目的: 建立双柱串联方式测定甘露醇有关物质的高效液相色谱-电喷雾检测器(HPLC-CAD)方法。方法: 采用2根钙型阳离子交换柱SHODEX SUGAR SC1011 (300 mm×8.0 mm)色谱柱串联的方式,柱温为80 ℃,水为流动相,流速为0.5 mL·min-1;雾化器温度:60 ℃,滤光片为10.0,采集频率:200 Hz,幂函数为1.15;进样体积20 μL。结果: 出峰顺序分别为异麦芽糖醇峰1、麦芽糖醇峰、异麦芽糖醇峰2、甘露醇峰、山梨醇峰,麦芽糖醇与异麦芽糖醇峰2分离度为1.2,其余各峰之间的分离度均大于1.5;以各物质浓度的自然对数对峰面积的自然对数建立标准曲线,r均＞0.9992;精密度良好(RSD均＜1.3%);加样回收率为97.3%~99.3%(RSD 为1.1%~1.2%,n=9);检测限为2.0~3.7 μg·mL-1,定量限为6.8~12.0 μg·mL-1;样品测定结果与现行标准方法测定结果一致。结论: 使用双柱串联后,麦芽糖醇峰和异麦芽糖醇峰2之间的分离度得到有效提高,优于现有国内外药典及文献方法;本方法准确、简便、可靠,可用于测定甘露醇有关物质。
Objective: To establish an HPLC-CAD method for the determination of mannitol related substances of mannitol with two columns in series. Methods: Two SHODEX SUGAR SC1011(300 mm×8.0 mm) columns were used in series, the column temperature was set at 80 ℃, water was used as mobile phase and the flow rate was 0.5 mL·min-1. The detector of CAD was applied with nebulization temperature at 80 ℃, filter frequency at 10.0, collection frequency at 200 Hz,and power function value(PFV)at 1.15. The injection volume was 20 μL. Results: The peak order of mixed solution was isomalt peak 1, maltitol peak, isomalt peak 2, mannitol peak, sorbitol peak respectively, the resolution between maltitol and isomalt peak 2 was 1.2, and the resolution between the other peaks was greater than 1.5. The linearity of the natural logarithm and the natural logarithm of the peak area of each substance was good (r>0.999 2). The precision of injection was good (RSD<1.3%). The recovery was 97.3%-99.3% (RSD 1.1%-1.2%, n=9). The LOD was 2.0-3.7 μg·mL-1 and the LOQ was 6.8-12.0 μg·mL-1. The measurement results of the samples are consistent with those of the current standard method. Conclusion: The new established HPLC-CAD method shows better resolution of determination than that of existing methods and can make up for the defects of present pharmacopoeia methods,which can be a reliable method for the quality control of mannitol.
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