Department of Biomedical Sciences, University of Sassari, Sassari, Italy.
PLoS One. 2013 Jul 29;8(7):e70374. doi: 10.1371/journal.pone.0070374. Print 2013.
Two sensitive and reproducible capillary electrophoresis and high-performance liquid chromatography-fluorescence procedures were established for quantitative determination of L-egothioneine in plasma. After derivatization of L-ergothioneine with 5-iodoacetamidofluorescein, the separation was carried out by HPLC on an ODS-2 C-18 sperisorb column by using a linear gradient elution and by HPCE on an uncoated fused silica capillary, 50 µm id, and 60 cm length. The methods were validated and found to be linear in the range of 0.3 to 10 µmol/l. The limit of quantification was 0.27 µmol/l for HPCE and 0.15 µmol/l for HPLC. The variations for intra- and inter-assay precision were around 6 RSD%, and the mean recovery accuracy close to 100% (96.11%).
建立了两种灵敏且重现性好的毛细管电泳和高效液相色谱-荧光检测法，用于定量测定血浆中的 L-谷氨酰基-L-半胱氨酸。L-谷氨酰基-L-半胱氨酸与 5-碘乙酰胺荧光素衍生化后，在 ODS-2 C-18 反相色谱柱上采用线性梯度洗脱进行 HPLC 分离，在未涂层的熔融石英毛细管（50 µm id，60 cm 长）上进行 HPCE 分离。方法经过验证，在 0.3 至 10 µmol/l 范围内呈线性。HPCE 的定量下限为 0.27 µmol/l，HPLC 的定量下限为 0.15 µmol/l。日内和日间精密度的变异系数约为 6%，平均回收率准确度接近 100%（96.11%）。
