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中国农学通报

中国农学通报 ›› 2013, Vol. 29 ›› Issue (12): 164-167.doi: 10.11924/j.issn.1000-6850.2012-2740

• 生物技术科学 • 上一篇    下一篇

反相高效液相色谱检测低浓度5-氨基乙酰丙酸

张龙涛 李昂 史怀 朱育菁 缪伏荣 冯玉兰 刘波   

  • 收稿日期:2012-08-11 修回日期:2012-10-11 出版日期:2013-04-25 发布日期:2013-04-25
  • 基金资助:
    福建省自然科学基金;福建省科技重大专项;福建省属公益类科研院所基本科研专项

Determination of Low Concentration 5-Aminolevulinic Acid by Reversed Phase High Performance Liquid Chromatography

  • Received:2012-08-11 Revised:2012-10-11 Online:2013-04-25 Published:2013-04-25

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摘要： 建立了一种柱前衍生反相高效液相色谱检测溶液中低浓度5-氨基乙酰丙酸的方法，填补了低浓度ALA的HPLC法检测的空白。使用Agilent SB-C18型色谱柱(250 mm×4.6 mm i.d.，5 μm)，以50 mM甲醇-醋酸钠缓冲溶液(pH 4.0)洗脱，流速为1 mL/min，柱温30℃，254 nm波长下检测10 min。在此条件下，5-氨基乙酰丙酸在浓度为1~1000 mg/L范围内与色谱峰面积值之间线性关系良好，回归系数均在0.9999以上。发酵液产物的加标回收率平均值为101.7%。将检测结果与分光光度法相比，符合较好。测试数据表明，此方法简单、可靠，对于低浓度的ALA测定效果良好。

关键词: 籽粒充实, 籽粒充实

Abstract: A reversed phase high performance liquid chromatography with precolumn derivatization was developed for the determination of low concentration 5-aminolevulinic acid (ALA). An Agilent SB-C18 column (250 mm ×4.6 mm i.d., 5μm) was used for separation, and the elution was performed with methanol and 50 mM sodium acetate solution (adjustment of pH to 4.0 with acetic acid). The flow rate was 1 mL.min-1, the temperature of column was 30℃, and the detection wavelength was at 254 nm with the detection time of 10 minutes. The dynamic linear range is 1-1000 mg.L-1, and the correlation coefficient is above 0.9999. The average recoveries of ALA in 3 samples of fermentation were 101.7%. The results agreed well with those of spectrophotometry. So the developed method is simple and reliable for the determination of low concentration ALA.