高聚物型色谱柱在高效液相色谱法分离测定有机酸中的应用

考察高聚物型色谱柱(聚苯乙烯-二乙烯基苯,PS-DVB)分离5种有机酸标准品以及琥珀酸发酵液中5种成分的适用性。采用MKF-YJS色谱柱(250mm×4.6mm,5μm),流动相为0.02mol·L-1磷酸二氢铵溶液(用磷酸调pH至2.5)与乙腈以体积比97比3组成的混合溶液;流量1.0mL.min-1;柱温25℃;检测波长为210nm。结果表明:5种有机酸标准品以及琥珀酸发酵液中5种成分均达到了良好的分离效果;琥珀酸质量浓度在0.05～12.0g.L-1范围呈线性,琥珀酸的检出限(3S/N)为0.005g.L-1;进样精密度0.63%(n=5);发酵液中琥珀酸平均回收率为91.8%。

Application of Chromatographic Column Packed with Copolymer to Separation and Determination of Organic Acids by HPLC

The feasibility of application of copolymer packed chromatographic column using copolymer of polystryrene and divinylbenzene resin(PS-DVB) as packing material,to the separation of 5 standards of organic acids and 5 components in fermentation broth of succinic acid was studied.The PS-DVB based MKF-YJS column(250 mm×4.6 mm,5 μm) with column temperature of 25 ℃ was used.The mobile phase containing 0.02 mol·L-1 NH4H2PO4 solution(adjust pH to 2.5 with H3PO4) and acetonitrile mixed in the ratio of 97∶3(by vol.),with flow rate of 1.0 mL·min-1 was used.The detection wavelength was set at 210 nm.Five standard organic acids and five compounds in succinic acid fermentation broth were well separated by the proposed procedure.The calibration curve was in good linearity in the range of 0.05-12.0 g·L-1.The limit of detection was 0.005 g·L-1.The precision of sample introduction was 0.63%(n=5).The average recovery of succinic acid in fermentation broth was 91.8%.