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HPLC

Description

Mind Map on HPLC, created by emgoy1 on 21/04/2014.
emgoy1
Mind Map by emgoy1, updated more than 1 year ago
emgoy1
Created by emgoy1 over 11 years ago
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Resource summary

HPLC
  1. Resolution
    1. Selectivity

      Annotations:

      • Change in: Elution order Elution rate Ionisation state Activity
      1. pH

        Annotations:

        • Change in pH of mobile phase. Usually around 2-3pH. Ionic interactions
        1. Temperature

          Annotations:

          • Only if the column is held in an oven (HPLC 3&4) Change of conformation results in less surface-surface interactions
          1. Organic Modifier

            Annotations:

            • Liquid Parameter
          2. Retention Factor
            1. Sample Solvent

              Annotations:

              • Can effect interactions when mixing with mobile phase occurs Liquid parameter
              1. Volume of Injection
                1. Concentration of Sample
                2. %B

                  Annotations:

                  • More polar mobile phase = decreased tR Liquid parameter
                3. Efficiency
                  1. Band Broadening
                    1. Eddy Diffusion
                        1. Column Length
                        2. Longitudal Diffusion

                          Annotations:

                          • Decreased longitudal diffusion with increased flow rate. Does not occur in gradient method
                          1. Pore Size
                              1. Particle Size
                              2. Mass transfer

                                Annotations:

                                • Decreased mass transfer with decreased flow rate
                                1. Column Length
                                  1. Particle size
                                    1. Pore Size
                              3. Run Time
                                1. Retention Time
                                  1. Flow Rate
                                  2. Efficiency
                                    1. Linear Velocity

                                      Annotations:

                                      • Effects back pressure
                                      1. Column diameter

                                        Annotations:

                                        • Effects linear velocity
                                        1. Flow rate
                                    2. Back Pressure

                                      Annotations:

                                      • Increasing pressure too much can effect the machine functioning
                                      1. Linear Velocity
                                      2. Equipment Parameters
                                        1. Injector
                                          1. Pump
                                            1. Sampler
                                              1. Sensitivity
                                                1. Wavelength/Detector
                                              2. Gradient or isocratic method
                                                1. Steady %B

                                                  Annotations:

                                                  • Liquid parameter
                                                  1. Increasing %B

                                                    Annotations:

                                                    • Liquid parameters
                                                    1. Resolution >1.5
                                                  2. Column Chemistry

                                                    Annotations:

                                                    • Changing the column chemistry ie the stationary phase composition will completely change the interactions of the analyte and the stationary phase.
                                                    1. Tailing factor
                                                      1. End Capping

                                                        Annotations:

                                                        • Incomplete capping will leave free acid silanols that lead to tailing
                                                        1. Free Silanols

                                                          Annotations:

                                                          • Free acids on the stationary phase will be able to ionically bond (depending on pH) to any ionised groups in the analyte
                                                        2. Stationary phase arms
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