Wang Jennie, Liu Mingtao, Yang Chun, Wu Xiaogang, Wang Euphemia, Liu Paul
SRI International, 333 Ravenswood Avenue, Menlo Park, CA 94025, USA.
SRI International, 333 Ravenswood Avenue, Menlo Park, CA 94025, USA.
J Pharm Biomed Anal. 2016 May 30;124:267-273. doi: 10.1016/j.jpba.2016.02.036. Epub 2016 Feb 27.
An HPLC method for the assay of a DNA topoisomerase inhibitor, LMP776 (NSC 725776), has been developed and validated. The stress testing of LMP776 was carried out in accordance with International Conference on Harmonization (ICH) guidelines Q1A (R2) under acidic, alkaline, oxidative, thermolytic, and photolytic conditions. The separation of LMP776 from its impurities and degradation products was achieved within 40 min on a Supelco Discovery HS F5 column (150 mm × 4.6 mm i.d., 5 μm) with a gradient mobile phase comprising 38-80% acetonitrile in water, with 0.1% trifluoroacetic acid in both phases. LC/MS was used to obtain mass data for characterization of impurities and degradation products. One major impurity was isolated through chloroform extraction and identified by NMR. The proposed HPLC assay method was validated for specificity, linearity (concentration range 0.25-0.75 mg/mL, r = 0.9999), accuracy (recovery 98.6-100.4%), precision (RSD ≤ 1.4%), and sensitivity (LOD 0.13 μg/mL). The validated method was used in the stability study of the LMP776 drug substance in conformance with the ICH Q1A (R2) guideline.
已开发并验证了一种用于测定DNA拓扑异构酶抑制剂LMP776（NSC 725776）的高效液相色谱法。LMP776的强制降解试验按照国际协调会议（ICH）Q1A（R2）指南在酸性、碱性、氧化、热解和光解条件下进行。在Supelco Discovery HS F5柱（150 mm×4.6 mm内径，5μm）上，使用流动相为水和乙腈的梯度洗脱（乙腈含量38%-80%，两相均含0.1%三氟乙酸），40分钟内实现了LMP776与其杂质及降解产物的分离。采用液相色谱-质谱联用（LC/MS）获取杂质和降解产物的质谱数据以进行表征。通过氯仿萃取分离出一种主要杂质，并通过核磁共振（NMR）进行了鉴定。所提出的高效液相色谱分析方法在特异性、线性（浓度范围0.25 - 0.75 mg/mL，r = 0.9999）、准确度（回收率98.6 - 100.4%）、精密度（相对标准偏差≤1.4%）和灵敏度（检测限0.13μg/mL）方面均得到了验证。经验证的方法用于按照ICH Q1A（R2）指南对LMP776原料药进行稳定性研究。
