反相高效液相色谱法直接测定水蛭中14种未衍生氨基酸的含量
反相高效液相色谱,,反相高效液相色谱，蒸发光散射检测器，水蛭，未衍生化，氨基酸,摘要,关键词,1引言,2实验部分,3结果与讨论,Refe
摘 要建立了一种反相高效液相色谱/蒸发光散射检测器方法，同时测定中药水蛭中14种未衍生氨基酸含量。以异亮氨酸等氨基酸为标准品、PrevailTM C18(250 mm×4.6 mm i.d., 5 μm)为色谱柱、梯度洗脱、漂移管温度115℃、气体流量2.5 L/min，在25 min内即可将水蛭中14种氨基酸分离测定。氨基酸质量浓度0.072～2.29 g/L时，其对数值与峰面积对数值线性关系良好;14种氨基酸的加样回收率为94.8%～104.4%;信噪比为3时，异亮氨酸检出限为20 mg/L。该法快速、简便、准确，可作为水蛭中氨基酸直接测定方法，亦可为其它中药中氨基酸分析提供参考。
关键词 反相高效液相色谱，蒸发光散射检测器，水蛭，未衍生化，氨基酸
Direct Determination of 14 Underivatized Amino Acids in Leech by Reversed Phase High Performance Liquid Chromatography
Yan Dan*, Li Guo
(Chengdu University of Tradional Chinese Medicine, Chengdu 610075)
Abstract A reversed phase high performance liquid chromatograph evaporative lightscattering detection method for the determination of 14 underivatized amino acids (glycine, serine, alanine, threonine, lysine, histindine arginine, valine, methionine, tyrosineisoleucine, leucine, phenylalanine and tryptophan) in leech.The assay was conducted on a PrevailTM C18 column (250 mm×4.6 mmi.d., 5 μm) with isoleucine, etc, as the standard substance.The 14 amino acids were separated within 25 min under the condition of gradient elution. The detection was performed at the temperature of drift tube of 115℃ and the gas flow rate of 2.5 L/min. The calibration curves were showed good linearity within the concentration range of 0.0715-2.2870 g/L. The average recoveries were 94.8%-104.4%. While the signalnoise ratio was 3, the detection limit of isoleucine was 20 mg/L. This method is rapid, simple and accurate, and it can be used for the direct determination of 14 underivatized amino acids in leech. ......
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