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CHEM108: Analytical Chemistry

Unit 4: Chromatography   Chromatography is the science of separation.  It is useful to purify a type of sample.  Many times in organic chemistry after a reaction is run,column chromatography must be performed to separate the desired product from by-products and/or starting materials.  Within a more analytical chemistry context, a sample may need to be purified before further analysis can be performed.  Chromatography is the method by which to purify the sample.

Note: As you may recall from organic chemistry, mass spectra is the most well known and most useful atomic spectroscopy.  It is a method of finding the atomic mass of a sample.  It uses the mass/charge ratio of fragments produced after a sample has been ionized.

Unit 4 Time Advisory
This unit will take approximately 14.5 hours to complete.

☐    Subunit 4.1: 14.0 hours

☐    Introduction: 8.5 hours

☐    Sub-subunit 4.1.1: 1.5 hours

☐    Sub-subunit 4.1.2: 2.0 hours

☐    Sub-subunit 4.1.3: 0.5 hours

☐    Sub-subunit 4.1.4: 1.5 hours

☐    Subunit 4.2: 0.5 hours

Unit4 Learning Outcomes
Upon successful completion of this unit, the student will be able to:

  • Define the basic principles of chemical separations and how to optimize them.
  • Compare and contrast the various methods of separations, including column chromatography, GC, GC/MS, and HPLC.
  • Identify other liquid chromatography methods, such as liquid-solid absorption, ion-exchange, size-exclusion, and supercritical-fluid.
  • Describe the method and application of electrophoretic separations.

4.1 Chromatography Overview   - Reading: David Harvey’s “Analytical Chemistry 2.0”: “Chapter 12: Chromatographic and Electrophoretic Methods” Link: David Harvey’s “Analytical Chemistry 2.0”: “Chapter 12: Chromatographic and Electrophoretic Methods” (PDF)

 Instructions: Click on the link to Chapter 12 to open the PDF. 
Read all of Chapter 12, pages 783-866 of the text.  This material
covers all sections of Unit 4.  

 Terms of Use: <span href="http://purl.org/dc/dcmitype/Text"
rel="dc:type">Analytical Chemistry 2.0</span> by [David
Harvey](http://web.me.com/dtharvey1213/Analytical_Chemistry_2.0/Welcome.html) is
licensed under a [Creative Commons
Attribution-NonCommercial-ShareAlike 3.0 Unported
License](http://creativecommons.org/licenses/by-nc-sa/3.0/). The
original version can be
found [here](http://www.asdlib.org/onlineArticles/ecourseware/Analytical%20Chemistry%202.0/Text_Files.html). 
  • Lecture: YouTube: MIT: Sarah Tabacco’s “TLC - The Basics” Link: YouTube: MIT: Sarah Tabacco’s “TLC – The Basics” (YouTube)

    Also available in:
    Adobe Flash, Mp4 video

    Instructions: Watch this 16-minute video about thin layer chromatography. Samples are often tested by TLC prior to attempting separations by column chromatography.  

    Terms of Use: This video is reproduced from Massachusetts Institute of Technology: MIT OpenCourseWare. License: Creative Commons BY-NC-SA 3.0. The original version can be found here

  • Reading: Sheffield Hallam University’s “Chromatography – Introductory Theory” Link: Sheffield Hallam University’s “Chromatography – Introductory Theory” (HTML)

    Instructions: Read the entire webpage.  This material explains the basic terminology of column chromatography and gives insight to optimization methods.

    Terms of Use: Please respect the copyright and terms of use displayed on the webpage above.

  • Assessment: Sheffield Hallam University’s “Chromatography – Introductory Theory Quiz” Link: Sheffield Hallam University’s “Chromatography – Introductory Theory Quiz” (HTML)

    Instructions: Complete the quiz, and then check your answers by clicking on the button at the bottom of the page. 

    Terms of Use: Please respect the copyright and terms of use displayed on the webpage above.

4.1.1 Column Chromatography   - Lecture: YouTube: MIT: Sarah Tabacco’s “Column Chromatography” Link: YouTube: MIT: Sarah Tabacco’s “Column Chromatography” (YouTube)

 Also available in:  
 [Adobe Flash or Mp4
Video](http://ocw.mit.edu/resources/res-5-0001-digital-lab-techniques-manual-spring-2007/videos/column-chromatography/)  

 Instructions: Please watch the video about column chromatography
(runtime: 22:24 minutes).  Column chromatography is usefulin
purification ofproducts from organic reactions.  

