高效液相色谱法同时检测食用菌中的4种嘌呤

Simultaneous detection of 4 kinds of purines in edible fungi by high performance liquid chromatography

中文关键词:  食用菌  嘌呤；高效液相色谱；色谱条件

英文关键词:edible fungi  purine  high performance liquid chromatography  chromatographic conditions

中文摘要:

      目的 建立高效液相色谱法同时检测食用菌中次黄嘌呤、鸟嘌呤、黄嘌呤、腺嘌呤的方法, 并将其应用于市售食用菌检测。方法 利用酸水解的方法提取食用菌中的次黄嘌呤、鸟嘌呤、黄嘌呤、腺嘌呤, 通过4种嘌呤的分离度和峰形筛选色谱柱, 优化流动性比例、流速、柱温和进样量等检测条件, 并对检测条件进行方法学验证。结果 当使用Insert Sustain AQ-C18色谱柱, 以pH为4.0、浓度为50 mmol/L的乙酸铵溶液与甲醇(V:V=97:3)为流动相, 流速为0.6 mL/min, 柱温30 ℃, 进样量10 μL, 在254 nm检测波长下4种嘌呤可以完全分离且峰形较好。经方法学验证, 得出此方法在检测范围内线性关系良好。结论 通过检测条件优化建立了一种可有效区分和同时检测食用菌中4种嘌呤的高效液相色谱检测方法, 所测市售食用菌中尿酸前体物质次黄嘌呤和黄嘌呤含量较低或不能检出, 根据《中国营养科学全书》中嘌呤摄入量控制, 确定了现有市售食用菌的膳食量。

英文摘要:

      Objective To establish a method for the simultaneous detection of hypoxanthine, guanine, xanthine, and adenine in edible fungi by high performance liquid chromatography, and use this method to the analysis of commercially available edible fungi. Methods The purines (hypoxanthine, guanine, xanthine, and adenine) from edible fungi were extracted by using acid hydrolysis. The chromatographic column was selected based on the resolution and peak shape of the 4 kinds of purines. Detection conditions, including the mobile phase ratio, flow rate, column temperature, and injection volume, were optimized, and the methodology was validated. Results The results showed that using Insert Sustain AQ-C18 column with a mobile phase consisting of 50 mmol/L ammonium acetate solution (pH 4.0) and methanol (V:V=97:3), at a flow rate of 0.6 mL/min and the column temperature of 30 ℃, with an injection volume of 10 μL and detected at 254 nm, the 4 kinds of purines could be completely separated with good peak shapes. And the methodological validation results showed that this method exhibited a good linear relationship within the detected range. Conclusion A high performance liquid chromatography method is established through the optimization of detection conditions, which effectively differentiates and simultaneously detects the 4 kinds of purines in edible fungi. The content of hypoxanthine and xanthine (uric acid precursor substances) are very low or undetectable in the detected edible fungi. Based on the purine intake guidelines in the Encyclopedia of Chinese nutritional science, the dietary intake of the currently available commercial edible mushrooms is determined.