海螵蛸中游离核苷与氨基酸类成分的同时检测与分析

目的 建立同时测定海螵蛸中12种核苷、18种氨基酸含量的检测方法。方法 使用超快速液相色谱-串联质谱（UFLC-MS/MS），采用Acquity BEH Amide色谱柱（2.1 mm×100 mm，1.7 μm），采用乙腈-0.1%甲酸水溶液作为流动相进行梯度洗脱，流速0.3 mL·min^–1，柱温30℃；采用电喷雾离子源正离子多反应监测模式测定。采用主成分分析和二维主成分分析对样品进行综合评价。结果 12种核苷、18种氨基酸在一定范围内线性关系良好（r²>0.999 0），稳定性、精密度、重复性均良好，RSDs<3.24%，加样回收率在95.14%~103.84%，RSDs在0.93%~3.82%。18个批次海螵蛸中核苷总量为82.38~565.65 μg·g^–1，氨基酸总量为1 233.12~9 913.49 μg·g^–1。通过主成分分析不同产地海螵蛸主成分包含9种氨基酸和7种核苷，并且具有地域性特征。结论 该方法准确、快速、高效，可同时完成对海螵蛸中12种核苷、18种氨基酸的含量测定，可为海螵蛸的质量评价与鉴定提供相关依据，为后续研究提供参考。

Simultaneous Determination and Analysis of Free Nucleosides and Amino Acids in Sepiae Endoconcha

OBJECTIVE To construct a method for simultaneous determination of 12 nucleosides and 18 amino acids in Sepiae Endoconcha. METHODS The analysis was performed on a Acquity BEH Amide column(2.1 mm×100 mm, 1.7 μm) with elution by mobile phase of acetonitrile-0.1% formic acid at a flow rate of 0.3 mL·min^-1 with the column temperature at 30℃ by an ultrafast liquid chromatography coupled with mass spectrometer(UFLC-MS/MS). In addition, the positive ion multiple reaction monitoring mode was used. Principal component analysis and two-dimensional principal component analysis were used to comprehensively evaluate the samples. RESULTS All 12 nucleosides and 18 amino acids showed good linearity (r²>0.999 0), the RSDs of the stability, precision and reproducibility tests were less than 3.24%. The recoveries were in the range of 95.14%-103.84% and RSDs were in the range of 0.93%-3.82%. The total nucleoside content and amino acid content in 18 batches of Sepiae Endoconcha were 82.38-565.65 μg·g^-1 and 1 233.12-9 913.49 μg·g^-1. PCA analysis showed that the main components of Sepiae Endoconcha from different areas contained 9 amino acids and 7 nucleosides, and showed regional characteristics. CONCLUSION This method is accurate, rapid and efficient, and can simultaneously determine 12 nucleosides and 18 amino acids in Sepiae Endoconcha, which can provide relevant basis for quality evaluation and identification of Sepiae Endoconcha, provide reference for subsequent studies.