 Terms of Use: This video is reproduced from Massachusetts Institute
of Technology: MIT OpenCourseWare. License: [Creative Commons
BY-NC-SA 3.0](http://creativecommons.org/licenses/by-nc-sa/3.0/us/).
The original version can be found
[here](http://www.youtube.com/watch?v=B_QyhG2-VBI).
  • Web Media: You Tube: Amrita University’s “Isolation of Plant Pigments by Column Chromatography” Link: You Tube: Amrita University’s “Isolation of Plant Pigments by Column Chromatography” (YouTube)

    Instructions: Please watch this brief video (runtime: 3:15 minutes).  This video demonstrates and describes the column chromatography.

    Terms of Use: Please respect the copyright and terms of use displayed on the webpage above.

  • Assessment: David Harvey’s Analytical Chemistry 2.0: “Chapter 12: Chromatographic and Electrophoretic Methods:” Practice Exercises 12.1-12.4 Link: David Harvey’s Analytical Chemistry 2.0: “Chapter 12: Chromatographic and Electrophoretic Methods:” Practice Exercises 12.1-12.4 (PDF)

    Instructions: Complete the following practice exercises: 12.1 (p. 792), 12.2 (p. 794), 12.3 (p. 795), and 12.4 (p. 797).  Solutions to these problems begin on page 881.   

    Terms of Use: Analytical Chemistry 2.0 by David Harvey is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported License.  The original version can be found here.   

4.1.2 Gas Chromatography   - Assessment: David Harvey’s Analytical Chemistry 2.0: “Chapter 12: Chromatographic and Electrophoretic Methods:” Practice Exercises 12.5 and 12.6 Link: David Harvey’s Analytical Chemistry 2.0: “Chapter 12: Chromatographic and Electrophoretic Methods:” Practice Exercises 12.5 and 12.6 (PDF)

 Instructions: Work through practice problems 12.5 (p. 821) and 12.6
(p. 823).  Solutions to these problems begin on page 882.  
     
 Terms of Use: <span href="http://purl.org/dc/dcmitype/Text"
rel="dc:type">Analytical Chemistry 2.0</span> by [David
Harvey](http://web.me.com/dtharvey1213/Analytical_Chemistry_2.0/Welcome.html) is
licensed under a [Creative Commons
Attribution-NonCommercial-ShareAlike 3.0 Unported
License](http://creativecommons.org/licenses/by-nc-sa/3.0/). The
original version can be
found [here](http://www.asdlib.org/onlineArticles/ecourseware/Analytical%20Chemistry%202.0/Text_Files.html). 
  • Reading: Sheffield Hallam University’s “Gas Chromatography” Link: Sheffield Hallam University’s “Gas Chromatography” (HTML)

    Instructions: Read the entire webpage.  This material covers the theory and instrumental components of gas chromatography.

    Terms of Use: Please respect the copyright and terms of use displayed on the webpage above.

  • Assessment: Sheffield Hallam University’s “Gas Chromatography Quiz” Link: Sheffield Hallam University’s “Gas Chromatography Quiz” (HTML)

    Instructions: Complete the quiz, and then check your answers by clicking on the button at the bottom of the page. 

    Terms of Use: Please respect the copyright and terms of use displayed on the webpage above.

4.1.2.1 Mass Spectroscopy   - Web Media: YouTube: The Royal Society of Chemistry’s “Mass Spectrometry MS” Link: YouTube: The Royal Society of Chemistry’s “Mass Spectrometry MS” (YouTube)

 Instructions: Please watch this brief video (runtime: 7:59
minutes).  This video demonstrates the laboratory technique of mass
spectrometry, including a review of the instrumentation and how data
is obtained.  

 Terms of Use: Please respect the copyright and terms of use
displayed on the webpage above.

4.1.2.2 GC/MS Chromatography   - Web Media: YouTube: C.R. Harrison’s “Gas Chromatography Demo” Link: YouTube: C. R. Harrison’s “Gas Chromatography Demo” (YouTube)

 Instructions: Please watch this short video (runtime: 2:12
minutes).  This video demonstrates the laboratory technique used for
gas chromatography.  

 Terms of Use: The linked material above has been reposted by the
kind permission of C.R. Harrison. This work is licensed under a
[Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported
License](http://creativecommons.org/licenses/by-nc-sa/3.0/). You can
find the original version
[here](http://www.youtube.com/watch?v=kw_07NKZUhY). 

4.1.3 High Performance Liquid Chromatography   - Assessment: David Harvey’s Analytical Chemistry 2.0: “Chapter 12: Chromatographic and Electrophoretic Methods:” Practice Exercises 12.7 and 12.8 Link: David Harvey’s Analytical Chemistry 2.0: “Chapter 12: Chromatographic and Electrophoretic Methods:” Practice Exercises 12.7 and 12.8 (PDF)

 Instructions: Try to complete the following practice problems: 12.7
(p. 831) and 12.8 (p. 841).  Solutions to these problems begin on
page 883.     

