Urbánek Lubor, Krcmová Lenka, Solichová Dagmar, Melichar Bohuslav, Opletalová Veronika, Solich Petr
Charles University in Prague, Department of Analytical Chemistry, Faculty of Pharmacy, Hradec Králové, Czech Republic.
J Sep Sci. 2006 Nov;29(16):2485-93. doi: 10.1002/jssc.200600153.
Among other side effects, administration of anticancer agents is accompanied by manifestations of gastrointestinal toxicity and disturbances of antioxidant balance. The monitoring of these toxic effects in clinical practice is impeded by a dearth of reliable laboratory methods. Therefore, a simple and rapid reversed-phase high-performance liquid chromatography procedure for selective and sensitive determination of retinol, a-tocopherol, and retinyl esters (retinyl-palmitate and retinyl-stearate) in blood serum has been developed and presented in this study. A Series 200 LC HPLC instrument from Perkin Elmer (Norwalk, USA) with diode-array detector (DAD) was used for the analysis. Separations of retinol, alpha-tocopherol, retinyl-palmitate, and retinyl-stearate were performed using a Chromolith Performance RP-18e, 100 x 4.6 mm monolithic column from Merck (Darmstadt, Germany). Gradient elution was used at a flow rate of 3 mL/min; the mobile phase was methanol-water (95:5, v/v) for 0-2.1 min and methanol-2-propanol (60:40, v/v) for 2.1-4.9 min. The total time of analysis was 6 min. The injection volume was 20 microL and the analysis was performed at ambient temperature. Detection of retinol, alpha-tocopherol, and retinyl esters was carried out at 325, 295, and 330 nm, respectively. For practical assessment of the method, the vitamin A absorption test was performed on seven healthy controls as well as on six patients with non-small cell lung carcinoma or head and neck carcinoma previously treated by chemotherapy and/or radiotherapy, six patients with rectal carcinoma before chemoradiotherapy, four patients with gastrointestinal stromal tumor (GIST) before treatment with imatinib, and a breast cancer patient with chemotherapy-induced diarrhea. Present data demonstrate the feasibility of large scale HPLC determination of vitamin E, vitamin A, and retinyl esters in human serum using a silica monolithic column, and this method may represent a valuable aid in the laboratory monitoring of the toxicity of anticancer therapy.
在抗癌药物的其他副作用中，其使用会伴随着胃肠道毒性表现以及抗氧化平衡紊乱。临床实践中对这些毒性作用的监测因缺乏可靠的实验室方法而受到阻碍。因此，本研究开发并介绍了一种简单快速的反相高效液相色谱法，用于选择性和灵敏地测定血清中的视黄醇、α-生育酚和视黄酯（棕榈酸视黄酯和硬脂酸视黄酯）。使用了美国珀金埃尔默公司（诺沃克）的200系列液相色谱仪及二极管阵列检测器（DAD）进行分析。视黄醇、α-生育酚、棕榈酸视黄酯和硬脂酸视黄酯的分离使用了德国默克公司（达姆施塔特）的Chromolith Performance RP - 18e 100×4.6 mm整体柱。采用梯度洗脱，流速为3 mL/min；流动相在0 - 2.1分钟为甲醇 - 水（95:5，v/v），在2.1 - 4.9分钟为甲醇 - 2 - 丙醇（60:40，v/v）。总分析时间为6分钟。进样体积为20微升，分析在室温下进行。视黄醇、α-生育酚和视黄酯的检测分别在325、295和330 nm波长处进行。为了对该方法进行实际评估，对7名健康对照者以及6名先前接受过化疗和/或放疗的非小细胞肺癌或头颈癌患者、6名直肠癌患者在放化疗前、4名胃肠道间质瘤（GIST）患者在接受伊马替尼治疗前以及1名患有化疗诱导性腹泻的乳腺癌患者进行了维生素A吸收试验。目前的数据表明，使用硅胶整体柱对人血清中的维生素E、维生素A和视黄酯进行大规模高效液相色谱测定是可行的，并且该方法可能在抗癌治疗毒性的实验室监测中具有重要帮助。
