固相萃取结合UPLC-MS/MS法测定陈皮中6种黄酮类成分

Determination of Six Flavonoids in Citri Reticulatae Pericarpiumby Solid Extraction Phase Ultra Performance Liquid Chromatography-tandem Mass Spectrometry

建立超高效液相色谱串联三重四极杆质谱测定陈皮中6种主要黄酮类物质的分析方法。采用HLB-D101复合固相萃取柱净化和富集样品,通过Hypersil Gold C18(100 mm×2.1 mm,1.9μm)色谱柱分离,以0.1%甲酸和乙腈为流动相,电喷雾离子化正负离子模式,多反应监测方式(MRM)检测。结果表明,选用的复合固相萃取小柱可有效分离纯化陈皮样品中6种黄酮类成分。通过优化好的色谱和质谱条件,在负离子模式下,芸香柚皮苷、柚皮苷、橙皮苷和新橙皮苷4种黄烷酮类成分在0.05~10μg/mL浓度范围内线性良好,检出限为5 mg/kg;在正离子模式下,川陈皮素和橘皮素在0.005~1.0μg/mL范围内线性良好,检出限为0.1 mg/kg。在不同浓度的加标水平下,样品中6种待测物的平均加样回收率为92.5%~107.5%,相对标准偏差(RSD)为1.7%~6.4%。本研究建立的方法快速高效、灵敏度高、回收率和重现性好。通过对不同品种陈皮样品检测结果表明,相同品种的陈皮中黄酮类的含量相对稳定,部分黄酮类成分在不同品种间存在明显差别。

An analytical method was developed for the determination of six flavonoids in Citri Reticulatae Pericarpium based on the solid phase extraction by UPLC-MS/MS.The samples were purified by HLB-D101 dual cartridge solid phase extraction.Chromatographic column-Hypersil Gold C18 column(100 mm 2.1 mm,1.9 m),was used with 0.1%formic acid-acetonitrile as the mobile phase.Six flavonoids were detected by electrospray ionization mass spectrometry in positive and negative switching with MRMmode.The results showed that the selected dual solid phase extraction packing could effectively separate and purify 6 flavonoids from CRP.By optimizing the chromatographic and mass spectrometry parameters,the four flavanones of naringin,narirutin,hesperidin and neohesperidin showed good linearity in the concentration range of 0.05~10 g/mL in negative ion mode,and the detection limit was 5 mg/kg.In the positive ion mode,the linear ranges of citrine and citrine were 0.005~1.0 g/mL,and the detection limit was 0.1mg/kg.The average recoveries were 92.5%~107.5%with relative standard deviations(RSD)of 1.7%~6.4%.The established method is rapid and efficient,with a high sensitivity,good recovery and reproducibility in this study.The results showed that the content of flavonoids in CRP of the same variety was relatively stable,and there were obvious differences in some flavonoids among different varieties.