Abstract：
BACKGROUND: Thunbergia laurifolia is used in traditional Thai medicine to reduce fever and treat mouth ulcers. However, the quantitative analysis of chemical markers has not yet been officially defined.
OBJECTIVE: The objective of this study is to develop a high-performance liquid chromatography (HPLC) method using a design of experiment (DoE) for the quantitative analysis of multicomponents by single marker (QAMS) and fingerprinting of the T. laurifolia aqueous extract.
METHODS: Critical variables were screened using a two-level fractional factorial design, followed by the optimization of the selected variables using a central composite design. The validated method was applied for quality assessment based on QAMS and fingerprinting of the extract.
RESULTS: Optimum conditions of DoE for the analysis of caffeic acid, vicenin-2, and rosmarinic acid were determined. The relative correction factors for caffeic acid and vicenin-2 were calculated using rosmarinic acid as an internal reference standard, and their contents in 30 samples were determined. The differences between the external standard method (ESM) and QAMS were compared. No significant difference was observed in the quantitative determination, proving the consistency QAMS and ESM. HPLC fingerprints of T. laurifolia were established with 8 of 12 characteristic peaks that were structurally characterized using HPLC-diode array detection-electrospray ionization/tandem mass spectrometry. The similarity of the fingerprints in all samples was ≥0.74, and the pattern recognition of the characteristic peaks was satisfied.
CONCLUSIONS: The proposed method efficiently detected multiple components of the T. laurifolia extract. Thus, the method is beneficial in providing references for enhancing the quality control of other herbal medicines.
OBJECTIVE: The objective of this study is to develop a high-performance liquid chromatography (HPLC) method using a design of experiment (DoE) for the quantitative analysis of multicomponents by single marker (QAMS) and fingerprinting of the T. laurifolia aqueous extract.
METHODS: Critical variables were screened using a two-level fractional factorial design, followed by the optimization of the selected variables using a central composite design. The validated method was applied for quality assessment based on QAMS and fingerprinting of the extract.
RESULTS: Optimum conditions of DoE for the analysis of caffeic acid, vicenin-2, and rosmarinic acid were determined. The relative correction factors for caffeic acid and vicenin-2 were calculated using rosmarinic acid as an internal reference standard, and their contents in 30 samples were determined. The differences between the external standard method (ESM) and QAMS were compared. No significant difference was observed in the quantitative determination, proving the consistency QAMS and ESM. HPLC fingerprints of T. laurifolia were established with 8 of 12 characteristic peaks that were structurally characterized using HPLC-diode array detection-electrospray ionization/tandem mass spectrometry. The similarity of the fingerprints in all samples was ≥0.74, and the pattern recognition of the characteristic peaks was satisfied.
CONCLUSIONS: The proposed method efficiently detected multiple components of the T. laurifolia extract. Thus, the method is beneficial in providing references for enhancing the quality control of other herbal medicines.
摘要：
背景：传统的泰国医学中使用了劳联Thunbergia来减轻发烧和治疗口腔溃疡。然而,化学标记物的定量分析尚未正式定义。
目的：本研究的目的是开发一种高效液相色谱（HPLC）方法，该方法使用实验设计（DoE）通过单标记（QAMS）定量分析多组分。
方法：使用两级分数阶乘设计筛选关键变量，然后使用中央复合设计对所选变量进行优化。验证的方法用于基于QAMS和提取物指纹的质量评估。
结果：分析咖啡酸的最佳DoE条件，vicenin-2和迷迭香酸进行测定。使用迷迭香酸作为内部参考标准，计算咖啡酸和vicenin-2的相对校正因子，并测定了30份样品中的含量。比较外标法(ESM)与QAMS的差异。在定量测定中没有观察到显著差异，证明QAMS和ESM的一致性。利用HPLC-二极管阵列检测-电喷雾电离/串联质谱对12个特征峰中的8个进行了结构表征。所有样本的指纹图谱相似度为≥0.74,特征峰的模式识别得到满足。
结论:所提出的方法有效地检测了蓝草提取物的多种成分。因此，该方法有利于为加强其他中药材的质量控制提供参考。
目的：本研究的目的是开发一种高效液相色谱（HPLC）方法，该方法使用实验设计（DoE）通过单标记（QAMS）定量分析多组分。
方法：使用两级分数阶乘设计筛选关键变量，然后使用中央复合设计对所选变量进行优化。验证的方法用于基于QAMS和提取物指纹的质量评估。
结果：分析咖啡酸的最佳DoE条件，vicenin-2和迷迭香酸进行测定。使用迷迭香酸作为内部参考标准，计算咖啡酸和vicenin-2的相对校正因子，并测定了30份样品中的含量。比较外标法(ESM)与QAMS的差异。在定量测定中没有观察到显著差异，证明QAMS和ESM的一致性。利用HPLC-二极管阵列检测-电喷雾电离/串联质谱对12个特征峰中的8个进行了结构表征。所有样本的指纹图谱相似度为≥0.74,特征峰的模式识别得到满足。
结论:所提出的方法有效地检测了蓝草提取物的多种成分。因此，该方法有利于为加强其他中药材的质量控制提供参考。
