Department of Pharmaceutical Science, Beijing Institute of Radiation Medicine, Beijing 100850, China.
China Z. Pharmaceutical Productivity Centre, Beijing 102600, China.
Int J Biol Macromol. 2019 Jan;121:89-96. doi: 10.1016/j.ijbiomac.2018.09.194. Epub 2018 Sep 30.
Chitosan has multiple biological activities, but a sensitive and rapid characterization method is yet to be developed for its further application. This study presented an optional mass spectrometry method for the characterization of chitosan. Nine kinds of chitosan (degree of deacetylation (DD), 63.08%-89.06% & MW, 106.1-485.0 kDa) were adopted for the method development. Most species of chitosan, detected by an ESI-MS technique, were observed below 1000 m/z, which seemed that only chito-oligosaccharide (COS) impurities were detected. Then, a sensitive UPLC-ESI-MS/MS method was established to assess the COS impurities in chitosan, and no COS impurities were detected. However, dissociation of chitosan and COSs in the ESI source were observed, and then the mass spectra patterns were deeply evaluated via an accurate Q-TOF mass spectrometer. Our research demonstrated that the mass spectra of COSs and chitosan resulted from the dissociation of glycosidic linkage and dehydration. Although the distribution of GlcN and GlcNAc units in these chitosan samples might be different, similar dissociation efficiencies were observed. Furthermore, good linearities were obtained between the intensities of product ions, detected by an UPLC-pseuedo-MS2 method, and DDs determined by conventional method. This method could be suitable for the DD determination and quantitative analysis of chitosan.
壳聚糖具有多种生物学活性,但仍需要开发一种敏感且快速的表征方法,以进一步应用。本研究提出了一种可选的质谱法用于壳聚糖的表征。采用了 9 种壳聚糖(脱乙酰度(DD),63.08%-89.06%&MW,106.1-485.0 kDa)进行方法开发。通过 ESI-MS 技术检测到的大多数壳聚糖物种的分子量低于 1000 m/z,似乎仅检测到壳寡糖(COS)杂质。然后,建立了一种灵敏的 UPLC-ESI-MS/MS 方法来评估壳聚糖中的 COS 杂质,但未检测到 COS 杂质。然而,在 ESI 源中观察到了壳聚糖和 COSs 的解离,然后通过精确的 Q-TOF 质谱仪对质谱图谱进行了深入评估。我们的研究表明,COSs 和壳聚糖的质谱图谱是由糖苷键的解离和脱水产生的。尽管这些壳聚糖样品中 GlcN 和 GlcNAc 单元的分布可能不同,但观察到相似的解离效率。此外,通过 UPLC-pseuedo-MS2 方法检测到的产物离子强度与通过常规方法测定的 DD 之间呈良好的线性关系。该方法可适用于 DD 测定和壳聚糖的定量分析。