Center for Advanced Biomolecular Recognition, Korea Institute of Science and Technology, Seoul 02792, Republic of Korea; Division of Bio-Medical Science and Technology, KIST School, University of Science and Technology, Daejeon 34113, Republic of Korea.
Department of Nutritional Science and Food Management, Graduate Program in System Health Science and Engineering, Ewha Womans University, Republic of Korea.
J Chromatogr B Analyt Technol Biomed Life Sci. 2024 Jan 1;1232:123957. doi: 10.1016/j.jchromb.2023.123957. Epub 2023 Dec 11.
Agastache rugosa contains phenolic compounds and flavonoids, and has been extensively used as a traditional herbal medicine. The major components in Agastache rugosa extract (ARE) are rosmarinic acid, tilianin, and acacetin, for which several analytical techniques have been reported. However, these substances have yet to be simultaneously quantified in human plasma. In this study, we aimed to simultaneously determine the three active components of ARE in human plasma by developing a reliable quantitative analytical method using ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). Chromatographic separation of the plasma samples was achieved using an ACQUITY UPLC® BEH C18 column with a gradient mobile phase of water and acetonitrile containing 0.1 % formic acid. Mass spectrometric detection was performed using a triple quadrupole tandem mass spectrometer in negative electrospray ionization (ESI-) and multiple reaction monitoring (MRM) modes. The developed quantitative method was validated for the three active components. All three analytes exhibited a linear response over the ranges of 0.5-50 ng/mL for rosmarinic acid, 0.1-20 ng/mL for acacetin, and 0.5-20 ng/mL for tilianin with a weighting factor of 1/x (where x is the concentration). At three quality control (QC) concentration levels (low, medium, and high), including the lower limit of quantitation (LLOQ), acceptable accuracy (±15 %) was achieved in the intra- and interday validations. The concentration of rosmarinic acid was highest in plasma. Tilianin and acacetin appeared and were eliminated earlier in the plasma than rosmarinic acid. This study provides a successfully validated method that can be used in further clinical applications of Agastache rugosa extracts.
藿香含有酚类化合物和类黄酮，被广泛用作传统草药。藿香提取物（ARE）的主要成分是迷迭香酸、荭草苷和乙酰丁香酮，已有几种分析技术对此进行了报道。然而，这些物质尚未在人血浆中同时定量。在这项研究中，我们旨在通过开发一种使用超高效液相色谱-串联质谱（UHPLC-MS/MS）的可靠定量分析方法，同时定量测定人血浆中 ARE 的三种活性成分。使用 ACQUITY UPLC®BEH C18 柱和包含 0.1%甲酸的水和乙腈梯度流动相实现血浆样品的色谱分离。采用三重四极杆串联质谱仪在负离子电喷雾（ESI-）和多反应监测（MRM）模式下进行质谱检测。对开发的定量方法对三种活性成分进行了验证。迷迭香酸、乙酰丁香酮和荭草苷在 0.5-50ng/mL、0.1-20ng/mL 和 0.5-20ng/mL 范围内均呈线性响应，权重因子为 1/x（其中 x 为浓度）。在三个质量控制（QC）浓度水平（低、中、高），包括定量下限（LLOQ），日内和日间验证中均达到了可接受的准确度（±15%）。迷迭香酸在血浆中的浓度最高。荭草苷和乙酰丁香酮在血浆中的出现和消除时间早于迷迭香酸。本研究提供了一种经过验证的方法，可用于藿香提取物的进一步临床应用。
