全自动二维高效液相色谱测定霉酚酸血药浓度的研究

目的：采用全自动二维液相色谱（2D-LC-UV）建立测定霉酚酸（MPA）血药浓度的方法，并应用于临床。方法：样品去除蛋白后直接进样，待测物在一维柱AstonSC2T （3.5 mm×50 mm，5 μm）上初步分离，通过中间柱Aston SBR（3.0 mm×10 mm，5 μm）截取保留，转移到二维柱Aston SBX4（4.6 mm×150 mm，5 μm）上进一步分离。一维流动相为MPA-1A移动相，流速0.8 mL·min^-1；二维流动相为MPA-2A移动相，流速1.2 mL·min^-1；柱温40℃；紫外检测波长304 nm。结果：在所建立的色谱条件下，霉酚酸与各杂质分离良好，在0.32~25.03 μg·mL^-1范围内与峰面积呈良好的线性关系（r=0.999 9），可在13 min内完整出峰，方法回收率高于95%；日内、日间的精密度RSD值均小于5%；稳定性试验RSD小于5%。结论：本方法简便快速、结果准确、稳定性良好、自动化程度高、检测成本低，适用于临床霉酚酸快速检测。

Measurement of mycophenolic acid in blood by two-dimensional liquid chromatography

OBJECTIVE To establish a method for rapid determination of mycophenolic acid (MPA) blood concentration by automated two-dimensional liquid chromatography (2D-LC-UV) and apply it in clinical practice.METHODS The samples were directly injected after protein removal. The analytes were preliminarily separated on a one-dimensional column Aston SC2T (3.5 mm×50 mm, 5 μm), intercepted and retained by an intermediate column Aston SBR (3.0 mm×10 mm, 5 μm), and transferred to a two-dimensional column Aston SBX4 (4.6 mm×150 mm, 5 μm) for further separation.The one-dimensional mobile phase was MPA-1A mobile phase at a flow rate of 0.8 mL·min^-1; the two-dimensional mobile phase was MPA-2A mobile phase at a flow rate of 1.2 mL·min^-1; the column temperature was 40 ℃; and the UV detection wavelength was 304 nm.RESULTS Under the established chromatographic conditions, mycophenolic acid was well separated from each impurity, showed a good linear relationship with the peak area in the range of 0.32-25.03 μg·mL^-1 (r=0.999 9), could completely peak within 13 min, the method recovery was higher than 95%; the intra-day and inter-day precision RSD was less than 5%; the RSD of stability test was less than 5%.CONCLUSION The method is simple, rapid, accurate, stable, automatic and low cost, and suitable for rapid detection of mycophenolic acid in clinic.