目的 利用超高效液相色譜-線性離子阱-靜電場軌道阱聯(lián)用質(zhì)譜（UPLC-LTQ-Orbitrap-MS）技術(shù)快速鑒別瓜拉那Paullinia cupana葉中的化學(xué)成分，建立高效液相色譜（HPLC）法測定瓜拉那葉中咖啡因的含量。方法 定性分析采用Waters Acquity UPLC HSS T₃色譜柱（100 mm×2.1 mm，1.8 μm），流動(dòng)相為0.1%甲酸水-乙腈，梯度洗脫，體積流量0.3 mL·min^-1，進(jìn)樣量3 μL，在電噴霧正負(fù)離子模式下采集數(shù)據(jù)。定量分析采用Prevail色譜柱（250 mm×4.6 mm，5 μm），柱溫30℃，流動(dòng)相為純水-乙腈，梯度洗脫，體積流量為1 mL·min^-1，進(jìn)樣量10 μL，檢測波長275 nm。結(jié)果 利用UPLCLTQ-Orbitrap-MS技術(shù)，根據(jù)質(zhì)譜一級(jí)精確相對(duì)分子質(zhì)量和二級(jí)特征碎片，從瓜拉那葉中共鑒定出46個(gè)化學(xué)成分，包括22個(gè)酚酸類，7個(gè)黃酮類，7個(gè)生物堿類，5個(gè)長鏈脂肪酸類和5個(gè)其他類化合物。建立HPLC-UV測定瓜拉那葉咖啡因含量的方法中，咖啡因在0.201 8～2.018 0 μg線性關(guān)系良好（R²=0.999 8），平均加樣回收率（n=9）為99.52%，RSD=1.16%，瓜拉那葉中咖啡因質(zhì)量分?jǐn)?shù)在2.8%左右。結(jié)論 首次采用UPLC-LTQ-Orbitrap-MS技術(shù)鑒定瓜拉那葉中的化學(xué)成分，提供了一種快速、高效的分析方法，所建立的HPLC-UV測定咖啡因含量的方法準(zhǔn)確、可靠，為瓜拉那葉后續(xù)的深入研究奠定了基礎(chǔ)。

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

厄瓜多爾植物泡林藤與寶樂果雄花資源開發(fā)及應(yīng)用（2020071720314）