Nierenberg D W
J Chromatogr. 1984 Nov 28;311(2):239-48. doi: 10.1016/s0378-4347(00)84717-9.
An isocratic high-performance liquid chromatographic method specifically developed to allow simple and rapid determination of retinol concentrations in serum and plasma is reported. Retinol and retinol acetate (the internal standard) are extracted into butanol-ethyl acetate, with no subsequent evaporation step. Separation is achieved on a reversed-phase C-18 column, with a mobile phase consisting of acetonitrile-1% ammonium acetate (89:11), and UV detection at 313 nm. Recoveries of both retinol and the internal standard were 100%, and both compounds were stable in the extraction solvent for at least 2.5 h. Three anticoagulants (oxalate, citrate, EDTA) and perchloric acid (used in some methods to denature protein) all caused losses of retinol. Each run required 9 min; same-day coefficient of variation (C.V.) for identical samples averaged 2.5%; between-day C.V. was 6.4%; sensitivity was better than 10 ng/ml, while clinical concentrations were 400-1200 ng/ml. This method permits simple, rapid, sensitive, precise, and accurate determination of retinol using 0.5 ml serum or heparinized plasma.
本文报道了一种专门开发的等度高效液相色谱法，用于简单快速地测定血清和血浆中的视黄醇浓度。视黄醇和视黄醇乙酸酯（内标）被萃取到丁醇 - 乙酸乙酯中，无需后续蒸发步骤。在反相C - 18柱上进行分离，流动相由乙腈 - 1%乙酸铵（89:11）组成，并在313 nm处进行紫外检测。视黄醇和内标的回收率均为100%，两种化合物在萃取溶剂中至少2.5小时内稳定。三种抗凝剂（草酸盐、柠檬酸盐、乙二胺四乙酸）和高氯酸（在某些方法中用于使蛋白质变性）都会导致视黄醇损失。每次运行需要9分钟；相同样品的同日变异系数（C.V.）平均为2.5%；日间C.V.为6.4%；灵敏度优于每毫升10纳克，而临床浓度为每毫升400 - 1200纳克。该方法允许使用0.5毫升血清或肝素化血浆简单、快速、灵敏、精确且准确地测定视黄醇。
