Huang Qianyang, Zhou Xiang, Liu Danting, Xin Baozhong, Cechner Karen, Wang Heng, Zhou Aimin
Clinical Chemistry Program, Department of Chemistry, Cleveland State University, Cleveland, OH 44115, USA.
Clinical Chemistry Program, Department of Chemistry, Cleveland State University, Cleveland, OH 44115, USA.
Anal Biochem. 2014 Jun 15;455:26-34. doi: 10.1016/j.ab.2014.03.014. Epub 2014 Mar 27.
Gangliosides are a family of glycosphingolipids characterized by mono- or polysialic acid-containing oligosaccharides linked through 1,3- and 1,4-β glycosidic bonds with subtle differences in structure that are abundantly present in the central nervous systems of many living organisms. Their cellular surface expression and physiological malfunction are believed to be pathologically implicated in considerable neurological disorders, including Alzheimer and Parkinson diseases. Recently, studies have tentatively elucidated that mental retardation or physical stagnation deteriorates as the physiological profile of gangliosides becomes progressively and distinctively abnormal during the development of these typical neurodegenerative syndromes. In this work, a reverse-phase liquid chromatography/tandem mass spectrometry (LC/MS/MS) assay using standard addition calibration for determination of GM2, GM3, GD2, and GD3 in human plasma has been developed and validated. The analytes and internal standard were extracted from human plasma using a simple protein precipitation procedure. Then the samples were analyzed by reverse-phase ultra-performance liquid chromatography (UPLC)/MS/MS interfaced to mass spectrometry with electrospray ionization using a multiple reaction monitoring mode to obtain superior sensitivity and specificity. This assay was validated for extraction recovery, calibration linearity, precision, and accuracy. Our quick and sensitive method can be applied to monitor ganglioside levels in plasma from normal people and neurodegenerative patients.
神经节苷脂是一类糖鞘脂，其特征在于通过1,3 - 和1,4 - β糖苷键连接的含单唾液酸或多唾液酸的寡糖，结构上存在细微差异，在许多生物体的中枢神经系统中大量存在。它们的细胞表面表达和生理功能紊乱被认为在包括阿尔茨海默病和帕金森病在内的多种神经疾病的病理过程中起作用。最近，研究初步阐明，在这些典型神经退行性综合征的发展过程中，随着神经节苷脂的生理特征逐渐且明显异常，智力发育迟缓或身体发育停滞会加剧。在这项工作中，已开发并验证了一种使用标准加入法校准的反相液相色谱/串联质谱（LC/MS/MS）分析法，用于测定人血浆中的GM2、GM3、GD2和GD3。使用简单的蛋白沉淀程序从人血浆中提取分析物和内标。然后通过反相超高效液相色谱（UPLC）/ MS/MS与质谱联用，采用电喷雾电离和多反应监测模式对样品进行分析，以获得卓越的灵敏度和特异性。该分析法在提取回收率、校准线性、精密度和准确度方面进行了验证。我们快速灵敏的方法可用于监测正常人和神经退行性疾病患者血浆中的神经节苷脂水平。
