HPLC法同时测定小根蒜中8种碱基和核苷类成分的含量

Simultaneous Determination of Eight Bases and Nucleosides Contents in Allium macrostemon Bunge. by HPLC

目的:建立同时测定不同产地、不同部位小根蒜中尿嘧啶、鸟嘌呤、尿苷、腺嘌呤、鸟苷、胸苷、腺苷和2'-脱氧腺苷8种碱基和核苷含量的HPLC法。方法:运用HPLC法测定38份不同部位小根蒜碱基和核苷的含量,并用聚类分析和主成分分析方法,对14份小根蒜鳞茎的测定结果进行分析。结果:38份不同产地、不同部位小根蒜中8种碱基和核苷类成分含量在线性范围内的线性关系良好(r>0.9995),检出限为10.78～37.62 ng·mL^(-1),定量限为35.81～115.28 ng·mL^(-1)及平均回收率为97.72%～100.29%,RSD<3%。小根蒜不同部位均可检测到8种碱基和核苷类成分,且不同产地、不同部位小根蒜中8种碱基和核苷类成分的质量分数及组成结构比存在明显的差异,部分非入药、非食用部位碱基和核苷含量高于其鳞茎。采用聚类分析能将不同产地的14份小根蒜鳞茎分为2类,且主成分分析法筛选出重庆市万州区小根蒜鳞茎质量的综合评价最好。结论:该实验方法操作快速、简便,重现性好,可用于不同产地、不同部位小根蒜中8种碱基和核苷类成分的含量测定。同时,测定结果为小根蒜不同部位的高效利用提供理论依据。

Objective：To establish a HPLC analytical method on the simultaneous determination of eight bases and nucleosides （ uracil, adenine, guanine, uridine, guanosine, thymidine, adenosine and deoxyadenosine 2＇- deoxyadenosine） in different parts of Allium macrostemon Bunge from different producing areas. Methods ： HPLC method was used to determine contents of 38 different parts of A.macrostemon samples, and the results of 14 bulbs were analyzed by cluster analysis and principal component analysis.Results：The eight bases and nueleosides in 38 different producing areas and parts of A.macrostemon showed a good linear relationship（r 〉 0.9995 ） within their linear ranges, with the detection limit for 10.78 -37.62 ng·mL^-1 , the limit of quantitation for 35.81 - 115.28 ng·mL^-1 , the average recovery for 97.72% - 100.29% and RSD less than 3% . The eight nucleosides and bases could be detected from different parts of A.macrostemon.There were obviously different mass fraction and composition structure in eight bases and nucleosides among different producing areas and parts of A.macrostemon, and some non -medicine and non-food parts were higher than the bulb.14 bulbs from different habitats could be divided into two categories for by cluster analysis. Moreover, principal component analysis showed that comprehensive quality of bulbs produced in Wanzhou in Chongqing province was best.Conclusion：This was a fast and simple method with good reproducibility, and it was appropriate for measuring contents of the eight nucleosides. Meanwhile, the results provided theoretical basis for efficient utilization on different parts of A.macrostemon.