目的: 建立超高效液相色谱串联质谱(UPLC-MS/MS)同时测定红豆树中12个黄酮类成分含量的方法,并对不同批次的红豆树药材进行质量评价。方法: 采用Waters CORTECS UPLC C18色谱柱(2.1 mm×100 mm, 1.6 μm),以0.1%甲酸水溶液(A)-乙腈(B)为流动相,梯度洗脱,流速0.2 mL·min-1,柱温45 ℃,进样量2 μL;质谱采用电喷雾离子源正离子模式扫描,在多反应监测模式下进行定量分析;采用SPSS软件对含量测定结果进行聚类分析。结果: 12个成分在各自线性范围内均呈良好的线性关系(r2≥0.998 6),平均加样回收率为94.8%~100.8%(RSD≤4.7%),其精密度、重复性、稳定性的RSD均小于4.7%。聚类分析将16批药材分为2类:红豆树叶(S1~S8)可归为Ⅰ类,红豆树枝(S9~S16)可归为Ⅱ类,各部位的成分含量存在较大差异。结论: 该方法简单快速,灵敏度高,专属性好,可用于红豆树的质量评价。
Objective: To evaluate quality of Ormosia hosiei Hemsl. Et Wils.(O. hosiei), 12 flavonoids were determined by ultra-high performance liquid chromatography tandem mass spectrometry method. Methods: Chromatographic separation was performed on a Waters CORTECS UPLC C18 column (2.1 mm×100 mm, 1.6 μm) with mobile phase consisting of 0.1% formic acid aqueous solution(A) and acetonitrile(B)in gradient elution (0-0.5 min,5%B;0.5-1.5 min,5%B→10%B;1.5-2.5 min,10%B→15%B;2.5-4.5 min,15%B→20%B;4.5-7.5 min,20%B;7.5-8.0 min,20%B→23%B;8.0-10.85 min,23%B;10.85-11.0 min,23%B→26%B;11.0-13.8 min,26%B→40%B;13.8-13.9 min,40%B→5%B;13.9-15.5 min,5%B). The flow rate was set at 0.2 mL·min-1 and the column temperature was 45 ℃. MS monitoring was performed by electrospray ionization in positive ionization mode, quantification was performed using multiple reaction monitoring mode, and SPSS software was employed to perform the hierarchical cluster analysis for the content determination results. Results: The calibration curves for 12 compounds had good linearity (r2≥0.998 6) in their linear ranges. The average recoveries were in the range of 94.8%-100.8% (RSD≤4.70%), the RSDs of precision,repeatability, and stability were all below 4.7%. Cluster analysis classified 16 batechesof O.hosiei into 2 categories with the leaves as categoryⅠand the twigs (S9-S16) as category Ⅱ. Great differencesexisted in the contents of componentsin different parts of O. hosiei. Conclusion: The established method is simple, rapid, sensitive and specific, and can be used for quality evaluation of O. hosiei.
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