Liu Yang, Chen Xi, Yuan Song, Liu Wanhui, He Lan, Zhang Qingsheng
National Institutes for Food and Drug Control, Beijing 102629, China.
YanTai University, Shandong 264005, China.
J Anal Methods Chem. 2020 Feb 7;2020:6201656. doi: 10.1155/2020/6201656. eCollection 2020.
Application of liquid chromatography-mass spectrometry (LC-MS) in analyzing the content of alfacalcidol tablets dissolution faces big challenges due to the low amount of alfacalcidol in each tablet and the low ionization efficacy of the compound with electrospray ionization (ESI) or atmospheric-pressure chemical ionization (APCI). Here, extraction, derivatization, and LC-MS quantitation method have been developed and validated for measuring alfacalcidol tablets dissolution content. After alfacalcidol dissolution solution was extracted with dichloromethane to remove surfactant and inorganic salts, alfacalcidol was then derivatized via a Cookson reagent, 4-phenyl-1, 2, 4-triazoline-3, 5-dione (PTAD), under ambient conditions. Alfacalcidol derivative was successfully analyzed by LC-MS. Limit of detection (LOD) of the derivatized alfacalcidol was improved 100 times (0.01 g/mL) compared with the nontreated compound (1 g/mL). The new method was then validated following International Conference on Harmonization (ICH) guidance. The method shows a good linearity with > 0.99. Interday and intraday reproducibility was 3.3% and 7.9%, respectively. This procedure can be used in quantification of alfacalcidol tablets dissolution content and corresponding pharmaceutical quality control.
由于阿法骨化醇片剂中每片的含量较低，且该化合物采用电喷雾电离（ESI）或大气压化学电离（APCI）时的电离效率较低，液相色谱-质谱联用（LC-MS）在分析阿法骨化醇片剂溶出度含量方面面临巨大挑战。在此，已开发并验证了用于测定阿法骨化醇片剂溶出度含量的提取、衍生化和LC-MS定量方法。用二氯甲烷萃取阿法骨化醇溶出液以去除表面活性剂和无机盐后，在环境条件下，阿法骨化醇通过库克森试剂4-苯基-1,2,4-三唑啉-3,5-二酮（PTAD）进行衍生化。阿法骨化醇衍生物通过LC-MS成功分析。与未处理的化合物（1 μg/mL）相比，衍生化阿法骨化醇的检测限（LOD）提高了100倍（0.01 μg/mL）。然后按照国际协调会议（ICH）指南对新方法进行验证。该方法显示出良好的线性关系，r²>0.99。日间和日内重现性分别为3.3%和7.9%。该方法可用于阿法骨化醇片剂溶出度含量的定量分析及相应的药品质量控制。
