Table of Contents

MCF Spring Quarterly Meeting

Title:
"Coupling Laboratory Animals to Liquid Chromatography Using Microdialysis Probes and Robotic Blood Sampling. Pharmacokinetics, Pharmacodynamics, LC/MSMS and LC/EC."

Abstract:

In the preclinical phase of drug development it is inevitable that animal studies must be accomplished to understand such issues as drug safety, drug metabolism, oral absorption (if any), blood brain barrier penetration (if any), the rate of excretion, the effect on behavior (CNS effects), and so forth. Bioanalytical chemistry is crucial to gaining an understanding. Samples must be taken and compounds must be quantitated, including the drug substance and any metabolites, as well as endogenous substances the drug candidate might influence. We have been developing sampling approaches which reduce animal stress, improve data quality, and reduce the number of animals required to get to a satisfactory result (this reduces cost and speeds the drug candidate to the next steps). This presentation will illustrate this integrated approach in which liquid chromatography, mass spectrometry, and electrochemistry are coupled to sampling in a more automated fashion than was possible even a just a few years ago. Components of the approach also include 96-well SPE, the Raturn animal containment system, the Culex robotic blood sampler, and epsilon microbore liquid chromatography levels. 

Dr. Peter Kissinger is a Professor of Chemistry at Purdue University and the founder/CEO of Bioanalytical Systems West Lafayette, IN. Peter Kissinger received his B.S. from Union College in 1966 and his Ph.D. from the University of North Carolina in 1970. After spending time at the University of Kansas and Michigan State University, he moved to Purdue University. Over the years Dr. Kissinger has given over 400 invited lectures and published over 200 manuscripts. When not doing chromatographic work you can find Peter working in other ventures, including bee keeping.

2001 MCF SPRING SYMPOSIUM 
UPDATE

Peter Johnson

Abstracts are still being accepted for Poster and Oral Presentations at the 2001 MCF Spring Symposium Abstract Deadline April 15, 2001

Instructions for Abstract submission are available at the MCF website - www.minnchrom.org

REGISTRATION

Registration information for the 2001 MCF Spring Symposium is available inside this newsletter and also at the MCF website - www.minnchrom.org.

Course registration deadline is April 30, 2001. Course information is available on the MCF website.

Preliminary Program will be available on-line at the MCF website in late April.

Vendor Seminars – Information will be available at the MCF website in early May.

INVITED SPEAKERS FOR THE 2001 MCF SPRING SYMPOSIUM

Keynote Speaker
Professor Csaba Horvath,Yale University
Topic – Capillary Electrochromatography

Focus Speakers
Dr. Steven Naylor,Mayo Clinic
Topic - Capillary Electrophoresis

Dr. Art Fitchett, Dionex Corporation
Topic - Ion Chromatography-MS

Dr. Frank Yang, Microtech
Topic - Capillary HPLC

Professor Richard Sacks, Univ. Michigan
Topic - Capillary Gas Chromatography

Pittcon 2001

Donald F. Decou, 3M Pharmaceuticals

The 52nd annual Pittsburgh Conference and Exposition on Analytical Chemistry and Applied Spectroscopy was held in New Orleans, LA from March 4 through March 9 2001. As usual, this year the conferees could stay busy by attending one of the 1440 oral presentations, viewing over 800 posters or expand their knowledge through one or more of the approximately 100 short courses.

Chromolith column from Merck KGAa of Darmstadt, Germany was the most interesting new thing I found. The monolithic silica technology used in Chromolith columns creates porous silica rods of polymerized silica that are mechanically stable. The Chromolith column, according to the manufacturer is a column with lower pressure drops than conventional silica columns that can be used with flow rates as fast as 9 mL/min, which allow short equilibration times and high throughput. I wasn’t the only one that found this product interesting. When I read the newsclip that the Chromolith column won the Gold Award for best product in the show, I could not help but pat myself on my back for picking this as the most interesting new thing! More information can be found at http://www.chromatography.co.uk.

