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

发布时间：2018-05-10 16:34

本文选题：稳定同位素稀释 + 高效液相色谱　；参考：《色谱》2016年05期

【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26.4μg/L,基本满足口服1 mg2H8-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。
[Abstract]:A stable isotope dilution technique combined with high performance liquid chromatography-gas chromatography / negative chemical ionization source mass spectrometry (HPLC-GC / NCI / MS) was developed for the determination of vitamin A (VA) reserves in human liver. After oral administration of deuterium labeled retinol acetate 2H 8-RAC, blood samples were collected and serum samples were isolated after 21 days of stable phase. The serum was extracted by n-hexane, separated and purified by HPLC, then dried by nitrogen for further derivation, the derivative products were detected by GC/NCI/MS, and the abundance ratio of labeled and unlabeled VA was obtained. Finally, the hepatic reserves of VA were calculated by Furr-Olson formula. Under the optimized conditions, the recovery rate of VA in serum samples was more than 85um and the precision was less than 100.The quantitative limit was 26.4 渭 g / L, which basically met the requirements of labeled and unlabeled VA after oral administration of 1 mg2H8-RAC. Compared with the current human VA evaluation method in China, this method can reflect the nutritional level of human VA more objectively. Combined with the VA intervention experiment, the dietary intake level of a certain population to maintain a stable level of VA reserves in a certain population can be obtained. At the same time, the application of stable isotope tracer technology plays an important role in promoting the biotransformation efficiency of oral VA sources in vivo.
【作者单位】：南京医科大学公共卫生学院卫生检验学系;南京医科大学公共卫生学院儿少与妇幼保健学系;
【基金】：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)~~
【分类号】：R446.1;O657.63

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