Application of Chromatographic Column Packed with Copolymer to Separation and Determination of Organic Acids by HPLC
摘 要
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(250 mm×4.6 mm,5 μm),流动相为0.02 mol·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1.0 mL·min-1;柱温25 ℃;检测波长为210 nm。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0.05~12.0 g·L-1范围呈线性,琥珀酸的检出限(3S/N)为0.005 g·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%。
Abstract
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%.
中图分类号 O652.63
所属栏目
基金项目 “863”项目(No.2007AA02Z200;2007AA06A402)
收稿日期 2010/12/16
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备注魏荣卿(1953-),女,山西汾阳人,教授,研究方向为色谱分离技术及应用。
引用该论文: WEI Rong-qing,WANG Jin,FANG Shuo,LIU Xiao-ning. Application of Chromatographic Column Packed with Copolymer to Separation and Determination of Organic Acids by HPLC[J]. Physical Testing and Chemical Analysis part B:Chemical Analysis, 2012, 48(2): 188~190
魏荣卿,王瑾,方硕,刘晓宁. 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[J]. 理化检验-化学分册, 2012, 48(2): 188~190
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参考文献
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【2】田宏,黄海欣,张玉洁.降脂宁颗粒的质量标准研究[J].时珍国医国药, 2003,14(4):214-215.
【3】石尚友,聂明华,王小东.厌食健脾颗粒的薄层色谱鉴别[J].中医药导报, 2005,11(2):59-60.
【4】张峻松,贾春晓,张文叶,等.山楂中有机酸的定量分析研究[J].中州大学学报, 2003,20(3):119-121.
【5】郭德华,夏琳.离子排斥色谱法同时测定果汁中11种有机酸[J].色谱, 2001,19(3):276-278.
【6】COLLINS R N. Separation of low-molecular mass organic acid-metal complexes by high-performance liquid chromatography[J]. Journal of Chromatography, 2004,1059:1-12.
【7】金高娃,章飞芳,薛兴亚,等.反相高效液相色谱法测定山楂中的有机酸[J].分析化学, 2006,34:987-990.
【8】白冬梅,杜国民,沈菲,等.反相HPLC同时测定产琥珀酸放线杆菌发酵液中的有机酸与葡萄糖[J].分析试验室, 2004,23(3):15-18.
【9】易娇,尹笃林,廉世勋,等.反相HPLC法同时测定烟草中的6种有机酸[J].烟草化学, 2006(7):36-39.
【10】TORMO M, IZCO J M. Alternative reversed-phase high-performance liquid chromatography method to analyse organic acids in dairy products[J]. Journal of Chromatography A, 2004,1033(2):305-310.
【11】CAWTHRAY G R. An improved reversed-phase liquid chromatographic method for the analysis of low-molecular mass organic acids in plant root exudates [J]. Journal of Chromatography A, 2003,1011:233-240.
【12】SYCHOV C S, ILYIN M M, DAVANKOV V A, et al. Elucidation of retention mechanisms on hypercrosslinked polystyrene used as column packing material for high-performance liquid chromatography. [J]. Journal of Chromatography, 2004,1030:17-24.
【13】LLOYD L L. Rigid macroporous copolymers as stationary phases in high-performance liquid chromatography[J]. Journal of Chromatography A, 1991,544:201-217.
【14】CHI G T, HUDDERSMAN K D. Novel ion chromatography technique for the rapid identification and quantification of saturated and unsaturated low molecular weight organic acids formed during the Fenton oxidation of organic pollutants[J]. Journal of Chromatography, 2007,1139:95-103.
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