基于液质联用法对白及中2-异丁基苹果酸葡萄糖氧基苄酯类成分的定性定量分析

目的： 建立白及中2-异丁基苹果酸葡萄糖氧基苄酯类成分的UPLC-ESI-Q/TOF-MS快速识别鉴定方法，以及主成分militarine的UPLC测定方法。方法： 液质联用方法采用Waters Acquity UPLC BEH C₁₈（1.7 μm，2.1 mm×100 mm）色谱柱，乙腈-0.1%甲酸溶液梯度洗脱，检测器为高分辨四极杆飞行时间质谱，ESI离子源，MS^E（^E代表碰撞能量）采集方式，采集质量数范围m/z 100~1 000。含量测定采用UPLC法，以乙腈-0.1%磷酸溶液（22：78）为流动相；检测波长为223 nm；柱温：35℃；流速：0.2 mL·min^-1。结果： 利用建立的白及化学成分数据库，结合2-异丁基苹果酸葡萄糖氧基苄酯类成分的诊断离子、准确分子量、质谱碎片及对照品信息，快速分析鉴定21个目标化合物。主成分militarine在19.059 0~381.180 8 μg·mL^-1范围内线性关系良好（r=0.998 6）。加样回收率为100.9%，RSD为1.5%。36批白及中militarine的含量范围是1.5%~4.6%。结论： UPLC-Q/TOF-MS法能快速、准确地鉴定白及中的化学成分，为其药效物质基础研究提供科学依据。建立的UPLC含量测定方法操作简便、准确可靠，可用于白及的质量控制。

Qualitative and quantitative analysis of 2-isopropyl-(glucosyloxy)benzyl malate in Bletilla striata by UPLC-ESI-Q/TOF-MS

OBJECTIVE To establish a UPLC-ESI-Q/TOF-MS method for rapid identification of 2-isobutylmalate gluconoxybenzyl esters in Bletilla striata and a UPLC method for the determination of militarine.METHODS LC-MS was performed on a Waters Acquity UPLC BEH C₁₈ (1.7 μm, 2.1 mm×100 mm) column with gradient elution of acetonitrile-0.1% formic acid solution. The detector was high-resolution quadrupole time-of-flight mass spectrometry, ESI ion source, MSE (E represents collision energy) acquisition mode, and the acquisition mass range was m/z 100-1 000.The assay was performed by UPLC with acetonitrile-0.1% phosphoric acid solution (22:78) as the mobile phase; the detection wavelength was 223 nm; column temperature:35℃; flow rate:0.2 mL·min^-1.RESULTS Twenty-one target compounds were rapidly analyzed and identified by using the established database of Baiji chemical components, combined with the diagnostic ions, accurate molecular weight, mass spectrometry fragments and reference substance information of 2-isobutylmalate gluconoxy-benzyl esters. The linear range was 19.059 0-381.180 8 μg·mL^-1 for militarine (r=0. 998 6); the average recoveries were 100.9%, and RSD was 1.5%. The contents of militarine in 36 batches were 1.5%-4.6%.CONCLUSION UPLC-Q/TOF-MS can rapidly and accurately identify the chemical constituents in Bletilla striata, providing a scientific basis for the study of its pharmacodynamic substance basis. The UPLC method is simple, accurate and reliable, and it is suitable for the quality control of Bletilla striata.