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

中文关键词:  龙眼肉  核苷酸  高效液相色谱-串联质谱法

英文关键词:longan meat  nucleotide  ultra performance liquid chromatography-tandem mass spectrometry

中文摘要:

      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, 1.8 μm)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0.305~6.100 μg/mL、鸟嘌呤在0.435~8.700 μg/mL、尿嘧啶在0.340~6.800 μg/mL、鸟苷在0.325~6.500 μg/mL、尿苷在0.320~6.400 μg/mL、次黄嘌呤在0.330~6.600 μg/mL、肌苷在0.305~6.100 μg/mL、胞苷在0.360~7.200 μg/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0.1 μg/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。

英文摘要:

      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, 1.8 μm), 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 0.305–6.100 μg/mL, guanine in 0.435–8.700 μg/mL, uracil in 0.340–6.800 μg/mL, guanosine in 0.325–6.500 μg/mL, uridine in 0.320–6.400 μg/mL, hypoxanthine in 0.330–6.600 μg/mL, inosine in 0.305–6.100 μg/mL, cytidine in 0.360–7.200 μg/mL had a good linear relationship, the correlation coefficient was greater than 0.991, the limit of detection was 0.1 μg/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.