Quantitative analysis of carbohydrate residues in Dextran 40 from various sources: A comparative study using high performance liquid chromatography coupled with charged aerosol detector

Abstract

Dextran 40, a typical high molecular weight carbohydrate drug refined through fermentation, is widely used in the clinical field in injectable form. The final product obtained through fermentation may contain by-products such as fructose and residual sucrose, which carry a risk of adverse reactions. Current quality standards do not effectively control for the possible presence of carbohydrate residues, and methods for detecting such residues are lacking. This gap in quality control is present in the vast majority of existing carbohydrate drugs. This study established and compared liquid chromatography methods equipped with three different detectors (RID, MS, CAD), selecting a convenient, rapid, and efficient HILIC-CAD method. This method combines the high sensitivity of HILIC-MS method with the high throughput of HILIC-RID method, using porous silica as the stationary phase and a highly-precision charged aerosol detector in tandem, achieving rapid separation and quantification of fructose and sucrose. Additionally, pre-treatment optimization was conducted to eliminate the impact of dextran 40 on the detection of fructose and sucrose. The method has been validated, showing good repeatability, recovery, robustness, and linearity, capable of quantifying carbohydrate residues at approximately 3.3 ppm. This study compared the residual levels of fructose and sucrose in dextran 40 obtained from different purification processes, analyzing key purification operations that influence the extent of carbohydrate residues. These findings provide a reference for optimizing the production process of dextran 40, ensuring the quality of the drug and public drug safety. Furthermore, the approach used in this study for detecting carbohydrate residues is applicable to the quality control of other carbohydrate drugs produced via fermentation.