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 μg/mL) compared with the nontreated compound (1 μg/mL). The new method was then validated following International Conference on Harmonization (ICH) guidance. The method shows a good linearity with r2 > 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.

中文翻译：

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通过化学衍生化和LC-MS定量测定阿法骨化醇片剂的溶解含量。

液相色谱-质谱法（LC-MS）在分析阿法骨化醇片溶出度中的应用面临着巨大的挑战，因为每片中阿法骨化醇的含量低，并且该化合物在电喷雾电离（ESI）或常压下的电离效率低。高压化学电离（APCI）。在这里，提取，衍生化和LC-MS定量方法已经开发和验证用于测量阿法骨化醇片的溶出度。用二氯甲烷萃取阿法骨化醇溶解溶液以除去表面活性剂和无机盐后，阿法骨化醇然后在环境条件下通过Cookson试剂4-苯基-1、2、4-三唑啉-3、5-二酮（PTAD）衍生化。用LC-MS成功地分析了阿法骨化醇衍生物。与未处理的化合物（1μg/ mL）相比，衍生的阿法骨化醇的检出限（LOD）提高了100倍（0.01μg/ mL）。然后，根据国际协调会议（ICH）的指导对新方法进行了验证。该方法显示出良好的线性，r2> 0.99。日间和日内重现性分别为3.3％和7.9％。该程序可用于阿法骨化醇片的溶出度定量和相应的药物质量控制。