Abahusain M A, Wright J, Dickerson J W, el-Hazmi M A, Aboul Enein H Y
Department of Family and Community Medicine, College of Medicine, King Saud University, Riyadh, Saudi Arabia.
Biomed Chromatogr. 1998 Mar-Apr;12(2):89-93. doi: 10.1002/(SICI)1099-0801(199803/04)12:2<89::AID-BMC732>3.0.CO;2-0.
In this report we describe a modified reverse phase HPLC method that avoids the solvent evaporation step and allows simple and rapid determination of retinol, alpha-tocopherol, alpha-carotene and beta-carotene and achieves complete separation of alpha- and beta-carotene. Retinyl acetate, alpha-tocopheryl acetate and retinyl palmitate in ethanol were added to serum as internal standards. Serum was then deproteinized with an equal volume of ethanol, and the lipid was extracted with ethyl acetate-butanol (1:1 v/v). A portion of this solution was injected into a C18 reverse phase chromatographic column and absorbencies of the vitamins and internal standards were measured at 292 nm for tocopherols, 325 nm for retinoids and 450 nm for carotenoids; peak-height ratios were used to quantify each vitamin. The analytical recoveries for retinol, alpha-tocopherol alpha- and beta-carotene at various concentrations tested were 95-103, 90-98, 92-99 and 94-96%, respectively. The intra- and interassay variations for low and high concentrations of retinol, alpha-tocopherol, alpha- and beta-carotene ranged from 2.4 to 6.7 for intraassay and from 4.3 to 8.5 for interassay replication. The detection limits were 1.25 (0.04), 19 (0.44), 0.35 (0.006) and 0.94 (0.017) micrograms/dL (delta mol/L) for retinol, alpha-tocopherol, alpha- and beta-carotene, respectively.
在本报告中，我们描述了一种改进的反相高效液相色谱法，该方法避免了溶剂蒸发步骤，能够简单快速地测定视黄醇、α-生育酚、α-胡萝卜素和β-胡萝卜素，并实现α-胡萝卜素和β-胡萝卜素的完全分离。将乙醇中的乙酸视黄酯、乙酸α-生育酚酯和视黄基棕榈酸酯作为内标加入血清中。然后用等体积的乙醇使血清脱蛋白，并用乙酸乙酯 - 丁醇（1:1 v/v）萃取脂质。将该溶液的一部分注入C18反相色谱柱，在292 nm处测定生育酚类维生素、在325 nm处测定类视黄醇、在450 nm处测定类胡萝卜素的维生素和内标的吸光度；峰高比用于定量每种维生素。在测试的各种浓度下，视黄醇、α-生育酚、α-胡萝卜素和β-胡萝卜素的分析回收率分别为95 - 103%、90 - 98%、92 - 99%和94 - 96%。低浓度和高浓度的视黄醇、α-生育酚、α-胡萝卜素和β-胡萝卜素的批内和批间变异，批内变异范围为2.4至6.7，批间重复变异范围为4.3至8.5。视黄醇、α-生育酚、α-胡萝卜素和β-胡萝卜素的检测限分别为1.25（0. .04）、19（0.44）、0.35（0.006）和0.94（0.017）微克/分升（δ摩尔/升）。
