Gas Chromatography - Mass Spectrometry Instrument for Multiple Chemistry Courses

Project Title: Gas Chromatography-Mass Spectrometry (GC-MS) Instrument for Multiple Chemistry Courses

Long Title (if desired): Gas Chromatography-Mass Spectrometry (GC-MS) Instrument for the Analytical Chemistry and Organic Chemistry Laboratory Courses

Project Lead's Name: Neil Danielson

Project Lead's Email:

Project Lead's Phone: 513-529-2872

Project Lead's Division: CAS

Primary Department: Chemistry and Biochemistry

Other Team Members and their emails:

  • Michael Crowder
  • Richard Taylor
  • Meredith Erb
  • Benjamin Gung

List Departments Benefiting or Affected by this proposal: Chemistry and Biochemistry

Estimated Number of Under-Graduate students affected per year (should be number who will actually use solution, not just who is it available to): 500

Estimated Number of Graduate students affected per year (should be number who will actually use solution, not just who is it available to): 6

Describe the problem you are attempting to solve and your approach for solving that problem: This proposal is to fund a gas chromatography-mass spectrometry (GC-MS) instrument with autosampler and Cerno Bioscience high-resolution Mass Works software primarily for our instrumental analysis analytical chemistry course (CHM 375), organic chemistry laboratory courses (CHM 244 and 255) and the laboratory component of Principles of Fermentation (CHM 436A/MBI 436A/CPB 436A). In addition, this instrument will be demonstrated in the advanced undergraduate/graduate course, Separation Science, CHM 460C/664.

Because of the powerful problem-solving capability of GC-MS, it is almost guaranteed that our chemistry majors will be expected to have this expertise for their future jobs, from BS to graduate degree level. Hands-on learning of GC-MS in a laboratory-type setting will provide an unparalleled means of emphasizing the importance of this technique with students.

GC-MS is the coupling of the separation technique gas chromatography (GC) with the detection method mass spectrometry (MS). After the sample containing a mixture of volatile organic compounds of interest is injected, the components are propelled by the helium carrier through a temperature-controlled open tubular capillary column, often 30m x 0.3mm ID in dimensions, coated with a polymeric liquid stationary phase. Based on differing interactions of the organic compounds with the stationary phase, separation occurs as a function of time and the isolated organic compounds are sent directly to the mass spectrometer detector. This detector ionizes and fragments each separated organic compound reproducibly to generate a characteristic spectrum of signal intensity (ion current) versus mass/charge ratio of the fragments. The total ion current determines how much of the compound is present and the spectrum identifies the structure of the organic compound based on a match through a search of a computer library of spectra. GC-MS is considered the primary analytical technique to separate volatile organic compounds at subpart per million levels in complex samples. For example, the components responsible for the scent of perfumes can all be quantitated and identified by GC-MS. Complex liquid samples such as gasoline or extracts of whiskey can be characterized by GC-MS. GC­ MS is the primary technique used in drug testing of athletes in professional and Olympic sports because the data is so definitive that it can stand up in a court of law.

The GC-MS instrument we are proposing is not only sophisticated in hardware but also in software. The high-resolution software permits mass/charge resolution of 0.001 units permitting the deconvolution of overlapping peaks but also positive identification of organic compound isomers. This is simply not possible with a standard GC-MS instrument. The concept for this spectral accuracy for mass spectrometry has been published by Cemo scientists Yongdong Wang and Ming Gu as a feature article in Analytical Chemistry 2010, 82, 7055-7062.

Our current CHM 375 GC-MS experiment uses a 1990s vintage refurbished Hewlett-Packard GC-MS instrument that demonstrates how organic compounds of widely different structures in a mixture prepared at percent level concentrations can be identified. The instrument is rugged and works well for this specific application. However, this instrument cannot detect easily even part per million (ppm) level concentrations and uses Windows 2000 software which is incompatible with the Cerno high-resolution software. We also use this instrument to look at extracts of hops as a "dry lab" or demo experiment for our Principles of Fermentation course. However, the sensitivity is so poor that most of the extract components are missed. This instrument does not have an autosampler and it is not feasible for it to be used by the high enrollment organic chemistry laboratory classes. We need an instrument that can clearly show both the analytical and organic chemistry capability of GC-MS. Although a quite new Thermo Scientific GC-MS instrument is present on the Miami Middletown campus for teaching analytical chemistry, no such state-of-the-art GC-MS instrument dedicated to teaching is available on the Miami University, Oxford campus.

How would you describe the innovation and/or the significance of your project: The significance of this project is to make clear to science students the importance of GC-MS as one of the primary instrumental methods to solve complicated analysis problems. Students majoring in science, not just chemistry/biochemistry majors, need to have hands-on experience with such a cutting edge research instrument. We can teach a wide variety of science students GC-MS through our organic chemistry laboratory course. Our chemistry/biochemistry majors need to understand the importance of GC-MS for both the characterization of organic reaction products (organic chemistry laboratory course) as well for the separation and quantitation of important complex mixtures such as gasoline or plant extracts (analytical chemistry course). In addition, chemical engineering and microbiology students through our Principles of Fermentation course will gain this vital importance of GC-MS to analytical chemistry.

