HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC METHOD FOR ROUTINE DETERMINATION OF VITAMIN-A AND VITAMIN-E AND BETA-CAROTENE IN PLASMA
HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC METHOD FOR ROUTINE DETERMINATION OF VITAMIN-A AND VITAMIN-E AND BETA-CAROTENE IN PLASMA
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DOI：
10.1016/0378-4347(92)80445-v
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发表时间：
1992-10-02
期刊：
影响因子：
--
通讯作者：
ONG, CN
中科院分区：
文献类型：
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作者：
LEE, BL;CHUA, SC;ONG, CN
建立了一种简便、可靠的反相高效液相色谱法，用于常规测定血浆(或血清)中维生素A、维生素E和β-胡萝卜素的含量。200份血清或血浆样品经乙醇脱蛋白后，以生育酚醋酸酯为内标，用正丁醇-乙酸乙酯提取。加入硫酸钠进行脱水。提取样本的分析物被发现至少稳定了四天。用10等份的有机提取物进行高效液相色谱分析。流动相为甲醇-丁醇-水(89.5：5：5.5，v/v)，流速为1.5ml/min。感兴趣的分析物在45℃下22分钟内与其他血浆成分很好地分离。维生素A、维生素E和β-胡萝卜素的最低检出限分别为0.02、0.5和0.1微克/毫升。本方法的回收率和重现性均在90%左右。该方法灵敏、特异，可用于流行病学研究和维生素缺乏症的常规检测。文中还讨论了可能影响分析的几个重要因素。
A simple and reliable reversed-phase high-performance liquid chromatographic (HPLC) method for the routine determination of vitamins A and E and beta-carotene in plasma (or serum) with wavelength-programmed ultraviolet-visible absorbance detection is described. A 200-mul aliquot of serum or plasma sample, after deproteinization with ethanol, and containing tocopherol acetate as internal standard, was extracted with butanol-ethyl acetate. Sodium sulphate was added for dehydration. Analytes of extracted samples were found to be stable for at least four days. A 10-mul aliquot of this organic extract was used for HPLC analysis. The mobile phase was methanol-butanol-water (89.5:5:5.5, v/v) and the flow-rate was set at 1.5 ml/min. The analytes of interest were well separated from other plasma constituents within 22 min at 45-degrees-C. The lowest detection limits of vitamins A and E and beta-carotene were 0.02, 0.5 and 0.1 mug/ml, respectively. The recovery and reproducibility of the present method were around 90%. The method is sensitive, specific and can be used for epidemiological studies and for routine determination of vitamin deficiency. Several important factors that may affect the analysis are also discussed in this paper.