Bortolotti A, Lucchini G, Barzago M M, Stellari F, Bonati M
Perinatal Clinical Pharmacology Unit, Istituto di Richerche Farmacologiche Mario Negri, Milan, Italy.
J Chromatogr. 1993 Aug 11;617(2):313-7. doi: 10.1016/0378-4347(93)80505-x.
A reversed-phase high-performance liquid chromatographic method is described for the simultaneous determination of retinol, alpha-tocopherol and retinyl palmitate in plasma. Plasma containing an internal standard (tocol) was deproteinized with ethanol, then extracted with n-hexane. The organic layer was removed and evaporated under a nitrogen stream, and chromatographed on a reversed-phase RP-18 column using a water/acetonitrile-ethyl acetate/2-propanol gradient solvent system over 15 min at 305 nm. The recovery exceeded 93%. The detection limit was 0.1 microgram/ml for retinol, 1.3 micrograms/ml for alpha-tocopherol and 0.95 micrograms/ml for retinyl palmitate. The reproducibility, precision (expressed as coefficients of variation) and accuracy were less than 8% for all analytes. The small sample requirement, the simplicity of extraction, the short run-time and the good reproducibility make this procedure particularly useful for monitoring retinol and alpha-tocopherol supplementation in premature newborns.
本文描述了一种反相高效液相色谱法，用于同时测定血浆中的视黄醇、α-生育酚和棕榈酸视黄酯。含有内标（生育三烯酚）的血浆用乙醇脱蛋白，然后用正己烷萃取。除去有机层并在氮气流下蒸发，然后在反相RP - 18柱上进行色谱分析，使用水/乙腈 - 乙酸乙酯/2 - 丙醇梯度溶剂系统，在305nm下运行15分钟。回收率超过93%。视黄醇的检测限为0.1微克/毫升，α-生育酚为1.3微克/毫升，棕榈酸视黄酯为0.95微克/毫升。所有分析物的重现性、精密度（以变异系数表示）和准确度均小于8%。该方法所需样品量少、提取简单、运行时间短且重现性好，特别适用于监测早产儿视黄醇和α-生育酚的补充情况。
