In order to establish a NP-HPLC method for the determination of enantiomer in vildagliptin intermediate, the determination is carried out on the column of Chiralpak(R)AD-H(250 mm×4.6 mm, 5 μm), with the mobile phase of n-hexane, ethanol and methanol(volume ratio of 65∶25∶10)at flow rate of 0.8 mL/min.The sample volume is 10 μL, the wavelength is 210 nm and the column temperature is 35 ℃.The result shows that the vildagliptin intermediate and its enantiomer could be well separated and detected effectively;blank solvent doesn't interfere with the enantiomer assaying;the detection limit is 27 ng/mL and the quantification limit is 81 ng/mL;in repetitive test, the RSD of enantiomer assaying of samples are no more than 2.0%;in stability test, the RSD are no more than 2.0% in 12 h;the vildagliptin intermediate and its enantiomer could be well separated in the test of durability with all RSDs below 2.0%.The method is simple, reliable, accurate and durable, and can be used for determination of enantiomer in vildagliptin intermediate.%为建立维格列汀中间体对映异构体含量的测定方法,采用正相高效液相色谱法,色谱柱为Chiralpak(R)AD-H型手性色谱柱(250mm×4.6mm,5μm),流动相为正己烷-乙醇-甲醇(三者体积比为65:25:10),流速为0.8mL/min,进样体积为10μL,检测波长为210nm,柱温为35℃.实验得知:维格列汀中间体与其对映异构体的分离度良好,空白溶剂不干扰对映异构体的含量测定;检测限为27 ng/mL,定量限为81ng/mL;重复性试验中,供试品所含对映异构体含量的RSD值均小于2.0%;稳定性试验中,12h内对映异构体峰面积及含量的RSD值均小于2.0%;耐用性试验中,维格列汀中间体与其对映异构体峰可实现完全分离,并且不同参数下对映异构体含量的RSD值小于2.0%.结果表明,此方法专属性强,灵敏度高,重复性、耐用性良好,可用于维格列汀中间体对映异构体含量的测定.
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