Northeast Regional
Chromatography
Discussion Group
Fall 2005 Meeting

November 1, 2005 MEETING

Summary of Accepted Papers
(click on the title to view the Abstract)

Author Title Affiliation

Luis A. Colon
 
Ultrahigh Pressure Chromatography (UPLC)

SUNY @ Buffalo, Dept. of Chemistry


Liquid chromatography (LC) continues to be an indispensable technique in most analytical laboratories, with a remarkable impact on chemical analysis.  It is difficult to imagine an analytical laboratory without state-of-the-art HPLC equipment.  Column technology is one of the most investigated research topics in separation sciences; this is not surprising since it is within the column that the chromatographic processes take place, making the column the “heart” of the separation system.  The use of HPLC columns packed with small particle diameter leads to an increase in the speed of the separation, with a concomitant increased in separation efficiency.  To reduce the back pressure requirements that accompany the decreased in particle diameter, the length of the column has traditionally been shortened.  Recently, however, the use of very high pressures, beyond the traditional 6,000 psi, has gained considerable attention as a means to drive the mobile phase through columns packed with small diameter particles (<2 um).  We are investigating the use of columns with inner diameters of about 1 mm packed with small particles, under relatively high-pressure conditions (>15,000 psi).  These columns have been operated at such high pressures and do not appear to be more problematic than using regular 4.6 mm i.d. columns under typical HPLC pressure conditions.  Our presentation will discuss the advantages and need for ultrahigh pressures, as well as our most recent investigation using ultrahigh pressure LC.

 


Dwight Tshudy
Bio-Diesel, From French Fries to Rabbits

Gordon College, Dept. of Chemistry


The talk of alternative energy is increasing, especially as gasoline and natural gas prices increase.  There are many options that are being investigated as ways of offsetting the dependency on fossil fuel based energy sources.  These range from energy sources such as solar, wind, fuel cells and alternative fuels.  Biodiesel is alternative fuel that comes from renewable sources.  What is considered biodiesel? How is it made?  How similar is it to petrol diesel?  Why are we interested in using biodiesel?  How are fuels such as biodiesel tested?  These questions and other will be presented and discussed.

 


Troy. D. Wood
LC-MS - Nuts and Bolts of Instrumentation

SUNY @ Buffalo, Dept. of Chemistry


The combination of liquid chromatography with mass spectrometry (LC-MS) has proved a revolutionary tool in the fields of biomedicine, pharmaceutical sciences, and environmental protection.  In this presentation, the history of the development of LC-MS from its roots to the present will be presented.  A focus on the fundamentals of the ionization sources, mass analyzers, and interfaces that have made LC-MS such a broadly applied technique will be discussed.

 


Robin DiFrancesco
Biological Matrix Suppression Concerns in LC-tandem MS Analytical Methods

SUNY @ Buffalo, Dept. of Pharmacy


Liquid chromatography tandem mass spectrometry (LCMS) analytical methods are practiced as common state-of-the-art laboratory methods in an array of industrial, government, and academic laboratories.  Our laboratory specializes in methods that support clinical trials in true patient populations.  Consequently, the heterogeneity of the samples analyzed as well as numerous concomitant medications taken create special issues to consider when developing and validating quantitative LCMS methods for measuring specific analytes of interest.  This short presentation will review the concepts of matrix ion suppression [and enhancement], highlight existing approaches to investigating matrix suppression effects, and present findings from our laboratory’s approach to exploring matrix effects while validating several LCMS methods in biological matrice

 


Daniel Sudlik
Using Enterprise Content Management to Integrate Laboratory and non-Laboratory Information

Agilent


Information management in the laboratory is becoming increasingly complex due to the need to sort through a stream of email, powerpoint slides, PDF files and other data sources, along with standard instrumentation data files like chromatograms and spectra.  ECM (Enterprise Content Management) offers a unique tool to collate multiple data sources into a usable format that allows a scientist to easily locate or store information.

 


Timothy C. Schunk
A Systematic Approach to the Development of Gradient HPLC Copolymer Composition Separation Methods

Eastman Kodak Company


HPLC copolymer composition separation methods can be difficult and time consuming to develop. Based on the available literature, it is often unclear to the non-expert what path will provide the needed separation in an efficient manner. The general experience of chromatographers often leads to confusing results because of the differences in solution thermodynamics and kinetics of polymers relative to small molecules. The work presented here provides the foundation for a systematic approach to the development of gradient HPLC copolymer composition methods emphasizing adsorption separations. An organized series of steps are used to define a linkage between the information required and the chemical structure and solubility behavior of copolymers. A method development plan is outlined to define an initial method, and it is refined to provide the needed separation. Throughout, the focus is maintained on the chemical information required rather than the separation type employed (reversed-phase, normal-phase, etc.)

