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GC-MS Lab

Chromatographic methods, in particular those coupled with mass spectrometry, are among the most frequently used analytical techniques. However, while a number of users increases to encompass their broader range, the understanding of chromatographic methods by these users declines. This training will cover basics and practical approaches to quantization using GC-MS. The first part of this training is learning to process data using specific software features within the homework on quantization. This will be followed by actual lab preparing set of calibration standards, and quantification in test and unknown sample. the preferred software is Masshunter.

Homework on Quantitation

The GC data for MassHunter (alternatively ChemStation) files should be copied to the desktop in the computer lab Abbott hall 343. The instructions and videos how to work with the data are provided below in the Lab assignment.

MassHunter

Instructions homework using massHunter

MassHunter Datafiles

Chemstation

Instructions

ChemStation Datafiles

Lab: GC-MS Quantitation

Gas chromatography coupled with mass spectrometry (GC-MS) is an analytical method used for the identification and quantification of numerous volatile compounds. The qualitative GC-MS analysis is conducted by matching the retention time of given unknowns and mass spectra of known chemicals, typically electron ionization (EI) mass spectra with known standards or libraries. The quantitative analysis in a total ion current mode (TIC) can be performed as a traditional GC analysis based on the total peak area, however the correct approach to quantification is based on the peak area of a specific ion. In addition to the instrumentation, quantification is also dependent on the type of the calibration method used. Various calibration methods such as external standard calibration, internal standard calibration, and method of standard addition may be performed. Today the most common in gas chromatography is the internal standard calibration.

The aim of this lab assignment is to pursue GC-MS analyzes using the internal standard calibration with selected compounds of interest to the trainee; determine their instrumental limits of quantification and validate method based on replicate determination of the targeted compound in an evaluation sample of a defined (known) and unknown composition.

Assignment Materials

  • GC-MS Quant Lab Assignment Instruction
  • Solution Preparation in Pictures
  • Prelab Test
  • Example: MS Excel spreadsheet of data organization for stock solutions
  • Example: MS Excel spreadsheet of calibration
  • Tutorial for calibration curve in MS Excel (2007)

Video Tutorials

Labs

  • Lab stock preparation from a chemical in a solid state
  • Lab stock preparation from a chemical in a liquid state
  • Performing serial dilution for internal standard calibration

MassHunter Data Processing

  • Navigating MassHunter Qual 10
  • Viewing data in MassHunter (scaling and zoom)
  • Identification in GC-MS analysis
  • General peak integration (TIC)
  • Quantification using extracted ion current (EIC) chromatograms 

ChemStation Data Processing

  • Storing and transferring the data for GC-MS dada processing
  • View chromatograms and mass spectra in MSD Chemstation
  • Extracted ion integration in MSD ChemStation

Advanced Processing

  • MassHunter - Advanced quantitative processing
  • ChemStation - Creating automatic quantification method
  • ChemStation - Creating database for peak areas (from quant method)

MS Excel Data Processing

  • How to organize data in MS Excel - Stock solutions
  • Serial dilution in MS Excel
  • Internal standard calibration in Excel Part 1
  • Internal standard calibration in Excel Part 2: Charts
  • LOD and LOQs with LINEST
Alena Kubatova
Office: Abbott Hall, Room 301
Lab: Abbott Hall, Room 309
Grand Forks, ND 58202
P 701.777. 0348
F 701.777.2331
alena.kubatova@UND.edu

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College of Arts & Sciences

Columbia Hall, Room 1930
501 N Columbia Rd Stop 8038
Grand Forks, ND 58202-8038

UND.artssci@UND.edu |  701.777.2749
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