KM Science Research Division, Korea Institute of Oriental Medicine, Daejeon 34054, Korea.
Molecules. 2022 Feb 11;27(4):1223. doi: 10.3390/molecules27041223.
Gungha-tang (GHT), a traditional herbal medicine, consists of nine medicinal herbs (Cnidii Rhizoma, Pinelliae Tuber, Poria Sclerotium, Citri Unshius Pericarpium, Citri Unshius Pericarpium Immaturus, Aurantii Fructus Immaturus, Atracylodis Rhizoma Alba, Glycyrrhizae Radix et Rhizoma, and Zingiberis Rhizoma Recens). It has been used for various diseases caused by phlegm. This study aimed to develop and verify the simultaneous liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis method, using nine marker components (liquiritin apioside, neoeriocitrin, narirutin, naringin, hesperidin, neohesperidin, liquiritigenin, glycyrrhizin, and 6-shogaol) for quality control of GHT. LC-MS/MS analysis was conducted using a Waters TQ-XS system. All marker analytes were separated on a Waters Acquity UPLC BEH C column (2.1 × 100 mm, 1.7 μm) using gradient elution with a distilled water solution (containing 5 mM ammonium formate and 0.1% [/] formic acid)-acetonitrile mobile phase. LC-MS/MS multiple reaction monitoring (MRM) analysis was carried out in negative and positive ion modes of an electrospray ionization source. The developed LC-MS/MS MRM method was validated by examining the linearity, limits of detection and quantification, recovery, and precision. LOD and LOQ values of nine markers were calculated as 0.02-8.33 ng/mL and 0.05-25.00 ng/mL. The recovery was determined to be 89.00-118.08% and precision was assessed with a coefficient of variation value of 1.74-8.64%. In the established LC-MS/MS MRM method, all markers in GHT samples were detected at 0.003-16.157 mg/g. Information gathered during the development and verification of the LC-MS/MS method will be useful for the quality assessment of GHT and other herbal medicines.
甘海胃康胶囊由九味中药（荜茇、瓜蒌、高良姜、延胡索、白术、白芍、甘草、海螵蛸、沙棘）组成，具有治疗痰饮病症的功效。本研究旨在建立并验证一种同时测定甘海胃康胶囊中九种指标性成分（甘草苷、新橙皮苷、柚皮苷、橙皮苷、柚皮苷、新橘皮苷、甘草素、甘草酸和 6-姜辣素）的液相色谱-串联质谱（LC-MS/MS）分析方法，用于甘海胃康胶囊的质量控制。LC-MS/MS 分析采用 Waters TQ-XS 系统，使用 Waters Acquity UPLC BEH C 柱（2.1×100mm，1.7μm）进行梯度洗脱，以含 5mM 甲酸铵和 0.1%甲酸的蒸馏水-乙腈为流动相，在正负离子模式下进行电喷雾电离源多反应监测（MRM）分析。通过线性、检测限和定量限、回收率和精密度考察对建立的 LC-MS/MS 方法进行验证。九种标志物的检测限和定量限分别计算为 0.02-8.33ng/mL 和 0.05-25.00ng/mL。回收率为 89.00%-118.08%，精密度以变异系数值 1.74%-8.64%表示。在建立的 LC-MS/MS MRM 方法中，甘海胃康胶囊样品中的所有标志物的含量均在 0.003-16.157mg/g 之间。该方法的建立为甘海胃康胶囊及其他中草药的质量评价提供了有用的信息。
