HPLC法测定富马酸替诺福韦二吡呋酯原料药和胶囊中的有关物质

Determination of Related Substances in Tenofovir Disoproxil Fumarate Raw Material and Capsule by HPLC

目的：建立测定富马酸替诺福韦二吡呋酯原料药和胶囊中有关物质的方法。方法：采用高效液相色谱法。色谱柱为Eclipse XDB-C18,流动相为甲醇-0.05%磷酸溶液（40∶60,V/V）和甲醇（梯度洗脱）,检测波长为260 nm,流速为1.0 ml/min,柱温为40℃,进样量为20μl。结果：已知杂质替诺福韦单吡呋酯浓度在2.515～20.120μg/ml范围内与峰面积呈良好的线性关系（r=0.999 9）;检测限为3.75μg/ml,定量限为12.50 ng/ml;精密度、稳定性、重复性试验的RSD分别为0.42%、2.00%、2.67%。按自身对照法测得替诺福韦单吡呋酯含量的结果偏高。结论：该方法操作简便、专属性强、灵敏度高,可用于富马酸替诺福韦二吡呋酯原料药和胶囊中有关物质的测定。

OBJECTIVE： To establish a method for the determination of related substances in tenofovir disoproxil fumarate raw material and capsules. METHODS： HPLC method was adopted. The determination was performed on Eclipse XDB-C18 column with mobile phase consisted of methanol-0.05% phosphoric acid （40：60, V/V） and methanol （gradient elution） at the flow rate of 1.0 ml/ min. The detection wavelength was set at 260 nm, and column temperature was 40 ℃. The volume of injection was 20μl. RE- SULTS： The linear range of tenofovir monosoproxil was 2.515-20.12 μg/ml（r=0.999 9）. RSDs of precision, stability and repeat- ability tests were 0.42%, 2.00% ,2.67%. The limits of detection and quantification were 3.75 μg/ml and 12.50 ng/ml. The content of disoproxil fumarate determined by self-control method. CONCLUSIONS： The method is convenient, specific, sensitive, and can be used for the determination of related substances in tenofovir disoproxil fumaratc raw material and capsule.