UPLC-MS/MS同时测定青钱柳中6种黄酮类成分

Simultaneous Determination of Six Flavonoids in Cyclocarya Paliurus by UPLC-MS/MS

关键词:  青钱柳  异槲皮苷  槲皮素-3-O-葡萄糖醛酸苷  黄酮  液质联用法

Key words:  Cyclocarya paliurus  isoquercitrin  quercetin-3-O-glucuronide  flavonoids  UPLC-MS/MS

摘要:

目的 建立UPLC-MS/MS同时测定青钱柳中芦丁、金丝桃苷、槲皮素-3-O-葡萄糖醛酸苷、异槲皮苷、槲皮苷、槲皮素6种黄酮类成分的分析方法。方法 超高效液相法采用色谱柱为Agilent ZORBAX RRHD SB-C₁₈（2.1 mm×100 mm，1.8 μm），流动相为乙腈-0.1%甲酸水溶液，梯度洗脱，流速为0.3 mL·min^-1，柱温40℃，进样量为1 μL；质谱法采用三重四级杆质谱，电喷雾离子源、负离子模式（ESI^-），通过多重反应监测同时对青钱柳中6种黄酮类成分进行定量分析。结果 测定青钱柳中6种黄酮类成分芦丁、金丝桃苷、槲皮素-3-O-葡萄糖醛酸苷、异槲皮苷、槲皮苷、槲皮素的线性范围分别为0.002~0.24 μg·mL^-1（R²=0.999 4），0.049~4.91 μg·mL^-1（R²=0.999 5），0.504~50.37 μg·mL^-1（R²=0.999 4），0.051~5.12 μg·mL^-1（R²=0.999 4），0.005~0.52 μg·mL^-1（R²=0.999 8），0.052~5.17 μg·mL^-1（R²=0.999 9），平均回收率分别为96.1%，98.4%，98.5%，97.3%，96.7%，96.5%，RSD分别为1.78%，1.22%，1.91%，1.08%，1.21%，1.29%； 16批青钱柳样品中6种黄酮的含量分别为0.019 8~0.253 7 ，0.923 5~15.568 0 ，12.987 8~130.510 9 ，0.122 9~12.101 1 ，0.007 4~0.573 9 ，0.387 7~12.456 7 mg·g^-1，其中槲皮素-3-O-葡萄糖醛酸苷的含量最高，约占总量的78%。结论 所建立的分析方法简单、快速、灵敏度高、专属性好，可应用于青钱柳中黄酮类成分的含量测定及质量控制。

Abstract:

OBJECTIVE To develop an UPLC-MS/MS method for simultaneously determination of six flavonoids(rutin, hyperoside, quercetin-3-O-glucuronide, isoquercitrin, quercitrin, quercetin) in Cyclocarya paliurus. METHODS The UPLC method was performed on an Agilent ZORBAX RRHD SB-C₁₈ column(2.1 mm×100 mm, 1.8 μm) with a gradient elution of acetonitrile and 0.1% formic acid in water at a flow rate of 0.3 mL·min^-1, the column temperature was 40℃, and the injection volume was 1 μL; mass spectrometry used the triple quadrupole mass spectrometry, electrospray ionization source, negative mode(ESI^-) and the multiple reaction monitoring mode was employed for the quantitative analysis of six flavonoids. RESULTS The linear ranges of rutin, hyperoside, quercetin-3-O-glucuronide, isoquercitrin, quercitrin and quercetin were 0.002-0.24 μg·mL^-1 (R²=0.999 4), 0.049-4.91 μg·mL^-1(R²=0.999 5), 0.504-50.37 μg·mL^-1(R²=0.999 4), 0.051-5.12 μg·mL^-1(R²=0.999 4), 0.005-0.52 μg·mL^-1(R²=0.999 8), 0.052-5.17 μg·mL^-1(R²=0.999 9); the average recoveries were 96.1%, 98.4%, 98.5%, 97.3%, 96.7%, 96.5%, respectively. The RSDs were 1.78%, 1.22%, 1.91%, 1.08%, 1.21%, 1.29%, respectively. The content of six flavonoids in 16 batches were 0.019 8-0.253 7, 0.923 5^-15.568 0, 12.987 8^-130.510 9, 0.122 9^-12.101 1, 0.007 4-0.573 9, 0.387 7^-12.456 7 mg·g^-1, respectively, and the content of quercetin-3-O-glucuronide was the highest, which accounted for about 78% of the total amount. CONCLUSION The developed method is simple, rapid, sensitive and specific, which can be used for the determination of flavonoids and the quality control of Cyclocarya paliurus.