学术
摘 要:
建立了反相高效液相色谱法测定微量全血样品中维生素A的方法。取全血50μL,用100μL无水乙醇沉淀蛋白后,加入400μL×2正己烷漩涡混匀提取维生素A,高速离心分层后取正己烷层在弱氮气流下挥干,加100μL甲醇溶解后用反相色谱柱C8（150 mm×4.6 mmi.d.,5μm）分离,紫外检测波长325nm,外标法定量。色谱条件：柱温,60℃;流动相为V（甲醇）∶V（水）=92∶8;流速：0.8 mL/min。用本法同时测定了27例成人微量全血及其血清中的维生素A。标准曲线的相关系数大于0.999;相对标准偏差（RSD）小于5%。对于50μL全血,方法检出限为0.02μg/mL。加标回收率为88%-115%。成人血清与其全血中维生素A含量之比为2.907±0.160（x-±s）。方法适合于微量全血中维生素A的测定,并可以通过测定全血中维生素A含量推算血清中维生素A的含量。[著者文摘]
文章出处:
《分析试验室》-2008年27卷3期 -76-79页
栏目信息:
分 类 号:
文献标识码:
A
文章编号:
1000-0720（2008）03-076-04
Determination of vitamin A in micro-amount of human blood by high performance liquid chromatography
CHEN Dong, ZHANG Rui-yun, YANG Liu-hua ,SUN Cheng-jun （West China School of Public Health, Sichuan University, Chengdu 610041）
Abstract:
A method for the determination of vitamin A in the micro-amount of human blood by high performance liquid chromatography was reported in the paper. 50 μL of whole blood sample was added to a 1.5 mL Eppendorf centrifuge tube and then 100 μL anhydrous alcohol was added to precipitate protein, the mixture was shaken by vortex and then vitamin A in the solution were extracted with 400 μL hexane for two times. After centrifuged at 12000 rpm for 3 min, the hexane layer was taken out and blown to dryness under gentle N2 gas. The residue was redissolved with 100 μL methanol and determined by HPLC. The analytical column used was Zorbax （ Cs, 4.6 mm×15 cm, 5μm） and detection wavelength was 325 nm. Mobile phase was methanol-water （92：8）, flow rate was 0.80 mL/min and the temperature was 60 %. The range of correlation coefficients for the standard curves were large than 0. 999; the detection limit of the method is 0.02 μg/mL. The recoveries of the method ranged from 88% to 115%. The method has been used for the determination of 27 adult whole blood and their corresponding sermn samples and found that the vitamin A content in their whole blood were 0.213 ± 0.062 μg/mL （x ± s） and the vitamin A content in their serum were 0.619± 0.189 μg/mL （x ± s）, with a ratio of 2.907 ± 0.160 μg/mL （x ± s）. The method was sensitive, accurate and has been applied to the determination of vitamin A in the micro-amount of human blood.[著者文摘]
Key words:
Vitamin A; High performance liquid chromatography; Human blood
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