Table of Contents

Call for Palmer Award Nominations

Luke Charpentier

The Minnesota Chromatography Forum (MCF) is seeking nominations for the Palmer Award to be presented at the 2002 Spring Symposium. The Palmer Award has been given out by the MCF for the past 22 years. The award recognizes individuals for their contributions to the art and science of chromatography. The award can also be given to an individual for his or her service to the MCF or the chromatography community. Past recipients have included internationally recognized chromatographers as well as long-term MCF volunteers. For listing of the past recipients of the Palmer Award please visit the MCF web site at www.minnchrom.org. The Past President of the MCF heads the Palmer Awards committee which reads and discusses all individuals nominated for the award, and then selects a winner. The nomination form can be found on the MCF web site at www.minnchrom.org.

Nominations Sought

Brian Leafblad

It's that time of year again! The 2002 Spring Symposium is just around the corner, and the Symposium Committee is busy getting ready. This may sound like a cliché, but in a volunteer organization such as MCF it really is the people that make the difference. When we all come to the Spring Symposium we get to enjoy the final product, and may not realize all the work that went into preparing for it. We get to participate in a fabulous conference thanks to the efforts of people who have volunteered their time, some year after year. So, when you come to the Symposium this year, take a moment to thank the Committee for their hard work. The high quality we have come to expect is truly a reflection of the people that are doing the work.

One of the annual activities at the Symposium is the election of some of our Board members. The MCF Board is the group of people who chart the course of MCF. If you would like to get involved, now is your chance! Or, if you know of someone to recommend, nominate them!

We will be electing a 1st year Director, President-elect and Secretary-elect. If you would like to find out what people in these positions actually do, feel free to contact us. You can find out who and how at www.minnchrom.org/board_members.htm. (Or, if you would like to get involved with one of the committees, contact the chairperson.) All of the contact names and numbers can be found on the MCF website.

We will compile a list of candidates for the three positions with brief bios for you to read at the Spring Symposium, where we will decide by ballot the new leadership. Please give thoughtful consideration to whether you might volunteer or know of someone to nominate. This is your chance to have an impact on how MCF is run and what we do.

You can volunteer or forward nominations to me at brian.leafblad@ecolab.com or at (651)306-4147.

Coming Soon!

Grant Paul

The last of the MCF quarterly presentations will be held on Monday April 22, 2002 at Wyndham Garden Hotel in Bloomington, MN. The speaker will be Mr. John C. Hudson of the Royal Canadian Mounted Police Forensic Laboratory in Regina, Saskatchewan Canada. The presentation will cover applications of Capillary Zone Electrophoresis in Forensics and Toxicology.

Watch your mail for a special announcement!

2002 MCF Undergraduate Research Award (UGA)

The MCF Undergraduate Research Award is awarded annually (this year in March!) and presented at the Spring Symposium of the Minnnesota Chromatography Forum.  This year's award winner is Ms. Molly Warnke of Hamline University.  She will be guided in her research by Prof. Eugene Smith.  The title of her research is Arson Analysis by GC and Simplex Optimization.

Education Corner:  Winter Course Reviews

"Beginning Gas Chromatography"

by DeWayne Townsend

John Freeburg and Gary Reineccius taught a stimulating and informative session on the basics of gas chromatography in January 2002.  The class was down a bit in attendance, not surprising considering the state of the economy at the time, but 14 individuals should understand their gas chromatographs much better than they did in 2001.

The class covered the basic chromatographic theory along with some history and launched into the details of a gas chromatograph following the flow of carrier gas.  Questions such as what gas should one use, how does a split injector work, why would one want to temperature program and why does my flame ionization detector give a higher response for octane than butane, were all answered during this three day class.  Beyond the gas chromatograph the issues of data collection and analysis were discussed in relationship to the peculiarities of gas chromatography.  The characteristics of various detectors, the issues regarding qualitative/quantitative  analysis and what to do if it doesn't work were also explored.  The class was organized with lecture in the morning and a hands on lab in the afternoon.  Installing capillary columns, testing for leaks, lighting a FID, and actually trying to identify and quantify unknowns were some of the topics uncovered by the class during the afternoons. 

"Use of Designed Experiments (DOE) in Chromatographic Applications"

by Patrick Crain

On February 5TH -7th, 2002, MCF offered a three day course titled "Use of Designed Experiments (DOE) in Chromatographic Applications". This course was prepared and presented by Dr. Lars Pekay and held at the Earl Brown Continuing Education Center on the University of Minnesota, St. Paul Campus.

Dr. Pekay’s background includes a Ph.D. in Analytical Chemistry from Ohio State University with abundant experience in SFC, GC, and HPLC. He is currently the Quality & Regulatory Operations Manager, for General Mills at their Lodi, California facility. However, before his move to California, Dr. Pekay was located here in the Twin Cities (with General Mills) and very involved in MCF. Even though he had experience with the local climate, we were still able to convince him to make this trip to Minnesota in early February.

