5种虾类嘌呤含量的测定及其不同组织嘌呤的分布研究

Study on the determination of purine content in 5 kinds of shrimp and the distribution of purine in different tissues

中文关键词:  嘌呤化合物  高效液相色谱法  虾  不同组织

英文关键词:purine compounds  high performance liquid chromatography  shrimp  different tissues

中文摘要:

      目的 建立一种同时测定腺嘌呤(adenine, A)、鸟嘌呤(guanine, G)、次黄嘌呤(hypoxanthine, H)和黄嘌呤(xanthine, X)的高效液相色谱法(high performance liquid chromatography, HPLC), 并研究5种虾类肌肉、虾黄及内脏组织中的嘌呤分布。方法 虾类中嘌呤化合物由CF3COOH/HCOOH/H2O混合酸处理, 于90℃水浴振荡12 min后65℃水浴旋转蒸发至干, 残余物用KH2PO4-H3PO4流动相复溶并过0.22 μm滤膜, 最后采用Waters Atlantics T3 (250 mm×4.6 mm , 5 μm)色谱柱, 0.01 mol/L KH2PO4-H3PO4溶液(pH 3.70)作为流动相, 流速1.0 mL/min等色谱条件在254 nm波长下检测分析。结果 4种嘌呤组分均在0.5~250.0 mg/L范围内有良好的线性关系, 相关系数(r2)均接近于1.0000, 检出限和定量限分别在0.042~0.096 mg/L和0.139~0.320 mg/L之间, 方法精密度相对标准偏差在0.04%~0.30%之间, A、G、H和X样品水解重复性相对标准偏差依次为1.87%、2.27%、1.42%和2.59%, 加标回收率在97.83%~102.21%之间。哈氏仿对虾、中华管鞭虾、日本对虾、口虾蛄、凡纳滨对虾5种虾类肌肉中总嘌呤含量在696.56~1862.16 mg/kg之间; 中华管鞭虾和口虾蛄的虾黄中总嘌呤含量分别为4720.34 mg/kg和3650.60 mg/kg; 虾类内脏中总嘌呤含量在936.67~3531.75 mg/kg之间。结论 该方法精密度高、回收率好且检出限低, 适用于虾类不同组织中4种嘌呤的测定。5种虾类中总嘌呤含量及不同部位的嘌呤含量差异较大, 且虾黄和虾内脏均高于同种虾类肌肉中的嘌呤含量。因此, 在食用虾类时, 建议彻底清除内脏和严格控制虾黄和虾仁的摄入量, 以减缓高尿酸血症和痛风等健康风险。

英文摘要:

      Objective To establish a high performance liquid chromatography (HPLC) method for the simultaneous determination of adenine (A), guanine (G), hypoxanthine (H) and xanthine (X) and study the purine distribution in the muscles, yellow, and viscera tissues of 5 kinds of shrimps. Method Purine compounds in shrimp were extracted using a mixed acid of CF3COOH/HCOOH/H2O, oscillated in a water bath at 90℃ for 12 minutes, and then evaporated and dried in a water bath at 65℃. The residue was redissolved in the KH2PO4-H3PO4 mobile phase and passed through 0.22 μm filter membrane. Finally, the samples were detected and analyzed by Waters Atlantic T3 (250 mm×4.6 mm , 5 μm) chromatographic column with 0.01 mol/L KH2PO4-H3PO4 buffer solution (pH 3.70) as the mobile phase at a flow rate of 1.0 mL/min and a wavelength of 254 nm. Results The 4 kinds of purine components had a good linear relationship in the range of 0.5?250.0 mg/L, and the correlation coefficients (r2) were all close to 1.0000. The limits of detection and limits of quantitation were 0.042?0.096 mg/L and 0.139?0.320 mg/L, respectively. The relative standard deviations of the method precision were between 0.04% and 0.30%. The relative standard deviations of water hydrolysis repeatability for A, G, H, and X in sample were 1.87%, 2.27%, 1.42%, and 2.59%, respectively. The spiked recoveries were between 97.83% and 102.21%. The total purine content in the muscles of 5 kinds of shrimp samples, including Parapenaeopsis hardwickii, Solenocera crassicornis, Penaeus japonicus, Oratosquilla oratoria, Litopenaeus vannamei and other shrimps, ranged from 696.56 mg/kg to 1862.16 mg/kg; the content of total purine in shrimp yolk of Solenocera crassicornis and Oratosquilla oratoria was 4720.34 mg/kg and 3650.60 mg/kg, respectively; the total purine content in shrimp viscera ranged from 936.67 mg/kg to 3531.75 mg/kg. Conclusion This method has high precision, good recovery and low detection limit, and is suitable for the determination of 4 kinds of purines in different tissues of shrimp. The total purine content and purine content in different parts of 5 kinds of shrimps are significant differences. The purine content in the yolk and viscera of shrimp was higher than that in the muscle of the same kind of shrimp. Therefore, it is recommended to completely remove the internal organs and strictly control the intake of shrimp yolk and muscle of shrimp to reduce health risks such as hyperuricemia and gout when eating shrimp.