Department of Chemistry, School of Advanced Sciences, Vellore Institute of Technology (VIT), Vellore 632014, India.
Department of Chemistry, School of Advanced Sciences, Vellore Institute of Technology (VIT), Chennai 600127, India.
J Chromatogr B Analyt Technol Biomed Life Sci. 2024 Sep 15;1245:124275. doi: 10.1016/j.jchromb.2024.124275. Epub 2024 Aug 15.
Dolutegravir (DLG) has become a distinctive first-line antiretroviral therapy for the treatment of HIV in most countries due to its affordability, high efficacy, and low drug-drug interactions. However, the evaluation of genotoxic impurities (GTIs) in DLG and their toxicity assessment has not been explored thoroughly. Thus, in this study, a simple, fast, and selective analytical methodology was developed for the identification and determination of 7 GTIs in the comprehensive, explicit route of synthesis for the dolutegravir sodium (DLG-Na) drug. A facile, fast ultrasonication-assisted liquid-liquid extraction procedure was adapted to isolate the GTIs in DLG-Na and then analyzed using the gas chromatography (GC)-electron impact (EI)/mass spectrometer (MS) quantification (using selective ion monitoring mode) technique. This EI-GC/MS method was validated as per the current requirements of ICH Q2 (R1) guidelines. Under optimal method conditions, excellent linearities were achieved with R between 0.9959 and 0.9995, and high sensitivity was obtained in terms of detection limits (LOD) between 0.15 to 0.63 µg/g, and quantification limits (LOQ) between 0.45 to 1.66 µg/g for the seven GTIs in DLG. The obtained recoveries ranged from 98.2 to 104.3 % at LOQ, 15 µg/g, and 18 µg/g concentration levels (maximum daily dose of 100 mg). This developed and validated method is rapid, easy to adopt, specific, sensitive, and accurate in estimating the seven GTIs in a relatively complex sodium matrix of the DLG-Na drug moiety. As a method application, two different manufactured samples of DLG-Na drug substances were analyzed for the fate of the GTIs and drug safety for the intended dosage applications. Moreover, an in-silico QSAR toxicity prediction assessment was carried out to prove scientifically the potential GTI nature of each impurity from the alerting functional groups.
多替拉韦（DLG）由于其价格合理、疗效高、药物相互作用低，已成为大多数国家治疗 HIV 的一线抗逆转录病毒治疗药物。然而，DLG 中遗传毒性杂质（GTIs）的评估及其毒性评估尚未得到充分探讨。因此，在这项研究中，开发了一种简单、快速、选择性的分析方法，用于鉴定和测定多替拉韦钠（DLG-Na）药物综合、明确合成路线中的 7 种 GTIs。采用简便、快速的超声辅助液-液萃取程序分离 DLG-Na 中的 GTIs，然后使用气相色谱（GC）-电子轰击（EI）/质谱（MS）定量（采用选择离子监测模式）技术进行分析。该 EI-GC/MS 方法符合 ICH Q2（R1）指南的现行要求。在最佳方法条件下，7 种 GTIs 的线性关系良好，相关系数（R）在 0.9959 至 0.9995 之间，检测限（LOD）在 0.15 至 0.63 µg/g 之间，定量限（LOQ）在 0.45 至 1.66 µg/g 之间，灵敏度高。在 LOQ、15 µg/g 和 18 µg/g 浓度水平（100 mg 最大日剂量）下，回收率范围为 98.2%至 104.3%。该方法快速、易于采用、特异性强、灵敏度高、准确度高，可用于估计 DLG-Na 药物部分相对复杂的钠基质中的 7 种 GTIs。作为方法应用，分析了两种不同制造的 DLG-Na 药物样品，以了解 GTIs 的命运和预期剂量应用的药物安全性。此外，还进行了基于定量构效关系（QSAR）的毒性预测评估，从警示官能团的角度科学证明了每种杂质的潜在 GTI 性质。
