Wang George, Tomasella Frank P
Analytical and Bioanalytical Development, Pharmaceutical Development, Bristol-Myers Squibb Company, New Brunswick, NJ 08903, USA.
J Pharm Anal. 2016 Jun;6(3):150-156. doi: 10.1016/j.jpha.2016.01.002. Epub 2016 Jan 21.
Ion-pairing high-performance liquid chromatography-ultraviolet (HPLC-UV) methods were developed to determine two commonly used chelating agents, ethylenediaminetetraacetic acid (EDTA) in Abilify® (a small molecule drug with aripiprazole as the active pharmaceutical ingredient) oral solution and diethylenetriaminepentaacetic acid (DTPA) in Yervoy® (a monoclonal antibody drug with ipilimumab as the active pharmaceutical ingredient) intravenous formulation. Since the analytes, EDTA and DTPA, do not contain chromophores, transition metal ions (Cu, Fe) which generate highly stable metallocomplexes with the chelating agents were added into the sample preparation to enhance UV detection. The use of metallocomplexes with ion-pairing chromatography provides the ability to achieve the desired sensitivity and selectivity in the development of the method. Specifically, the sample preparation involving metallocomplex formation allowed sensitive UV detection. Copper was utilized for the determination of EDTA and iron was utilized for the determination of DTPA. In the case of EDTA, a gradient mobile phase separated the components of the formulation from the analyte. In the method for DTPA, the active drug substance, ipilimumab, was eluted in the void. In addition, the optimization of the concentration of the ion-pairing reagent was discussed as a means of enhancing the retention of the aminopolycarboxylic acids (APCAs) including EDTA and DTPA and the specificity of the method. The analytical method development was designed based on the chromatographic properties of the analytes, the nature of the sample matrix and the intended purpose of the method. Validation data were presented for the two methods. Finally, both methods were successfully utilized in determining the fate of the chelates.
开发了离子对高效液相色谱 - 紫外（HPLC - UV）方法，以测定两种常用螯合剂，即阿立哌唑口服溶液（商品名：阿立哌唑，活性药物成分为阿立哌唑的小分子药物）中的乙二胺四乙酸（EDTA）和Yervoy®（商品名：易普利姆玛，活性药物成分为伊匹单抗的单克隆抗体药物）静脉制剂中的二乙烯三胺五乙酸（DTPA）。由于分析物EDTA和DTPA不含发色团，因此在样品制备中加入与螯合剂形成高度稳定金属配合物的过渡金属离子（Cu、Fe），以增强紫外检测。金属配合物与离子对色谱联用，使得在方法开发中能够实现所需的灵敏度和选择性。具体而言，涉及金属配合物形成的样品制备实现了灵敏的紫外检测。测定EDTA时使用铜，测定DTPA时使用铁。对于EDTA，梯度流动相将制剂成分与分析物分离。在DTPA的测定方法中，活性药物成分伊匹单抗在死体积处洗脱。此外，还讨论了离子对试剂浓度的优化，以此作为增强包括EDTA和DTPA在内的氨基多羧酸（APCA）保留率及方法特异性的一种手段。基于分析物的色谱性质、样品基质的性质以及方法的预期用途设计了分析方法开发。给出了这两种方法的验证数据。最后，两种方法均成功用于测定螯合物的去向。