The key innovation of this project is the introduction of high-resolution GC-MS through the use of the Cerno Bioscience Mass Works software. The standard GC-MS instrument has a resolution of about 0.1 mass units permitting organic compounds with a significant structural difference to be positively identified. However, compounds like caffeine and dimethyl phthalate with a molar mass similarity to 0.001 would need to be identified through mass spectral fragmentation patterns. More problematic is that isomers of organic compounds such as o-, m-, and p-substituted aromatics cannot be positively identified by standard GC­ MS either by molar mass or fragmentation. This one major limitation of MS is something that is commonly taught in all analytical chemistry courses. However, the Cerno Mass Works software integrates carefully each mass spectral peak and can give mass values to the 0.001 place and also distinguish organic isomers. In addition, it can deconvolute at least three overlapping components in a single peak providing complete and positive mixture component identification. Further information about the Cerno Bioscience Mass Works is described in the attached Cerna Mass Works file.

How will you assess the success of the project: Impact on Existing Chemistry Courses

CHM 244 is the first-semester non-majors organic chemistry laboratory course for second-year students, taught by Meredith Erb. We typically have about 300-350 students, most of them are pre-med, but there is a decent chunk of engineers as 'Nell. Most of CHM 244 Is spent teaching organic techniques and instrumentation such as extraction, thin layer chromatography, mass spectrometry, and distillation. Infrared Spectroscopy (IR), nuclear magnetic resonance (NMR), polarimetry and UV spectrophotometry. At this time, we do not teach our non-majors students GC or MS. We typically have 90 students per section spread among three laboratory rooms, 4 sections total. We commonly group students into larger groups (4-6 students) in order to run their samples on other instrumentation (such as the NMR). This would be the case for GC-MS analysis as well indicating that an autosampler is a must as part of the GC-MS instrument.

The present essential oils lab involves the steam distillation of various crude plant materials such as anise seed, cloves, and spearmint and the collection of the primary essential oil, respectively anethole, eugenol, and carvone. These essential oil isolations illustrate approaches in ethnobotany and in the perfume Industry. Currently, we restrict ourselves to plants that provide large amounts of their oils and confirm their structure by TLC and IR spectroscopy, a technique that mostly involves matching rather than structure determination. All of these essential oils mentioned previously are structurally compatible with GC­ MS. It would be easy to add in the GC-MS experiment in our Essential Oils lab because it is a two-week experiment and the second-week is when we analyze our sample. We currently only analyze the isolated oils by TLC which can be problematic by relative retention to standards. GC­ MS would be much more informative showing the degree of purity for the distillation process.

CHM 255 is our second-semester laboratory course for second-year chemistry and biochemistry majors, taught by Ben Gung. We typically have about 60 students in two sections. MS is taught in this course but only in a lecture and student problem-solving format; there is no corresponding laboratory experience after the lecture. Free radical chemistry is a very important class of reactions but not often considered in the organic laboratory course. We envision the entire class doing the free radical halogenation experiment involving the chlorination of chlorobutane (see Free Radical Halogenation section in the Organic Chemistry Lab attached file) and then dividing the class to work on different substitution reactions (see Substitution Reactions section in the Organic Chemistry file). Previously such laboratory experiments were investigated by Richard Taylor but all three of these reactions generate isomeric products that could not be positively identified by GC with flame ionization detection. These isomers would not be distinguishable by electron impact MS found on a standard GC­ MS instrument. However, the high mass resolution of the Cerna Bioscience Mass Works software will allow positive isomer identification (see Cerno Mass Works attached file). This software overcomes the major limitation of MS and would represent an invaluable update to the students' learning of the power of MS.

CHM 375, analytical chemistry, teaches the use of modern instrumentation (molecular spectroscopy, atomic spectroscopy, chromatography, mass spectrometry, and electrochemistry) for the determination of both organic compounds such as quinine and caffeine and ions such as potassium, manganese, and fluoride. This course was developed by N. D. Danielson about five years ago and he has been the only instructor. It is a required course for all chemistry/biochemistry majors and is usually taken by juniors or seniors. Students majoring in science education with an emphasis on chemistry are also required to take this course and chemical engineering students will also occasionally elect this course. This course is offered both semesters with a yearly enrollment of about 70 students. Our current MS experiment uses a 1990s vintage refurbished Hewlett-Packard instrument that demonstrates how organic compounds of widely different structures in a mixture prepared at percent level concentrations can be identified. The first part of the GC-MS experiment emphasizes MS fundamentals such as the Mclafferty rearrangement and halogen isotope effects and we clearly show electron impact MS cannot distinguish o-, m-, and p- xylene isomers. The second part of the experiment would be the headspace (vapor above the liquid or solid sample) analysis of an unknown such as the main solvents in an auto touch up paint. However, this instrument cannot detect easily even part per million (ppm) level concentrations and uses Windows 2000 software which is incompatible with the Cerno high-resolution software. We need an instrument that can clearly show the analytical capability of GC-MS. Positive identification of these xylene isomers has been shown in the Cerno Bioscience literature. The petroleum industry is one of the main users of GC-MS. We need to add the separation and characterization of gasoline by GC-MS as a student special project to ensure students are aware of the critical role GC-MS plays in this huge part of the chemical industry.

