石龙芮HPLC指纹图谱及5种成分含量测定

目的 利用HPLC建立石龙芮药材的指纹图谱及5种成分含量测定的方法。方法 采用Agilent ZORBAX SB C18(4.6 mm×250 mm,5μm)色谱柱，以甲醇(A)-0.1%磷酸水溶液(B)为流动相，梯度洗脱，建立石龙芮药材HPLC指纹图谱。结合相似度评价，聚类、主成分分析、正交偏最小二乘判别分析等方法对13批石龙芮药材进行质量评价。结果13批石龙芮样品共确定了20个共有峰，指认出其中5个峰，相似度为0.874～0.984。聚类分析和主成分分析均将13批药材按不同产区分为3类，表明不同地区石龙芮药材化学成分含量可能存在差异。对13批石龙芮样品中的原儿茶醛、咖啡酸、阿魏酸、金丝桃苷和异槲皮苷进行定量研究，其含量分别为0.016～0.035,0.010～0.070,0.010～0.029,0.016～0.051,0.028～0.086 mg·g^–1。结论 建立的石龙芮HPLC指纹图谱和含量测定方法简便、稳定、可靠，可为石龙芮质量评价及资源利用提供参考。

HPLC Fingerprint and Content Determination of Five Components in Ranunculus Sceleratus L.

OBJECTIVE To establish the HPLC fingerprint and content determination of five components in Ranunculus sceleratus L.. METHODS The separation was developed on an Agilent ZORBAX SB C₁₈ chromatographic (4.6 mm×250 mm, 5 μm)column by gradient elution with methanol(A)-0.1 % phosphoric acid aqueous solution(B) as mobile phase to establish HPLC fingerprint of Ranunculus sceleratus L.. Combined with similarity evaluation, cluster analysis, principal component analysis, and orthogonal partial least squares-discriminant analysis, the quality of 13 batches of Ranunculus sceleratus L. was evaluated. RESULTS Thirteen batches of Ranunculus sceleratus L. samples were calibrated with 20 common peaks, of which 5 common peaks were identified, and the similarity ranged from 0.874 to 0.984. The results of cluster analysis and principal component analysis were basically the same, indicating that there might be differences in the content of chemical components of Ranunculus sceleratus L. in different regions. Protocatechuic aldehyde, caffeic acid, ferulic acid, hyperoside and isoquercitrin were determined in thirteen batches of Ranunculus sceleratus L., and their contents were 0.016-0.035, 0.010-0.070, 0.010-0.029, 0.016-0.051, 0.028-0.086 mg·g^–1, respectively. CONCLUSION The established HPLC fingerprint and content determination method is simple, stable, and reliable, which can be used for qualitative analysis and provide reference to quality evaluation and resource utilization of Ranunculus sceleratus L..