HPLC测定格列美脲有关物质的改进

Improvement of HPLC Determination Method for Related Substances in Glimepiride

关键词:  格列美脲  有关物质  杂质  高效液相色谱

Key words:  glimepiride  related substance  impurity  HPLC

摘要:

目的 改进格列美脲有关物质检测的高效液相色谱法。方法 采用Agilent Zorbax SB-C₁₈色谱柱(250 mm×4.6 mm，5 μm)，以磷酸二氢钠溶液(取0.5 g磷酸二氢钠，加水500 mL溶解，用磷酸调pH至2.5)-乙腈(50∶50)为流动相，流速1.0 mL· min^-1，检测波长为228 nm。结果 溶剂对杂质检测无影响，格列美脲与已知杂质分离度良好，杂质1与杂质2的定量限均为10 ng·mL^-1，杂质1在0.304 2~2.028 μg·mL^-1，杂质2在0.316 2~2.108 μg·mL^-1内呈良好线性关系(r分别为0.999 5和0.999 7)，平均回收率分别为 100%和99%，RSD为1.4%和1.3%。结论 改进的方法消除了溶剂对杂质检测的干扰，可用于格列美脲中有关物质的检测。

Abstract:

OBJECTIVE To improve the method for determining related substances in glimepiride by HPLC. METHODS The determination was performed on an Agilent Zorbax SB-C₁₈ column(250 mm×4.6 mm, 5 μm), and the mobile phase consisted of 500 mL sodium dihydrogen phosphate solution (dissolve 0.5 g of sodium dihydrogen phosphate in 500 mL of water and adjust to pH 2.5 with phosphoric acid) and 50 mL of acetonitrile at the flow rate 1.0 mL·min^-1. The detection wavelenth was 228 nm. RESULTS The solvents had no effects on the impurity determination. An excellent separation was achieved. The LOQs of impurity 1 and 2 were both 10 ng·mL^-1. The linearity were found in the rang of 0.304 2-2.028 μg·mL^-1 and 0.316 2-2.108 μg·mL^-1 for impurity 1 and 2 respectively. The corresponding average recoveries were 100% with RSD 1.4% for impurity 1 and 99% with RSD 1.3% for impurity 2. CONCLUSION The improved method eliminated the solvent disturbance to impurity determination, and is suitable for the determination of impurities in glimepirede.