In the graph the X axes is the temperature the Y axes is retention time unit

# In the graph the x axes is the temperature the y axes

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In the graph, the X axes is the temperature, the Y axes is retention time (unit: min). We can clearly find that the retention time shows an obviously decreased trend by the temperature increase. That means that the higher temperature the column is, the less retention time and t0 will be. Temperature influences the energy that molecule has. When we increase the temperature, we increase the flow rate, which reduce the time that the sample stay in mobile phase and stationary phase. Hence, the retention time decreases with the temperature increase. Conclusions When temp increase, molecule move faster, Tr will reduce, and the efficiency will increase. The pk7's AUC will stay the same with the temperature change. W on the other hand when temperature increases the molecules will move faster. Tr will reduce and the efficiency will increase making W to decrease. When temp increase, molecule move faster, Tr will reduce, and the efficiency will increase. The H will increase by the temperature increase. In the hypothesis, the back pressure decreases along with the growth of temperature, because the temperature increase the flow rate and decrease the viscosity of mobile phase. In the
HPLC CHROMATOGRAPHY 14 hypothesis, the retention time decreases along with the growth of temperature, because the temperature increase the flow rate. Then the selectivity and efficiency of the HPLC will be influenced. 20 25 30 35 40 45 50 55 60 65 0.0000 0.5000 1.0000 1.5000 2.0000 2.5000 3.0000 3.5000 Temperature Vs RT URA Tr Pk 7 tR Temperature RT 20 25 30 35 40 45 50 55 60 65 125.00 126.00 127.00 128.00 129.00 130.00 131.00 132.00 133.00 Temperature Vs µ URA Tr Temperature RT
HPLC CHROMATOGRAPHY 15 20 25 30 35 40 45 50 55 60 65 0.000 1000.000 2000.000 3000.000 4000.000 5000.000 6000.000 Temperature Vs AUC,H,W URA AUC URA H URA W PK7 AUC PK7 H PK7 W Temperature RT 0.9 1 1.1 1.2 1.3 1.4 1.5 1.6 0.0000 0.5000 1.0000 1.5000 2.0000 2.5000 3.0000 3.5000 4.0000 4.5000 5.0000 Flow rate Vs RT URA Tr Pk 7 tR Flow rate RT 0.9 1 1.1 1.2 1.3 1.4 1.5 1.6 0.000 1000.000 2000.000 3000.000 4000.000 5000.000 6000.000 7000.000 8000.000 9000.000 Flow rate Vs AUC,H,W URA AUC URA H URA W PK7 AUC PK7 H PK7 W Flow rate AUC/H/W
HPLC CHROMATOGRAPHY 16 0.9 1 1.1 1.2 1.3 1.4 1.5 1.6 0.00 20.00 40.00 60.00 80.00 100.00 120.00 140.00 Flow rate Vs µ URA Tr Flow rate μ
HPLC CHROMATOGRAPHY 17 References Smith, I. (Ed.). (2013). Chromatography . Elsevier. Giddings, J. C. (2017). Dynamics of chromatography: principles and theory . CRC Press. Lenca, N., & Poole, C. F. (2017). Liquid chromatography with room temperature ionic liquids. JPC-Journal of Planar Chromatography-Modern TLC , 30 (2), 97-105. Bobaly, B., Beck, A., Veuthey, J. L., Guillarme, D., & Fekete, S. (2016). Impact of organic modifier and temperature on protein denaturation in hydrophobic interaction chromatography. Journal of pharmaceutical and biomedical analysis , 131 , 124-132.

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