目的 鉴定狗脊中的化学成分并建立原儿茶酸的UPLC含量测定方法，比较狗脊药材与饮片中原儿茶酸的含量差异。方法 采用超高效液相色谱-串联四极杆飞行时间质谱（UPLC-Q-TOF/MS）法，ESI电离源，负离子扫描模式采集狗脊药材质谱数据，根据质谱精确相对分子质量、二级碎片离子、紫外特征吸收光谱，并与文献数据或标准品比对鉴定狗脊药材中的主要化学成分；UPLC色谱条件为采用ACQUITY UPLC CORTECS T3（100 mm×2.1 mm，1.6 μm）色谱柱，以0.1%甲酸水溶液（A）-乙腈-甲醇（70∶30，B）为流动相进行梯度洗脱，柱温40 ℃，体积流量0.4 mL/min，进样量4 μL。结果 从狗脊药材中共鉴定出26个化学成分，主要包括糖苷类成分16个、酚酸类成分2个、皂苷类成分1个、苯丙素类成分1个、脂类成分1个、其他类成分5个；原儿茶酸在0.94～202.00 μg/mL（R²＝0.999 9）线性关系良好，平均回收率为101.90%，检测限为0.20 μg/mL，定量限为0.79 μg/mL。狗脊饮片中原儿茶酸的含量均符合药典规定标准（大于0.020%）；狗脊药材中原儿茶酸的含量（质量分数为0.003 8%～0.005 3%）显著低于饮片（0.556 0%～0.850 9%），虽然经酸水解处理后，原儿茶酸的含量显著升高（0.205 2%～0.450 1%），但仍低于饮片含量。结论 建立的UPLC-Q-TOF/MS方法可快速识别狗脊药材中的主要化学成分，可为狗脊药材的质量控制和药效物质基础研究奠定基础；建立的原儿茶酸UPLC含量测定方法灵敏度高，准确度好，具有可行性，可用于狗脊药材的质量控制。

Objective To identify the chemical constituents of Cibotium barometzand establish a UPLC method for the determination of protocatechuic acid, and to compare the content differences of protocatechuic acid between crude drug and decoction pieces. Methods Ultra-performance liquid chromatography-tandem quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS) combined ESI ionization source, and negative ion scanning mode were used to collect the mass spectrometry data of C. barometz extract. According to the precise molecular mass, secondary fragment ions and comparison with literature data or standards, the major constituents of C. barometz were identified. The UPLC method was performed on a ACQUITY UPLC CORTECS T3 (100 mm × 2.1 mm, 1.6 μm) column with 40 ℃, and gradient elution with 0.1% formic acid in water (A)-acetonitrile-methanol (70∶30) (B) was used at the flow rate of 0.4 μg/mL, and the injection volume was 4 μL. Results A total of 26 constituents were identified from C. barometz, including 16 glycosides, 2 phenolic acids, 1 saponin, 1 phenylpropanoid, 1 lipid and 5 other constituents. The linearity of protocatechuic acid was good in the range of 0.94—202.00 μg/mL, (R²＝0.999 9). The average recovery was 101.90%, the limit of detection was 0.20 μg/mL, and the limit of quantification was 0.79 μg/mL. The content of protocatechuic acid in the decoction pieces of Cibotium barometz achieved the standard of pharmacopoeia (＞0.020%, n＝6). The content of protocatechuic acid in crude drug was lower the standard of pharmacopoeia (content was 0.003 8%—0.005 3%, n＝4); But after acid hydrolysis, the content of protocatechuic acid significantly increased (0.205 2%—0.450 1%), while also less than in the decoction pieces of C. barometz.Conclusion The established UPLC-Q-TOF/MS method in this study can quickly identify the major constituents of C. barometz, which can lay a foundation for the quality control and the pharmacodynamic material basis of C. barometz. The established UPLC method for the determination of protocatechuic acid has high sensitivity, good accuracy, and feasibility, and can be used for the quality control of Cibotium barometz.

R286.2

2021年度国家药品标准制修订研究课题（2021Z004）