Instrumental Analysis - Exam Preparation an der Hochschule Bonn-Rhein-Sieg | Karteikarten & Zusammenfassungen

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What does DAD stand for?

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TESTE DEIN WISSEN

Diode array detector

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TESTE DEIN WISSEN

Which advantage has the retention factor k compared to the retention time?

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TESTE DEIN WISSEN
  • k is standardized to the dead time and thus independent of column dimensions and flow rate
  • k allows to compare results based on different experimental conditions
Lösung ausblenden
TESTE DEIN WISSEN

Which wavelength is commonly used for the UV detector of HPLC analysis to detect protein?

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TESTE DEIN WISSEN

HPLC UV-detector: 190-400nm (full range)

For proteins: 280 nm

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TESTE DEIN WISSEN

What does SDS-PAGE stand for?

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TESTE DEIN WISSEN

Sodium Dodecylsulfate Polyacrylamide Gel Electrophoresis

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TESTE DEIN WISSEN

In isoelectric focusing, why do you expect to find the proteins on the gel at their pI?

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At pI, the structure of a protein tends to be most compact (least repulsion of charges)

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What is the isoelectric point pI?

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TESTE DEIN WISSEN

The isoelectric point is the pH at which the overall charge of a peptide or protein is zero.

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TESTE DEIN WISSEN

Why is using an internal standard for HPLC quantification recommended?

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TESTE DEIN WISSEN

Quantifications based upon external standards may lead to inaccurate results

  • If the sample volume is imprecisely known due to manual injection
  • If some of the original target analyte is lost during sample preparation steps (such as extractions and derivations 
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TESTE DEIN WISSEN

What is the function of DTT?

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TESTE DEIN WISSEN

cleaves covalent disulfide bridges (by reducing them to thiol groups)

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TESTE DEIN WISSEN

Name three different analyzers for mass spectrometry.

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TESTE DEIN WISSEN
  • Quadrupole Analyzers
  • Time of Flight Analyzers
  • Ion Traps
Lösung ausblenden
TESTE DEIN WISSEN

Explain briefly the separation principle of HPLC.

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TESTE DEIN WISSEN

HPLC uses polar and non-polar properties of an analyte to separate it. With the flow rate of the mobile phase, the analyte is transported through the column and its particle pores, where the analyte can get separated by their affinity to the stationary phase.

Lösung ausblenden
TESTE DEIN WISSEN

Why is DAD preferable to a simple UV detector?

Lösung anzeigen
TESTE DEIN WISSEN
  • Simultaneous analysis of all wavelengths
  • Accuracy of wavelength increases with number of diodes


But: Should not be used for light-sensitive samples

Lösung ausblenden
TESTE DEIN WISSEN

Which range of k values is recommended and why?

Lösung anzeigen
TESTE DEIN WISSEN

Recommended values are in the range of 0.5-20

  • k < 0.5 too skinny peaks (could not determine e.g. peak width)
  • k > 20 produces too broad peaks 
Lösung ausblenden
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Q:

What does DAD stand for?

A:

Diode array detector

Q:

Which advantage has the retention factor k compared to the retention time?

A:
  • k is standardized to the dead time and thus independent of column dimensions and flow rate
  • k allows to compare results based on different experimental conditions
Q:

Which wavelength is commonly used for the UV detector of HPLC analysis to detect protein?

A:

HPLC UV-detector: 190-400nm (full range)

For proteins: 280 nm

Q:

What does SDS-PAGE stand for?

A:

Sodium Dodecylsulfate Polyacrylamide Gel Electrophoresis

Q:

In isoelectric focusing, why do you expect to find the proteins on the gel at their pI?

A:

At pI, the structure of a protein tends to be most compact (least repulsion of charges)

Mehr Karteikarten anzeigen
Q:

What is the isoelectric point pI?

A:

The isoelectric point is the pH at which the overall charge of a peptide or protein is zero.

Q:

Why is using an internal standard for HPLC quantification recommended?

A:

Quantifications based upon external standards may lead to inaccurate results

  • If the sample volume is imprecisely known due to manual injection
  • If some of the original target analyte is lost during sample preparation steps (such as extractions and derivations 
Q:

What is the function of DTT?

A:

cleaves covalent disulfide bridges (by reducing them to thiol groups)

Q:

Name three different analyzers for mass spectrometry.

A:
  • Quadrupole Analyzers
  • Time of Flight Analyzers
  • Ion Traps
Q:

Explain briefly the separation principle of HPLC.

A:

HPLC uses polar and non-polar properties of an analyte to separate it. With the flow rate of the mobile phase, the analyte is transported through the column and its particle pores, where the analyte can get separated by their affinity to the stationary phase.

Q:

Why is DAD preferable to a simple UV detector?

A:
  • Simultaneous analysis of all wavelengths
  • Accuracy of wavelength increases with number of diodes


But: Should not be used for light-sensitive samples

Q:

Which range of k values is recommended and why?

A:

Recommended values are in the range of 0.5-20

  • k < 0.5 too skinny peaks (could not determine e.g. peak width)
  • k > 20 produces too broad peaks 
Instrumental Analysis - Exam Preparation

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