有机酸类化合物的反相高效液相色谱法的分离条件研究

 研究了在用RP HPLC法测定有机酸时如何对流动相的酸度进行控制 ,给出了流动相最佳pH的选择通式 :色谱柱允许的最低 pH值≤pH≤pKa- 2 ;研究了流动相中甲醇、乙腈存在时对此通式的影响。用研究得出的结论选择了测定甲酸、乙酸、丙酸、丁酸和丙二酸、丁二酸、戊二酸、己二酸时的色谱条件 ,并且研究了每种酸在各自浓度范围内测定的线性关系、回归方程、相关系数以及相对标准偏差。实验显示 ,在选定的色谱条件下进行分析测试时 ,各种酸的线性相关系数r >0 9995 ,相对标准偏差RSD <0 90 %。

Study on Reversed-Phase High Performance Liquid Chromatography Separation Condition and Determination Method of Organic Acids

The pH values of the mobile phase which affect the chromatographic behavior of organic acids in RP-HPLC have been studied. It was considered that the problem of organic acids on an RP-HPLC column was dissociation. The equation for finding the optimum pH of mobile phase was pHlimit of colunm < or = pH < or = pKa -2. In addition the effects of contents of methanol and acetonitrile in mobile phase were studied. By using the formula the mobile phases used for the analysis of organic monobasic acids--formic, acetic, propionic and butyric acid and of organic dibasic acids--propane diacid, butane diacid, pentane diacid and hexane diacid were proposed. For the analysis of the monobasic acids, the wavelength of UV-detector was set at 210 nm and the mobile phase was a mixture of phosphate buffer solution(15 mmol/L, pH 2.1)-methanol(85:15, V/V) and at a rate of 1.5 mL/min and the column was Diamond C18, 150 mm x 4.6 mm i.d.. The average coefficient of correlation was 0.9997 and RSD was less than 0.90%. The mobile phase for organic dibasic acids was a mixture of phosphate buffer solution (20 mmol/L, pH 2.1)-methanol(75:25, V/V). The other conditions were the same as for organic monobasic acids. The average coefficient of correlation was 0.9998 and RSD was less than 0.70%.