混合模式色谱柱全水相测定阿仑膦酸钠片的含量和有关物质

Determination of the Content and Related Substances of Alendronate Sodium Tablet Using the Mixed-mode Chromatographic Column in All Aqueous System

关键词:  阿仑膦酸钠  混合模式色谱柱  电雾式检测器  全水相体系  有关物质

Key words:  alendronate sodium  mixed-mode chromatographic column  charged aerosol detector  all aqueous system  related substances

摘要:

目的 建立一种混合模式色谱柱联用电雾式检测器测定阿仑膦酸钠片的含量和有关物质的方法。方法 采用Acclaim Trinity P2色谱柱（2.1 mm×100 mm，3 μm），柱温35℃，以水（A）-100 mmol·L^-1甲酸铵溶液（用甲酸调至pH 3.3，B）为流动相进行梯度洗脱（0~4 min，20%→50% B；4~6 min，50%→40% B；6~8 min，40%→20% B），流速为0.3 mL·min^-1，进样量为20 μL；采用电雾式检测器测定，雾化温度为50℃，采样频率为5 Hz，过滤常数为3.6 s。结果 阿仑膦酸钠在0.70~1.40 mg·mL^-1内线性良好（r²=0.999 2），方法重复性的RSD为0.53%，平均加样回收率（n=9）为99.8%（RSD=1.25%）；有关物质测定项下，主成分阿仑膦酸钠，杂质磷酸盐、亚磷酸盐及4-氨基丁酸分别在19.04~76.16，3.95~49.38，3.97~49.69，3.94~49.23 μg·mL^-1内线性良好（r²>0.998），且各杂质的校正因子为0.67，0.56，0.55，各杂质定量限约为主成分浓度的0.05%，杂质测定方法的平均回收率（n=9）分别为86.2%，85.4%，91.0%，且RSD<2%；3家企业生产的阿仑膦酸钠片剂中阿仑膦酸的标示含量分别为92.0%，91.5%，95.2%，各杂质含量均小于中国药典阿仑膦酸钠项下规定的限度0.5%。结论 该方法操作简便，灵敏度和准确性较高，可用于阿仑膦酸钠片的含量测定和有关物质检查。

Abstract:

OBJECTIVE To establish a mixed-mode chromatographic column coupled with charged aerosol detector method for the determination of related substances and content of the alendronate sodium tablets. METHODS The chromatographic separation was achieved on Acclaim Trinity P2 column(2.1 mm×100 mm, 3 μm) with column temperature of 35℃ by a gradient elution using the mobile phase of water(A)-100 mmol·L^-1 ammonium formate(B, adjusted with formic acid to pH 3.3)(0-4 min, 20%→50%B; 4-6 min, 50%→40%B; 6-8 min, 40%→20%B). The flow rate was 0.3 mL·min^-1, and the injection volume was 20 μL. Charged aerosol detector was used. Nebulization temperature was 50℃, collection frequency was 5 Hz, and the filter constant was 3.6 s. RESULTS The method for content determination of alendronate sodium had a good linearity in the range of 0.70-1.40 mg·mL^-1(r²=0.999 2), the repeatability was good(RSD=0.53%). The average recovery(n=9) of alendronate sodium was 99.8%(RSD=1.25%). Under the terms of dertermination of related substances, the main component of alendronate sodium, impurity of phosphate, phosphite and 4-aminobutyric acid had good linear range in 19.04-76.16, 3.95-49.38, 3.97-49.69, 3.94-49.23 μg·mL^-1, respectively(r²>0.998), and the corrected factors for each impurity were 0.67, 0.56, 0.55. The limit of quantitation concentration of each impurity was about 0.05% of the concentration of the main component. In addition, the average recoveries(n=9) of impurity test method were 86.2%, 85.4%, 91.0%, and RSD<2%, respectively; the labeled content of alendronate in alendronate sodium tablets produced by three companies were 92.0%, 91.5% and 95.2%, respectively, and the content of each impurity was less than the limit of 0.5% under alendronate sodium in the Chinese Pharmacopoeia. CONCLUSION The method is simple, accurate, sensitive, and applicable for the content determination and related substances inspection of alendronate sodium tablets.