骨化三醇前体对照品的制备及其在软胶囊中检查方法的研究

目的:制备骨化三醇前体对照品,进行方法研究,并建立高效液相色谱法测定骨化三醇软胶囊中骨化三醇前体的含量.方法:采用C8色谱柱,以乙腈-Tris缓冲液(配制1 g·L-1的3-羟甲基氨基甲烷溶液,用磷酸调节pH为7.0～7.5)为流动相,梯度洗脱,流速1 mL·min-1,检测波长265 nm,柱温40℃,进样体积20μ...

Study on the preparation of pre-calcitriol reference substance and its examination method in calcitriol soft capsules

Objective:To study the preparation method of pre-calcitriol reference substance,and establish an high performance liquid chromatography method to determin^-1e the content of pre-calcitriol in calcitriol soft capsules.Methods:The determin^-1ation was performed on a C8 column with acetonitrile-Tris buffer as mobile phase,wherein a Tris buffer solution was obtained by preparing 1 g·L^-1 of 3-(hydroxymethyl)amin^-1omethane solution and then adjusting pH to 7.0-7.5 with phosphoric acid.The flow rate was set at 1 mL^-1·min^-1 and the detection wavelength was set at 265 nm.The column temperature was set at 40℃and the injection volume was 20μL.Results:The specificity of this quantitative method was good,the linearity of calcitriol was within the concentration range of 1.275-255.0μg·mL^-1,and the linearity of pre-calcitriol was within the concentration range of 0.142-2.126μg·mL^-1.The limit of detection of calcitriol and pre-calcitriol were 0.0064μg·mL^-1 and 0.035μg·mL^-1 respectively,and the quantitative limits were 0.032μg·mL^-1 and 0.12μg·mL^-1 respectively.The precision was good,and the average recovery rate of added samples was 100.0%.An area normalization method with correction factors was selected for calculation,calcitriol soft capsules of different batches were determin^-1ed,and pre-calcitriol contents of the calcitriol soft capsules were 6.06%,5.08%and 6.12%respectively.Conclusion:This method is simple and accurate,and therefore can be used for determin^-1ing pre-calcitriol in calcitriol soft capsules.It can be used as a reference for the control of precursor limits of calcitriol drugs with similar structure.