Simultaneous determination of 10 nucleosides and nucleobases from different cultivation years of Fritillaria unibracteata var. wabuensis by HPLC-DAD  

HPLC-DAD法对不同生长年限瓦布贝母10种核苷和碱基含量的测定（英文）

作　　者：潘峰[1] 吴卫[1] 董品利 胡博[1] 官玲亮[3] 舒玲[1] 

机构地区：[1]四川农业大学农学院,四川成都611130 [2]四川省茂县农业局,四川茂县623200 [3]中国热带农业科学院热带作物品种资源研究所,海南儋州571737

出　　处：《Journal of Chinese Pharmaceutical Sciences》2017年第5期346-354,共9页中国药学（英文版）

基　　金：The Specialized Research Fund for the Doctoral Program of Higher Education of China(Grant No.20115103110009);"211"Project Double-Support Plan of Sichuan Agricultural Un iversity(Grant No.03570313);Modernization of Chinese Traditional Medicines in Hainan Province(Grant No.ZY201410)

摘　　要：Determination of nucleosides and nucleobases is important for the quality control of Fritillaria unibracteata Hsiao et K.C. Hsia var. wabuensis （FUW） due to their physiological and pharmacological actions. In the present study, we developed a sensitive and reliable HPLC-diode-array detection method to simultaneously determine ten nucleosides and nucleobases, including cytosine, uracil, cytidine, uridine, thymine, adenine, inosine, guanosine, thymidine and adenosine. Complete separation of all the analytes was achieved on a Zorbax 300 A 300 Extend C18 column with a gradient of methanol-ultrapure water at a flow rate of 1 mL/min in less than 30 min. The diode-array detector wavelength was set at 260 nm for the UV detection of nucleosides and nucleobases. The optimized method provided good linearity （R2〉0.9993 for all the analytes）, satisfactory precision （RSD〈3.715%）, good repeatability （RSD_〈3.748%） and good recovery （RSD from 97.688% to 102.923%）. In addition, the developed method was successfully applied to simultaneous determination of ten nucleosides and nucleobases from FUW, and their content changes of various cultivation time （1-7 years） were further analyzed for the first time. Our findings were useful for ensuring the cultivation time choice of artificial cultivation, quality control, pharmaceutical studies and clinical efficacy of FUW.核苷和碱基具有多种生理生化活性,定量测定瓦布贝母的含量对其质量控制也很重要。本文建立了高效液相色谱-二极管阵列检测器联用技术(HPLC-DAD)同时测定瓦布贝母鳞茎中6种核苷(鸟苷、胞嘧啶核苷、胸苷、腺苷、尿苷、肌苷)和4种碱基(胞嘧啶、尿嘧啶、胸腺嘧啶、腺嘌呤)成分含量的测定方法。采用Zorbax 300?300 Extend C_(18)色谱柱(150 mm×4.6 mm,5μm),检测波长为260 nm,以甲醇和水为流动相梯度洗脱,以1.0 mL/min的流速在30 min以内能够实现所选用的核苷和碱基实现完全分离。采用优化色谱条件后,所有10种待测成分具有良好的线性关系(R^2≥0.9993),精密度(RSD≤3.715%),重复性(RSD≤3.748%),稳定性(RSD≤5.545%)和加样回收率(97.688%–102.923%)。利用所建立的色谱方法成功地应用于不同生长年限瓦布贝母核苷和碱基的测定,并且首次分析了不同生长年限(1–7年)瓦布贝母的10种核苷和碱基的含量变化。本研究将有助于对瓦布贝母的栽培时间选择、质量控制、药理研究和临床应用。

关 键 词：Quantitative determination Nucleosides and nucleobases Fritillaria unibracteata var. wabuensis HPLC-DAD 

分 类 号：R284.1[医药卫生—中药学]