Mass Spectrometry Principles

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Questions and Answers

What determines the degree of compound separation in LC-MS?

  • The interaction of compounds with the stationary and mobile phases (correct)
  • The pressure of the mobile phase
  • The type of mobile phase used
  • The temperature of the column

What is the primary difference between HPLC and LC?

  • The pressure of the mobile phase (correct)
  • The temperature of the column
  • The type of mobile phase used
  • The type of column used

What is the advantage of using LC-MS over other methods?

  • It provides extensive sample coverage (correct)
  • It provides low sample coverage
  • It is less expensive
  • It is more time-consuming

What is the original name of HPLC?

<p>High-pressure liquid chromatography (D)</p> Signup and view all the answers

What is the name of the mass spectrometer mentioned in the text?

<p>LTQ-0001 (B)</p> Signup and view all the answers

How do the compounds' interactions with the stationary and mobile phases affect the sample's components?

<p>They affect the degree of compound separation (B)</p> Signup and view all the answers

What is the pressure range in which HPLC operates?

<p>Between 50 and 350 bar (B)</p> Signup and view all the answers

What is the main difference in how the mobile phase moves through the column between LC and HPLC?

<p>LC uses gravity, while HPLC uses pumps (A)</p> Signup and view all the answers

What is the advantage of using multiple column chemistries, including reversed phase liquid chromatography?

<p>It provides extensive sample coverage (D)</p> Signup and view all the answers

What is the relationship between HPLC and LC?

<p>HPLC is an advancement over LC (C)</p> Signup and view all the answers

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Study Notes

Basic Principle

  • Mass spectrometry is a powerful quantitative and qualitative analytical technique.
  • It is used to study atoms, molecules, and macromolecules.
  • It requires the sample to evaporate or sublimate without thermal decomposition.

Equipment

  • All mass spectrometers consist of:
    • Ion source
    • Vacuum system
    • Mass analyzers
    • Detector
    • Computer

How it Works

  • Introduction of the sample
  • Derivatization of compounds (if necessary)
  • Ionization (production of ions)
  • Acceleration (ions sorted and separated according to their mass and charge)
  • Detection (results detected and recorded)

Applications

  • Identification of drug usage markers in urine, blood, and saliva
  • Identification of structures of biomolecules
  • Molecular mass determination of known and unknown samples
  • Specific medical procedures (e.g., gas composition in breathing)

Instrumentation Comparison

  • Liquid Chromatography (LC):
    • Separates liquids with multiple components
    • Analyzes polar and nonpolar analytes
  • Gas Chromatography (GC):
    • Analyzes biological compounds
    • Used for drug screening, pesticide detection, and inborn errors of metabolism

Troubleshooting

  • Signal instability causing inaccuracy of results
  • Decrease in sensitivity (contamination, etc.)
  • Discrepancy of mass to charge ratio (m/z)
  • High degree of expertise required for performance and maintenance
  • Elevated cost of machinery

QC Type/Calibration

  • Regular calibration is necessary for accurate results

Who Performs this Assay?

  • Tandem mass spectrometry should be performed byMLS only
  • Liquid Chromatography-Mass Spectrometry can be performed by MLS or MLT with MLS supervision

Tests Types

  • Tandem mass spectrometry: MLS only
  • Liquid Chromatography-Mass Spectrometry: MLS/MLT

Tandem Mass Spectrometer

  • Uses the mass to charge (m/z) ratio to separate and identify analytes
  • Excellent for plasma with a large number of backdrop elements
  • Benefits of MS/MS include:
    • Enhances the quantity of assays that may be run on dried blood spot type samples
    • Minimal false positive rate
    • Fast results (20 tests in about 2 minutes)

Liquid Chromatography

  • Separates chemicals from a sample before analysis
  • Compounds are separated based on their interactions with the stationary and mobile phases
  • Provides extensive sample coverage with various column chemistries (e.g., reversed phase LC)
  • High Performance Liquid Chromatography (HPLC) is an advancement over LC, operating at higher pressures (50-350 bar)

Basic Principle

  • Mass spectrometry is a powerful quantitative and qualitative analytical technique.
  • It is used to study atoms, molecules, and macromolecules.
  • It requires the sample to evaporate or sublimate without thermal decomposition.

Equipment

  • All mass spectrometers consist of:
    • Ion source
    • Vacuum system
    • Mass analyzers
    • Detector
    • Computer

How it Works

  • Introduction of the sample
  • Derivatization of compounds (if necessary)
  • Ionization (production of ions)
  • Acceleration (ions sorted and separated according to their mass and charge)
  • Detection (results detected and recorded)

Applications

  • Identification of drug usage markers in urine, blood, and saliva
  • Identification of structures of biomolecules
  • Molecular mass determination of known and unknown samples
  • Specific medical procedures (e.g., gas composition in breathing)

Instrumentation Comparison

  • Liquid Chromatography (LC):
    • Separates liquids with multiple components
    • Analyzes polar and nonpolar analytes
  • Gas Chromatography (GC):
    • Analyzes biological compounds
    • Used for drug screening, pesticide detection, and inborn errors of metabolism

Troubleshooting

  • Signal instability causing inaccuracy of results
  • Decrease in sensitivity (contamination, etc.)
  • Discrepancy of mass to charge ratio (m/z)
  • High degree of expertise required for performance and maintenance
  • Elevated cost of machinery

QC Type/Calibration

  • Regular calibration is necessary for accurate results

Who Performs this Assay?

  • Tandem mass spectrometry should be performed byMLS only
  • Liquid Chromatography-Mass Spectrometry can be performed by MLS or MLT with MLS supervision

Tests Types

  • Tandem mass spectrometry: MLS only
  • Liquid Chromatography-Mass Spectrometry: MLS/MLT

Tandem Mass Spectrometer

  • Uses the mass to charge (m/z) ratio to separate and identify analytes
  • Excellent for plasma with a large number of backdrop elements
  • Benefits of MS/MS include:
    • Enhances the quantity of assays that may be run on dried blood spot type samples
    • Minimal false positive rate
    • Fast results (20 tests in about 2 minutes)

Liquid Chromatography

  • Separates chemicals from a sample before analysis
  • Compounds are separated based on their interactions with the stationary and mobile phases
  • Provides extensive sample coverage with various column chemistries (e.g., reversed phase LC)
  • High Performance Liquid Chromatography (HPLC) is an advancement over LC, operating at higher pressures (50-350 bar)

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