高效液相色谱法测定水产品中5种核苷酸含量

Determination of 5 kinds of nucleotides content in aquatic products by high performance liquid chromatography

中文关键词:  高效液相色谱法  水产品  五中核苷酸  定性定量分析:

英文关键词:high performance liquid chromatography  aquatic products  nucleotides

中文摘要:

      目的 建立高效液相色谱法测定水产品中5种核苷酸的分析方法。方法 通过色谱条件优化和前处理条件选择, 样品在4 ℃条件下, 用5%高氯酸溶液均质提取2次, 以0.01 mol/L KH2PO4(含1.45 mmol/L四丁基硫酸氢铵, pH=5.0)-乙腈(95:5, V:V)为流动相洗脱, 经C18 (250 mm×4.6 mm, 5 μm)色谱柱分离, 紫外检测器检测, 标准曲线外标法定量。结果 5种核苷酸在0.1~100.0 μg/mL范围内线性关系良好, 相关系数大于0.999, 检出限和定量限分别为1.0~2.0 mg/kg和3.0~6.0 mg/kg, 方法精密度为2.00%~6.87%, 加标回收率在92.3%~105.6%之间, 批内相对标准偏差为0.63%~7.90%, 批间相对标准偏差为1.72%~7.34%。结论 该方法灵敏度高、重复性好, 可用于水产品中5种核苷酸的定性定量分析。

英文摘要:

      Objective To establish a method for the determination of 5 kinds of nucleotides in aquatic products by high performance liquid chromatography. Methods After optimization of chromatographic conditions and selection of pretreatment conditions, the sample was extracted twice with 5% perchloric acid solution at 4 ℃, and then extracted with 0.01 mol/L KH2PO4 (containing 1.45 mmol/L tetrabutyl ammonium bisulfate, pH=5.0)-acetonitrile (95:5, V:V) was eluted by mobile phase, separated by C18 (250 mm×4.6 mm×5 μm) column, detected by ultraviolet detector, and quantified by external standard method with standard curve. Results The 5 kinds of nucleotides had a good linear relationship in the range of 0.1–100.0 μg/mL, and the correlation coefficient was greater than 0.999. The limits of detection and limits of quantitation were 1.0–2.0 mg/kg and 3.0–6.0 mg/kg, respectively, and the precisions were 2.00%–6.87%. The spike recoveries were 92.3%–105.6%, the intra-assay standard deviations were 0.63%–7.90%, and the inter-assay standard deviations were 1.72%–7.34%. Conclusion This method has high sensitivity and good repeatability, and can be used for the qualitative and quantitative analysis of 5 kinds of nucleotides in aquatic products.