摘要
目的建立超高效液相色谱-四极杆静电场轨道阱质谱(UPLC-Q-Exactive Orbitrap-MS)法同时测定扶正和肤止痒方透皮成分的含量,并探讨其体外透皮扩散规律。方法采用Franz扩散池法进行体外释放与透皮试验,分别以BALB/c雄性小鼠空白皮肤和银屑病样皮肤作为透过屏障,采用UPLC-Q-Exactive Orbitrap-MS技术测定扶正和肤止痒方及其接收液的成分含量;采用零级、一级、Higuchi动力学方程评价扶正和肤止痒方的透皮成分的释放与经皮渗透效果。结果共鉴定出扶正和肤止痒方中落新妇苷、茵芋碱、阿魏酸、花椒毒酚、白鲜碱、柠檬苦素、花椒毒素、吴茱萸苦素、蛇床子素、异茴芹内酯等10个透皮成分。建立的UPLC-Q-Exactive Orbitrap-MS多成分同时测定方法中,10个透皮成分在相应的浓度范围内呈现良好的线性关系(r>0.9960),且精密度、稳定性、准确度及基质效应均符合标准。以空白小鼠皮肤作为半透膜,落新妇苷、蛇床子素的透皮扩散规律遵循零级动力学方程;阿魏酸、花椒毒素、白鲜碱、吴茱萸苦素、柠檬苦素的透皮扩散规律遵循一级动力学方程;花椒毒酚、茵芋碱、异茴芹内酯的透皮扩散规律遵循Higuchi动力学方程。以模型小鼠银屑病样皮肤作为半透膜,花椒毒酚、花椒毒素、白鲜碱、异茴芹内酯、吴茱萸苦素的透皮扩散规律遵循一级动力学方程;落新妇苷、阿魏酸、茵芋碱、柠檬苦素、蛇床子素透皮扩散规律遵循Higuchi动力学方程。结论该研究建立的扶正和肤止痒方透皮成分定量检测方法准确、灵敏、稳定,明确了扶正和肤止痒方10个透皮成分的体外扩散规律,可为扶正和肤止痒方治疗银屑病的药效研究奠定基础。
Objective To establish an ultra-performance liquid chromatography coupled with quadrupole-Orbitrap mass spectrometry(UPLC-Q-Exactive Orbitrap-MS)method for the simultaneous determination of transdermal components in Fuzheng Hefu Zhiyang Formula and to investigate their in vitro transdermal diffusion kinetics.Methods In vitro release and transdermal experiments were conducted using Franz diffusion cells,with both control skin and psoriatic-like skin from male BALB/c mice serving as barriers.The contents of components in Fuzheng Hefu Zhiyang Formula and its receptor solutions were determined by UPLC-Q-Exactive Orbitrap-MS.Zero-order,first-order,and Higuchi kinetic models were used to evaluate the release and percutaneous penetration of the transdermal components.Results Ten transdermal components in Fuzheng Hefu Zhiyang Formula were identified:astilbin,skimmianine,ferulic acid,xanthotoxol,dictamnine,limonin,xanthotoxin,evodiamine,osthole,and isopimpinellin.The established UPLC-Q-Exactive Orbitrap-MS method for multi-component simultaneous determination demonstrated good linearity for all ten components within their respective concentration ranges(r>0.9960),with precision,stability,accuracy,and matrix effects meeting validation criteria.Using blank mouse skin as the semi-permeable membrane,the transdermal diffusion of astilbin and osthole followed zero-order kinetics;ferulic acid,xanthotoxin,dictamnine,evodiamine,and limonin followed first-order kinetics;xanthotoxol,skimmianine,and isopimpinellin followed the Higuchi model.Using model mouse(psoriatic-like)skin as the barrier,the transdermal diffusion of xanthotoxol,xanthotoxin,dictamnine,isopimpinellin,and evodiamine followed first-order kinetics,while astilbin,ferulic acid,skimmianine,limonin,and osthole followed the Higuchi model.Conclusion The quantitative method established in this study for determining transdermal components of Fuzheng Hefu Zhiyang Formula is accurate,sensitive,and stable.It elucidates the in vitro transdermal diffusion kinetics of ten transdermal components,laying a foundation for understanding the pharmacodynamic study of Fuzheng Hefu Zhiyang Formula in treating psoriasis.
作者
钟海心
罗倩倩
刘炎杰
唐斌
黄晓冰
谢建辉
陈永根
喻靖傑
邓浩
卢传坚
陈海明
ZHONG Haixin;LUO Qianqian;LIU Yanjie;TANG Bin;HUANG Xiaobing;XIE Jianhui;CHEN Yonggen;YU Jingjie;DENG Hao;LU Chuanjian;CHEN Haiming(The Second School of Clinical Medicine,Guangzhou University of Chinese Medicine,Guangzhou 510120 Guangdong,China;Shenzhen Bao’an Traditional Chinese Medicine Hospital Group,Shenzhen 518100 Guangdong,China;State Key Laboratory of TCM Wet Syndrome/The Second Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510120 Guangdong,China;The Third Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510378 Guangdong,China;State Key Laboratory of Traditional Chinese Medicine Syndrome/The Second Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510120 Guangdong,China)
出处
《中药新药与临床药理》
北大核心
2026年第2期312-322,共11页
Traditional Chinese Drug Research and Clinical Pharmacology
基金
国家自然科学基金项目(U23A6012)
广州市科技计划项目(202201020353,2024A03J0707)
广东省中医院“拔尖人才”计划项目(BJ2022KY02)
深圳市“医疗卫生三名工程”项目(SZZYSM202106008)。
