反相高效液相色谱法同时测定罗氏海盘车中的7种核苷化合物

Simultaneous determination of 7 nucleosides in Asterias rollestoni using reversed-phase high performance liquid chromatography

建立了罗氏海盘车中7种核苷化合物的反相高效液相色谱分析测定方法。采用超声波辅助提取,选用两根不同的C18色谱柱串联,以甲醇和0.2%(体积分数)乙酸/水溶液为流动相梯度洗脱分离。优化的色谱条件为:柱温为室温,检测波长为260nm,流速为0.8mL/min,进样量为20μL。结果表明,7种核苷化合物在一定的浓度范围内线性关系良好,次黄嘌呤和胸苷的线性范围为0.65～40mg/L,尿苷、黄嘌呤和肌苷的线性范围为0.80～40mg/L,胸腺嘧啶的线性范围为1.15～40mg/L,鸟苷的线性范围为0.50～40mg/L。样品中7种核苷化合物的加标回收率为90.00%～105.00%,相对标准偏差为0.72%～3.23%。该方法操作简便、灵敏度高、重复性好,回收率高,适用于罗氏海盘车中7种核苷类成分的同时分析,也可用于罗氏海盘车的质量控制和综合评价。

A method for the simultaneous determination of 7 nucleosides in Asterias rollestoni was devel-oped 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 se-ries,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 μL. Under the optimized conditions,good linear relationships between the values of mass concentrations and the peak areas of hypoxanthine,uridine,xanthine,thy-mine,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 devia-tions 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.