[关键词]
[摘要]
目的 建立板蓝根高效液相色谱(HPLC)指纹图谱,并结合多成分定量测定方法及主成分分析对板蓝根 质量进行评价。方法 采用 ZORBAX Eclipse Plus C 18 (250 mm×4.6 mm,5 μm)色谱柱,以甲醇-0.3% 乙酸水 溶液为流动相,梯度洗脱,流速为 0.8 mL·min -1 ,检测波长为 254 nm,柱温为 30 ℃,进样量为 10 μL,建立 板蓝根 HPLC 指纹图谱;同时快速测定板蓝根中 6 个活性成分的含量,并结合主成分分析方法评价板蓝根的质 量。结果 10 批板蓝根 HPLC 指纹图谱确定了 10 个共有峰,经对照品匹配指认出尿苷、鸟苷、腺苷、(R,S)- 告依春、靛蓝、靛玉红 6 个色谱峰。10 批板蓝根样品一致性好,指纹图谱相似度为 0.993~1.000。HPLC 法同 时快速测定板蓝根中 6 个活性成分含量方法的精密度、稳定性、重复性及加样回收率结果均符合要求。板蓝根 中(R,S)-告依春含量最高,靛玉红含量最低,靛蓝含量样品间波动最大,尿苷含量波动最小。主成分分析结 果显示,甘肃省民乐县、平凉市及河南省洛阳市的板蓝根药材质量相对较好。结论 该研究所建立的 HPLC 指 纹图谱结合化学计量学方法能够全面、真实地反映板蓝根药材的质量,对板蓝根资源的深入开发和综合利用具 有重要的指导意义和参考价值。
[Key word]
[Abstract]
Objective To establish an high performance liquid chromatography (HPLC) fingerprint for Isatidis Radix and evaluate its quality by combining multi-component quantification and principal component analysis (PCA). Methods An HPLC fingerprint method was developed using a ZORBAX Eclipse Plus C 18 column (250 mm × 4.6 mm, 5 μm) with a mobile phase of methanol -0.3% acetic acid solution in a gradient elution mode. The flow rate was 0.8 mL·min -1 ,the detection wavelength was set at 254 nm,the column temperature was maintained at 30 °C,and the injection volume was 10 μL. Simultaneously,the contents of six active components were rapidly determined. The quality of Isatidis Radix was evaluated by combining these results with PCA. Results Ten common peaks were identified in the HPLC fingerprints of 10 batches of Isatidis Radix. Six of these peaks were identified by comparison with reference standards as uridine,guanosine,adenosine,(R,S)-goitrin,indigo,and indirubin. The 10 batches showed good consistency,with fingerprint similarities ranging from 0.993 to 1.000. The method for simultaneous quantification of the six active components demonstrated satisfactory results for precision,stability,repeatability,and recovery. Among the quantified components,(R,S)-goitrin was found to have the highest content,while indirubin had the lowest. The indigo content showed the greatest variation among the samples,whereas uridine content exhibited the least variation. PCA indicated that the quality of Isatidis Radix from Minle County and Pingliang City in Gansu Province,and Luoyang City in Henan Province,were relatively superior. Conclusion The established HPLC fingerprint method combined with chemometric analysis can comprehensively and accurately reflect the quality of Isatidis Radix. This approach provides significant guidance and reference value for the in-depth development and comprehensive utilization of Isatidis Radix resources.
[中图分类号]
R284.1
[基金项目]
甘肃省高校青年博士基金项目(2025QB-103);甘肃省省级人才项目(青年团体)(2024QNTD30);平凉市科技人才专项计划项目 (PL-STK-2023A-039)。