高等学校化学学报

高等学校化学学报 ›› 2020, Vol. 41 ›› Issue (2): 262.doi: 10.7503/cjcu20190483

• 分析化学 • 上一篇    下一篇

基于氧化镍沉积硅胶固相萃取与液相色谱-质谱联用技术的2型糖尿病血清中咪唑丙酸的检测

王天琦,余琼卫(),冯钰锜()   

  1. 生物医学分析化学教育部重点实验室, 武汉大学化学与分子科学学院, 武汉 430072
  • 收稿日期:2019-09-10 出版日期:2020-02-10 发布日期:2019-11-04
  • 通讯作者: 余琼卫,冯钰锜 E-mail:qwyu@whu.edu.cn;yqfeng@whu.edu.cn
  • 基金资助:
    国家自然科学基金资助(31671929);国家自然科学基金资助(31670373)

Analysis of Imidazole Propionic Acid in Serum of Patients with Type 2 Diabetes Based on NiO@SiO2 Solid-phase Extraction Coupled with Liquid Chromatography-Mass Spectrometry †

WANG Tianqi,YU Qiongwei(),FENG Yuqi()   

  1. Key Laboratory of Analytical Chemistry for Biology and Medicine, Ministry of Education, College of Chemistry and Molecular Sciences, Wuhan University, Wuhan 430072, China
  • Received:2019-09-10 Online:2020-02-10 Published:2019-11-04
  • Contact: Qiongwei YU,Yuqi FENG E-mail:qwyu@whu.edu.cn;yqfeng@whu.edu.cn
  • Supported by:
    ? Supported by the National Natural Science Foundation of China(31671929);Supported by the National Natural Science Foundation of China(31670373)

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摘要：

咪唑丙酸可以通过哺乳动物雷帕霉素靶蛋白复合物1(mTORC1)路径引起人的胰岛素抵抗, 从而导致2型糖尿病(T2D), 因此, 咪唑丙酸的准确定量可辅助2型糖尿病的诊断. 本文利用NiO与咪唑基团之间的配位作用, 采用氧化镍沉积硅胶(NiO@SiO2)萃取材料对咪唑丙酸进行选择性富集和萃取. 首先对NiO@SiO2 固相萃取(SPE)条件进行优化: 吸附剂用量为200 mg, 上样液为20 mmol/L的磷酸盐缓冲溶液(pH=3.0), 解吸液为1.0 mL含1%(质量分数)NH3·H2O的水溶液; 然后, 对萃取液进行高效液相色谱-质谱(HPLC-MS)分离分析, 建立了血清中咪唑丙酸的检测方法. 结果表明, 咪唑丙酸浓度在0.05~10 ng/mL范围内对质谱响应值具有良好的线性关系(R 2≥0.996), 检出限和定量限分别为0.02和0.05 ng/mL, 加标回收率为84.0%~119%, 相对标准偏差RSD<17.2%. 将该方法用于检测2型糖尿病患者与正常人血清样品中的咪唑丙酸, 发现在2型糖尿病患者与正常人的血清中咪唑丙酸含量存在显著性差异, 说明咪唑丙酸的准确定量对2型糖尿病具有医学诊断上的潜力.

关键词: 咪唑丙酸, NiO@SiO2, 2型糖尿病, 固相萃取, 高效液相色谱-质谱联用

Abstract:

Imidazole propionic acid can cause insulin resistance in human through mammalian target of rapamycin complex 1(mTORC1) pathway, leading to type 2 diabetes(T2D). Therefore, the accurate quantification of imidazole propionic acid benefits for the diagnosis of type 2 diabetes. Based on the coordination between NiO and imidazole groups, NiO@SiO2 was used for the selective extraction of imidazole propionic acid in serum. Firstly, the NiO@SiO2 solid phase extraction(SPE) conditions were optimized as follows: the amount of absorbent was 200 mg, the loading solution was 20 mmol/L of phosphate buffer(pH=3.0) and 1.0 mL of aqueous solution containing 1% NH3·H2O was used as the elution. Then, a simple and effective method for the analysis of imidazole propionic acid in serum was established by coupling NiO@SiO2 solid phase extraction with high performance liquid chromatography-mass spectrometry(HPLC-MS). The linearity of the developed method was in the range of 0.05—10 ng/mL with a good linearity(R 2≥0.996). The LOD and LOQ were 0.02 and 0.05 ng/mL, respectively. The recovery was 84.0%—119%, and the relative standard deviation(RSD) was less than 17.2%. The established method was applied to the detection of imidazole propionic acid in patients with type 2 diabetes and normal subjects. There was a significant difference in imidazole propionic acid content between type 2 diabetic patients and normal subjects, indicating that accurate quantification of imidazole propionic acid has great potential in medical diagnosis.

Key words: Imidazolepropionic acid, NiO@SiO2, Type 2 diabetes, Solid phase extraction(SPE), High performance liquid chromatography-mass spectrometry(HPLC-MS)

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