高效液相色谱法测定米诺膦酸的含量及有关物质

摘要

Objective: To establish a method for determination of the content and related substances of minodronic acid. Methods: HPLC method was adopted. Diamonsil C18 column ( 250 mm × 4. 6 mm, 5 μm) was used, and the mobile phase was composed of 0. 01 mol·L-1 sodium pyrophosphate solution and 0. 001 mol·L-1 tetrabutylammonium bromide solution ( p H adjusted to 6. 5 by phosphoric acid) -methanol ( 95∶ 5) at the flow rate of1. 0 m L·min-1. The column temperature was 25 ℃, and the injection volume was 20 μL. The detection wavelength of related substances of minodronic acid was 220 nm and the content was 280 nm. Results: Minodronic acid and all the related compounds could be well separated. The calibration curves were linear in appropriate concentration ranges, and the recovery of the compounds were between 98. 0％ ～ 102. 0％ ( RSD ＜ 2. 0％). The contents of total related substances of 3 batches of samples were 0. 23％, 0. 24％ and 0. 25％, respectively, and the contents of minodronic acid were 99. 72％, 99. 45％ and 99. 51％, respectively. Conclusion: The method is specific and accurate, which can be applied in the determination of the content and related substances of minodronic acid, and it can provide a theoretical basis for the following quality control research of minodronic acid products.%目的:建立米诺膦酸原料药的含量及其有关物质的检测方法.方法:采用高效液相色谱法,色谱柱为Diamonsil C18 (250 mm×4.6 mm,5μm) ;流动相为0.01 mol·L-1的焦磷酸钠与0.001 mol·L-1四丁基溴化铵混合溶液 (用磷酸调p H值至6.5) :甲醇 (95∶5) ;流速为1.0 m L·min-1;柱温为25℃;进样体积为20μL;有关物质检测波长为220 nm;含量测定检测波长为280 nm.结果:在该色谱条件下,米诺膦酸及其杂质能完全分离,在相应的浓度范围内,有关物质A和有关物质B线性关系良好,回收率在98.0％～102.0％之间 (RSD＜2.0％) ;3批样品有关物质总杂质测定结果分别为0.23％,0.24％,0.25％,含量测定结果分别为99.72％,99.45％,99.51％.结论:本方法专属性好、灵敏度高、准确性高,可用于米诺膦酸原料药的含量测定和有关物质的检测,为后续米诺膦酸制剂的质量控制提供了理论依据.