摘要
利用高效液相色谱法(HPLC)测定橘核中柠檬苦素、黄柏酮、诺米林等3个主要有效成分含量;利用色差分析仪测定橘核盐炙、清炒过程中样品的色度值,将盐炙橘核颜色与成分含量进行相关性分析;采用油浴模拟炮制技术结合高效液相色谱法,分析橘核中诺米林及其转化产物的相对含量变化,明确诺米林在炮制时的结构转化规律;此外,利用脂多糖(LPS)诱导RAW264.7细胞建立炎症模型,比较诺米林及其转化产物黄柏酮的抗炎活性。结果表明,随着炮制时间增加,E^(*)ab、L^(*)整体呈下降趋势;a^(*)在一定时间内缓慢上升,之后无明显变化;b^(*)在一定时间内变化不明显,之后开始下降;柠檬苦素含量变化不明显,诺米林含量显著下降,黄柏酮含量显著升高,盐炙和清炒在炮制过程中含量与色度值参数变化趋势基本一致;诺米林和黄柏酮2个成分含量与色度值(L^(*)、a^(*)、b^(*)、E^(*)ab)有显著相关性,柠檬苦素含量与色度值相关性不明显;通过分析诺米林在不同加热温度、时间的HPLC图,发现在200~250℃,加热5~60 min时,诺米林含量显著降低,黄柏酮含量显著升高;体外抗炎活性实验结果表明,与模型组相比,诺米林高剂量组、黄柏酮不同浓度剂量组均可显著抑制细胞NO的释放(P<0.01),且相同剂量浓度下,黄柏酮抑制NO释放的能力更强。该研究明确橘核炮制过程中颜色与主要有效成分含量有明显相关性,揭示橘核炮制过程的质量动态变化规律,发现并证实了橘核炮制后由诺米林向黄柏酮转化,且转化产物黄柏酮体外抗炎活性明显强于诺米林,为橘核炮制工艺优化、饮片质量控制及临床应用提供科学依据。
High-performance liquid chromatography(HPLC)was used to determine the content of three major components in Citri Reticulatae Semen(CRS),including limonin,nomilin,and obacunone.The chromaticity of the CRS sample during salt processing and stir-frying was measured using a color difference meter.Next,the relationship between the color and content of the salt-processed CRS sample was investigated through correlation analysis.By integrating the oil bath technique for processing simulation with HPLC,the changes in the relative content of nomilin and its transformation products were analyzed,with its structural transformation pattern during processing identified.Additionally,RAW264.7 cells were induced with lipopolysaccharides(LPSs)to establish an inflammatory model,and the anti-inflammatory activity of nomilin and its transformation product,namely obacunone was evaluated.The results indicated that as processing progressed,E^(*)ab and L^(*)values showed a downward trend;a^(*)values exhibited a slow increase over a certain period,followed by no significant changes,and b^(*)values remained stable with no significant changes over a certain period and then started to decrease.The limonin content remained barely unchanged;the nomilin content decreased,and the obacunone increased significantly.The changing trends in content and color parameters during salt-processing and stir-frying were basically consistent.The content of nomilin and obacunone was significantly correlated with the colorimetric values(L^(*),a^(*),b^(*),and E^(*)ab),while limonin content showed no significant correlation with these values.By analyzing HPLC patterns of nomylin at different heating temperatures and time,it was found that under conditions of 200-250℃for heating of 5-60 min,the content of nomilin significantly decreased,while the obacunone content increased pronouncedly.The in vitro anti-inflammatory activity results indicated that compared to the model group,the group with a high concentration of nomilin and the groups with varying concentrations of obacunone showed significantly reduced release of nitric oxide(NO)(P<0.01).When both were at the same concentration,obacunone showed better performance in inhibiting NO release.In this study,the obvious correlation between the color and content of major components during the processing of CRS samples was identified,and the dynamic patterns of quality change in CRS samples during processing were revealed.Additionally,the study revealed and confirmed the transformation of nomilin into obacunone during processing,with the in vitro anti-inflammatory activity of obacunone significantly greater than that of nomilin.These findings provided a scientific basis for CRS processing optimization,tablet quality control,and its clinical application.
作者
朱奎霖
邹金莲
邓徐丽
邓茂鑫
王海名
尹蕊
陈章贤
张云涛
何红平
董发武
ZHU Kui-lin;ZOU Jin-lian;DENG Xu-li;DENG Mao-xin;WANG Hai-ming;YIN Rui;CHEN Zhang-xian;ZHANG Yun-tao;HE Hong-ping;DONG Fa-wu(Yunnan Key Laboratory of Southern Medicinal Utilization,College of Traditional Chinese Medicine,Yunnan University of Chinese Medicine,Kunming 650500,China)
出处
《中国中药杂志》
北大核心
2025年第9期2382-2390,共9页
China Journal of Chinese Materia Medica
基金
国家自然科学基金项目(82260758,31860092)
云南省高层次人才培养支持计划“青年拔尖人才”专项(YNWR-QNBJ-2020-255)
云南省基础研究计划面上项目(202101AT070254)。
关键词
盐橘核
色度值
高效液相色谱法
模拟炮制
诺米林
黄柏酮
相关性
salt-processed Citri Reticulatae Semen
colorimetric value
high-performance liquid chromatography
simulated processing
nomilin
obacunone
correlation
