色谱

色谱 ›› 2016, Vol. 34 ›› Issue (5): 533-537.DOI: 10.3724/SP.J.1123.2015.12019

• 研究论文 • 上一篇    下一篇

稳定同位素稀释技术在人体肝脏维生素A储备量检测中的应用

王丽1, 邵莹莹2, 李磊1, 汪之顼2   

  1. 1. 南京医科大学公共卫生学院卫生检验学系, 江苏 南京 211166;
    2. 南京医科大学公共卫生学院儿少与妇幼保健学系, 江苏 南京 211166
  • 收稿日期:2015-12-17 出版日期:2016-05-08 发布日期:2012-12-18
  • 通讯作者: 汪之顼
  • 基金资助:

    国家自然科学基金项目(3097248,81172654);江苏省高校优势学科建设工程项目(PAPD2010).

Determination of vitamin A reserved in human liver by stable isotope dilution technique

WANG Li1, SHAO Yingying2, LI Lei1, WANG Zhixu2   

  1. 1. Department of Hygienic Analysis and Detection, School of Public Health, Nanjing Medical University, Nanjing 211166, China;
    2. Department of Maternal, Child and Adolescent Health, School of Public Health, Nanjing Medical University, Nanjing 211166, China
  • Received:2015-12-17 Online:2016-05-08 Published:2012-12-18
  • Supported by:

    Natural Science Foundations of China (Nos. 3097248, 81172654); Project of the Priority Academic Program Development of Jiangsu Higher Education Institutions (No. PAPD2010).

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被引次数

1

摘要：

建立了稳定同位素稀释技术结合高效液相色谱-气相色谱/负化学电离源质谱法(HPLC-GC/NCI/MS)测定人体肝脏维生素A(VA)储备量的方法。志愿者在口服氘标记的视黄醇醋酸酯(2H8-RAC)后,经21天的稳定期,采集血样,分离血清。血清经正己烷提取,高效液相色谱分离纯化后,氮气吹干供进一步衍生化,衍生化产物用GC/NCI/MS检测,获得标记与未标记VA的丰度比,最后利用Furr-Olson公式计算VA肝脏储备量。在优化的条件下,血清样本中VA的回收率大于85%,精密度(RSD, n=6)小于10%,定量限为26.4 μ g/L,基本满足口服1 mg 2H8-RAC后标记与未标记VA的检测要求。相比国内现行的人体VA评价方法,该方法能更客观地反映人体VA营养水平。该项测定技术与VA干预实验相结合,可获得某一人群维持体内VA稳定储备水平的膳食摄入量水平。同时,稳定同位素示踪技术的运用对促进经口摄入的VA源在体内生物转化效率的研究也起着重要作用。

关键词: 肝脏储备量, 高效液相色谱, 气相色谱/负化学电离源/质谱, 维生素A, 稳定同位素稀释

Abstract:

An analytical method for the determination of vitamin A (VA) reserved in human liver by stable isotope dilution technique and high performance liquid chromatography-gas chromatography/negative ion chemical ionization/mass spectrometry (HPLC-GC/NCI/MS) was developed. Before the test, deuterium labelled retinol acetate (2H8-RAC) (1 mg) was ingested by the volunteers. After 21 days, the blood samples were collected and the serum was separated. The VA was extracted by n-hexane and purified by HPLC. Then the purified VA was dried under nitrogen for further derivatization. The derivative was finally detected by GC/NCI/MS. The concentration of the VA in the liver was calculated by using the formula of Furr-Olson. Under the optimized conditions, more than 85% of the VA in the serum samples could be collected, purified and detected. The detection precision (relative standard deviation, RSD, n=6) was less than 10% and the quantitative limit reached 26.4 μ g/L, which met the marked and unmarked VA detection requirements. Compared with the current domestic VA evaluation method, this method can more objectively reflect human VA nutrition level. When this determination technology combines with VA intervention experiment, it could evaluate a dietary intake level of VA to maintain a stability reserve level. At the same time, it also plays an important role in the research on biological conversion efficiency of carotene ingestion.

Key words: gas chromatography/negative ion mode of chemical ionization/mass spectrometry (GC/NCI/MS), high performance liquid chromatography (HPLC), liver reserve, stable isotope dilution, vitamin A (VA)

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