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

贺平; 张利; 张奎虎; 朱美容

doi:10.13898/j.cnki.issn.1000-2200.2015.08.036

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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

贺平 , 张利 , 张奎虎 , 朱美容

文章导航 > 蚌埠医学院学报 > 2015, 40 > 8: (1093-1096)

引用本文:

Citation:

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

安徽省阜阳市疾病预防控制中心, 236000

作者简介: 贺平(1976-),女,主管技师.

中图分类号: R562

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

The Center for Disease Control and Prevention in Fuyang, Fuyang Anhui 236000, China
CLC number: R562

摘要: 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(VitA)水平的方法,以了解学龄儿童VitA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100 μl血清加入100 μl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,200 μl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,1.7 μm);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2 μl。结果:360例学龄儿童血清VitA含量在0.072 8～0.728 μg/ml范围内呈良好的线性关系(r>0.999),VitA的平均值为39.60 μg/dl,CV<5%,VitA的最低检测限为0.020 μg/ml,测定在4.0 min内完成。360例学龄儿童中VitA含量≤20 μg/dl占总人数的1.11%,VitA水平20～30 μg/dl占总人数的19.17%,VitA水平≥30 μg/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VitA的水平,方法快速、灵敏、准确,重复性好,样品处理简单宜行。
- 维生素A /
- 反相超高效液相色谱 /
- 内标法
Abstract: 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 than 20 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.
- vitamin A /
- reversed-phase ultra performance liquid chromatography /
- internal standard

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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

作者简介: 贺平(1976-),女,主管技师.

安徽省阜阳市疾病预防控制中心, 236000

收稿日期: 2015-04-13

关键词:

摘要: 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(VitA)水平的方法,以了解学龄儿童VitA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100 μl血清加入100 μl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,200 μl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,1.7 μm);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2 μl。结果:360例学龄儿童血清VitA含量在0.072 8～0.728 μg/ml范围内呈良好的线性关系(r>0.999),VitA的平均值为39.60 μg/dl,CV<5%,VitA的最低检测限为0.020 μg/ml,测定在4.0 min内完成。360例学龄儿童中VitA含量≤20 μg/dl占总人数的1.11%,VitA水平20～30 μg/dl占总人数的19.17%,VitA水平≥30 μg/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VitA的水平,方法快速、灵敏、准确,重复性好,样品处理简单宜行。

English Abstract

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

The Center for Disease Control and Prevention in Fuyang, Fuyang Anhui 236000, China

Received Date: 2015-04-13

Abstract: 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 than 20 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.