柱前衍生高效液相色谱法分析海洋褐藻多糖药物的糖醛酸组成  

Analysis of Uronic Acid Compositions in Marine Brown Alga Polysaccharides by Precolumn Derivatization High Performance Liquid Chromatography

作　　者：王金霞[1] 赵峡[1] 于广利[1] 李广生[1] 郝翠[1] 

机构地区：[1]中国海洋大学医药学院,教育部海洋药物重点实验室,青岛266003

出　　处：《分析化学》2009年第5期648-652,共5页Chinese Journal of Analytical Chemistry

基　　金：国家863计划项目(No.2007AA09Z445)资助

摘　　要：采用三氟乙酸(TFA)分别将藻酸双酯钠(PSS)、甘糖酯(PMS)和古糖酯(PGS)3种海洋褐藻多糖药物进行降解,再与1-苯基-3-甲基-5-吡唑啉酮(PMP)进行柱前衍生,建立了3种药物糖醛酸组成分析的高效液相色谱方法。结果表明:PSS、PMS、PGS的最适降解条件:在110℃下降解6 h,TFA浓度3 mol/L;最适衍生条件:反应温度70℃,反应90 min,PMP与供试品的摩尔比为12∶1,NaOH与供试品的摩尔比为2∶1;色谱条件为:0.1 mol/L磷酸盐(pH6.7)缓冲液-乙腈(82∶18,V/V),检测波长:245 nm,流速:1 mL/min。在此色谱条件下,甘露糖醛酸(M)和古罗糖醛酸(G)分离良好,测得PSS、PMS、PGS的M/G分别为2.37±0.05、6.60±0.22和0.22±0.03,该方法具有良好的精密度和重现性,灵敏度高,适合于海洋褐藻多糖类药物的微量分析。A high performance liquid chromatography method has been established to determine the uronic acid comopositions of three marine brown alga polysaccharide drugs, namely propylene glycol alginate sodium sulfate（PSS）, polymannuronate sulfate （PMS） and polyguluronate sulfate （PGS）. This method is based on precolumn derivatization with 1-phenyl-3-methyl-5-pyrazolone （PMP） , after three drugs are degraded by trifluoroacetic acid（TFA）. The results show that the optimum degradation conditions of PSS, PMS and PGS are as follows： the reaction temperature is 110 ℃, the reaction time is 6h and the concentration of TFA is 3 mol/L. The optimum derivation conditions are as follows： the reaction temperature is 70 ℃, the reaction time is 90 min, the molar ratio of PMP to uronic acid is 12：1 and the molar ratio of NaOH to uronic acid is 2： 1. The chromatographic conditions are as follows： the mobile phase consists of 0.1 mol/L phosphate buffer-acetonitrile（82：18, V/V） at a flow rate of 1 mL/min, and the detection wavelength at 245 nm. Mannuronic acid （M） and guluronic acid （G） are well separated under the optimum chromatographic conditions. The M/G ratios of PSS, PMS and PGS determined by HPLC are 2.37±0.05, 6.60 ±0. 22 and 0.22 ± 0. 03, respectively. This method is suitable for the microanalysis of marine brown alga polysaccharide drugs with high accuracy, reproducibility and sensitivity.

关 键 词：海洋褐藻多糖 柱前衍生 高效液相色谱法 1-苯基-3-甲基-5-吡唑啉酮 

分 类 号：R917[医药卫生—药物分析学]