| Title: | Application of heart-cutting two-dimensional liquid chromatography-mass spectrometry to the characterization of highly polar impurities in calcium gluconate injection |
| Author(s): | Gu X; Yang L; Tao Q; Ai J; Yan C; Zheng J; Hong L; |
| Address: | "Zhejiang Institute for Food and Drug Control (Key Laboratory of Core Technology for Generic Drug Evaluation, China Drug Administration), Hangzhou, Zhejiang, China. Green Pharmaceutical Collaborative Innovation Center of Yangtze River Delta Region & College of Pharmaceutical Science, Zhejiang University of Technology, Zhejiang, China. Zhejiang Institute for Food and Drug Control (Key Laboratory of Core Technology for Generic Drug Evaluation, China Drug Administration), Hangzhou, Zhejiang, China. Electronic address: zjq@zjyj.org.cn. Zhejiang Institute for Food and Drug Control (Key Laboratory of Core Technology for Generic Drug Evaluation, China Drug Administration), Hangzhou, Zhejiang, China. Electronic address: hongliya@zjyj.org.cn" |
| DOI: | 10.1016/j.chroma.2022.463632 |
| ISSN/ISBN: | 1873-3778 (Electronic) 0021-9673 (Linking) |
| Abstract: | "The separation and characterization of small polar impurities in polar drugs such as calcium gluconate products are always challenging, due to their poor retention on traditional reversed phase (RP) columns. Although ion-pair reversed-phase liquid chromatography (IPRP-LC) and hydrophilic interaction liquid chromatography (HILIC) are commonly used methods for polar compound analysis, both methods have some drawbacks. For example, IPRP-LC is incompatible with mass spectrometry (MS) due to the presence of non-volatile salts in its mobile phase and HILIC has limited sensitivity due to the poor solubility of polar drugs in the organic-rich sample diluents used in HILIC separations. In order to characterize the highly polar impurities in calcium gluconate injections, a heart-cutting two-dimensional liquid chromatography (2D-LC) method coupled with quadrupole time-of-flight mass spectrometry (Q-TOF/MS) was developed in this study. An IPRP-LC method in the first dimension ((1)D) provided the selectivity for the separation of polar analytes, using a 100% aqueous mobile phase containing phosphate buffer and ion-pair reagent. Heart cuts of target peaks were collected with sample loops and transferred to the second dimension ((2)D) HILIC column using an organic-rich mobile phase. In order to solve the mobile phase mismatch problem between the two dimensions, a make-up flow module was introduced in the 2D-LC system to dilute the (1)D-water-rich fractions with acetonitrile before entering the sample loops. By optimizing the loop size and dilution factor, good retention and peak shape of the highly polar impurities were obtained on the (2)D-HILIC column, and the ion suppression effect for MS detection from the ion-pair reagent and non-volatile salt in the (1)D-effluent was minimized. A total of five impurities were identified through fragmentation studies by Q-TOF/MS analysis and their fragmentation pathways were proposed. Four of them were further confirmed by reference substances. This study not only provided useful information for quality control of calcium gluconate injections, but also provided an alternative method for polar impurity characterization in pharmaceuticals" |
| Keywords: | "*Calcium Gluconate Chromatography, Liquid/methods Mass Spectrometry/methods *Chromatography, Reverse-Phase/methods Hydrophobic and Hydrophilic Interactions Water/chemistry Calcium gluconate injection Heart-cutting 2D-LC Highly polar impurity Iprp-hilic-ms;" |
| Notes: | "MedlineGu, Xiao Yang, Lishi Tao, Qiaofeng Ai, Jie Yan, Chenxiao Zheng, Jinqi Hong, Liya eng Netherlands 2022/11/09 J Chromatogr A. 2022 Dec 6; 1685:463632. doi: 10.1016/j.chroma.2022.463632. Epub 2022 Nov 1" |
