葡萄糖酸钙注射液有关物质测定与高温降解杂质结构鉴定

Determination of Related Substances of Calcium Gluconate Injection and Thermal Degradation Impurity Identification

关键词:  葡萄糖酸钙注射液  有关物质  高效液相色谱法  结构鉴定

Key words:  Calcium Gluconate injection  related substances  HPLC  structure identification

摘要:

目的 建立葡萄糖酸钙注射液有关物质的测定方法，对高温降解杂质[相对保留时间（relative retention time，RRT）4.18]进行结构鉴定。方法 采用HPLC进行有关物质测定，使用Xbridge C₈色谱柱（250 mm×4.6 mm，5 μm），流动相为含0.1 mol·L^-1磷酸氢二钾与1 mmol·L^-1四丁基氢氧化铵的混合溶液，用磷酸调节pH值至5.5，检测波长为210 nm。采用HPLC-Q-TOF/MS方法，对RRT 4.18杂质做结构鉴定。结果 有关物质测定方法条件下主成分与杂质分离度良好，RRT 4.18杂质鉴定为5-羟甲基-2-糠酸。5-羟甲基-2-糠酸的线性范围为0.18~2.26 μg·mL^-1（r=0.999 0），检测限为0.06 μg·mL^-1。结论 建立的方法能准确测定葡萄糖酸钙注射液中的有关物质，可为本品的工艺优化和质量控制提供参考依据。

Abstract:

OBJECTIVE To develop a method for related substances of Calcium Gluconate injection, and to identify the structure of impurity whose relative retention time(RRT) was 4.18. METHODS The related substances were determined by HPLC, which was performed on Xbridge C₈ column(250 mm×4.6mm, 5 μm), the moblie phase was 0.1 mol·L^-1 dipotassium phosphate and 1 mmol·L^-1 tetrabutylammonium hydroxide(pH adjusted to 5.5 by phosphoric acid), the detection wavelength was 210 nm. HPLC-Q-TOF/MS was used to identify the structure of impurity (RRT 4.18). RESULTS The separation of gluconic acid and its related substances was good. Impurity RRT 4.18 was supposed to be 5-hydroxymethyl furoic acid. The calibration curves of 5-hydroxymethyl furoic acid was linear in range of 0.18-2.26 μg·mL^-1(r=0.999 0), and its detection limit was 0.06 μg·mL^-1. CONCLUSION The HPLC method established is accutrate to determine the related substances in Calcium Gluconate injection, which can provide references for the process optimization and quality control of Calcium Gluconate injection.