超高效液相色谱内标法测定学龄儿童血清中维生素A水平

The detection of serum vitamin A level in school-age children

目的：建立反相超高效液相色谱（UPLC）内标法测定儿童血清中维生素A（Vit A）水平的方法,以了解学龄儿童Vit A营养状况,为制定膳食营养计划提供参考。方法：阜阳市颍上县学龄儿童100μl血清加入100μl内标后,振荡混匀,正己烷1.0 ml萃取后离心,取上清液用N2吹干,200μl无水乙醇复溶,UPLC法测定。色谱条件为色谱柱：Waters C18柱（2.1 mm×100 mm,1.7μm）;二级阵列管检测器;流动相：乙腈∶水（94∶6）临用前脱气;检测波长325 nm;柱温：40℃;流速：0.3 ml/min;进样2μl。结果：360例学龄儿童血清Vit A含量在0.072 8-0.728μg/ml范围内呈良好的线性关系（r〉0.999）,Vit A的平均值为39.60μg/dl,CV〈5%,Vit A的最低检测限为0.020μg/ml,测定在4.0 min内完成。360例学龄儿童中Vit A含量≤20μg/dl占总人数的1.11%,Vit A水平20-30μg/dl占总人数的19.17%,Vit A水平≥30μg/dl占总人数的79.72%。结论：调查结果总体满足膳食营养计划的要求。反相UPLC内标法检测儿童血清中Vit A的水平,方法快速、灵敏、准确,重复性好,样品处理简单宜行。

Objective： To establish the reversed-phase ultra performance liquid chromatography（ UPLC） with internal standard method for detecting the serum vitamin A level of children,in order to understand the school-age children vitamin A nutritional status,and provide reference in formulating the dietary nutrition plan. Methods： The 100 μl standard was added to the 100 ml serum of school aged children in Fuyang city Yingshang county,which was oscillated,mixed,extracted using 1. 0 ml of hexane extraction and centrifugated.The supernatant was dried with N2,redissoluting using 200 μl of anhydrous ethanol,and which was determined by UPLC. The chromatographic column was the chromatographic conditions,which included Waters C18 column（ 2. 1 mm × 100 mm,1. 7 μm）,secondary array detector; mobile phase,acetonitrile： water（ 96∶ 4） prodegassing before using. The detection wavelength was 325 nm,the column temperature was 40 ℃,the flow rate was 0. 3 ml / min and the adding sample was 2 μl. Results： The serum vitamin A concentration in 360 school-age children was 0. 072 8 to 0. 728 μg / ml,the range of which showed a good linear relationship（ r 0. 999）. The mean value of vitamin A was 39. 6 μg / dl,CV was less than 5%. The lowest detection limit of vitamin A was 0. 020 μg /ml,which could be detected within 4. 0 min. Among 360 school aged children,the children with vitamin A level equal to or less than20 μg / dl,equal to 30 μg / dl and more than or equal to 30 μg / dl were accounted for 1. 11%,19. 17% and 79. 72%,respectively.Conclusions： The overall survey results meet the requirements of dietary nutrition plan. The reversed-phase ultra performance liquid chromatography with internal standard method in detecting the serum vitamin A level of children is fast,sensitive,accurate and reproducible.