可靠的超高效液相色谱-电喷雾电离-串联质谱法（UHPLC-ESI-MS/MS），该方法以2-氯腺苷为内标，可快速同时测定冬虫夏草中的13种核苷和碱基。样品在冰浴中超声提取三次，最佳分析物分离在ACQUITY UPLC(TM) HSS C18柱（100 mm×2.1 mm，1.8μm）上进行梯度洗脱。所有目标分析物在5.5分钟内分离。此外，所有校准曲线在测试范围内均显示出良好的线性回归（r>0.9970），13种分析物的定量限和检测限分别小于150和75 ng/mL。日内和日间精密度的相对标准偏差（RSD）<6.23%。定量分析物的回收率在85.3%-117.3%范围内，RSD<6.18%。所建立的UHPLC-ESI-MS/MS方法成功应用于测定来自中国不同地区的11批冬虫夏草样品中的核苷和碱基。分析样品中的总含量范围为1329-2057μg/g。
