Department of Food Science and Technology, Jinan University, Guangzhou, 510632, China.
Division of Research and Development, Kingmed Diagnostics, Guangzhou, 510330, China.
Sci Rep. 2020 Feb 25;10(1):3378. doi: 10.1038/s41598-020-60426-3.
Acanthopanax trifoliatus (L.) Merr. (A. trifoliatus) belongs to the family Araliaceae, which is called "Le Cai", and is an indigenous plant to Guangdong Province that has been prevalently planted for years. A. trifoliatus is used in folk medicine and has ginseng-like activity. Kaurenoic acid ((-)-kaur-16-en-19-oic acid, KA) is a kaurane-type diterpenoid that is regarded as a major compound in A. trifoliatus. Early studies have reported the determination of KA by HPLC capillary electrophoresis. However, KA could not be completely separated from other components in the plant extract by HPLC because of their similar molecular structures and physical and chemical properties. UHPLC-MS/MS could be a useful tool to identify and quantify KA. In the present work, a UHPLC-ESI-MS/MS method for determining KA in A. trifoliatus was developed and validated. KA was extracted from lyophilized A. trifoliatus leaves by ultrasound-assisted extraction and further purified by solid phase extraction (SPE). KA was quantified and separated on an Accucore C LC column. Mass spectrometry with multi-reaction monitoring (MRM) and quantitative fragment ion/product ion (m/z: 301.3/301.3) in ESI negative mode was used for quantification. The intra-assay and inter-assay relative standard deviation (R.S.D.) were 2.8% and 3.2%, respectively. The inter-person R.S.D. on the same day was 3.6%. The inter-instrument R.S.D. with the same model on the same day was 2.9%. The recoveries evaluated upon spiking three different concentrations of KA were above 97%. A minor matrix effect of 94% was observed. This method has been applied successfully for the determination of KA in A. trifoliatus leaves.
三萜酸((-)-乌头酸-16-烯-19-酸，KA)是一种贝壳杉烷型二萜类化合物，被认为是三裂叶参的主要化合物。早期的研究报道了通过高效液相毛细管电泳法(HPLC-CE)测定 KA。然而，由于其相似的分子结构和理化性质，KA 不能通过 HPLC 完全从植物提取物中分离出来。UHPLC-MS/MS 可以作为一种有用的工具来鉴定和定量 KA。本研究建立并验证了一种用于测定三裂叶参中 KA 的 UHPLC-ESI-MS/MS 方法。KA 采用超声辅助提取法从冻干的三裂叶参叶片中提取，然后通过固相萃取(SPE)进一步纯化。KA 在 Accucore C LC 柱上进行定量和分离。采用电喷雾负离子模式下的多反应监测(MRM)和定量碎片离子/产物离子(m/z:301.3/301.3)进行质谱分析。内标和日间相对标准偏差(RSD)分别为 2.8%和 3.2%。同日内不同人员的 RSD 为 3.6%。同一日内同一型号仪器的 RSD 为 2.9%。在三种不同浓度 KA 的加标回收率均在 97%以上。观察到 94%的基质效应较小。该方法已成功应用于三裂叶参叶片中 KA 的测定。
