摘要
目的建立高效液相色谱-多波长荧光法同时快速检测人血清中维生素A和不同构型维生素E含量的方法。方法血清样品加入内标维生素A醋酸酯后,加入乙腈沉淀蛋白,经正己烷萃取,一定量的正己烷层经氮气挥干,用甲醇定容后进样,采用WATERS Symmetry C18色谱柱(4.6 mm×250 mm,5μm)进行分离,甲醇等度洗脱,控制柱温30℃,使用荧光检测器在线变换波长的方式进行定量检测。结果维生素A、α-维生素E、β-维生素E和δ-维生素E分别在质量浓度为0.050~2.0、0.50~50、0.050~5.0和0.050~5.0μg/mL的范围内呈良好的线性关系(r≥0.996);维生素A和各构型维生素E的方法检出限均为0.020μg/mL;各组分的日内、日间相对标准偏差在高、中、低浓度下均小于3%;样品在3个浓度点的回收率为91.2%~107.5%,相对标准偏差均小于10%。结论方法简单准确,相比高效液相-紫外检测法灵敏度更高,可实现血清中维生素A和不同构型维生素E含量的同时测定,适合大量血清样本的快速检测。
OBJECTIVE To establish a method for the simultaneous and rapid determination of vitamin A and vitamin E of different configurations in human serum by high performance liquid chromatography(HPLC)with multi-wavelength fluorescence detector.METHODS The serum was mixed after adding internal standard,and acetonitrile was added for protein precipitation.The mixture was centrifuged after extraction with n-hexane.The n-hexane layer was dried by N2 flow,the residue was dissolved with methanol.The HPLC system was consisted of WATERS Symmetry C18 column(4.6 mm×250 mm,5μm)and the mobile phase was methanol.The column temperature was 30℃and fluorescence detector with online wavelength conversion method was carried out for the quantitative detection.RESULTS The liner range of determination of vitamin A,α-vitamin E,β-vitamin E andδ-vitamin E were 0.050-2.0μg/mL,0.50-50μg/mL,0.050-5.0μg/mL and 0.050-5.0μg/mL,respectively(r≥0.996).The minimum detection limits of the method for vitamin A and vitamin E were all 0. 02 μg /mL. The intraday and interday relative standard deviations ( RSDs) were less than 3% at high,medium and low concentrations. The recoveries of the samples at the three concentrations were 91. 2% -107. 5%,and the RSDs were less than 10%. CONCLUSION This method is simple and accurate,with higher sensitivity than using UV detector and can be used for the simultaneous determination of vitamin A and vitamin E of different configurations in serum,and is suitable for rapid detection of batch serum samples.
作者
赵莹
胡佳薇
赵静珺
尹丹阳
Zhao Ying;Hu Jiawei;Zhao Jingjun;Yin Danyang(Center for Disease Control and Prevention of Shaanxi Province,Xi’an 710054,China)
出处
《卫生研究》
CAS
CSCD
北大核心
2020年第4期585-590,共6页
Journal of Hygiene Research
关键词
高效液相色谱法
荧光检测器
血清
维生素
A
维生素
E
high performance liquid chromatography
fluorescence detector
serum
vitamin A
vitamin E
