An Efficient and Rapid HPLC-QQQ-MS Method for the Quantitation of Tropane Alkaloids in Medicinal Plants

Anisodus tanguticus is an important Tibetan medicinal plant. The tropane alkaloids (TAs), such as hyoscyamine, anisodamine, and scopolamine, exhibit significant analgesic and sedative pharmacological activities, making this species a valuable natural source for extracting these bioactive compounds. However, the chemical composition of A. tanguticus is highly complex. It includes a large number of compounds without elucidated structures, rendering systematic identification and structural characterization of its active constituents particularly important. Although high-performance liquid chromatography (HPLC) has been widely used for the separation of extracts from this plant, many unknown components remain challenging to accurately identify, even with spectral database matching. This study established an analytical method based on high-performance liquid chromatography coupled with triple quadrupole mass spectrometry (HPLC-QQQ-MS) for the determination of tropane alkaloids in A. tanguticus. The method encompasses a series of systematic procedures, including standardized sample preparation, mass spectrometric parameter configuration, LC pre-equilibration, method establishment, acquisition of MS data, multistage mass spectrometry (MSⁿ) scanning, and manual data interpretation. Using this strategy, we successfully identified three representative tropane alkaloids in A. tanguticus: hyoscyamine, anisodamine, and scopolamine. Additionally, it validates the precision of the established HPLC-QQQ-MS method, confirming that the method is rapid, sensitive, feasible, and accurate for the quantitative determination of tropane alkaloids (TAs) in A. tanguticus. The established approach demonstrates good versatility and reliability, and is suitable for the high-throughput identification and quantification of tropane alkaloids in A. tanguticus and other medicinal plants.