Li Xuequn, Shen Xinnan, Wu Min
Department of Nutrition and Food Hygiene, School of Public Health, Fudan University, Key Laboratory of Ministry of Education, Public Health and Safety, Shanghai 200032, China.
Wei Sheng Yan Jiu. 2016 Jul;45(4):658-662.
To establish LC-MS method to determine quercetin in human serum.
Serum samples were extracted by acetone- methanol and hydrolyzed by β- glucuronidase. The Agilent Poroshell 120 EC C_(18)( 50 mm × 4. 6 mm, 2. 7 μm) was used and acetonitrile( B) and 1% formic acid( A) as the mobile phase with gradient elution. Flow rate was 1. 0 m L / min and column temperature 40℃. Liquid chromatography-tandem mass spectrometry( LC-MS) with electrospray ionization( ESI~-)was performed in a negative mode and using selective negative ion detection. Detected ion m / z was 301. 1( quercetin) and 285. 1( fisetin).
The method had good linearity in the concentration ranges of 1- 200 ng / m L. The correlation coefficient( r) was0. 9998. The detection limits of quercetin was 0. 025 ng / m L. Interday precision( RSD)was 4. 98%-8. 35%. Intraday precision( RSD) was 7. 62%-9. 73%. The recoveries of quercetin in serum was 97. 4%-112. 8%.
The established method is sensitive and reproducible, and can be used in determination of quercetin in human serum.
建立液相色谱-质谱法测定人血清中槲皮素的含量。
血清样品用丙酮-甲醇提取，并用β-葡萄糖醛酸酶水解。采用安捷伦Poroshell 120 EC C₁₈（50 mm×4.6 mm，2.7 μm）色谱柱，以乙腈（B）和1%甲酸（A）为流动相进行梯度洗脱。流速为1.0 mL/min，柱温为40℃。采用电喷雾电离（ESI⁻）的液相色谱-串联质谱法（LC-MS）在负离子模式下进行，采用选择性负离子检测。检测离子的m/z为301.1（槲皮素）和285.1（非瑟酮）。
该方法在1-200 ng/mL浓度范围内具有良好的线性关系。相关系数（r）为0.9998。槲皮素的检测限为0.025 ng/mL。日间精密度（RSD）为4.98%-8.35%。日内精密度（RSD）为7.62%-9.73%。血清中槲皮素的回收率为97.4%-112.8%。
所建立的方法灵敏、可重复，可用于测定人血清中槲皮素的含量。
