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目的建立柱前衍生化高效液相色谱法测定2-去氧葡萄糖(2-DG)含量的方法。方法采用1-苯基-3-甲基-5-吡唑酮(PMP)为柱前衍生化试剂,将2-DG在碱性条件下衍生化后直接进样测定。分离柱为HypersilODS2色谱柱(4.6 mm×250 mm,5μm),流动相为100 mmol·L-1醋酸铵缓冲液(pH 5.5)-乙腈(78∶22),流速1.0 mL·min-1,波长249 nm。结果 2-DG在19.68~393.6μg·mL-1浓度范围内与峰面积具有良好的线性关系(r=0.9997);其定量限(S/N=10)和最低检出限(S/N=3)分别为7.8和3.1 ng;平均回收率为101.21%,RSD 0.63%。结论该方法简便实用、检测灵敏度高、测定结果准确,适用于2-DG的质量控制。
Objective To establish a method for the determination of 2-deoxyglucose (2-DG) by precolumn derivatization high performance liquid chromatography. Methods 1-Phenyl-3-methyl-5-pyrazolone (PMP) was used as a precolumn derivatization reagent to derivatize 2-DG under basic conditions and then directly injected into the sample. The separation column was Hypersil ODS column (4.6 mm × 250 mm, 5 μm) with a mobile phase of 100 mmol·L-1 ammonium acetate buffer (pH 5.5) -acetonitrile (78:22) at a flow rate of 1.0 mL · min- 249 nm. Results 2-DG had a good linear relationship with the peak area (r = 0.9997) in the concentration range of 19.68-393.6μg · mL-1. The limit of quantification (S / N = 10) and the lowest detection limit (S / N = 3) were 7.8 and 3.1 ng respectively; the average recovery was 101.21% and RSD 0.63%. Conclusion The method is simple and practical, high detection sensitivity, accurate measurement results and is suitable for quality control of 2-DG.