The objective of the course was to provide both a fundamental and practical knowledge of (factorial) design of experiments. It was shown how DOE could be used as a powerful tool to improve experimental efficiency and increase the amount of information obtained from a set of experiments. This methodology is applicable to all types of experiments; however, the course was able to focus on experiments geared toward optimization of a chromatographic separation. By carefully setting up the experiment, several parameters can be varied simultaneously and the effect from each variable, as well as the interaction between variables, can be determined.

Along with the hardcopy, the course manual was also provided on CD, courtesy of Dr. Pekay. This makes for easy printing of the DOE tables and forms and had the added benefit of easing the demand on our limited shelf space. The course was very well received by those attending and the Earl Brown Continuing Education Center was a great venue for holding such a course.

2002 Spring Symposium Program

2002 Spring Symposium Registration

Invited Talks in 2002 MCF Spring Symposium

High Throughput Chemical and biochemical Measurements

Edward Yeung

Abstract: In the next decade, high-throughput measurements of all types will be in great demand. Not only will it be necessary to keep up with the many samples and the many tests in clinical, environmental and processing applications, it will also be possible to enhance the chances for new scientific discoveries by doing many more experiments in parallel. New instrumental approaches include 96-capillary electrophoresis with absorption detection and in situ laser-excited fluorescence imaging of heterogeneous catalysts. Examples will be presented in mutation detection, proteomics, combinatorial synthesis, drug screening and the development of new catalysts.

Proteomics Without Polyacrylamide:  Qualitative and Quantitative Approaches to Proteomics via Mass Spectrrometry

David R. Goodlett

Abstract: Proteomics can be described as the study of proteins expressed by a given cellular state, and like genomics, it is on first pass a global rather than hypothesis driven science. Questions for study are not asked in series, such as which single protein causes a given biological activity or effect, but rather in parallel, such as how is the proteome diagnostic of the environment a cell experienced before analysis. Mass spectrometry is currently the most important discovery based tool in Proteomics. In an effort to move the role of mass spectrometry role in Proteomics from descriptive to quantitative, several research groups recently published mass spectrometric methods for determining the relative abundance of proteins expressed between two different states of a similar biological system (Aebersold and Goodlett Chem. Rev. 2001, 101, 269). These methods allow the changes in protein expression between cells in different states or environments to be studied on a global scale so that information on protein identity and expression may be gathered on multiple proteins in a single experiment. Given the high variability in electrospray ionization (ESI) efficiency between peptides of different sequence and the variability in ESI across a chromatographic separation, the advantage of these methods is that each peptide entering the mass spectrometer will have a suitable internal standard for reliable "relative" quantification. We will present qualitative and quantitative mass spectrometric approaches for studying proteomes and discuss the limitations of current methods. Specifically, we will present data describing generic mass spectrometric methods for determining high proteome coverage without the use of polyacrylamide and the use of isotope coded affinity tags (ICATÔ ) to determine: 1) the relative abunance of proteins between two different cellular states and 2) de novo sequencing.

Automated Multidimensional HPLC For Proteomics Sample Preparation Prior to Nanoscale LC-MS/MS Protein Identification 

Kerry D. Nugent

Abstract: Now that the human geonome has been sequenced, attention is shifting to proteomics and the role that proteins play in the structure, function and control of biological systems. In comparison to gene expression analysis at the m-RNA level, proteome analysis provides more accurate biological system information by directly measuring the actual molecules responsible for biological effects. With recent developments in methods and instrumentation for automated, data dependent ESI-MS/MS in conjunction with nanoscale HPLC and database searching, simple protein mixtures (tens of proteins) can be quickly identified without the need to purify each protein to homogeneity prior to analysis at the femtomole level. More complex protein mixtures (hundreds to thousands of proteins) still require some degree of fractionation prior to nanoLC-MS/MS of protein digests, since a 1D HPLC separation is limited to resolution of 50-200 components. Orthogonal 2D HPLC methods dramatically improve the resolution of highly complex mixtures of proteins and peptides versus a 1D separation. Although 2D HPLC may not be able to achieve the same degree of resolution as 2D PAGE, it is sufficient to fractionate complex protein mixtures into simple mixtures for analysis by nanoLC-MS/MS. 2D HPLC is easily automated to give reproducible separations with quantitative recoveries at femtomole levels. With proper conditions, 2D HPLC can also be non selective, yielding good recovery of all classes of proteins including membrane proteins and lower abundance regulatory proteins (transcription factors, protein kinases, etc). Several schemes for orthogonal HPLC of proteins and peptides were explored to find optimum conditions for low level protein identification in complex proteomic samples, using a new, fully automated multidimensional HPLC system.

Development and Validation of CE and HPLC Screening Methods For Early Stage Drug Development Projects

Kurt Sedo

Abstract: Suitable methods for the assessment of purity, potency and stability of drug substances and drug products are required to be rapidly developed and validated to provide appropriate data for early project development decisions. In order to routinely provide methods of consistent quality to meet aggressive timelines, Pharmacia has begun to utilize systematic approaches to both develop and validate analytical technologies that are appropriate to the stage of project development. Method development and validation approaches for CE and HPLC methodologies supporting an early stage project will be discussed. A comparison of the validation and authentic sample data between the two methodologies will be presented. Finally, conclusions in regards to implementation and routine use of the systematic approaches will be discussed.