夏天无提取物UPLC指纹图谱及成分定性研究

目的 ：建立夏天无提取物的超高效液相色谱(UPLC)指纹图谱，并采用电喷雾(ESI)-飞行时间质谱(TOF-MS^E)对夏天无提取物指纹图谱中特征化学成分进行鉴定。 方法：采用ACQUITY UPLC^® BEH-C₁₈色谱柱(2.1 mm×100 mm，1.7 μm)，以乙腈(A)-乙酸铵甲酸水溶液(每1000 mL水中含1 mL甲酸和10 mmol乙酸铵)(B)为流动相，梯度洗脱(0~2 min，5%A；2~30 min，5%A→23%A；30~37 min，23%A→60%A)，流速0.3 mL·min^-1，检测波长280 nm，柱温35 ℃，进样量1μL；质谱采用电喷雾正离子模式采集数据，毛细管电压为3.5 kV，锥孔电压为35 kV，离子源温度为150 ℃，扫描范围m/z 100~800，碰撞低能量为6 eV，碰撞高能量15~45 eV。 结果：37 min内完成夏天无提取物的UPLC指纹图谱分析，各主要成分色谱峰分离良好，方法精密度、重复性、稳定性的RSD值均小于1.0%。高分辨质谱数据结合对照品信息及相关文献，鉴定了夏天无提取物UPLC指纹图谱中12个主要色谱峰，分别认定为hydrohydrastinine、延胡索单酚碱、球紫堇碱、比枯枯灵、四氢药根碱、原阿片碱、α-别隐品碱、延胡索乙素、药根碱、非洲防己碱、巴马汀和去氢紫堇碱。 结论：本方法为夏天无提取物的质量控制及成分的全面阐述奠定了基础。

Objective: To establish the UPLC fingerprint of Corydalis Decumbentis Rhizoma extract and identify the characteristic chemical components in the fingerprint by electrospray(ESI)-time of flight tandem mass spectrometry with collision energy(TOF-MS^E). Methods: The chromatographic separation was performed on an ACQUITY UPLC^® BEH-C₁₈ column(2.1 mm×100 mm,1.7 μm) by gradient elution(0-2 min,5%A;2-30 min,5%A→23%A;30-37 min,23%A→60%A) using acetonitrile(A)-ammonium acetate-formic acid solution(each 1000 mL of water containing 1 mL formic acid and 10 mmol ammonium acetate)(B)the mobile phase at the flow rate of 0.3 mL·min^-1,the detection wavelength was 280 nm,the column temperature was maintained at 35 ℃,the injection volume was 1 μL; the mass spectrometer was operated in the positive ion mode,the capillary voltage was 3.5 kV,the cone voltage was 35 kV,the ion source temperature was set at 150 ℃ at a scanning range of m/z 100-800,the low collision energy was 6 eV,the high collision energy ramp of 15-45 eV. Results: The fingerprint of Corydalis Decumbentis Rhizoma extract was established within 37 min,the main peaks were separated clearly,the RSDs of precision,reproducibility,stability were less than 1.0 %.Based on the TOF-MS^E data,information from reference substances and published literatures,twelve main chromatographic peaks in fingerprint of Corydalis Decumbentis Rhizoma extract were identified as peaks for hydrohydrastinine,kikemanine,bulbocapnine,bicuculline,corypalmine,protopine,α-allocryptopine,tetrahydropalmatine,jatrorrhizine,columbamine,palmatine and dehydrocorydaline. Conclusion: The method is useful for the quality control of Corydalis Decumbentis Rhizoma extract and the elucidation of its chemical constituents.