目的 利用超高效液相色谱-线性离子阱-静电场轨道阱联用质谱（UPLC-LTQ-Orbitrap-MS）技术快速鉴别瓜拉那Paullinia cupana叶中的化学成分，建立高效液相色谱（HPLC）法测定瓜拉那叶中咖啡因的含量。方法 定性分析采用Waters Acquity UPLC HSS T₃色谱柱（100 mm×2.1 mm，1.8 μm），流动相为0.1%甲酸水-乙腈，梯度洗脱，体积流量0.3 mL·min^-1，进样量3 μL，在电喷雾正负离子模式下采集数据。定量分析采用Prevail色谱柱（250 mm×4.6 mm，5 μm），柱温30℃，流动相为纯水-乙腈，梯度洗脱，体积流量为1 mL·min^-1，进样量10 μL，检测波长275 nm。结果 利用UPLCLTQ-Orbitrap-MS技术，根据质谱一级精确相对分子质量和二级特征碎片，从瓜拉那叶中共鉴定出46个化学成分，包括22个酚酸类，7个黄酮类，7个生物碱类，5个长链脂肪酸类和5个其他类化合物。建立HPLC-UV测定瓜拉那叶咖啡因含量的方法中，咖啡因在0.201 8～2.018 0 μg线性关系良好（R²=0.999 8），平均加样回收率（n=9）为99.52%，RSD=1.16%，瓜拉那叶中咖啡因质量分数在2.8%左右。结论 首次采用UPLC-LTQ-Orbitrap-MS技术鉴定瓜拉那叶中的化学成分，提供了一种快速、高效的分析方法，所建立的HPLC-UV测定咖啡因含量的方法准确、可靠，为瓜拉那叶后续的深入研究奠定了基础。

Objective UPLC-LTQ-Orbitrap-MS technology was used to identify the chemical constituents in the leaves of Paullinia cupana, and a method of HPLC was established for the determination of caffeine in Paullinia cupana leaves. Methods The Waters Acquity UPLC HSS T₃ column (100 mm×2.1 mm, 1.8 μm) was used for qualitative analysis. The mobile phase was 0.1% formic acid water (A)-acetonitrile (B) with gradient elution and the flow rate was 0.3 mL·min^-1, the injection volume was 3 μL, and the data were collected under the electrospray positive and negative ion mode. Quantitative analysis was carried out at 30℃ on a prevail column (250 mm×4.6 mm, 5 μm), the mobile phase consisted of pure water (A)-acetonitrile (B) with gradient elution and the flow rate was 1.0 mL·min^-1, the injection volume was 10 μL and the detection wavelength was 275 nm. Results Using UPLC-LTQOrbitrap-MS technology, 46 chemical constituents including 22 phenolic acids, seven flavonoids, seven alkaloids, five long-chain fatty acids and five other compounds were identified from Paullinia cupana leaves according to the first accurate molecular weight and secondary characteristic fragments of mass spectrometry. For the quantitative analysis method, caffeine had good linear relationship in the range of 0.201 8-2.018 0 μg (R²=0.999 8). The average recoveries (n=9) was 99.52% and the RSD was 1.16%. The content of caffeine in Guarana leaves is about 2.8%. Conclusion For the first time, UPLC-LTQ-Orbitrap-MS technology was used to identify the chemical components in Paullinia cupana leaves, which provided a fast and efficient analysis method. The HPLC-UV method for the determination of caffeine content was accurate and reliable, which laid a foundation for the further study of Paullinia cupana leaves.

R284.1

厄瓜多尔植物泡林藤与宝乐果雄花资源开发及应用（2020071720314）