Instrumental Analysis - Exam Preparation at Hochschule Bonn-Rhein-Sieg

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Exemplary flashcards for Instrumental Analysis - Exam Preparation at the Hochschule Bonn-Rhein-Sieg on StudySmarter:

What does DAD stand for?

Exemplary flashcards for Instrumental Analysis - Exam Preparation at the Hochschule Bonn-Rhein-Sieg on StudySmarter:

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

Exemplary flashcards for Instrumental Analysis - Exam Preparation at the Hochschule Bonn-Rhein-Sieg on StudySmarter:

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

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Exemplary flashcards for Instrumental Analysis - Exam Preparation at the Hochschule Bonn-Rhein-Sieg on StudySmarter:

What does SDS-PAGE stand for?

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

Exemplary flashcards for Instrumental Analysis - Exam Preparation at the Hochschule Bonn-Rhein-Sieg on StudySmarter:

What is the isoelectric point pI?

Exemplary flashcards for Instrumental Analysis - Exam Preparation at the Hochschule Bonn-Rhein-Sieg on StudySmarter:

Why is using an internal standard for HPLC quantification recommended?

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Exemplary flashcards for Instrumental Analysis - Exam Preparation at the Hochschule Bonn-Rhein-Sieg on StudySmarter:

What is the function of DTT?

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Name three different analyzers for mass spectrometry.

Exemplary flashcards for Instrumental Analysis - Exam Preparation at the Hochschule Bonn-Rhein-Sieg on StudySmarter:

Explain briefly the separation principle of HPLC.

Exemplary flashcards for Instrumental Analysis - Exam Preparation at the Hochschule Bonn-Rhein-Sieg on StudySmarter:

Why is DAD preferable to a simple UV detector?

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Which range of k values is recommended and why?

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Exemplary flashcards for Instrumental Analysis - Exam Preparation at the Hochschule Bonn-Rhein-Sieg on StudySmarter:

Instrumental Analysis - Exam Preparation

What does DAD stand for?

Diode array detector

Instrumental Analysis - Exam Preparation

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

  • 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

Instrumental Analysis - Exam Preparation

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

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

For proteins: 280 nm

Instrumental Analysis - Exam Preparation

What does SDS-PAGE stand for?

Sodium Dodecylsulfate Polyacrylamide Gel Electrophoresis

Instrumental Analysis - Exam Preparation

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

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

Instrumental Analysis - Exam Preparation

What is the isoelectric point pI?

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

Instrumental Analysis - Exam Preparation

Why is using an internal standard for HPLC quantification recommended?

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 

Instrumental Analysis - Exam Preparation

What is the function of DTT?

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

Instrumental Analysis - Exam Preparation

Name three different analyzers for mass spectrometry.

  • Quadrupole Analyzers
  • Time of Flight Analyzers
  • Ion Traps

Instrumental Analysis - Exam Preparation

Explain briefly the separation principle of HPLC.

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.

Instrumental Analysis - Exam Preparation

Why is DAD preferable to a simple UV detector?

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


But: Should not be used for light-sensitive samples

Instrumental Analysis - Exam Preparation

Which range of k values is recommended and why?

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 

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