目的 通过HPLC指纹图谱结合化学计量学构建了宁夏5个主要种质基地50批宁夏枸杞L. barbarum叶（S1～S50）指纹图谱，并对其中主要生物碱成分定量测定。方法 利用HPLC技术和中药色谱指纹图谱相似度评价系统（2012.1版）绘制指纹图谱，进行相似度评价和共有峰确认，并结合化学计量学综合分析。同时采用HPLC法测定7种生物碱成分（胆碱、甜菜碱、葫芦巴碱、枸杞素A、枸杞素B、N-反式阿魏酰酪胺、N-阿魏酰-3-甲氧基酪胺）含量，对宁夏不同产地宁夏枸杞叶予以评价。结果 所获指纹图谱共18个共有峰，50批宁夏枸杞叶生物碱指纹图谱与对照指纹图谱相似度为0.692～0.981，中宁、固原产地宁夏枸杞叶指纹图谱与对照指纹图谱共有模式相似度较高，而银川产宁夏枸杞叶指纹图谱与对照指纹图谱共有模式相似度较低；聚类分析（hierarchical clustering analysis，HCA）将50批样品聚为4类；主成分分析（principal component analysis，PCA）筛选出4个主成分，累积方差达89.014%；正交偏最小二乘法-判别分析（orthogonal partial least squares-discriminant analysis，OPLS-DA）筛选出组间差异贡献率较大的成分，按VIP大小顺序依次为5号峰＞3号峰＞10号峰＞11号峰＞13号峰＞2号峰＞4号峰＞9号峰；含量测定结果表明固原产地的宁夏枸杞叶中7种生物碱含量最高。结论 构建了稳定性强的不同产地宁夏枸杞叶HPLC指纹图谱以及7种生物碱的测定方法，结合化学计量学可用于宁夏枸杞叶产地差异分析与质量评价。

Objective To evaluate the quality of Lycium barbarum leaves from various production areas in Ningxia, an HPLC fingerprinting technique combined with chemometrics and quantitative determination of multiple alkaloids constituents with LC-MS/MS was used to construct the alkaloidal fingerprints of L. barbarum leaves (S1—S50) from five bases in three production areas in Ningxia. Methods The HPLC method and Similarity Evaluation System for Chromatographic Fingerprinting in Chinese Medicine (version 2012) were used to generate fingerprints, and similarity evaluation and common peak confirmation were carried out. Meanwhile, LC-MS/MS was used to establish a method for the determination of the contents of seven alkaloids constituents (choline, betaine, trigonelline, lyciumin A, lyciumin B, N-trans-feruloyltyramine, N-feruloyl-3-methoxytyramine) to evaluate the leaves of L. barbarum from different origins in Ningxia. Results The obtained fingerprints had a total of 18 common peaks. The similarity between the alkaloid fingerprints of 50 batches of L. barbarum leaves. The control fingerprints ranged from 0.692 to 0.981. Zhongning and Guyuan origin L. barbarum leaves fingerprints have higher similarity to the control fingerprints, while Yinchuan origin L. barbarum leaves fingerprints have lower similarity to the control fingerprints. Hierarchical clustering analysis (HCA) clustered the 50 batches of samples into four categories; principal component analysis (PCA) screened out four principal components, with a cumulative variance of 89.014%; orthogonal partial least squares-discriminant analysis (OPLS-DA) screened out the components with larger contributions to inter-group variance, which were, in the order of the size of the VIP, peak 5 > peak 3 > peak 10 > peak 11 > peak 13 > peak 2 > peak 4 > peak 9; content determination showed that seven alkaloids from Guyuan origin had the highest content. The results of content determination showed that the seven alkaloids of Guyuan origin had the highest content. Conclusion A stable HPLC fingerprint of L. barbarum leaves of different origins and a quantitative LC-MS/MS method for the determination of seven alkaloids were constructed, which can be used for the analysis of the difference in the origins of L. barbarum leaves and quality evaluation by combining with chemometrics.

R282.6

宁夏重点研发项目（2024BBF02009）