Jaishetty Nagadeep, Palanisamy Kamaraj, Maruthapillai Arthanareeswari, Jaishetty Rajamanohar
Department of chemistry, SRM University, Kattankulathur, Channai-603203, Tamilnadu, India.
Department of chemistry, Osmania University, Hyderabad, Telanagana-500007, India.
Sci Pharm. 2015 Oct 18;84(3):456-466. doi: 10.3390/scipharm84030456.
Efavirenz is a non-nucleoside reverse transcriptase inhibitor used in the treatment of human immunodeficiency virus type-1 (HIV). (2S)-(2-Amino-5-chlorophenyl)-4-cyclopropyl-1,1,1-trifluoro-3-butyn-2-ol (AMCOL), used as an intermediate in the synthesis of efavirenz and a degradation impurity, has an aminoaryl derivative which is a well-known alerting function for genotoxic activity. Upon request from a regulatory agency, a selective and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for trace level quantitative determination of AMCOL related compound of efavirenz, for a risk assessment and comparison of impurity levels with the commercially available innovator product (brand name: Sustiva). The method provided excellent sensitivity at a typical target analyte level of <2.5 ppm, an established threshold of toxicological concern (TTC), when the drug substance and drug product samples were prepared at 15.0 mg/mL. The AMCOL sample was analyzed on a Luna C18 (2) (100 mm × 4.6 mm, 3 µm) column interfaced with a triple quadrupole tandem mass spectrometer operated in a multiple reaction monitoring (MRM) mode. Positive electrospray ionization (ESI) was employed as the ionization source and the mobile phase used was 5.0 mM ammonium acetate-methanol (35:65, v/v). The calibration curve showed good linearity over the concentration range of 0.2-5.0 ppm with a correlation coefficient of >0.999. The limit of detection (LOD) and limit of quantification (LOQ) were found to be 0.07 and 0.2 ppm, respectively. The developed method was validated as per international council on harmonization (ICH) guidelines in terms of LOD, LOQ, linearity, precision, accuracy, specificity, and robustness.
依非韦伦是一种用于治疗人类免疫缺陷病毒1型（HIV）的非核苷类逆转录酶抑制剂。(2S)-(2-氨基-5-氯苯基)-4-环丙基-1,1,1-三氟-3-丁炔-2-醇（AMCOL）用作依非韦伦合成的中间体和降解杂质，其氨基芳基衍生物具有众所周知的遗传毒性警示功能。应监管机构要求，开发了一种选择性和灵敏的液相色谱-串联质谱（LC-MS/MS）方法，用于痕量水平定量测定依非韦伦的AMCOL相关化合物，以进行风险评估并将杂质水平与市售创新产品（商品名：施多宁）进行比较。当原料药和制剂样品以15.0 mg/mL制备时，该方法在典型目标分析物水平<2.5 ppm（已确定的毒理学关注阈值（TTC））下具有出色的灵敏度。AMCOL样品在Luna C18(2)（100 mm×4.6 mm，3 µm）柱上进行分析，该柱与以多反应监测（MRM）模式运行的三重四极杆串联质谱仪相连。采用正电喷雾电离（ESI）作为电离源，所用流动相为5.0 mM醋酸铵-甲醇（35:65，v/v）。校准曲线在0.2 - 5.0 ppm浓度范围内显示出良好的线性，相关系数>0.999。检测限（LOD）和定量限（LOQ）分别为0.07和0.2 ppm。所开发的方法按照国际协调理事会（ICH）指南在LOD、LOQ、线性、精密度、准确度、特异性和稳健性方面进行了验证。
