Presenters: Phil DeLand, Doug Cyr, David Neyer, Jason Rehm, and Phillip H Paul
Authors: Justin Kittell
Current trends in HPLC system development have embraced higher pressures, higher temperatures and lower flow rates to increase separation efficiency and analytical throughput. By combining recent advances in microscale fluid delivery, small particle (~ 3μm) stationary phases, high temperature separations and chip-based UV absorbance detection to produce a fully integrated microflow gradient HPLC system, the overall chromatographic performance has been optimized for short cycle times, high resolution and good detection sensitivity. Results of tests using several pharmaceutical formulations indicate that the system design optimization enables separation efficiencies approaching that of UHPLC systems without the requisite ultra high pressures.
Fast and robust microflow HPLC method for open-access product analysis in discovery medicinal chemistryPresenter: Hung-Yuan Cheng
Authors: Hung-Yuan Cheng, Sung Baek and Remco van Soest
The advent of high throughput synthesis and purification has created a need for more efficient ways to do reaction scouting, route development and product profiling. Microflow HPLC, defined as having chromatographic columns in the 0.3 to 0.5 mm range and flow rates in the 2 to 60 µL/min range, can significantly shorten the HPLC analysis time attributable to its characteristic rapid gradient mixing and fast column re-equilibration. Additionally, the substantial reduction in solvent usage and waste generation can contribute to the increasingly important "Green Chemistry" effort. Using a novel microflow HPLC system, we have developed a robust short-cycle time (< 2 minutes) rapid gradient method for the analysis of crude synthesis products in discovery chemistry labs. Chromatographic columns of different phases, particle sizes and dimensions are evaluated on a set of compounds covering a wide range of lipophilicity. The capabilities of the microflow HPLC method to deal with real world "dirty" samples are critically investigated. The results compare favorably to those obtained using a conventional 4.6 mm column rapid gradient HPLC system. The benefits and issues of using microflow HPLC in an open access medicinal chemistry environment will be discussed.
Green LC techniques for solvent waste reductionPresenter: Justin Kittell
Authors: Justin Kittell
HPLC is a nearly universal analytical technique applied on a routine basis around the world for separation, quantitation and identification of chemical compounds by industry and academia. Approximately 130,000 analytical HPLC instruments are in use today, with another 16,000 instruments sold yearly. The majority of these instruments use a typical column diameter of 4.6 mm internal diameter (I.D.) with most analytical methods using a flow rate of 1 ml/min for this sized column. If the typical HPLC is used 20 hours a week, a single conventional HPLC would require 60 L of solvent (typically acetonitrile, methanol, isopropanol, hexane, and/or aqueous buffers) to operate and generate 60 L of mixed organic/aqueous waste per year. This translates into 8 million liters of solvent produced each year to supply mobile phase for analytical HPLC while producing the equivalent of nearly 40,000 55-gallon drums of waste per year. In this poster, we will analyze HPLC solvent waste trends on a geographic basis and look at the potential impact of various HPLC technologies for corporate green initiatives. Conventional HPLC, ultra high pressure LC, SFC, and micro-flow HPLC all provide the same information but different impacts on waste generation. Besides the analytical performance characteristics of each LC technology we will compare their environmental impact.
Rapid identification of metabolites of haloperidol in rat liver microsomes using microflow LC/MS/MSPresenter: Hung-Yuan Cheng
Authors: Hung-Yuan Cheng and Remco van Soest
When coupled to a triple quadrupole mass spectrometry, microflow HPLC offers significant advantages in analysis speed and chromatographic resolution for the identification of drug metabolites in biological media. We investigate systematically the detection and identification of metabolites of haloperidol in rat liver microsomal preparations with a series of reverse phase gradient elution methods from 15 min to 1 min runtime with the mass spec operating in the multiple reaction monitoring mode. The chromatographic resolution and quality of the mass spec data are evaluated with respect to the number of identifiable and quantifiable metabolites. The value of using rapid gradient microflow LC/MS/MS for a quick profiling and semi-quantitative estimate of metabolites from candidates in early drug discovery will be discussed.
