色谱

色谱 ›› 2014, Vol. 32 ›› Issue (11): 1197-1200.DOI: 10.3724/SP.J.1123.2014.07002

• 研究论文 • 上一篇    下一篇

液相色谱-串联质谱法检测人血浆及脂蛋白唾液酸的含量

郭守东, 桑慧, 杨娜娜, 阚玉杰, 李富裕, 李煜, 李方圆, 秦树存   

  1. 山东省高校动脉粥样硬化重点实验室, 泰山医学院, 山东 泰安 271000
  • 收稿日期:2014-07-02 修回日期:2014-08-14 出版日期:2014-11-08 发布日期:2014-10-31
  • 通讯作者: 秦树存,E-mail:shucunqin@hotmail.com.
  • 基金资助:

    国家自然科学基金(31300639,81170785);山东省青年基金项目(ZR2013HQ014);山东省中医药科学技术研究项目(2013-257);泰山医学院博士启动基金项目(2234).

Measurement of sialic acid from lipoproteins and human plasma by liquid chromatography-tandem mass spectrometry

GUO Shoudong, SANG Hui, YANG Nana, KAN Yujie, LI Fuyu, LI Yu, LI Fangyuan, QIN Shucun   

  1. Key Laboratory of Atherosclerosis in Universities of Shandong, Taishan Medical University, Taian 271000, China
  • Received:2014-07-02 Revised:2014-08-14 Online:2014-11-08 Published:2014-10-31

PDF

217

被引次数

9

摘要：

建立了液相色谱-串联质谱(LC-MS/MS)检测人血浆脂蛋白唾液酸的方法,并比较了糖尿病患者与健康人血浆脂蛋白唾液酸的差异。采用pH=2的醋酸水解唾液酸,高速离心后采用优化的条件进行LC-MS/MS分析。结果表明:在负离子模式下,唾液酸的检出限和定量限分别为7.4和24.5 pg。唾液酸在2.5~80 ng/mL范围内呈良好的线性关系,相关系数R2大于0.998。糖尿病患者(平均年龄51.6岁)和健康人(平均年龄50.7岁)血浆中唾液酸的含量分别为(548.3±88.9)和(415.3±55.5)mg/L;两组实验人群中极低密度脂蛋白、低密度脂蛋白和高密度脂蛋白唾液酸的含量分别为(4.91±0.19)、(6.95±0.28)、(3.61±0.22)μg/mg和(2.90±0.27)、(7.03±0.04)、(2.40±0.09)μg/mg。糖尿病患者极低密度脂蛋白和高密度脂蛋白唾液酸含量显著高于同龄健康人水平(P<0.01)。该法可快速检测血浆中脂蛋白唾液酸含量,省时省力。

关键词: 人血浆, 糖尿病, 唾液酸, 液相色谱-串联质谱, 脂蛋白

Abstract:

A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was established to quantify sialic acid (N-acetylneuraminic acid, NANA) from lipoproteins and human plasma. The method was used to investigate the different contents of NANA from lipoproteins between diabetic with an average age of 51.6 years and healthy participants with an average age of 50.7 years. The NANA from lipoprotein samples was hydrolyzed by acetic acid (pH=2) at 80 ℃ for 2 h and analyzed by the optimized LC-MS/MS method after high speed centrifugation and filtration. The limits of detection and quantification of NANA were 7.4 and 24.5 pg, respectively. The linear range was 2.5-80 ng/mL for NANA and the correlation coefficient (R2) was more than 0.998. The levels of NANA in the plasma of type Ⅱ diabetics and healthy participants were (548.3±88.9) and (415.3±55.5) mg/L, respectively; and the levels of NANA from very low density lipoproteins (VLDL), low density lipoproteins (LDL), and high density lipoproteins (HDL) of the type Ⅱ diabetics and the healthy participants were (4.91±0.19), (6.95±0.28), (3.61±0.22) μg/mg and (2.90±0.27), (7.03±0.04), (2.40±0.09) μg/mg, respectively. The sialic acid levels of VLDL and HDL from the type Ⅱ diabetics were markedly higher than those of the corresponding healthy participants with the similar ages (P<0.01). The method can rapidly determine the sialic acid from lipoproteins, and is reproducible and time saving.

Key words: diabetes mellitus, human plasma, lipoprotein, liquid chromatography-tandem mass spectrometry (LC-MS/MS), sialic acid

中图分类号: