[目的]阐明栀子水提浸膏中的化学成分并对主要成分进行定量分析,为栀子配方颗粒及其药效研究提供物质基础。[方法]用纯净水作为溶剂提取制备栀子水提浸膏,采用超高效液相高分辨质谱联用技术(ultra high performance liquid chromatography coupled with mass spectrometry,UHPLC-MS)对栀子水提浸膏主要化学成分进行分析,并采取高效液相色谱法(high performance liquid chromatography,HPLC)测定出栀子水提浸膏中主要成分栀子苷、京尼平1-β-D-龙胆双糖苷的含量。[结果]根据栀子中主要成分的裂解规律和质谱裂解碎片,栀子水提浸膏中共鉴定化合物16种,包括环烯醚萜类7种、有机酸类3种、黄酮类2种、其他类4种。选择其中两个主要成分栀子苷和京尼平1-β-D-龙胆双糖苷进行含量测定,结果显示栀子水提浸膏中栀子苷和京尼平1-β-D-龙胆双糖苷的平均含量分别为13.42%和4.88%。[结论]采用HPLC测定栀子水提浸膏中栀子苷和京尼平1-β-D-龙胆双糖苷含量,方法简便、效果稳定,为后续栀子配方颗粒的药效研究提供了物质基础。
Huang-Lian-Jie-Du-Decoction (HLJDD) has been widely used for the treatment of Alzheimer's disease (AD) in clinic. However, the relationship between its chemical profile and neuroprotective bioactivity was not clearly clarified yet. In present study, the water extract of HLJDD and subsequent three polarity fractions divided by different reagents were investigated. A total of 17 chromatographic peaks were confirmed by comparison with standards and their UV, MS spectra. Among them, 11 major compounds were determined by HPLC-DAD method with good linear regression relationship (r2, 0.9994-0.9999), precisions (inter-day precision RSD, 0.79%-1.07%; intra-day precision RSD, 1.59%-2.10%), repeatability (RSD, 1.66%-3.67%), stability (RSD, 1.26%-4.77%) and recovery (95.24%-105.41%, RSD, 0.29%-2.69%). Furthermore, PC12 cells and primary neurons cells were used for the neuroprotective effective assessment of aforementioned four samples from HLJDD. 3"he total aqueous extract and n-butanol extract of HLJDD presented more significant effects than the other two parts. According to their quality and quantity determination results, iridoids and alkaloids have a positive correlation with the neuroprotective effectiveness of HLJDD.