 

Witold Lawrynowicz Ion Chromatography Analysis of Insoluble Samples

Xerox Corporation


Many types of samples can be difficult to analyze by ion chromatography.  While some aqueous-insoluble samples can be prepared by solvent/non-solvent preparations such as in dimethylformamide and water, others are not soluble at all.  Although surface-extracted ions are relevant and can be determined, it is still often desired to determine the total ion content of some samples.  In these cases, insoluble samples such as pigments can be combusted using an oxygen bomb apparatus.  Following combustion, the residue is dissolved in typical aqueous IC solutions.  Using this technique, the levels of halogens, sulfate and phosphate can be quantitatively determined.

 


Jill Hochreiter
Conversion of a Traditional HPLC Method to UPLC Method

SUNY @ Buffalo, Dept. of Pharmacy


Ultra Performance Liquid Chromatography (UPLC) is a new technology touting increased speed of analyses along with superior resolution and sensitivity.  Our laboratory desired to incorporate a published method for several nucleoside antiretrovirals that required an analysis time of 25 minutes and a lower limit of quantitation of 15 ng/mL.  Initial experiments using the UPLC equipment provided a decreased run-time of ~10 minutes.  Several comparisons within the UPLC system’s options were performed to determine optimum parameters to gain the sensitivity desired:  use of a UV detector and a photodiode array detector, sample injection volume using different loop sizes, and use of different UPLC analytical columns.  In addition, we were concerned about the many concomitant medications HIV patients might also be taking.  Specifically, would the decreased retention times create more opportunities for interferences by other analytes?  Would the increased resolution compensate adequately?  This short presentation will provide an overview of our findings while investigating the applicability of the UPLC technology in our laboratory.

 

Lee Marotta Optimizing Detection Limits Using Headspace Trap Technology

PerkinElmer LAS


The advantages of determining volatile compounds in difficult sample matrices via headspace have been reported for a wide range of applications, making the technique a popular choice in many of today’s laboratories. The necessity to enhance detection limits from what “classical headspace” can provide is increasing. 
This seminar will discuss a novel headspace sampling system that will enable up to 100 times better detection than that provided by classical headspace. This is possible without the need for liquid cryogen, and will retain the convenience and performance offered by headspace analysis systems.
Therefore, the analyst will still enjoy the excellent precision, recovery, ease of use, and productivity (virtually no sample prep) offered with classical headspace, and achieve up to 100 times better detection limits.  In addition, this will enhance precision since responses will be greater; therefore, peak integration will be more repeatable.
In addition to enhanced detection limits, this headspace trapping technique uses a dry purge, which eliminates the majority of water and eliminates all of the air being transfer to the analytical system while retaining the analytes of interest for analysis.
We will demonstrate the ability to obtain enhanced detection limits of volatile compounds in several matrices.

 

Dan Bazavan Fast HPLC Methodology which Increases Productivity

Dionex

 

POSTERS

 

Julia Kohn

Proteomic analysis of Pseudomonas Putida KT2440 in the presence of Diethylstilbestrol

RIT Proteomics lab


The RIT Proteomics lab has been using a 2D-Gel Electrophoresis analysis of proteins to study different carcinogens and their effects on the protein expression in the bacteria Pseudomonas Putida.  The current screening test for a carcinogen is called the Ames Salmonella Assay.  The problem with this assay is that it looks only at cell growth and not at protein expression, and it also gives false negatives and false positives.  Looking at the protein expression allows for a more precise view of whether the compounds being tested are carcinogens by seeing its effects on the protein level.  The strain being studied is KT2440 and is grown on a 5mM Succinic acid carbon source.  The carcinogen being studied is Diethylstilbestrol (DES).  DES is a synthetic estrogen that was given to women between the 1930s and 1970s and was believed to prevent premature births as well as miscarriages.  Later results determined that DES actually caused a rare vaginal cancer in the women who were born from a mother that was given DES.  DES is also one of the carcinogens that give a false negative in the Ames Test.  Analysis of the effect of the concentration of DES on KT2440 was performed and analyzed, showing an effect on the length of the growth curve as well as an effect on the amount of cells that were able to grow.  Growth curves for KT2440 in the presence of .0025mM DES and Succinic acid have been performed and also the mid-log phase of growth was determined. Initial gels are being run to look at the changes in the protein expression.

 

 

 

 

 

 

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