Química Bioanalítica, QFA, ESTCE, Campus Riu Sec, Universitat Jaume I, 12071 Castelló, Spain.
Talanta. 2011 Apr 15;84(2):314-8. doi: 10.1016/j.talanta.2011.01.022. Epub 2011 Jan 15.
A simple micellar liquid chromatographic procedure is described to determine tamoxifen and endoxifen in plasma. For the analysis, tamoxifen and endoxifen solutions were diluted in water and UV-irradiated for 20 min to form the photocycled derivative with a phenanthrene core which shows intense fluorescence. Samples were then directly injected, thus avoiding long extraction and experimental procedures. The resolution from the matrix was performed using a mobile phase containing 0.15 mol L(-1) SDS-7% n-butanol at pH 3, running at 1.5 mL min(-1) through a C18 column at 40°C. Detection was carried out by fluorescence, and the excitation and emission wavelengths were 260 and 380 nm, respectively. The chromatographic analysis time was 20 min. The analytical methodology was validated following the International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH) guidelines. The response of the drugs in plasma was linear in the 0.5-15 μg mL(-1) range, with r(2)>0.99. Accuracy and precision were <14% in both cases. Limits of detection and quantification (ng mL(-1)) in plasma were 75 and 250 for endoxifen, and 50 and 150 in tamoxifen. The method developed herein does not show interferences by endogenous compounds. Finally the analytical method was used to determine the amount of tamoxifen and endoxifen in several plasma samples of breast cancer patients from a local hospital.
本文描述了一种简单的胶束液相色谱法，用于测定血浆中的他莫昔芬和依西美坦。在分析中，他莫昔芬和依西美坦溶液在水中稀释，并经紫外线照射 20 分钟，形成具有菲核心的光循环衍生物，该衍生物具有强烈的荧光。然后直接进样，从而避免了长时间的提取和实验过程。采用含有 0.15mol/L SDS-7%正丁醇的流动相，在 pH 3 下，以 1.5mL/min 的流速通过 C18 柱，在 40°C 下进行分离，从而实现与基质的分离。检测采用荧光法，激发和发射波长分别为 260nm 和 380nm。色谱分析时间为 20min。分析方法按照人用药物注册技术要求国际协调会议（ICH）指南进行验证。药物在血浆中的响应在 0.5-15μg/ml 范围内呈线性，r(2)>0.99。两种情况下的准确度和精密度均<14%。在血浆中的检测限和定量限（ng/ml）分别为依西美坦 75 和 250，他莫昔芬 50 和 150。本文所建立的方法不显示内源性化合物的干扰。最后，该分析方法用于测定当地医院几位乳腺癌患者血浆中他莫昔芬和依西美坦的含量。
