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ANA CHEM > HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY > Flashcards

HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY Flashcards

1
Q

• 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

A

High Performance Liquid Chromatography (HPLC)

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2
Q

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

A

Solvents

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3
Q

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

A

Absorption
Partition

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4
Q

2 commonly used type of HPLC

A

Absorption and Partition chromatography

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5
Q

Type of chromatography where separation is based on absorption

A

Absorption chromatography

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6
Q

Type of chromatography where separation is based on partition

A

Partition chromatography

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7
Q

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

A

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

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8
Q

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

A

Retention time

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9
Q

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

A

identification
quantification

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10
Q

(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

A

Polar stationary phase

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11
Q

Higher polarity = ?

A

Slower movement through column

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12
Q

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

A

retention
reference

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13
Q

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

A

Chromatogram

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14
Q

The more concentrated, the ?

A

stronger the signal

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15
Q

General components of HPLC

A

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

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16
Q

Are a group of organic liquid used in HPLC

A

Solvents

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17
Q

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

A

Pump

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18
Q

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

A

Mobile phase

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19
Q

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

A

Degasser

20
Q

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

A

Gases

21
Q

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

A

Injector

22
Q

Device used in chromatography for the separation of chemical compounds

A

HPLC column

23
Q

• 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

A

Stationary phase

24
Q

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

A

Adsorbent

25
Q

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

A

Temperature

26
Q

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

A

Column heater

27
Q

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

A

Detector

28
Q

Detector could be

A

UV detector
IR detector
refractive index detector
mass spectrometer

29
Q

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

A

Recorder

30
Q

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

A

Chromatogram

31
Q

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

A

Waste reservoir

32
Q

The composition of the eluent is consistent without the presence of

A

Analyte

33
Q

Two common types of hplc

A

Normal phase hplc
Reverse phase hplc

34
Q

• 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

A

Normal phase hplc

35
Q

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

A

Polar
Less polar

36
Q

• 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)

A

Reverse phase hplc

37
Q

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

A

Hydrophobicity

38
Q

The stationary phase is ___ and the mobile phase is __

A

Nonpolar (C-18 silica beads
Polar (methanol)

39
Q

In normal phase hplc nonpolar moves ___

A

Quickly

40
Q

In reverse phase hplc nonpolar moves___

A

Slowly

41
Q

Journal article

A

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

42
Q

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

A

hysterectomy
cervical cancer

43
Q

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

A

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)

44
Q

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

A

cervical tissues

45
Q

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

A

100 %

46
Q

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 __

A

Possible errors

ANA CHEM (8 decks)

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