Huang Yifei, Hu Jing
Technology Center, China Tobacco Guangdong Industrial Co., Ltd., Guangzhou 510145, China.
Se Pu. 2010 Jun;28(6):615-22. doi: 10.3724/sp.j.1123.2010.00615.
A method for simultaneous analysis of 20 free amino acids in tobacco was developed using liquid chromatography-electrospray ionization/ion trap tandem mass spectrometry (LC-ESI-IT-MS/MS). Tobacco samples were extracted and filtered, and analyzed without derivatization or solid phase extraction. The LC separation was carried on a reversed-phase HyPURITY C18 column (200 mm x 2.1 mm, 5 microm) with the mobile phase of 1% (v/v) acetonitrile/water solution containing 0.1% nonafluoropentanoic acid and 90% acetonitrile/water solution containing 0.1% nonafluoropentanoic acid in gradient mode. The results showed that the limits of detection (LODs) of 20 amino acids were in the range of 0.01-0.05 micromol/L (S/N = 3), the linear correlation coefficients were above 0. 997 7 in all cases, and the relative standard deviations (RSDs) of peak areas of the extracted ion chromatograms were in the range of 0.78%-4.93%. The developed method is of good efficiency, sensitivity and selectivity, and has been successfully applied to determine the free amino acids in varieties of tobacco samples.
建立了一种使用液相色谱-电喷雾电离/离子阱串联质谱（LC-ESI-IT-MS/MS）同时分析烟草中20种游离氨基酸的方法。烟草样品经提取和过滤后，无需衍生化或固相萃取即可进行分析。液相色谱分离在反相HyPURITY C18柱（200 mm×2.1 mm，5μm）上进行，流动相为含0.1%九氟戊酸的1%（v/v）乙腈/水溶液和含0.1%九氟戊酸的90%乙腈/水溶液，采用梯度模式。结果表明，20种氨基酸的检测限（LOD）在0.01 - 0.05μmol/L范围内（S/N = 3），所有情况下线性相关系数均高于0.997 7，提取离子色谱图峰面积的相对标准偏差（RSD）在0.78% - 4.93%范围内。所建立的方法具有良好的效率、灵敏度和选择性，并已成功应用于多种烟草样品中游离氨基酸的测定。
