上海交通大学学报（医学版）

上海交通大学学报（医学版）

• 论著（基础研究） •    下一篇

新型神经保护剂N-硬脂酰酪氨酸的HPLC-ED法质量监测与稳定性评价

乐可佳，史一鸣，谢一凡，陆 阳   

  1. 上海交通大学  基础医学院药学系， 上海 200025
  • 出版日期:2013-08-28 发布日期:2013-09-16
  • 通讯作者: 陆阳, 电子信箱: luyangssmu@yahoo.com。
  • 作者简介:乐可佳（1985—），女，硕士生; 电子信箱: lekejia@hotmail.com。
  • 基金资助:

    国家自然科学基金项目（30873057，81171245）；上海市科委基础重点课题（11JC1406600）；上海市科委科技支撑项目（12431900604）

Quality control and stability determination with HPLC-ED of N-stearoyltyrosine

LE Ke-jia, SHI Yi-ming, XIE Yi-fan, LU Yang   

  1. Department of Pharmacy, Basic Medical College, Shanghai Jiaotong University, Shanghai 200025, China
  • Online:2013-08-28 Published:2013-09-16
  • Supported by:

    National Natural Science Foundation of China, 30873057, 81171245；Shanghai Science and Technology Committee Foundation, 11JC1406600, 12431900604

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摘要：

目的 建立N-硬脂酰酪氨酸(NsTyr)含量的高效液相色谱电化学（HPLC-ED）测定法以监测合成过程中的产物纯度，优化制备工艺，并考察NsTyr溶液的稳定性。方法 HPLC色谱条件：安捷伦Eclipse XDB-C8色谱柱（5 μm，4.6×150 mm）；流动相为甲醇∶磷酸二氢钠（NaH2PO4）水溶液（80∶20）；检测电势600 mV；流速0.7 mL/min；柱温25 ℃；进样量10 μL。利用所建立的方法测定不同重结晶次数NsTyr样品的含量，并考察不同贮藏条件下NsTyr溶液的稳定性。结果 该方法专属性良好，NsTyr在0.03~10 μg/mL内峰面积与浓度呈线性相关（r2＞0.999），灵敏度、精密度和回收率均符合检测要求。经过3次重结晶后的NsTyr纯度高于99%，短期室温、长期冷藏以及反复冻融条件下NsTyr在溶液中的含量变化在5%以内。结论 建立了简便可靠、可用于(NsTyr)质量控制与含量分析的HPLC-ED检测方法。NsTyr样品经3次重结晶后，其纯度较高、溶液稳定，可用于细胞和动物实验。

关键词: 高效液相色谱法, 电化学检测器, N-硬脂酰酪氨酸, 纯度, 稳定性

Abstract:

Objective To establish the method of high performance liquid chromatography with electrochemical detection (HPLC-ED) for determination of content of N-stearoyltyrosine (NsTyr) to monitor the purity of product during reactions, optimize the preparation process and evaluate the stability of NsTyr solution. Methods The separation was performed on an Agilent Eclipse XDB-C8 column (5 μm, 4.6×150 mm). The mobile phase was methanol-water (80∶20). Detection potential: 600 mV; flow rate: 0.7 mL/min; column oven temperature: 25 ℃; sample volume: 10 μL. The stability trials of NsTyr in methanol were tested under certain concentrations and varied storage circumstances, based on current methodology verified from content assays on NsTyr after numerous recrystallization. Results Amid 0.03-10 μg/mL of NsTyr, the area of peak was in linear correlation with concentration (r2＞0.999). The methodology was fully validated of rational precision, sensitivity and selectivity. The purity of NsTyr after recrystallization for three times surpassed 99%, and the concentration alternation in solution under short-term ambient temperature, long-term refrigeration storage and freezing-thaw circle test was within 5%. Conclusion Endowed with fairly high convenience and reliability, the established method is feasible for both quality control and quantity determination of NsTyr. The high purity and solubility of NsTyr may facilitate the cell culture and animal experiment.

Key words: high performance liquid chromatography, electrochemical detection, N-stearoyltyrosine, purity, stability