采用超高效液相色谱法快速同时测定培养虫草中的14种核苷和碱基。
Fast simultaneous determination of 14 nucleosides and nucleobases in cultured Cordyceps using ultra-performance liquid chromatography.
作者信息
Yang F Q, Guan J, Li S P
机构信息
Institute of Chinese Medical Sciences, University of Macau, Taipa, Macao SAR, PR China.
出版信息
Talanta. 2007 Sep 15;73(2):269-73. doi: 10.1016/j.talanta.2007.03.034. Epub 2007 Mar 24.
Determination of nucleosides and their metabolic compounds is important for physiological and pharmacological studies. Herein, a rapid ultra-performance liquid chromatography (UPLC) method was developed for the simultaneous determination of 14 nucleosides and nucleobases, namely adenine, adenosine, cytosine, cytidine, uracil, uridine, guanine, guanosine, hypoxanthin, inosine, thymine, thymidine, 2'-deoxyuridine and cordycepin. The separation was performed on Waters Acquity UPLC system with Acquity UPLC BEH C(18) column and gradient elution of 0.5mM acetic acid and acetonitrile in 5min. The correlation coefficients of 14 analytes were high (R(2)>0.9995) within the test ranges. The LOD and LOQ were lower to 11.9 and 47.0ng/ml with 1mul of injection volume, respectively. The overall R.S.D. for intra- and inter-day of 14 analytes were less than 1.8%. The developed method was applied for the analysis of nucleosides and nucleobases in cultured Cordyceps, which also could be used for the fast determination of the analytes in pharmaceutical products and biological fluids.
核苷及其代谢化合物的测定对于生理学和药理学研究很重要。在此，开发了一种快速超高效液相色谱（UPLC）方法，用于同时测定14种核苷和碱基，即腺嘌呤、腺苷、胞嘧啶、胞苷、尿嘧啶、尿苷、鸟嘌呤、鸟苷、次黄嘌呤、肌苷、胸腺嘧啶、胸苷、2'-脱氧尿苷和虫草素。在Waters Acquity UPLC系统上，使用Acquity UPLC BEH C(18) 色谱柱，以0.5mM乙酸和乙腈在5分钟内进行梯度洗脱，实现分离。14种分析物在测试范围内的相关系数很高（R(2)>0.9995）。进样量为1μl时，检测限和定量限分别低至11.9和47.0ng/ml。14种分析物日内和日间的总体相对标准偏差均小于1.8%。所开发的方法用于分析培养的冬虫夏草中的核苷和碱基，也可用于快速测定药品和生物流体中的分析物。
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