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目的:建立了苯及其同系物体内代谢产物粘糠酸、苯乙醛酸、马尿酸的弱阴离子交换-HPLC测定方法,并进行了方法学研究。方法:采用Acclaim Mixed-Mode WAX-1(150 mm×2.1 mm,5μm)弱阴离子交换色谱柱,以甲醇-50 mmol/L磷酸氢二钾(18∶82)为基础流动相,以波长时间程序分别在254 nm下检测粘糠酸、苯乙醛酸,224nm下检测马尿酸,探讨了弱阴离子交换色谱柱分离粘糠酸、苯乙醛酸及马尿酸的影响因素。结果:流动相的pH值、离子强度、有机相比例对上述三种代谢物产生的分离产生明显影响。结论:在确定的色谱条件下,从分离度、对称性、重复性、线性4个方面来评价,弱阴离子交换色谱柱分析酸性可离子化化合物要优于常规C18色谱柱。
OBJECTIVE: To establish a weak anion exchange-HPLC method for the determination of the metabolites mucoferric acid, phenetidinol and hippuric acid in benzene and its homologues. The methodology was also studied. METHODS: Weak anion exchange chromatography with Acclaim Mixed-Mode WAX-1 (150 mm × 2.1 mm, 5 μm) was used. The mobile phase was methanol-50 mmol / L dipotassium hydrogen phosphate (18:82) The detection of muconic acid and phenetidinic acid at 254 nm and the detection of hippuric acid at 224 nm were carried out. The factors affecting the separation of muconic acid, phenetidinic acid and hippuric acid by weak anion exchange chromatography were discussed. Results: The mobile phase pH value, ionic strength and organic phase ratio had a significant effect on the separation of the above three metabolites. CONCLUSION: The weak anion exchange chromatography is superior to the conventional C18 in the determination of acid ionizable compounds under certain chromatographic conditions from the aspects of resolution, symmetry, repeatability and linearity.