That should be enough to attract any chemist, but it is not what attracts most conferees to the Pittcon. The biggest draw is the exposition floor. This year more then 1200 instrument manufacturers, laboratory suppliers and contract laboratories set up shop in over 3200 booths. Companies from Ace Glass to the Zymark Corporation were willing to pull you over to demonstrate the latest advances in analytical chemistry and applied spectroscopy. The Thermo Corporation must be trying to become the single largest instrument supplier on the planet. By my count, there were 20 "Thermo-" booths. Thermo-Finnigan was selling mass spectrometers right next to Thermo-Nicolet selling FT infrared equipment, which was right next to Thermo-Orion selling pH and conductivity meters. It was an interesting sight!

Pittcon is not all business however. There were 13 tours and cruises of the New Orleans area. An added bonus is the fact that Bourbon Street is only about eight blocks from the convention center. Let me tell you that the fact that this year’s event was held the week after Mardi Gras did not stop people from having a good time in the evenings. More then one Pat O’Briens original Hurricane was consumed in the name of science! There was also the traditional Sunday night mixer. This is an event where old friends and acquaintances can meet and reminisce about the good old days as I did with 10 fellow Kansas State University alumni and professors.

If there is one problem with the Pittcon it is that it is too big! Seriously! The Morial Convention Center has 1.1 million square feet of contiguous exhibit space on one level in one building. It is one of only four convention centers that routinely hold a convention the size of the Pittcon. The others are in Atlanta, Chicago and Orlando. There is literally too much to see and do, and I always take it as a personal challenge to do everything possible. The first two days and nights were so busy that by Wednesday I was exhausted and needed a break! Thursday was a little less hectic, but by the time I returned, I found that I actually needed to go to a hospital to pass a kidney stone, but that’s another story!

Is spring here yet? I am sure you are all asking the same question. Certainly, the spring symposium is less than 8 weeks away. Please send in those abstracts. For those who are planning to stay in the Twin Cities near the convention center, Blocks of rooms are available at the Country Suites and AmericInn near the EBHC. It can be very difficult to secure accommodations at the last minute.

Those of you who returned from Pittcon, please send your comments or thoughts like my colleague Don Decou did. We will find space to print your impressions and observations about Pittcon or any other meeting you may have attended as long as it has the flavor of "Separation Sciences".

I received a comment via the web which can be found in the full unedited form under the new column Reader’s Comments. By the way, whoever sent that comment will see that his/her suggestion has been faithfully implemented although I disagree with the reader’s calling the placement of the items "inappropriate". We, the editors, really do appreciate such comments. Only one request. However critical your comments, please do not sign the letter "Anonymous". This is your news letter and we want to improve it and make it useful to all the members of MCF.

Ward Swanson, President-elect of MCF is soliciting nominations for President-elect, Secretary-elect and 1st year Director. For further details please contact Ward at 651-832-2660 via email at wswanson@barr.com.

Readers’ Comments

MessageType: MCF Website Comment - Suggestion

Subject: Newsletter

Username: anonymous

After receiving this month’s newsletter, I finally had to comment.

I think it a bit inappropriate for the "From the Editors Desk" to be on the front page. Articles or announcements of greater importance should be there. For example, this month - the Spring Quarterly meeting announcement.

Large Volume Injection in Gas Chromatography

Rick Rossiter,
3M Corporate Analytical Technology Center

The promise:

"Inject 100無 into your gas chromatograph instead of 1無 and get two more orders of magnitude sensitivity". Who among us would not be lured by such a promise. I know that I was. Of course, it turns out not to be that simple. Getting one factor of ten is quite easy, but getting that second one is a lot more challenging.

The first thing you need, besides a large volume injector, is a strong nerve - to inject 100無 of solvent into a GC/MS system. Mass spectrometers are not very forgiving if you get it wrong, I discovered.

The mechanisms:

There are three main types of injectors on the market today. All of them contrive to vent enough of the injection solvent so that the separation column is not flooded, but retain the analytes in the system. They do, however, have different ways to accomplish this.

Early solvent vapor exit(Type 1).

The solution is injected cool into a retention gap onto the front of the column, and the solvent vapor exits at a T- before the analytical column.

