Mauri Lucio, Boccardi Giovanni, Torri Giangiacomo, Karfunkle Michael, Macchi Eleonora, Muzi Laura, Keire David, Guerrini Marco
Institute for Chemical and Biochemical Research G. Ronzoni, via G. Colombo 81, 20133 Milan, Italy.
Division of Pharmaceutical Analysis, Office of Testing and Research, Center for Drug Evaluation and Research, U.S. Food and Drug Administration, 645 S. Newstead Ave., St. Louis, MO 63110, USA.
J Pharm Biomed Anal. 2017 Mar 20;136:92-105. doi: 10.1016/j.jpba.2016.12.031. Epub 2016 Dec 26.
An NMR HSQC method has recently been proposed for the quantitative determination of the mono- and disaccharide subunits of heparin and low molecular weight heparins (LMWH). The focus of the current study was the validation of this procedure to make the 2D-NMR method suitable for pharmaceutical quality control applications. Pre-validation work investigated the effects of several experimental parameters to assess robustness and to optimize critical factors. Important experimental parameters were pulse sequence selection, equilibration interval between pulse trains and temperature. These observations were needed so that the NMR method was sufficiently understood to enable continuous improvement. A standard validation study on heparin then examined linearity, repeatability, intermediate precision and limits of detection and quantitation; selected validation parameters were also determined for LMWH.
最近有人提出了一种核磁共振异核单量子相干（NMR HSQC）方法，用于定量测定肝素和低分子肝素（LMWH）的单糖和二糖亚基。本研究的重点是对该方法进行验证，以使二维核磁共振方法适用于药物质量控制应用。验证前的工作研究了几个实验参数的影响，以评估稳健性并优化关键因素。重要的实验参数包括脉冲序列选择、脉冲串之间的平衡间隔和温度。需要这些观察结果，以便充分理解核磁共振方法，从而实现持续改进。然后对肝素进行了标准验证研究，考察了线性、重复性、中间精密度以及检测限和定量限；还为低分子肝素确定了选定的验证参数。
