Simultaneous Determination of Free Amino Acid Content in Different Teas using Supercritical Fluid Chromatography Coupled with Single Quadrupole Mass Spectrometry

Tea (Camellia sinensis L.) is a complex mixture containing a wide range of biological activities and has been used as widely consumed beverages and natural medicine for over thousand years [1-2]. In this study, a novel method for high-performance liquid chromatography with mass spectrometry (SFC-MS) has been developed to simultaneously determine the contents of 11 free amino acids in different types of teas (pu-erh tea, green tea, black tea and oolong tea). The separation conditions for the selected amino acids were carefully optimized such as the column type, temperature and backpressure, the type of additive. The best compromise for tested analytes in terms of chromatographic performance was obtained when water (5%) and trifluoroacetate acid (0.4%) were added to the supercritical carbon dioxide/methanol mobile phase. Finally, the developed SFC-MS method was successfully applied to the analysis of the 11 amino acids present in the teas and fully validated as well. The results indicated a good linearity (r ≥0.995), precision (RSD≤ 2.99%), stability (RSD≤ 2.88%) and accuracy (91.95%~107.99%). The limits of detection ranged from 1.42 to 14.69 ng/mL, respectively, while the limits of quantification were between 4.53 and 47.0 ng/mL. The content of the amino acids in six different tea samples were also determined and presented some difference basing on the fermentation processes. The proposed SFC-MS method showed a great potential in further investigations to differentiate tea varieties