不同生长年限粉沙参游离氨基酸和核苷类成分分析

Analysis of Free Amino Acids and Nucleosides in Different Growth Years of Changii Radix

关键词:  粉沙参  明党参根皮  氨基酸  核苷类  生长年限  液质联用  主成分分析  聚类分析

Key words:  Changii Radix  root barks of Changium smyrnioides Wolff  amino acids  nucleosides  growth years  liquid chromatography-mass spectrometry  principal component analysis  hierarchical cluster analysis

摘要:

目的 探讨野生粉沙参中氨基酸和核苷类成分的含量与其生长年限的关系。方法 采用超快速液相色谱和三重四级杆-线性离子阱串联质谱（UFLC-QTRAP-MS/MS），对6个不同生长年限粉沙参及其明党参根皮中的氨基酸类和核苷类成分进行含量测定，并采用主成分分析（PCA）与聚类分析（HCA）对其不同生长年限的差异性及其代表性成分进行了分析。色谱柱为氨基柱（2.1 mm×100 mm，1.7 μm），流动相乙腈-0.2%甲酸水（梯度洗脱），流速为0.4 mL·min^-1，柱温为30℃；质谱检测采用电喷雾正离子（ESI⁺）及多反应监测（MRM）模式。结果 野生粉沙参中含量最高的是具有化痰止咳功效的天门冬酰胺，其2~3年生者含量高达2.97%，且>3年各生长年限粉沙参中含量均恒定在同一水平[（1.77±0.048）%]；不同生长年限粉沙参差异代表性成分为2~3年生为天门冬酰胺，4~5年生和9~15年生均为丙氨酸和鸟苷，6~8年生为鸟苷和腺苷，16~18年生为缬氨酸和精氨酸，19~20年生为甲硫氨酸和天冬氨酸。明党参根皮中氨基酸和核苷类成分含量则随生长年限增加而下降。结论 天门冬酰胺为粉沙参主要功效成分之一；依据差异代表性氨基酸和核苷可以大致判别粉沙参的生长年限；明党参根皮含有粉沙参同样的氨基酸和核苷类成分，具有开发利用的价值。

Abstract:

OBJECTIVE To explore the relationship between the amino acid and nucleoside contents in Changii Radix with their growth years. METHODS Ultra-fast liquid chromatography and triple quadrupole-linear ion trap tandem mass spectrometry(UFLC-QTRAP-MS/MS) was used to determine the contents of amino acids and nucleosides in Changii Radix and the root barks of Changium smyrnioides with six different growth years, then principal component analysis(PCA) and hierarchical cluster analysis(HCA) were used to analyze the differences of these contents in different growth years and their representative components. The amino column(2.1 mm×100 mm, 1.7 μm) and the mobile phase with acetonitrile-0.2% formic acid were used for gradient elution. The flow rate was 0.4 mL·min^-1 while the column temperature was 30℃. Mass spectrometry was performed by positive electrospray ionization(ESI⁺) and multiple reaction monitoring(MRM) modes. RESULTS The highest content in Changii Radix was asparagine which had been proved to reduce phlegm and relieve cough, and this content in Changii Radix aged 2-3 was as the highest(2.97%), while that of samples aged above 3 was at the same level[(1.77±0.048)%]. The representative components in different growth years of Changii Radix were:asparaginase in 2-3 years, alanine and guanosine in 4-5 years and 9-15 years, guanosine and adenosine in 6-8 years, proline and arginine in 16-18 years, and finally methionine and aspartic acid in 19-20 years. The content of amino acids and nucleosides in the root barks of Changium smyrnioides decreased with increasing growth years. CONCLUSION Asparagine is one of the main functional ingredients in Changii Radix; based on the difference of representative amino acids and nucleosides, the growth period of Changii Radix can be roughly determined; the root barks of Changium smyrnioides contain the same amino acids and nucleosides as Changii Radix, which means the root barks can be exploited rather than discarded.