基于超高效液相色谱-串联质谱法的一测多评法同时测定龙眼肉中8种核苷类成分含量

Simultaneous determination of 8 kinds of nucleosides in longan meat by multi-components by single-marker method based on ultra performance liquid chromatography-tandem mass spectrometry

目的 基于超高效液相色谱-串联质谱法,建立一测多评法同时测定龙眼肉中腺苷、鸟嘌呤、尿嘧啶、鸟苷、尿苷、次黄嘌呤、肌苷、胞苷含量的方法 。方法 待测样品经甲醇提取,由Agilent SB-C18色谱柱(150 mm×2.1 mm, 1.8μm)分离,以0.1%甲酸-甲醇溶液为流动相梯度洗脱,流速为0.2 mL/min,采用电喷雾正电离源模式(electronic spray ion, ESI+)和多反应监测模式(multiple reactions monitoring, MRM)监测,分别采用外标法和一测多评法进行定量。结果 腺苷在0.305~6.100μg/mL、鸟嘌呤在0.435~8.700μg/mL、尿嘧啶在0.340~6.800μg/mL、鸟苷在0.325~6.500μg/mL、尿苷在0.320~6.400μg/mL、次黄嘌呤在0.330~6.600μg/mL、肌苷在0.305~6.100μg/mL、胞苷在0.360~7.200μg/mL范围内具有良好的线性关系,相关系数大于0.991,检出限为0.1μg/g,回收率为99.5%,相对标准偏差为0.46%(n=9)。采用外标法和一测多评法计算鸟嘌呤、腺苷、尿嘧啶、鸟苷、尿苷、胞苷的含量,两种方法 计算结果 差异极小。结论 一测多评法稳定可靠,可用于龙眼肉中核苷类成分的质量控制。

Objective To establish a method for simultaneous determination of adenosine,guanine,uracil,guanosine,uridine,hypoxanthine,inosine and cytidine in longan meat by multi-components by single-marker based method on ultra performance liquid chromatography-tandem mass spectrometry.Methods The samples were extracted by methanol and separated by Agilent SB-C18 column(150 mm×2.1 mm,1.8μm),then eluted with 0.1%formic acid-methanol solution as mobile phase gradient at a flow rate of 0.2 mL/min.Electronic spray ion(ESI+)and multiple reactions monitoring(MRM)were used for monitoring.External standard method and multi-components by single-marker method were used for quantitative analysis respectively.Results Adenosine in 0.305-6.100μg/mL,guanine in 0.435-8.700μg/mL,uracil in 0.340-6.800μg/mL,guanosine in 0.325-6.500μg/mL,uridine in 0.320-6.400μg/mL,hypoxanthine in 0.330-6.600μg/mL,inosine in 0.305-6.100μg/mL,cytidine in 0.360-7.200μg/mL had a good linear relationship,the correlation coefficient was greater than 0.991,the limit of detection was 0.1μg/g,and the recovery was 99.5%.The relative standard deviation was 0.46%(n=9).The content of guanine,adenosine,uracil,guanosine,uridine and cytidine were calculated by external standard method and multi-components by single-marker method.The difference between the two methods was very small.Conclusion Multi-components by single-marker method is stable and reliable,and can be used for the quality control of nucleosides components in longan meat.