金银花抗炎质量标志物筛选鉴定及化学指纹图谱的研究

Screening and Identification of Lonicerae japonicae Flos Anti-inflammatory Quality Markers and Study on Chemical Fingerprint

目的研究金银花抗炎质量标志物筛选鉴定方法以及化学指纹图谱用于金银花药材品质判定及其伪品鉴定。方法利用人胚胎肾293T细胞及核转录因子-κB(NF-κB)双荧光素酶报告系统进行抗炎活性实验,选择活性最优指标进行活性抗炎单体的筛选,并通过超高效液相色谱/四级杆-飞行时间质谱法(UPLC/Q-TOF)进行抗炎质量标志物的鉴定,对金银花进行化学指纹图谱分析,通过相似度软件处理,判定指纹图谱相似度差异性。结果筛出4个活性单体,并对这4个峰进行解谱,鉴定分别为绿原酸、木犀草苷、獐芽菜苷和獐牙菜苦苷作为金银花抗炎活性的质量标志物,同时结合异绿原酸A、异绿原酸C、马钱苷、芦丁、金丝桃苷等功能性成分进行化学指纹图谱识别研究,结果显示,122批不同产地金银花指纹图谱相似度均大于0.9,具有良好一致性。结论鉴定出4个成分作为金银花抗炎质量标志物评价的物质基础,金银花化学指纹图谱研究可用于金银花药材品质判定及伪品鉴定。

OBJECTIVE To study the screening and identification methods of anti-inflammatory quality markers of Lonicerae and the identification of the quality and counterfeit products of Lonicerae by chemical fingerprints.METHODS Human embryonic kidney 293T cells and nuclear factor-κB(NF-κB)dual luciferase reporter system were used for the anti-inflammatory activity test.The optimal index of activity was selected for the screening of active anti-inflammatory monomers,and the anti-inflammatory markers were identified by ultra-high performance liquid chromatography/four-stage rod-time-of-flight mass spectrometry(UPLC/Q-TOF).The chemical fingerprint analysis of honeysuckle was carried out,and the similarity difference of the fingerprint was determined by the similarity software.RESULTS Four active monomers screened out were identified as chlorogenic acid,mignonette glycosides,roe sprouts glycosides and swertia,respectively,and they were considered to be the anti-inflammatory activity markers of honeysuckle.Chemical fingerprint was studied on the basis of four anti-inflammatory marker ingredients and other five functional components,isochlorogenic acid A,isochlorogenic acid C,margenin,rutin and hyperin.The similarity of 122 batches of Lonicerae from different producing areas was greater than 0.9,showing good consistency.CONCLUSION Four anti-inflammatory active components identified should be used to evaluate the quality of honeysuckle,and the fingerprints of Lonicerae Japonicae Flos could be used for the authenticity of honeysuckle.