Zhang Daolai, Chen Junhui, Zhou Ming, Shi Qian, Zhao Hengqiang, Cheng Hongyan, Yang Huanghao, Wang Xiaoru
Research Centerfor Marine Ecology, The First Institute of Oceanography, State Oceanic Administration, Qingdao 266061, China.
Se Pu. 2010 Aug;28(8):795-9. doi: 10.3724/sp.j.1123.2010.00795.
A method for the simultaneous determination of 7 nucleosides in Asterias rollestoni was developed using reversed-phase high performance liquid chromatography (RP-HPLC). Analytes were extracted by ultrasonic-assisted extraction and separated on two different C18 columns, which were connected in series, under the gradient elution with the mobile phases of methanol and 0.2% (v/v) acetic acid/water at room temperature. The chromatographic conditions were as follows: flow rate, 0.8 mL/min; detection wavelength, 260 nm; injection volume, 20 microL. Under the optimized conditions, good linear relationships between the values of mass concentrations and the peak areas of hypoxanthine, uridine, xanthine, thymine, inosine, guanosine and thymidine were observed in the ranges of 0.65 -40, 0.80 -40, 0.80 -40, 1.15 - 40, 0.80 - 40, 0.50 - 40, and 0.65 - 40 mg/L, respectively. The relative standard deviations were around 0.72% - 3.23% and the recoveries were around 90.00% - 105.00%. The results showed that the developed method is sensitive, accurate and reproducible. It is suitable for the analysis of nucleosides in Asterias rollestoni with high recoveries and it is expected to be used for the quality control and evaluation of Asterias rollestoni.
建立了一种利用反相高效液相色谱法（RP-HPLC）同时测定罗氏海盘车中7种核苷的方法。采用超声辅助萃取法提取分析物，并在两根串联的不同C18柱上进行分离，于室温下用甲醇和0.2%（v/v）乙酸/水的流动相进行梯度洗脱。色谱条件如下：流速0.8 mL/min；检测波长260 nm；进样量20 μL。在优化条件下，次黄嘌呤、尿苷、黄嘌呤、胸腺嘧啶、肌苷、鸟苷和胸苷的质量浓度值与峰面积之间在0.65 - 40、0.80 - 40、0.80 - 40、1.15 - 40、0.80 - 40、0.50 - 40和0.65 - 40 mg/L范围内分别呈现良好的线性关系。相对标准偏差约为0.72% - 3.23%，回收率约为90.00% - 105.00%。结果表明，所建立的方法灵敏、准确且具有重现性。该方法适用于罗氏海盘车中核苷的分析，回收率高，有望用于罗氏海盘车的质量控制与评价。
