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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 effect of pH value of plasma sample pretreatment environment on the matrix effects of stilbene compounds by HPLC-MS / MS. Methods Three commonly used extractants (methyl tert-butyl ether, ethyl ether and ethyl acetate) were prepared under the conditions of pH4.01, 5.00, 6.86, 8.00 and 9.18 pH values. 8 kinds of stilbene compounds in human plasma. The mobile phase consisted of acetonitrile and 0.2 mmol·L-1 ammonium formate (HCOONH4) with Ultra C18 column (2.1 mm × 50 mm, 5.0 μm) as stationary phase and gradient elution mode , MRM scanning mode for quantitative determination of eight kinds of stilbene compound HPLC-MS / MS detection method. Results Under the selected chromatographic conditions, the eight stilbenes separated from the plasma were well separated. The matrix effects of the eight stilbenes were positively correlated with the amount of residual phospholipids in the pretreated samples. Plasma samples extracted under alkaline conditions showed less residual phospholipids and less matrix effects. At pH 8.0, the matrix effect is minimal. Conclusion In the pretreatment of biological samples, the moderate alkalization of samples will help to reduce the residual phospholipid content after plasma sample pretreatment, thus reducing the matrix effect.