高效液相色谱法检测固体饮料中4种嘌呤含量

Determination of 4 kinds of purine content in solid beverages by high performance liquid chromatography

中文关键词:  高效液相色谱法  固体饮料  嘌呤  高尿酸血症  痛风

英文关键词:high performance liquid chromatography  solid beverage  purine

中文摘要:

      目的 建立高效液相色谱法测定预包装食品中固体饮料的嘌呤含量, 并将其应用于市售预包装固体饮料检测的方法。方法 利用酸水解提取固体饮料中的4种嘌呤, 通过分离度和峰形筛选色谱柱, 优化流动相pH、流速、柱温等检测条件, 并进行方法学验证。结果 采用Sunniest C18色谱柱, 以pH为3.68的10 mmol/L甲酸铵溶液-甲醇(99:1, V:V)为流动相, 流速1.0 mL/min, 柱温30 ℃, 进样量10 μL, 检测波长254 nm, 4种嘌呤完全分离且峰形良好。鸟嘌呤、腺嘌呤、次黄嘌呤在1.00~40.00 mg/L范围内线性关系良好(r2≥0.9990), 黄嘌呤在0.10~4.00 mg/L范围内线性关系良好(r2=0.9998); 检出限0.01~0.14 mg/L, 定量限0.04~0.46 mg/L; 加标回收率88.00%~113.00%, 相对标准偏差为0.05%~5.90%。11种奶茶嘌呤含量为6.66~30.46 mg/100 g, 11种咖啡嘌呤含量为5.28~26.11 mg/100 g。结论 该方法具有良好的精密度和准确度, 可用于固体饮料中4种嘌呤的同时检测, 为高尿酸血症及痛风人群的膳食选择提供科学依据。

英文摘要:

      Objective To establish a method for determining the purine content in prepackaged solid beverages by high performance liquid chromatography and to apply it to the analysis of commercially available ones. Methods The 4 kinds of purines were extracted from solid beverages using acid hydrolysis. Chromatographic columns were screened based on resolution and peak shape. Detection conditions, including mobile phase pH, flow rate and column temperature, were optimized, and method validation was performed. Results Using a Sunniest C18 column, a mobile phase of 10 mmol/L ammonium formate solution at pH 3.68 methanol (99:1, V:V), a flow rate of 1.0 mL/min, a column temperature of 30 ℃, an injection volume of 10 μL, and a detection wavelength of 254 nm, the 4 kinds of purines were completely separated with good peak shapes. Guanine, adenine and hypoxanthine exhibited good linearity in the range of 1–40 mg/L (r2≥0.9990), while xanthine showed good linearity in the range of 0.1–4.0 mg/L (r2=0.9998); the limits of detection were 0.01–0.14 mg/L, and the limits of quantitation were 0.04–0.46 mg/L; spiked recovery rates were 88.00%–113.00%, with relative standard deviations were 0.05%–5.90%. The purine content in the 11 kinds of milk tea samples ranged from 6.66 mg/100 g to 30.46 mg/100 g, and in the 11 kinds of coffee samples, it ranged from 5.28 mg/100 g to 26.11 mg/100 g. Conclusion This method exhibits good precision and accuracy and can be used for the simultaneous determination of 4 kinds of purines in solid beverages, providing a scientific basis for dietary choices among individuals with hyperuricemia and gout.