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 mg 2 H8-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。
An analytical method for the determination of vitamin A( VA)reserved in human liv-er by stable isotope dilution technique and high performance liquid chromatography-gas chro-matography/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 separa-ted. 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 col-lected,purified and detected. The detection precision( relative standard deviation,RSD,n=6) was less than 10% and the quantitative limit reached 26. 4 μg/L,which met the marked and unmarked VA detection requirements. Compared with the current domestic VA evaluation meth-od,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.
参考文献
| [1] | Furr HC;Green MH;Haskell M;Mokhtar N;Nestel P;Newton S;Ribaya-Mercado JD;Tang G;Tanumihardjo S;Wasantwisut E.Stable isotope dilution techniques for assessing vitamin A status and bioefficacy of provitamin A carotenoids in humans[J].Public Health Nutrition,20056(6):596-607. |
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