UPLC法同时测定西红花有效成分含量及鉴别非法染色

目的：联合使用UPLC定时波长自动切换技术及梯度洗脱法，快速测定西红花有效成分西红花苷-Ⅰ、西红花苷-Ⅱ含量及鉴别柠檬黄、胭脂红、金胺O和新品红4个常见非法染色物。方法：采用UPLC定时波长自动切换技术，色谱柱为ACQUITY UPLC HSS T3柱（2.1 mm×150 mm， 1.8 μm）；流动相为乙腈-0.05 mol·L^-1乙酸铵水溶液（含0.05%乙酸），梯度洗脱，体积流量0.3 mL·min^-1；柱温35℃。结果：西红花苷-Ⅰ、西红花苷-Ⅱ和4个非法染色物可同时检出，均达到基线分离；西红花苷-Ⅰ、西红花苷-Ⅱ均有较宽的线性范围和良好的线性关系（r>0.999 9），平均回收率（n=6）分别为100.1%、101.6%， 12批样品中西红花苷-Ⅰ、西红花苷-Ⅱ的检测含量范围分别为2.75%~9.92%、1.00%~4.28%；柠檬黄、金胺O、新品红、胭脂红的检测下限分别为0.041、0.088、0.11、0.14 ng，定量下限分别0.17、0.23、0.32、0.42 ng。结论：本方法简便快速，准确灵敏，重复性好，能在短时间内较全面地评价西红花药材的质量，有助于提高企业对原药材的检测效率。

Objective: To develop an UPLC method with wavelength switching and gradient elution for simultaneous determination of effective constituents(crocin-Ⅰand crocin-Ⅱ)and four pigments(tartrazine,neucoccin,auramine O and new fuchsin)which were illegally added into Croci Stigma. Methods: An UPLC method was established with ACQUITY UPLC HSS T3 column(2.1 mm×150 mm, 1.8 μm)and ultraviolet wavelength switching. The mobile phase was acetonitrile(A)and 0.05 mol·L^-1 ammonium acetate(0.05% acetic acid)(B)with gradient elution at a flow rate of 0.3 mL·min^-1. And the column temperature was 35℃. Results: Six components were separated and detected on the same chromatogram by changing the wavelength at different time periods. Crocin-Ⅰ and crocin-Ⅱ had wide linear range and good linear relationship(r>0.999 9). The average recoveries(n=6)of crocin-Ⅰand crocin-Ⅱwere 100.1% and 101.6%,respectively. The contents of crocin-Ⅰand crocin-Ⅱin 12 batches of samples were 2.75%-9.92% and 1.00%-4.28%,respectively. The LODs of tartrazine,auramine O,new fuchsin and neucoccin were 0.041,0.088,0.11 and 0.14 ng,and the LOQs were 0.17,0.23,0.32 and 0.42 ng,respectively. Conclusion: This method is rapid,accurate,sensitive with good reproducibility,which can evaluate the quality of Croci Stigma in short time to improve inspection efficiency.

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