Devrukhakar Prashant S, Shiva Shankar M, Shankar G, Srinivas R
School of Advance Sciences, Vellore Institute of technology (VIT), Katpadi, Vellore, Tamil Nadu 632014, India.
National Centre for Mass Spectrometry, CSIR-Indian Institute of Chemical Technology, Hyderabad 500007, India.
J Pharm Anal. 2017 Aug;7(4):231-236. doi: 10.1016/j.jpha.2017.01.006. Epub 2017 Jan 18.
Zidvovudine (AZT) is a nucleoside analogue reverse transcriptase inhibitor (NRTI), a class of anti-retroviral drug. A stability-indicating assay method for AZT was developed in line with ICH guideline. Successful separation of AZT and its degradation products was achieved by gradient elution mode on reverse phase C column using 10 mM ammonium acetate: acetonitrile as the mobile phase at 0.8 mL/min flow rate, 25 µL injection volume, 30 °C column temperature and 285 nm detection wavelength. Two major acid degradation products were identified and characterized by liquid chromatography-electrospray ionization mass spectrometry (LC-ESI/MS/MS) and accurate mass measurements. The probable mechanisms for the formation of degradation products were identified based on a comparison of the fragmentation pattern of the [M + H] ions of AZT and its degradation products. One of the degradation products, DP-1, was isolated by semi-preparative high performance liquid chromatography (HPLC) using Waters XBridge Prep C (250 mm×10 mm, 5 µm). Degradation products showed higher toxicity compared to the drug in some models assessed by TOPKAT software. The method validation was performed with respect to robustness, specificity, linearity, precision and accuracy as per ICH guideline Q2 (R1).
齐多夫定（AZT）是一种核苷类似物逆转录酶抑制剂（NRTI），属于一类抗逆转录病毒药物。根据国际人用药品注册技术协调会（ICH）指南开发了一种齐多夫定的稳定性指示测定方法。采用梯度洗脱模式，以10 mM醋酸铵：乙腈为流动相，流速0.8 mL/min，进样量25 μL，柱温30°C，检测波长285 nm，在反相C柱上成功分离了齐多夫定及其降解产物。通过液相色谱-电喷雾电离质谱（LC-ESI/MS/MS）和精确质量测量鉴定并表征了两种主要的酸降解产物。基于对齐多夫定及其降解产物的[M + H]离子的碎裂模式的比较，确定了降解产物形成的可能机制。其中一种降解产物DP-1通过使用沃特世XBridge Prep C（250 mm×10 mm，5 µm）的半制备高效液相色谱（HPLC）进行分离。在TOPKAT软件评估的一些模型中，降解产物显示出比药物更高的毒性。根据ICH指南Q2（R1），对该方法的稳健性、特异性、线性、精密度和准确度进行了验证。