The T- limb is closed at the right moment just before all the solvent is gone and the chromatography proceeds through the analytical column.

Programmable Temperature Vaporizing Injector [PTV]
(Type2):

This looks very much like a standard split/splitless injector, with an absorbent solid packed into the liner. The solution is injected as a liquid onto the cool absorbent solid, so the solvent evaporates off in split mode via the split line – evaporative cooling keeps the temperature down and the analytes adsorbed onto the solid. When the solvent has gone the injector is switched to splitless mode and heated rapidly to release the trapped analytes and distill them onto the column.

GC-GC ( Type 3):

Here the split/splitless inlet is much longer, and functions as a short packed GC column. The injector temperature is held just above the boiling point of the solvent at injection, and the mixture passes through the GC column as a vapor. The solvent elutes first while the injector is in the split mode and is vented through the split vent, while the analytes are retained on the GC packing. When the solvent has gone, the pre-column is heated rapidly to transfer the analytes to the column in the splitless mode.

A listing of the main manufacturers, which I do not claim to be complete, is given below:

Type 1 Early solvent vapor exit

SVE kit from Agilent www.agilent.com

Type 2 PTV

Optic 2 from Atas – www.atas-int.com/htm/usapro.htm
Gerstel CIS – www.gerstel.com
Perkin Elmer ELVI – www.perkinelmer.com
Agilent PTV – www.agilent.com
Trace GC from Thermo Finnigan – www.tmquastin.com

Type 3 GC/GC

Apex ProSep – www.apex-technet.com

Manufacturers claim different capabilities – as this is not a "consumer digest report" check them out for your particular analyte / solvent / matrix system. I am not making any recommendations as to which system to buy!

The realities:

My own comments that follow are based on my own experience with the last two types of systems. The first type reminds me of on-column injections – those work beautifully for nice clean standards but I worry about the messier matrices.

All systems need some careful preparation, and you need to follow a logical set of steps to come up with the right conditions – these are not "plug and play" injectors.

Timing is everything – switch the solvent venting off too soon, and your column is flooded – but change too late and you risk some of your analyte heading off down the split line. Some systems can be fitted with a thermal conductivity detector in the split line to automatically trigger the purge change as the solvent peak dies down – such add-ons are a great help if you must attempt quick analyses of unknowns.

Obviously the bigger the difference in volatility between solvent and analyte, the more foolproof the separation. To give the best chance of success it helps to have some selective adsorptivity of the analyte onto the port packing. This is often a siloxane coated support, or Tenax^TM , so these possibilities have been reawakening an interest in all those old column packings which may be forgotten in a dusty corner of your lab – we have a set that have been drifting homeless around our organization like a silanized Flying Dutchman, and has now washed up in my lab!

Generally the solvent is split off, not vented altogether, although additional gas flows, as in the PE and Trace systems, can be brought in to divert all the solvent away from the column. Also, some solvent is often left in the injector at the time of switching and transfers with the analyte.

So most systems still transfer quite a lot of solvent to the column – you need solvent and temperature focusing as in any other splitless injection, so this is not easy to do with water and polar organic solvents. Like a splitless injection, the transfer to the column is slow, even more so given the extra packing in the liner, and the increased dimensions of a pre-column. It can take up to three times longer than a normal splitless injection although a pressure pulse can help – not exactly compatible with fast GC.

I am convinced that you need mass selective detection if you are going to do low level work, even high purity solvents contain many peaks at this level, and you will want to be sure that the particular little blip you are looking at is in fact your analyte.

Selectivity is important in the analytical column also. Two orders of magnitude more analyte may also bring with it two orders more of close co-elutants. For very busy chromatograms, injecting more may be counter productive without other ways of improving peak capacity. So mass selective detection and / or column switching may be important here.

In conclusion:

For me then, large volume injection usually means 20-30無, to get the benefits of larger volume without having to do so much optimization. The biggest syringes that fit most autosamplers are 50無 or 100無, and often the most you can inject is 50% of the capacity. My "compromise" between large volume and ease of use means that I get the benefits from a larger volume injection more often than if I were always to spend the time required to push these systems to their limits.

