HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY

Question 1

• Technique used in analytical chemistry to separate, identify, and quantify specific components in a mixture
• Have high resolution and separation capacity
• Used in quantitative and qualitative analysis

Answer 1

High Performance Liquid Chromatography (HPLC)

Question 2

High performance liquid Chromatography (HPLC) can only analyze mixtures dissolve in ___

Answer 2

Solvents

Question 3

(basic principles)
High performance liquid Chromatography (HPLC) depends on the nature of stationary phase whether if it is an ____ chromatography or ____ chromatography

Answer 3

Absorption
Partition

Question 4

2 commonly used type of HPLC

Answer 4

Absorption and Partition chromatography

Question 5

Type of chromatography where separation is based on absorption

Answer 5

Absorption chromatography

Question 6

Type of chromatography where separation is based on partition

Answer 6

Partition chromatography

Question 7

(basic principle)
The separation principle of HPLC is based on the distribution of sample (analyte) between the ____ and ____

Answer 7

Mobile phase (eluent)
Stationary phase (packing material in column)

Question 8

The time which is required for the compounds to be transported from the injection site to the detector unit is called ___

Answer 8

Retention time

Question 9

Retention time the data for ____ and ___ of the compounds in the mixture

Answer 9

identification
quantification

Question 10

(basic principle)
HPLC can separate compounds in the mixture through their interaction with ____, wherein the higher polarity, the slower it will move through the column

Answer 10

Polar stationary phase

Question 11

Higher polarity = ?

Answer 11

Slower movement through column

Question 12

(Basic principle)
Compounds are identified through the difference in their peak’s ___ time, with prior injected ____ standards

Answer 12

retention
reference

Question 13

____ and related data from the detector helps calculate the concentration of each compound (or quantify specific compounds

Answer 13

Chromatogram

Question 14

The more concentrated, the ?

Answer 14

stronger the signal

Question 15

General components of HPLC

Answer 15

Solvent
Pump
Injector
HPLC column
Detector
PC (for data acquisition)
Waste container

Question 16

Are a group of organic liquid used in HPLC

Answer 16

Solvents

Question 17

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

Answer 17

Pump

Question 18

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

Answer 18

Mobile phase

Question 19

Remove gases from a liquid such as the mobile phase in HPLC

Answer 19

Degasser

Question 20

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

Answer 20

Gases

Question 21

It allows the introduction of a precise sample volume onto the HPLC column

Answer 21

Injector

Question 22

Device used in chromatography for the separation of chemical compounds

Answer 22

HPLC column

Question 23

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

Answer 23

Stationary phase

Question 24

Stationery phase is made up of an ____ material that has a very small particle size

Answer 24

Adsorbent

Question 25

HPLC are largely influenced by _____. In order to obtain repeatable results it is important to be consistent in this condition

Answer 25

Temperature

Question 26

• Designed to allow to keep columns at a constant temperature or (isothermal) • utilize a series of temperature increases and constant hold temperatures ( gradient elution)

Answer 26

Column heater

Question 27

• Used to monitor the compounds themselves in order to quantify and identify the compound separated in the column • called "eyes" for hplc

Answer 27

Detector

Question 28

Detector could be

Answer 28

UV detector IR detector refractive index detector mass spectrometer

Question 29

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

Answer 29

Recorder

Question 30

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

Answer 30

Chromatogram

Question 31

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

Answer 31

Waste reservoir

Question 32

The composition of the eluent is consistent without the presence of

Answer 32

Analyte

Question 33

Two common types of hplc

Answer 33

Normal phase hplc Reverse phase hplc

Question 34

• Utilizes a polar stationary phase (usually silica) and less polar (non-aqueous) diluting solvents •used to separate hydrophobic compounds and matrices that are retained too strongly by reversed phase and have minimal solubility in aqueous mobile phase

Answer 34

Normal phase hplc

Question 35

Normal phase hplc utilizes a ___ stationary phase and less ___ eluting solvents

Answer 35

Polar Less polar

Question 36

• Technique can be used to separate identify and or quantitative components in mixtures of soluble organic components based on their hydrophobicity • the stationary phase is nonpolar (C-18 silica beads) and the mobile phase is polar (methanol)

Answer 36

Reverse phase hplc

Question 37

Reverse phase hplc separate, identify or quantitate components in mixtures of soluble organic components based on their ___

Answer 37

Hydrophobicity

Question 38

The stationary phase is ___ and the mobile phase is __

Answer 38

Nonpolar (C-18 silica beads Polar (methanol)

Question 39

In normal phase hplc nonpolar moves ___

Answer 39

Quickly

Question 40

In reverse phase hplc nonpolar moves___

Answer 40

Slowly

Question 41

Journal article

Answer 41

Applications of hplc combined with laser induced fluorescence for protein profile analysis of tissue homogenates in cervical cancer

Question 42

A total of 44 samples including normal cervical biopsy samples from ______ patients or patients whose uterus was removed and the patient suffering from different stages of ___ ___ were recorded using the hplcif

Answer 42

hysterectomy cervical cancer

Question 43

The collection of samples from normal cervical tissue and those with cervical cancer were obtained:

Answer 43

15 samples were collected from normal tissues 7 samples were collected from stage 2B cancer 19 samples were collected from stage 3b cancer 1 in stage 4 cancer 1 in CIN1(or the cells that line the cervix have abnormal changes wherein it is not yet considered as cancerous but if left and treated it will develop into one) 1 in CIS (or pre cancerous cells which still rest on the surface of the cervix and have not yet invaded to the basement membrane into the deeper tissues)

Question 44

the high performance liquid chromatography with laser induced fluorescence or hplc-lif technique gives very good diagnostic results for ____ ___

Answer 44

cervical tissues

Question 45

specificity and sensitivity of the analysis are found to be very high nearly ____%

Answer 45

100 %

Question 46

this technique can provide objective diagnosis of cervical cancer if compared to histopathology wherein the heterogeneity of the tissue samples and the subjectivity of the operator in histopathology may lead to __

Answer 46

Possible errors