血浆样品预处理环境的pH值对芪类化合物HPLC-MS/MS检测方法基质效应的影响  

Effect of pH Value of Pretreatment Environment of Plasma Samples on Matrix Effects in HPLC-MS /MS Analysis of Stilbene Compounds

作　　者：赵立波[1] 王宇航[1] 司霞[1] 马卓[1] 陈月[1] 冯婉玉[1] 

机构地区：[1]北京大学人民医院药剂科,北京100044

出　　处：《中国药学杂志》2014年第15期1358-1363,共6页Chinese Pharmaceutical Journal

基　　金：国家自然青年基金资助项目(81102877)

摘　　要：目的评估血浆样品预处理环境的pH值对芪类化合物HPLC-MS/MS检测方法基质效应的影响。方法分别在pH4.01、5.00、6.86、8.00、9.18 5种不同的pH条件下,使用甲基叔丁基醚、乙醚、乙酸乙酯3种常用的萃取剂,利用液液萃取的方法处理含8种芪类化合物的人血浆;以Ultra C18柱(2.1 mm×50 mm,5.0μm)为固定相,以乙腈、0.2 mmol·L-1甲酸铵(HCOONH4)为流动相,采用梯度洗脱模式,以MRM扫描模式进行定量,建立8种芪类化合物HPLC-MS/MS检测方法。结果在选定的色谱质谱条件下,8种芪类化合物与血浆中的杂质分离良好;8种芪类化合物的基质效应与预处理后样品中残存磷脂的量正相关。在碱性条件萃取处理的血浆样品磷脂残存少,基质效应小。在pH 8.0时,基质效应最小。结论在进行生物样品预处理时,适度碱化样品,有助于降低血浆样品预处理后残存的磷脂含量,从而降低基质效应。OBJECTIVE To evaluate the effects of the pH condition in which the plasma samples were prepared on the matrix effects of HPLC-MS/MS analysis of stilbene compounds. METHODS Eight stilbene compounds were extracted at five different pH levels （pH 4. 01,5. 00,6. 86,8. 00,9. 18）, with the most usual extractants including methyl tertiary butyl ether, diethyl ether, and ethyl acetate, respectively. Separation of the eight stilbene compounds was performed on an Agilent 1200 HPLC system. Detection was carried out on an API 5500 Qtrap mass spectrometry system with the MRM mode. The samples were injected into an Ultra column （2. 1 mm × 50 mm ,5. 0 μm）. The mobile phase was composed of acetonitrile and 0. 2 mmol · L^-1 ammonium formate delivered in gradient elution mode. The matrix effects were assessed by the post-extraction method. RESULTS The correlation between matrix effects and residual phospholipids in the sample extract was clearly shown. The effective removal of phospholipids and reduction of matrix effect demonstrated that pH 8.00 was the best condition for extraction of stilbenes in plasma samples. CONCLUSION Preparation of plas- ma samples in alkaline environment is helpful to remove residual phospholipids, thus beneficial to reduce matrix effects in HPLC-MS/ MS analysis of stilbene compounds.

关 键 词：PH值 芪类化合物 白藜芦醇 苯烯莫德 高效液相色谱-质谱联用 基质效应 

分 类 号：R917[医药卫生—药物分析学]