One last thing to watch out for – we add syringe washes to a programmed sequence with little thought – but several washes of a 100無 syringe will soon empty that little solvent vial !

A Message from The President!

Luke Charpentier

Hello Chromatographers! I am sure everyone is longing for spring and starting to think about our annual Symposium. Yes, it is just around the corner. The Symposium Committee has done an excellent job again this year of putting together a great lineup of speakers The Symposium offers us a chance to learn about new developments, discuss different techniques, and above all renew some of our old acquaintances. The Earle Brown Center is an excellent location for the Symposium, offering beautiful surroundings very conducive for informal discussions. Take a look at the classes organized by the Education Committee to be held during the Symposium. If past experience is any indication, the classes tend to fill up quickly so if you are considering signing up, I recommend you hurry. I sincerely hope that all members will try to make it to the Symposium this year.

Since I have space to write, I would like to request the members for some help. The MCF has evolved over my 12 years of involvement, with Environmental as the focus at one time, then Food Chemistry, and now Pharmaceuticals. What will come next is anyone's guess.

I believe, as does the Board, that there are other sectors and people out there that we need to contact. Many of them may not know what MCF is. My mind goes back to my first phone conversation with Sarah Cherney (our Secretary in the wings), when she called the MPCA about a chromatography issue. I casually asked her if she was a member of MCF. I was surprised to find out that she had never heard of the MCF. Can we minimize such occurrences? Of course we can.

I encourage all of you to share the newsletter with your peers, colleagues and supervisors to raise or increase the awareness about MCF. If there is a bulletin board where you can post notifications about MCF seminars, please do so. MCF has one of the best maintained websites. Please encourage your peers and colleagues to visit the website www.minnchrom.org

I also (I am using all my favors) ask that if you know other chemists interested in separations, invite them to attend the next Spring Quarterly Meeting on April 19, 2001. These meetings are the best place to network. When you attend a MCF quarterly meeting, you get the opportunity to socialize with other chromatographers and learn about the new developments in the area of Separation Science. We need to keep our organization growing as we continue to evolve with the field in the years ahead. If each of you can recruit just one new member, the organization will double! I am very optimistic this is an achievable goal.

Thanks for your time and your membership. Have a good month!

Call for Palmer Award Nominations

Ravi Ravichandran

MCF is seeking nominations for the Palmer Award to be presented at the 2001 Spring Symposium. The Palmer Award has been given out by the MCF for the past 21 years

The Palmer Award was created to recognize and to encourage the art and science of chromatography, in particular as it is related to the membership of the MCF. It is given to those individuals whose professional as well as scientific accomplishments have fostered progress in chromatography. The award consists of a plaque, suitably inscribed, which is presented at the MCF Spring Symposium. The first Palmer Award was received by Leslie Ettre in 1980 (appropriately, since Dr. Ettre was primarily responsible for calling Palmer's career achievements to our attention). Other past recipients include internationally recognized chromatographers as well as long time MCF volunteers. For listing of the past recipients of the Palmer Award please visit the MCF web site at www.minnchrom.org.

A committee that reads and discusses all individuals nominated for the award selects a winner. The Past President of the MCF heads the committee. The nomination form can be found on the MCF web site at www.minnchrom.org.

I encourage all of you to consider taking this opportunity to nominate that one special person you know, associated with chromatography and or MCF. Who knows? Your nominee may win the Palmer Award!

Completed nominations should be sent to

Dr. Ravi Ravichandran
3M Pharmaceuticals, 3M center 260-4S-13
St. Paul, MN 55144

Education Corner

Course Back by Popular Demand!

Mark your Calendars!

November 6th - 8th, 2001: "Beginning HPLC",

Instructors: Larry Felice (Medtox) and David Johnson (3M)

November 27th -28th, 2001: " Advanced HPLC",

Instructor: Derek Southern (LC Resources)

February 5th - 7th, 2002: "Use of Designed Experiments in Chromatography",

Instructor: Lars Pekay (General Mills)

Watch your mail or visit the MCF Website for registration and course details