柱后衍生化高效液相色谱法测定门冬氨酸鸟氨酸颗粒剂的有关物质及含量

Assay for the content and related substances of ornithine aspartate granules by postcolumn derivatization HPLC

目的:建立采用邻苯二甲醛(O-phthaldialdehyde,OPA)作为柱后衍生试剂的HPLC法测定门冬氨酸鸟氨酸颗粒的有关物质与含量。方法:采用色谱柱Synergi Hydro RP,C18(150 mm×3 mm,4μm),流动相A为0.005 mol/L辛烷磺酸钠溶液-高氯酸(100∶0.16),流动相B为0.01 mol/L辛烷磺酸钠溶液-乙腈-高氯酸(50∶50∶0.16),进行梯度洗脱;检测波长为336 nm。结果:门冬氨酸和鸟氨酸在37.5～112.5μg/ml范围内,浓度与峰面积呈良好的线性关系(门冬氨酸r=0.999 8,鸟氨酸r=0.999 9);门冬氨酸回收率为101.2%,RSD为0.92%(n=9),鸟氨酸回收率为101.0%,RSD为0.97%(n=9);门冬氨酸鸟氨酸主成分峰与3种杂质峰分离度均符合要求,门冬氨酸和鸟氨酸最低检出限均为5ng。结论:该方法方便、快速、灵敏度高、重现性好,适用于测定门冬氨酸鸟氨酸颗粒剂中的有关物质及含量。

Objective： A HPLC with 0 -phthaldialdehyde （OPA） postcolumn derivatization was developed to determine thecontent of ornithine aspartate granules and its related substances. Methods： The HPLC method was applied with a Synergi Hydro RP C18 column（ 3 mm ×150 mm, 4 μm） by a gradient elution. The mobile phase A was consisted of 0.005 mol/L 1 - Octanesulfonic acid sodium salt- perchloric acid （100： 0.16）, the mobile phase B was consisted of 0.01 mol/L 1 -Octanesulfonic acid sodium salt- acetonitrile - perchloric acid C 50： 50： 0.16）. The detection wavelength was set at 336 nm. Results： The linear range of ornithine and aspartate was 37.5 - 112.5 μg/ml （ r = 0. 999 8 and 0. 999 9 for aspartate and omithine, respectively ）. The recovery rate of aspartate was 101.2%, RSD was 0.92% （ n = 9 ） , and the recovery rate of ornithine was 101.0% , RSD was 0.97% （ n = 9 ）. The detection limit for ornithine and aspartate were both 5 ng. Conclusion： This method is simple, rapid, sensitive, reproducible, accurate and reliable, and it is suitable for the detection of the content and related substances of ornithine aspartate granules.