Fanali Salvatore, Catarcini Paolo, Quaglia Maria Giovanna, Camera Emanuela, Rinaldi Mariarosa, Picardo Mauro
Istituto di Cromatografia del C.N.R., Area della Ricerca di Roma, Via Salaria Km 29,300-C.P., P.O. Box 10, Monterotondo Scalo, Rome, Italy.
J Pharm Biomed Anal. 2002 Aug 1;29(6):973-9. doi: 10.1016/s0731-7085(02)00137-1.
In this study capillary electrochromatography (CEC) was used for the separation of three tocopherols (TOHs), namely delta-, gamma- and alpha-TOH and the antioxidant compound, butylated hydroxytoluene (BHT). The CEC experiments were carried out using an octadecylsilica (ODS) stationary phase packed, in our laboratory, in a fused-silica capillary (100 microm I.D., 365 microm O.D. x 33 cm of total length and 24.6 or 8.4 cm effective length). The mobile phase was composed by a mixture of methanol (MeOH) and acetonitrile (ACN), at different concentrations and 0.01% (w/v) of ammonium acetate. Retention time (t(R)), retention factor (k), resolution (R(s)) of the three TOHs were strongly influenced by the organic solvent composition of the run buffer and by the effective length of the capillary. Optimum experimental conditions were found even employing the short effective length of the capillary achieving the baseline separation of the studied analytes in a relatively short time (less than 5 min). The optimized method was applied to the qualitative analysis of vitamin E (alpha-TOH) present in a human serum extract.
在本研究中，采用毛细管电色谱法（CEC）分离三种生育酚（TOHs），即δ-、γ-和α-生育酚以及抗氧化化合物丁基羟基甲苯（BHT）。CEC实验使用在我们实验室填充了十八烷基硅胶（ODS）固定相的熔融石英毛细管（内径100微米，外径365微米，总长33厘米，有效长度24.6或8.4厘米）进行。流动相由甲醇（MeOH）和乙腈（ACN）的混合物组成，浓度不同，还有0.01%（w/v）的醋酸铵。三种生育酚的保留时间（t(R)）、保留因子（k）、分离度（R(s)）受运行缓冲液的有机溶剂组成和毛细管有效长度的强烈影响。即使采用较短的毛细管有效长度，也能找到最佳实验条件，在相对较短的时间内（不到5分钟）实现所研究分析物的基线分离。该优化方法应用于人体血清提取物中维生素E（α-生育酚）的定性分析。
