目的: 建立大黄䗪虫丸UPLC指纹图谱,通过研究不同剂量60Co射线辐照前后UPLC指纹图谱的变化,评价60Co射线灭菌对大黄䗪虫丸指纹图谱的影响。方法: 采用Waters Acquity UPLC HSS T3色谱柱(100 mm×2.1 mm,1.8 μm),以乙腈-0.2%甲酸水溶液为流动相,梯度洗脱,流速0.3 mL·min-1,柱温35 ℃,切换检测波长。采用辐照剂量分别为0、3、5、7、9 KGy对大黄䗪虫丸进行辐照灭菌,采用指纹图谱相似度评价分析、多元统计分析等方法,区分不同来源的大黄䗪虫丸,并分析60Co不同辐照剂量对大黄䗪虫丸化学成分的影响。结果: 经过方法学考察及14份样品的测定,建立了大黄䗪虫丸UPLC指纹图谱,共确认了26个共有峰,不同来源的大黄䗪虫丸样品指纹图谱与对照指纹图谱相似度为0.723~0.990,9种化学成分是引起大黄䗪虫丸样品差异性的主要成分;60Co辐照灭菌的辐照剂量大时,会影响大黄䗪虫丸中8个化学成分。结论: 建立的UPLC指纹图谱方法准确可靠、具有良好的重复性,能区分不同来源的大黄䗪虫丸,且可分析60Co辐照灭菌对大黄䗪虫丸质量的影响。
Objective: To establish an UPLC fingerprint of Dahuang Zhechong pills, and evaluate the effect of 60Co-γ radiation sterilization on the fingerprint by studying the changes of UPLC fingerprint before and after irradiation by 60Co-γ ray. Methods: A Waters Acquity UPLC HSS T3 column (100 mm×2.1 mm, 1.8 μm) was used, and the mobile phase was consisted of acetonitrile-0.2% formic acid solution at a flow rate of 0.3 mL·min-1. The detection wavelengths were 210 nm, 230 nm, 254 nm and 270 nm and the column temperature was 35 ℃. Dahuang Zhechong Pills was irradiated by 60Co-γ ray with the dosage of 0, 3, 5, 7 and 9 KGy, respectively. Similarity analysis and multivariate statistical analysis were used to distinguish Dahuang Zhechong Pills from different sources, and to analyze the effects of different doses of 60Co-γ on the chemical constituents of Dahuang Zhechong Pills. Results: Through methodological investigation and the determination of 14 batches of samples, the UPLC fingerprint of Dahuang Zhechong Pills was established and 26 common peaks were confirmed. The similarities of samples to the reference fingerprint were 0.723-0.990, and 9 components were found to cause the difference of Dahuang Zhechong pills. When the irradiation dose of 60Co-γ is high, the 8 components in the Dahuang Zhechong Pills might be affected. Conclusion: The established method for UPLC fingerprint of Dahuang Zhechong pills is accurate, reliable and reproducible. It can distinguish the different sources of Dahuang Zhechong pills, and analyze the influence of 60Co-γ irradiation sterilization on the quality of Dahuang Zhechong pills.
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