高效液相色谱法测定血清中的VA、VD3.docVIP

高效液相色谱法测定血清中的VA、VD3 VE和β-胡萝卜素* [摘要] 目的 建立同时测定血清中维生素A、维生素D3、维生素Eβ-胡萝卜素的高效液相色谱（high performance liquid chromatography, HPLC）法。方法 以无水乙醇沉淀蛋白质后，血清中的脂溶性维生素乙醚/石油醚混合溶液提取，吹氮浓缩后HPLC测定。结果 VA、VD、VE和β-胡萝卜素的线性范围分别为0.012μg/ml ~500 μg/ml、0.03μg/ml ~500μg/ml、0.1μg/ml ~500μg/ml 和0.015μg/ml~500μg/ml；检出限分别为0.012μg/ml、0.03μg/ml、0.1μg/ml和0.015μg/ml；分别为0.75%、%、%和%。血清样VA、VD、VE和β-胡萝卜素的回收率分别为：结论本法简便快速，适于血清样的测定。 [关键词] HPLC 维生素A 维生素D 维生素E β-胡萝卜素 血清 β-carotene in Serum with HPLC Zou Xiaoli Zeng Hongyan Li Yuanqian Li Xiaoxia Zheng Bo Huang Chengyu Department of Sanitary technology,West China School of Public Health of Sichuan University. (Chengdu 610041) [Abstract] Objective The determination method of retinol(vitamin A), vitaminD3, α-tocopherol(vitamin E) and β-carotene in serum with HPLC was developed. Methods After proteins having been precipitated with anhydrous ethanol, fat-soluable vitamins in serum were extracted with aether/petroleum ether mixture and determined with HPLC. Results The linear ranges were 0.012(g/ml~100(g/ml, 0.030(g/ml ~500μg/ml, 0.12(g/ml~500μg/ml, 0.015(g/ml ~500μg/ml and the detection limits were 0.012μg/ml, 0.030μg/ml, 0.12μg/ml and 0.015μg/ml for retinol, VD3, α-tocopherol and β-carotene respectively. The relative standard derivation (RSD) for retinol, VD3, α-tocopherol and β-carotene were 0.75%、%、% and2.74%. The proposed method was used for serum determination. The recoveries were 92%~116%，98%~112%，84.8%~106%和90%~105% for retinol, VD3, α-tocopherol and β-carotene respectively. Conclusion The method was simple, quick and adapt for serum analysis. 维生素A、维生素D、维生素E和β-胡萝卜素是维持人体正常生理活动和机能的脂溶性维生素，缺乏这些维生素会导致多种疾病。*国家自然科学基金资助 发生有着相应的关系[1]。食道癌是世界八大癌症之一，而我国是食管癌的高发地区，每年因食管癌死亡者约15万人通过病人和正常人血清中脂溶性维生素的含量，了解与脂溶性维生素的关系，为提供实验室依据目前我国尚无测定血清中脂溶性维生素的标准方法。HPLC因其具有高分离效能、灵敏快速、可自动化等优点，已经广泛用于脂溶性维生素的测定 []，同时测定VA、VD3、VE和β-胡萝卜素四种脂溶性维生素的报道。因此，本课题建立HPLC同时测定VA、VD3、VE和β-胡萝卜素的方法，应用于血清样品的测定，脂溶性维生素之间的关系提供简单可行的技术1 材料与方法 1.1 仪器与试剂 HP1100高效液相色谱仪附紫外检测器（HP1100，Agilent Technolo