Farn Shiou-Shiow, Yeh Yuen-Han, Lin Wuu-Jyh, Shen Lie-Hang
Isotope Application Division, Institute of Nuclear Energy Research, Taoyuan County, Taiwan (ROC).
Nucl Med Biol. 2009 Feb;36(2):225-31. doi: 10.1016/j.nucmedbio.2008.11.008.
A suitable three potential-time waveforms for the electrochemical detection of 2-deoxy-2-chloro-d-glucose (ClDG) by gold working electrode and palladium reference electrode have been developed and method validation was performed on Waters 2796 Bioalliance HPLC system coupled with pulsed amperometric detection (HPLC/PAD). FDG and ClDG could be completely separated by 50 mM NaOH mobile phase at a flow rate of 1.0 ml/min; 30 degrees C analytical column temperature; and E1 of 200 mV, T1 of 900 mS; E2 of -770 mV, T2 of 10 mS; E3 of 1400 mV, T3 of 10 mS; acquisition delay (AD) of 300 mS. The validation results were shown as follows: (1) in specificity study, mannose, FDG and ClDG could be completely separated and the retention times of these were 6.2, 11.1 and 13.5 min, respectively, with a total run time of 20 min; (2) the intraday repeatable precision expressed with the CV% in six successive analysis was 0.52% (for FDG) and 0.83% (for ClDG); (3) the interday variability precision expressed with the CV% value of the repeatable precision of 3 days was 0.99%, 0.52% and 0.58% for FDG and 0.71%, 0.83% and 1.24% for ClDG; both the CV% of intraday and interday reproducibilities of FDG and ClDG were better than 1.5%; (4) the accuracy and recovery of FDG and ClDG expressed with the percentage of mean value of three successive analysis were 99.75% (for FDG) and 100.68% (for ClDG) which were all greater than 95%; (5) under optimum conditions, the limit of detection of FDG and ClDG was 0.41 and 0.68 microg/ml, and the limit of quantization of FDG and ClDG was 1.24 and 2.04 microg/ml; (6) the correlation coefficient (r) value of linearity is over 0.999 by 5-50 microg/ml ranges of both compounds, respectively; (7) no interference peak effects by composition of mobile phase or increasing/decreasing flow rate or change of temperature was observed.
已开发出适用于通过金工作电极和钯参比电极对2-脱氧-2-氯-D-葡萄糖（ClDG）进行电化学检测的三种电位-时间波形，并在配备脉冲安培检测（HPLC/PAD）的沃特世2796生物联盟高效液相色谱系统上进行了方法验证。在流速为1.0 ml/min的50 mM NaOH流动相、3摄氏度的分析柱温度、E1为200 mV、T1为900 mS、E2为–770 mV、T2为10 mS、E3为1400 mV、T3为10 mS、采集延迟（AD）为300 mS的条件下，FDG和ClDG能够完全分离。验证结果如下：（1）在特异性研究中，甘露糖、FDG和ClDG能够完全分离，它们的保留时间分别为6.2、11.1和13.5分钟，总运行时间为20分钟；（2）在六次连续分析中，以CV%表示的日内重复性精密度对于FDG为0.52%，对于ClDG为0.83%；（3）以3天重复性精密度的CV%值表示的日间变异性精密度，对于FDG为0.99%（第一次）、0.52%（第二次）和0.58%（第三次），对于ClDG为0.71%（第一次）、0.83%（第二次）和1.24%（第三次）；FDG和ClDG的日内和日间重现性的CV%均优于1.5%；（4）以三次连续分析平均值的百分比表示的FDG和ClDG的准确度和回收率分别为99.75%（对于FDG）和100.68%（对于ClDG），均大于95%；（5）在最佳条件下，FDG和ClDG的检测限分别为0.41和0.68微克/毫升，定量限分别为1.24和2.04微克/毫升；（6）两种化合物在5-50微克/毫升范围内的线性相关系数（r）值均超过0.999；（7）未观察到流动相组成、流速增加/降低或温度变化产生的干扰峰效应。