 Terms of Use: <span href="http://purl.org/dc/dcmitype/Text"
rel="dc:type">Analytical Chemistry 2.0</span> by [David
Harvey](http://web.me.com/dtharvey1213/Analytical_Chemistry_2.0/Welcome.html) is
licensed under a [Creative Commons
Attribution-NonCommercial-ShareAlike 3.0 Unported
License](http://creativecommons.org/licenses/by-nc-sa/3.0/). The
original version can be
found [here](http://www.asdlib.org/onlineArticles/ecourseware/Analytical%20Chemistry%202.0/Text_Files.html). 

4.1.4 Other Liquid Chromatography Methods   4.1.4.1 Liquid-Solid Absorption Chromatography   - Reading: Separationprocesses.com: S.M. Cheah’s: Chromatography Mechanisms Link: Separationprocesses.com: S.M. Cheah’s: Chromatography Mechanisms (HTML)

 Instructions: Read the webpage for supplemental information on
liquid-solid adsorption chromatography.  This page also contains
additional information on ion-exchange chromatography (IEC) and
size-exclusion chromatography (SEC).  

 Terms of Use: Please respect the copyright and terms of use
displayed on the webpage above. 

4.1.4.2 Ion-Exchange Chromatography (IEC)   - Web Media: YouTube: C.R. Harrison’s “Ion Exchange Column” Link: YouTube: C. R. Harrison’s “Ion Exchange Column” (YouTube)

 Instructions: Please watch this video (runtime: 4:24 minutes). 
This video demonstrates the laboratory technique used in ion
exchange chromatography.  

 Terms of Use: The linked material above has been reposted by the
kind permission of C.R. Harrison. This work is licensed under a
[Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported
License](http://creativecommons.org/licenses/by-nc-sa/3.0/). You can
find the original version
[here](http://www.youtube.com/watch?v=VUKxTJMUE4E). 

4.1.4.3 Size-Exclusion Chromatography (SEC)   - Web Media: YouTube: Dr. Melissa Moss’: “Size-Exclusion Chromatography” Link: YouTube: Dr. Melissa Moss’: “Size-Exclusion Chromatography” (YouTube)

 Instructions: Please watch this video (runtime: 3:20 minutes). 
This video demonstrates the application and theory of size-exclusion
(or gel filtration) chromatography.  Please note that SEC is used in
combination with a scattering technique to gain information about
the analyte.  

 Terms of Use: Please respect the copyright and terms of use
displayed on the webpage above.

4.1.4.4 Supercritical-Fluid Chromatography   - Reading: Virginia Tech’s “Supercritical Fluid Chromatography (SFC)” Link: Virginia Tech’s “Supercritical Fluid Chromatography (SFC)” (HTML)

 Instructions:  Read the information on the page.  This material
covers the theory, instrumentation, and application of SFC.   

 Terms of Use: Please respect the copyright and terms of use
displayed on the webpage above.
  • Web Media: YouTube: Waters’ Dr. Curtis White: “SFC Explained” Link: YouTube: Waters’ Dr. Curt White: “SFC Explained” (YouTube)

    Instructions: Please watch this video (runtime: 3:25 minutes).  Note that SFC theory is similar to liquid chromatography.  This video illustrates the instrumentation.  Also note that SFC is used in combination with a mass spectroscopy to gain information about the analyte.

    Terms of Use: Please respect the copyright and terms of use displayed on the webpage above.

4.2 Electrophoresis   - Web Media: You Tube: Amrita University’s “Agarose Gel Electrophoresis” Link: You Tube: Amrita University’s “Agarose Gel Electrophoresis” (YouTube)

 Instructions: Please watch this brief video (runtime: 2:54
minutes).  This video demonstrates and describes the process of gel
electrophoresis.  

 Terms of Use: Please respect the copyright and terms of use
displayed on the webpage above.
  • Assessment: David Harvey’s Analytical Chemistry 2.0: “Chapter 12: Chromatographic and Electrophoretic Methods:” Practice Exercise 12.9 Link: David Harvey’s Analytical Chemistry 2.0: “Chapter 12: Chromatographic and Electrophoretic Methods:” Practice Exercise 12.9 (PDF)

    Instructions: Please complete practice problem 12.9 (p. 859).  The solution begins on page 884.   

    Terms of Use: Analytical Chemistry 2.0 by David Harvey is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported License. The original version can be found here