High-Performance Liquid Chromatography

Question 1

where does HPLC depend

Answer 1

nature of stationary phase

Question 2

liquid-solid extraction

Answer 2

Adsorption chromatography

Question 3

liquid-liquid extraction

Answer 3

Partition chromatography

Question 4

based on the distribution of analyte between the mobile pase and stationary phase

Answer 4

separation principle

Question 5

analyte

Answer 5

sample

Question 6

mobile phase

Answer 6

eluent

Question 7

stationary phase

Answer 7

packing material in column

Question 8

how are compounds identified

Answer 8

difference in their peak’s retention time with prior injected reference standards

Question 9

helps in quantifying specific compounds

Answer 9

chromatogram and related data

Question 10

group of organic liquids used in HPLC

Answer 10

solvents

Question 11

generates a flow of eluent from the solvent reservoir into the system

Answer 11

pump

Question 12

• liquid that dissolves the target compound
• mixture of solvent can be used (polar and non-polar)

Answer 12

mobile phase

Question 13

designed to remove gases from liquid such as the mobile pase in HPLC

Answer 13

degasser

Question 14

main cause of noise and cycling in the baseline, and poor compressibility of the eluent

Answer 14

Gases

Question 15

allows the introduction of a precise sample volume onto the column

Answer 15

injector

Question 16

used in chromatography for separation of chemical compounds

Answer 16

HPLC column

Question 17

• made up of an adsorbent material that has very small particle size
• particle size are kept uniform to obtain better performance
• part of column that interacts with the target compound

Answer 17

stationary phase

Question 18

designed to allow to keep your columns at a constant temperature or utilize a series of temperature increases and constant hold temperatures

Answer 18

column heater

Question 19

constant temperature

Answer 19

isothermal

Question 20

temperature is raised from below (IC) to above (SEC) the CAP temperature if the sorption of the polymer solute to the stationary phase is a typical, exothermic process

Answer 20

temperature gradient elution

Question 21

• monitor the compounds to quantify and identify the compounds separated in the column
• called as “eyes” for HPLC

Answer 21

detector

Question 22

examples of detector in HPLC

Answer 22

• UV detector
• IR detector
• refractive index detector
• mass spectrometer

Question 23

translates the signal generated by the detector into a graph called a chromatogram

Answer 23

recorder

Question 24

safely collects all the mobile phase and sample components after they pass through the HPLC system

Answer 24

waste reservoir

Question 25

Two common types of HPLC

Answer 25

1. Normal phase HPLC 2. Reverse phase HPLC

Question 26

- utilizes a polar stationary phase (usually silica) and nonpolar (nonaqueaous) eluting solvents - used to separate hydrophobic compounds and matrices

Answer 26

normal phase HPLC

Question 27

what is usually used in polar stationary phase

Answer 27

silica

Question 28

normal phase HPLC is used to separate what?

Answer 28

hydrophobic compounds and matrices

Question 29

stationary phase is non-polar and the mobile phase is polar

Answer 29

reversed phase HPLC

Question 30

example of application of HPLC

Answer 30

profile analysis of tissue homogenates in cervical cancer