State Key Laboratory of Bioactive Substances and Functions of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, PR China.
State Key Laboratory of Bioactive Substances and Functions of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, PR China.
Phytomedicine. 2021 Dec;93:153776. doi: 10.1016/j.phymed.2021.153776. Epub 2021 Sep 27.
Traditional Chinese herbal patches (TCHPs) characterized by good patient adherence and few side effects have been widely used in the clinic. However, their bioactive transdermal components, which are key to the quality and efficacy, have rarely been investigated.
Establishing an approach to probe the bioactive transdermal components of TCHPs, using Gutong patch (GTP) as a case study.
GTP constituents were identified using ultrahigh-pressure liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-MS). The identified GTP constituents were transferred to an ultrahigh-pressure liquid chromatography coupled with triple quadrupole mass spectrometry (UHPLC-QqQ-MS/MS) method in a dynamic multiple reaction monitoring (dMRM) mode for sensitively targeted analysis of transdermal components present at low concentrations. The ex vivo transdermal components were rapidly probed using a percutaneous permeation model and the release kinetics of marker components was assessed to evaluate the release behavior of GTP. The in vivo transdermal components were then studied in a rat model of arthritis and the changes in concentration of 15 representative bioactive transdermal components with time were well revealed.
A total of 120 phytochemical constituents have been identified in GTP extracts and 61 ex vivo transdermal components were targetedly detected. The release kinetics of GTP were evaluated by eight marker components and fitted to a zero-order kinetic model. In vivo, 62 and 40 transdermal components were observed in the skin and subcutaneous tissues of arthritic rats, respectively. The concentration-time changes of 15 representative transdermal components with documented bioactivities have been successfully exhibited after GTP administration.
We established a feasible approach to probe the bioactive transdermal components of TCHPs efficiently. The integration of mass spectrometry profiling and targeted detection in dMRM mode enabled a comprehensive investigation of phytochemical constituents and their transdermal delivery, thus addressing the challenge of direct probing of diverse transdermal components present at low concentrations. This approach could be used to rapidly probe the bioactive components and understand the mechanism of TCHPs.
传统中药贴剂（TCHP）以良好的患者依从性和较少的副作用而广泛应用于临床。然而，其生物活性透皮成分是 TCHP 质量和疗效的关键，却很少被研究。
建立一种方法来探测 TCHP 的生物活性透皮成分，以 Gutong 贴剂（GTP）为例进行研究。
采用超高效液相色谱-四极杆飞行时间质谱联用仪（UHPLC-QTOF-MS）鉴定 GTP 成分。将鉴定出的 GTP 成分转移到超高效液相色谱-三重四极杆质谱联用仪（UHPLC-QqQ-MS/MS）中，采用动态多重反应监测（dMRM）模式，对低浓度存在的透皮成分进行灵敏靶向分析。采用经皮渗透模型快速探测体外透皮成分，并评估标记成分的释放动力学，以评估 GTP 的释放行为。然后在关节炎大鼠模型中研究体内透皮成分，随着时间的推移，很好地揭示了 15 种代表性生物活性透皮成分的浓度变化。
共鉴定出 GTP 提取物中的 120 种植物化学物质成分，靶向检测到 61 种体外透皮成分。通过 8 种标记成分评估 GTP 的释放动力学，并拟合到零级动力学模型。体内，在关节炎大鼠的皮肤和皮下组织中分别观察到 62 种和 40 种透皮成分。成功展示了 GTP 给药后具有生物活性的 15 种代表性透皮成分的浓度-时间变化。
我们建立了一种高效探测 TCHP 生物活性透皮成分的可行方法。质谱分析和 dMRM 模式下的靶向检测相结合，全面研究了植物化学物质及其透皮传递，从而解决了直接探测低浓度多种透皮成分的挑战。该方法可用于快速探测生物活性成分并了解 TCHP 的作用机制。
