不同产地桑叶氨基酸、核苷、生物碱成分的含量测定及多元统计分析

Determination and Multivariate Statistical Analysis of Amino Acids, Nucleosides and Alkaloid in Mulberry Leaves from Different Origins

关键词:  桑叶  氨基酸  核苷  1-脱氧野尻霉素  主成分分析

Key words:  mulberry leaves  amino acids  nucleosides  1-deoxynojirimycin  principal component analysis(PCA)

摘要:

目的 建立超高效液相串联三重四极杆质谱（UPLC-TQD）同时测定桑叶样品中氨基酸、核苷及生物碱[1-脱氧野尻霉素（1-deoxynojirimycin，DNJ）]含量的方法，并采用多元统计分析方法分析比较不同产地桑叶中成分含量差异，为桑叶资源的品质评价提供科学依据。方法 采用Waters Acquity UPLC BEH Amide（100 mm×2.1 mm，1.7 μm）色谱柱，以0.2%甲酸乙腈（A）-0.2%甲酸水（B）梯度洗脱，流速0.4 mL·min^-1，柱温35℃；使用多反应监控正离子模式测定；采用多元统计分析如主成分分析对不同产地桑叶进行综合性评价。结果 22种成分的浓度与峰面积呈良好的线性关系（r² ≥ 0.990 3）；方法精密度、重复性及稳定性的RSD值均≤ 7.5%；平均加样回收率为86.7%~106.4%，RSD为2.2%~8.9%。不同产地桑叶中氨基酸、核苷及DNJ存在一定差异，结合多元统计分析，共找出8个差异性成分，可将样品分成2组。结论 所建立方法准确可靠，结合DNJ及8个差异性成分含量进行比较，湖北及江西产地的桑叶品质较优。

Abstract:

OBJECTIVE To establish a method for simultaneous determination of amino acids, nucleosides and alkaloid[1-deoxynojirimycin(DNJ)] in mulberry leaves from different origins by UPLC-TQD, multivariate statistical analysis is employed to analysis discrepancies in compositions from these samples, to provide scientific basis for quality evaluation of mulberry leaf resources. METHODS Waters Acquity UPLC BEH Amide(100 mm×2.1 mm, 1.7 μm) column was employed, 0.2% formic acid-acetonitrile(A)-0.2% formic acid-water(B) was served as the mobile phase, gradient elution, flow rate was 0.4 mL·min^-1, column temperature was set at 35℃. Multiple reaction monitoring and positive ions mode was used to detect ions. Comprehensive evaluation of mulberry leaves from different areas were subjected to multivariate statistical analysis such as principal component analysis, etc. RESULTS The concentration of 22 components showed a good linear relationship consistent with the peak areas(r² ≥ 0.990 3). The RSD values of the precision, reproducibility and stability of the method were ≤ 7.5%. Moreover, the average sample recovery rate was 86.7%-106.4% and the corresponding RSD was 2.2%-8.9%. Mulberry leaves from different origins revealed great differences in amino acids, nucleosides and DNJ, combined with multivariate statistical analysis, these samples could be divided into two groups based on the discovered 8 most inhomogeneous componeous. CONCLUSION The method is accurate and reliable, combined with DNJ and 8 different components, mulberry leaves from Hubei and Jiangxi are deemed to be better.