高效液相色谱法测定米诺膦酸的含量及有关物质

Determination of the content and related substances of minodronic acid by HPLC

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。

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.