农吉利有效成分的HPLC指纹图谱研究

HPLC fingerprints for effective constituents in Crotalaria sessiliflora L.

目的建立农吉利中生物碱和黄酮化合物两类主要有效成分的指纹图谱。方法采用C18色谱柱,分别以甲醇-0.5%乙酸溶液和甲醇-0.04%氨水溶液作为流动相梯度洗脱,用二极管阵列检测器(DAD)检测,研究了农吉利中黄酮类化合物和生物碱两种主要有效成分的色谱分离条件,利用中药色谱指纹图谱相似度评价系统(2004版)处理。结果各批药材有9个黄酮化合物和4个生物碱共有峰,分别以牡荆素和农吉利甲素作为黄酮化合物和生物碱的参照峰,建立了10批农吉利药材的指纹图谱,其相似度分别在0.975～0.995和0.908～0.990之间,各色谱峰精密度、稳定性和测定重复性的RSD均小于3%,主要成分牡荆素和农吉利甲素含有量分别在0.221～0.428 mg/g和0.012～0.082 mg/g之间。结论本法简便、特征性强、可用于不同产地农吉利药材的指纹图谱的鉴定和质量评价。

AIM To establish HPLC fingerprints for the quality evaluation of Crotalaria sessiliflora L..METHODS Using methanol-0.5% acetic acid solution and methanol-0.04% ammonia solution as mobile phases,the flavonoids and alkaloids extracted from the samples were separated on C18 column in gradient elution modes,and detected by diode array detector（DAD）,respectively.The HPLC data of ten batches of samples from four different origins were processed by the software ＂Similarity Evaluation System for Chromatographic Fingerprint of TCM（2004 version）＂.And the flavonoids and alkaloids were also identified by HPLC-ESI-MS/MS.RESULTS Based on obtained HPLC chromatograms,nine flavonoids and four alkaloids presented the common characteristic peaks.Vitexin and monocrotaline as major constituents were selected as reference substances of alkaloid and flavonoid,respectively.RSDs of above constituents in chromatograms for precision,stability and reproducibility were less than 3%.The similarities of ten batches of samples of Crotalaria sessiliflora L.were between 0.975-0.995 for flavonoids and 0.908-0.990 for alkaloids.CONCLUSIONS The method developed is convenient and specific,and could further serve for quality identification and comprehensive evaluation of Crotalaria sessiliflora L.from different habitats.