Disdier B, Bun H, Catalin J, Durand A
Laboratoire Hospitalo-Universitaire de Pharmacocinétique et Toxicocinétique, UFR Pharmacie et CHU Timone, Marseille, France.
J Chromatogr B Biomed Appl. 1996 Aug 30;683(2):143-54. doi: 10.1016/0378-4347(96)00112-0.
A gradient reversed-phase high-performance liquid chromatographic technique is described for the easy separation and quantification of some retinoids; all-trans-retinoic acid, 13-cis-retinoic acid, 9-cis-retinoic acid and their corresponding 4-oxometabolites, in plasma. The method involved a diethyl ether-ethyl acetate (50:50, v/v) mixture extraction at pH 7 with acitretin and 13-cis-acitretin as internal standards. A Nova-Pak C18 steel cartridge column was used. The mobile phase was methanol-acetonitrile (65:35, v/v) and 5% tetrahydrofuran (solvent A) and 2% aqueous acetic acid (solvent B) at 1 ml/min. The gradient composition was (only the percentages of solvent B are mentioned): I, 25% solvent B at the time of injection; II, 12% solvent B at 11 min until min; III, 25% solvent B and maintenance of 25% solvent B for 10 min until a new injection. Total time between injections was 40 min. Detection was by absorbance at 350 nm. The precision calculated for plasma concentrations ranging from 2 to 250 ng/ml was better than 15% and the accuracy was less than 12%. The linearity of the method was in the range of 2 to 400 ng/ml of plasma. The limit of quantification was 2 ng/ml for each of the compounds. The HPLC method was applied to plasma specimens collected from animals receiving single dose administrations of all-trans-retinoic acid, 13-cis-retinoic acid and 9-cis-retinoic acid.
描述了一种梯度反相高效液相色谱技术，用于轻松分离和定量血浆中的某些类视黄醇；全反式视黄酸、13-顺式视黄酸、9-顺式视黄酸及其相应的4-氧代代谢物。该方法包括在pH 7下用阿维A和13-顺式阿维A作为内标，用乙醚-乙酸乙酯（50:50，v/v）混合液萃取。使用Nova-Pak C18钢柱。流动相为甲醇-乙腈（65:35，v/v）、5%四氢呋喃（溶剂A）和2%乙酸水溶液（溶剂B），流速为1 ml/min。梯度组成（仅提及溶剂B的百分比）：I，进样时25%溶剂B；II，11分钟至分钟内12%溶剂B；III，25%溶剂B并保持25%溶剂B 10分钟直至新的进样。进样之间的总时间为40分钟。通过在350 nm处的吸光度进行检测。对于血浆浓度范围为2至250 ng/ml计算的精密度优于15%，准确度小于12%。该方法的线性范围为血浆2至400 ng/ml。每种化合物的定量限为2 ng/ml。该高效液相色谱法应用于从接受全反式视黄酸、13-顺式视黄酸和9-顺式视黄酸单剂量给药的动物收集的血浆标本。
