高效液相色谱荧光法检测南瓜中色氨酸的含量

Determination of tryptophan in pumpkin by high performance liquid chromatography with fluorescence detection

中文关键词:  高效液相色谱荧光法  南瓜  酶解法  色氨酸

英文关键词:high performance liquid chromatography with fluorescence detection  pumpkin  enzymatic hydrolysis  tryptophan

中文摘要:

      目的 建立高效液相色谱荧光法测定南瓜中色氨酸的检测方法。方法 南瓜样品经蛋白酶水解后, 经C18色谱柱分离, 以甲醇-0.0085 mol/L乙酸钠溶液作为流动相等度洗脱。采用荧光检测器检测, 激发波长283 nm, 发射波长343 nm。结果 色氨酸在质量浓度为1.0~100 mg/L范围内线性关系良好, 相关系数(r2)＞0.999, 检出限和定量限分别为17、50 mg/kg。在低、中、高3个加标水平下的回收率为95.9%~100.8%, 相对标准偏差为1.11%~2.44% (n=6)。结论 本方法操作简单、便捷, 准确度高, 重现性好, 适用于南瓜中色氨酸的检测。

英文摘要:

      Objective To establish a method for determination of tryptophan in pumpkin by high performance liquid chromatography with fluorescence detection. Method Samples were hydrolyzed by protease completely, and separated by C18 column using methanol?0.0085 mol/L sodium acetate as mobile phase by isocratic elution. Fluorescence detector was used for detection, with the excitation wavelength 283 nm and the emission wavelength 343 nm. Results Tryptophan had a good linear relationship within the mass concentration range of 1.0?100 mg/L, with the correlation coefficient (r2)＞0.999. The limit of detection and limit of quantification of tryptophan were 17 and 50 mg/kg. The recovery rates at the 3 spiked levels of low, medium and high were 95.9%?100.8%, the relative standard deviations (RSD) were 1.11%?2.44% (n=6). Conclusion The method is simple, convenient, accurate and reproducible, which is suitable for the effective detection of tryptophan in pumpkin.