RP-HPLC测定骨化三醇胶丸中的骨化三醇及反式骨化三醇

Determination of Calcitriol and 5,6-Trans-Calcitriol in Soft Capsules by RP-HPLC

目的采用高效液相色谱串联二极管阵列检测（reversephase—highperformanceliquidchromatography—diodearraydetector，RP—HPLC-DAD）测定市售骨化三醇胶丸中主要成分与其异构体。方法采用SymmetryC18色谱柱（4．6mm×250mm，5μm），以水-乙腈-甲醇混合溶液为流动相，梯度洗脱，采用二极管阵列检测器。应用ChemStation色谱工作站进行分析。结果骨化三醇与其异构体线性范围分别为0．1714～1．36和0．1613—1．28μg·mL-1，最低检测限分别为39．75和40．90ng·mL-1，最低定量限分别为141．6和136．4ng·mL-1，方法专属性良好，精密度与稳定性的RSD值均小于1．2％，加样回收率大于98％，对于3个厂家的骨化三醇药品进行测定，平均含量为72．71％、80．35％和78．18％。结论本方法简便、快速、准确、易于推广，适合于骨化三醇胶丸中骨化三醇和反式骨化三醇的测定。

OBJECTIVE To establish a new method for determination of the main active ingredient and its isomer in commercial- ly available calcitriol capsules by reverse phase-high performance liquid chromatography-diode array detector （RP-HPLC-DAD）. METHODS The chromatographic separation was performed on a Symmetry C ls column （4. 6 mm x 250 mm, 5 μm） with gradient e- lution of water-acetonitrile-methanol. The mass spectrometer was equipped with diode array detector. Using Chem Station chromatogra- phy workstation, the main ingredient and its isomer in commercially available calcitriol capsules were analyzed. RESULTS The line- ar ranges of calcitriol and its isomer were 0. 171 4 - 1.36 and 0. 161 3 - 1.28 μg · mL -1 , the detection limits were 39. 75 and 40. 90 ng · mL-1 and the limits of quantification were 141.6 and 136. 4 ng · mL-1 , respectively. The specificity of the method was good, the RSD values of the precision and stability tests were less than 1.2%, and the recoveries were greater than 98%. The average contents of calcitriol capsules from three manufacturers were 72. 71%, 80. 35%, and 78.18%, respectively. CONCLUSION The method is simple, rapid, accurate, and easy to be popularized, which is suitable for the determination of calcitriol and trans-calcitriol in calcitriol capsules.