Zhou Xianjing, Shi Yanping
Key Laboratory of Chemistry of Northwestern Plant Resources, Gansu Province, Lanzhou Institute qf Chemical Physics, Chinese Academy of Sciences, Lanzhou 730000, China.
Se Pu. 2013 Jul;31(7):661-6. doi: 10.3724/sp.j.1123.2013.05027.
A method of capillary electrophoresis with indirect ultraviolet (UV) detection was developed for the separation and determination of nine amino acids such as lysine, tryptophan, glutamic acid, etc. The effects of sodium dihydrogen phosphates concentration, pH of buffer and sample injection type and time on the reproducibility and efficiency were investigated. The optimum injection time was 5 s at 5 kPa. The optimum electrophoretic conditions were as follow: 10 mmol/L sodium dihydrogen phosphates (pH 10. 2) containing 0. 5 mmol/L cetrimonium bromide, 20 mmol/L nicotinic acid and 10% (v/v) methanol as running buffer, applied voltage of - 15 kV, detection wavelength of 220 nm. The base line separation of the nine amino acids was achieved successfully within 11 min. The lowest detection limit was 0. 3 mg/L. All of the nine analytes showed good linearities within 1. 0 - 1000 mg/L. The relative standard deviations of migration time and peak area were 0. 64% - 5. 83%. The recoveries of the eight amino acids spiked in a real sample were between 60. 00% and 118.37%. The method was applied in the determination of the amino acids in honey samples from different nectar plants and origins. Prolin, serine and aspartic acid were found in five honey samples, and tryptophan was only found in a litchi honey sample. This method can provide good reference to the evaluation of the quality and nectar origin of honey.
建立了一种采用间接紫外(UV)检测的毛细管电泳方法,用于分离和测定赖氨酸、色氨酸、谷氨酸等9种氨基酸。研究了磷酸二氢钠浓度、缓冲液pH值、进样方式和时间对重现性和分离效率的影响。最佳进样时间为5 s,进样压力为5 kPa。最佳电泳条件如下:以含0.5 mmol/L溴化十六烷基三甲基铵、20 mmol/L烟酸和10%(v/v)甲醇的10 mmol/L磷酸二氢钠(pH 10.2)为运行缓冲液,施加电压-15 kV,检测波长220 nm。9种氨基酸在11 min内成功实现基线分离。最低检测限为0.3 mg/L。9种分析物在1.0 - 1000 mg/L范围内均表现出良好的线性关系。迁移时间和峰面积的相对标准偏差为0.64% - 5.83%。加标到实际样品中的8种氨基酸的回收率在60.00%至118.37%之间。该方法应用于测定来自不同蜜源植物和产地的蜂蜜样品中的氨基酸。在5个蜂蜜样品中检测到脯氨酸、丝氨酸和天冬氨酸,仅在一个荔枝蜂蜜样品中检测到色氨酸。该方法可为蜂蜜质量和蜜源产地的评价提供良好参考。