Nierenberg D W
J Chromatogr. 1985 May 3;339(2):273-84. doi: 10.1016/s0378-4347(00)84654-x.
An isocratic high-performance liquid chromatographic method specifically developed to allow simple and rapid determination of beta-carotene concentrations in serum and plasma is reported. Using a method modified from a previously published technique, serum and plasma proteins are denatured by exposure to perchloric acid, and beta-carotene is subsequently extracted into an organic matrix consisting of ethyl acetate-tetrahydrofuran (1:1); no evaporation step is required. Separation is achieved using isocratic elution from a reversed-phase C18 column with UV detection at 436 nm. Recovery of beta-carotene from water and plasma was greater than 98.1%; beta-carotene was stable in the extraction matrix for at least 4 h. Three anticoagulants (oxalate, citrate, and EDTA) caused losses of beta-carotene; perchloric acid and tetrahydrofuran could also destroy beta-carotene under certain conditions. Each run required less than 15 min; within-day coefficient of variation for identical samples averaged 2.3%, between-day coefficient of variation was 4.4% and sensitivity was better than 10 ng/ml. Stability of beta-carotene in plasma was also examined. This method permits a simple, rapid, sensitive, precise, and accurate determination of beta-carotene using 0.5 ml of serum or heparinized plasma.
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β-胡萝卜素的浓度。采用一种对先前发表技术进行改进的方法，血清和血浆蛋白通过暴露于高氯酸而变性，随后β-胡萝卜素被萃取到由乙酸乙酯 - 四氢呋喃（1:1）组成的有机基质中；无需蒸发步骤。使用反相C18柱进行等度洗脱，并在436nm处进行紫外检测来实现分离。β-胡萝卜素从水和血浆中的回收率大于98.1%；β-胡萝卜素在萃取基质中至少4小时内稳定。三种抗凝剂（草酸盐、柠檬酸盐和乙二胺四乙酸）会导致β-胡萝卜素损失；在某些条件下高氯酸和四氢呋喃也会破坏β-胡萝卜素。每次运行时间少于15分钟；相同样品的日内变异系数平均为2.3%，日间变异系数为4.4%，灵敏度优于10ng/ml。还研究了β-胡萝卜素在血浆中的稳定性。该方法允许使用0.5ml血清或肝素化血浆简单、快速、灵敏、精确且准确地测定β-胡萝卜素。