CHM 436A/MBI 436A/CPB 436A is our Principles of Fermentation course taught every fall to chemistry/biochemistry, chemical engineering, and microbiology students (about 10 from each department giving a total enrollment of 30). The course is team-taught by Michael Crowder, Neil Danielson, and Rob Mccarrick, all from Chemistry/Biochemistry, Luis Actis from Microbiology, Andrew Jones from Chemical Engineering, and Matt McMurray from Psychology. The course objectives are as follows. Through a combination of lectures from faculty and experts in the fermentation industry, hands-on laboratory experiences, and site visits, students will develop an understanding of the importance of fermentation in the food, beverage, and drug industry. Students will have the opportunity to learn how microbiology, biology, chemistry/biochemistry, and engineering are interrelated in the fermentation Industry. Students will have a basic understanding of biological processes involved in fermentation, the theory and design principles behind the equipment and facilities used in different fermentation/distillation processes, analysis, and preservation of yeast strains and quality control of fermentation products. We will also explore the psychology and physiology of alcohol consumption. It is expected that students who successfully complete this course will be well-positioned to enter the fermentation industry and make relevant contributions to the different aspects and steps involved in this complex process that provides critical products for human consumption. The GC-MS experiment of extracts from hops is attached. However, comparison of our GC-MS chromatograms with ones in the literature shows many missing peaks due to issues in instrument sensitivity. In addition, we would like the students to use GC-MS to fingerprint different types of whiskey or bourbon that has been aged under similar or different conditions. This is not possible with our current 1990s vintage GC-MS Instrument.

CHM 460C/664 is an advanced undergraduate/graduate course entitled Separation Science. This course has been taught by Neil Danielson every other year during his entire career. It is divided Into three parts: Chromatography Theory, Practice of Gas Chromatography, and Practice of High-Performance Liquid Chromatography. Although it is primarily a lecture course, exposure of these students to modem chromatography instrumentation is important. We envision students doing an actual extraction of a plant substance such as hops or cinnamon (different from their prior experience) and characterizing these samples by GC-MS. The students would initiate the runs but would not have to stay because the autosampler would take case of the injections. The chromatographic output could then be sent to the students for a short report write-up.

Learning Objectives

  1. The ability to separate, identify, and quantitate compounds at very low concentrations in complex mixtures. There is no more sensitive analytical instrumentation that can be generally used in a teaching laboratory situation than GC-MS. This important instrument deserves to be reinforced in different ways throughout the chemistry/biochemistry curriculum.
  2. The ability to identify and quantitate isomeric compounds after separation using the Cerno high-resolution mass software. To the best of our knowledge, this approach to teaching MS has not entered the college curriculum. We are confident that this can be presented and published as original chemistry education research (see the next section below).
  3. For the organic chemistry experiments, a reaction can be interrogated at a low level of conversion, before the initial products undergo further reaction. As a result, reactivity data can be gained straightforwardly and interpreted clearly.

Presentation at the American Chemical Society (ACS) Meeting and Subsequent Publication

The principal scientists at Cerno Bioscience (Steve Best and Yongdong Wang) have been encouraging one of us (Neil Danielson) for the past several years to obtain an Agilent GC-MS with their software and develop a set of teaching experiments both for organic and analytical laboratory courses describing high-resolution GC-MS for identification of isomeric compounds. Presentation at a national ACS meeting and subsequent publication In the Journal of Chemical Education of these experiments will be done. This will make Miami University the leader in instrumental analysis education using GC-MS.

Financial Information

Total Amount Requested: $33,193.54

Budget Details: We are requesting the Agilent 8860 Turbo GC-MS with FID and 50 Position Autosampler instrument with the Cerna Bioscience Mass Works software (see attached quote). The Cerna software is designed specifically for the Agilent GC-MS instrument because Agilent is considered the leader in quality GC instruments for decades. The autosampler is needed for the organic chemistry experiments to provide fast turnaround time of sample analysis due to the large number of students meeting simultaneously. The flame ionization detector (FID) is an important option to allow the characterization of new sample analysis methods without risking contamination of the MS detector. A sample analysis method can be developed using GC with FID detection to check if nonvolatile sample matrix issues could be of concern to the MS detector. The list price for such an outstanding GC­ MS instrument is for $93,322; we are getting a 43% academic discount bringing the cost down to $53,193.54. This price is still significantly less than a standard GC-MS instrument without the Cerno software or autosampler. Michael Crowder, Chair of the Department of Chemistry and Biochemistry, believes so strongly that a new GC-MS instrument is needed for our chemistry laboratory courses that he has committed $20,000 of departmental money as a cost-share (see attached letter). Therefore our proposal request for funding is only $33,193.54.

Is this a multi-year request: No

Please address how, if at all, this project aligns with University, Divisional, Departmental or Center strategic goals: One Departmental goal is to ensure that the students' laboratory experiences include working with modern instrumentation, similar to what they will encounter in industrial, government, and other academic labs.