基于分子网络图结合液质联用技术的莲子皮中生物碱类成分分析及含量测定

Alkaloids in Lotus Seed Epicarp and Their Content Determination Based on Molecular Network Combined with LC-MS Technology

目的:利用分子网络图结合超高效液相色谱-四极杆-静电场轨道阱高分辨质谱法(UHPLC-Q-Exactive Orbitrap MS/MS)鉴定莲子皮中生物碱类成分,采用高效液相色谱法对莲子皮中甲基莲心碱含量进行测定。方法:选用ACQUITY UPLC HSS T3色谱柱(100 mm×2.1 mm,1.8μm),以乙腈(A)-0.1%氨水(B)为流动相,梯度洗脱(0~30 min,75%~40%B;30~40 min,40%~20%B),流速为0.2 mL·min^(–1),柱温为30℃,进样量为2μL,正离子模式下进行数据采集,将采集得到的MS/MS数据导入全球天然产物社会分子网络(GNPS)构建分子网络图并进行可视化分析。选用Aglient ZORBAX XDB-C18色谱柱(250 mm×4.6 mm,5μm),以乙腈-0.1%三乙胺(62∶38)为流动相等度洗脱,流速为1.0 mL·min^(–1),检测波长为282 nm,柱温为25℃,进样量为20μL,对莲子皮中甲基莲心碱进行含量测定。结果:从莲子皮中共鉴别出14个生物碱类化合物,其中甲基莲心碱质量分数为0.033 mg·g^(-1)。结论:莲子皮中含有丰富的生物碱类成分,为莲子炮制带皮入药提供了一定的科学依据。

Objective:To identify the alkaloids in lotus seed epicarp by molecular network coupled with UHPLC-QExactive Orbitrap MS/MS and determine the content of neferine in lotus seed epicarp by high-performance liquid chromatography(HPLC).Methods:An ACQUITY UPLC HSS T3(100 mm×2.1 mm,1.8μm)column was selected with a gradient elution of acetonitrile(A)-0.1%ammonium hydroxide(B)as the mobile phases(0-30 min,75%-40%B;30-40 min,40%-20%B)at a flow rate of 0.2 mL·min^(–1),column temperature of 30℃,and the injection volume of 2μL.Data acquisition was performed in the positive ion mode.The acquired MS/MS data were imported into the Georgia Native Plant Society(GNPS)website to construct molecular network diagrams for visualized analysis.An Agilent ZORBAX XDB-C18(250 mm×4.6 mm,5μm)column was selected for the determination of neferine in lotus seed epicarp with mobile phases of acetonitrile-0.1%triethylamine(62:38)for gradient elution at a flow rate of 1.0 mL·min^(–1),detection wavelength of 282 nm,column temperature of 25℃,and injection volume of 20μL.Results:Fourteen alkaloids were identified in lotus seed epicarp,of which the content of neferine was 0.033 mg·g^(–1).Conclusion:The lotus seed epicarp is rich in alkaloids,providing a scientific basis for the use of lotus seeds with epicarp as medicine.