Addressing the Challenges of Complex Mixture Analysis with Sub-2 µm Particle Liquid Chromatography: Long Columns and High Temperatures

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Robert Plumb, Paul Rainville, and Uwe Neue (Waters)
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Application Notes
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Application Notes
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Q-Tof Premier
Q-Tof Premier

The challenge of analyzing complex mixtures requires an analytical system and methodology that, together, extract the maximum amount of information from the sample in the shortest time to optimize productivity. The ACQUITY UPLC System has been specifically designed to produce chromatograms of the highest resolution, allowing scientists to fully interrogate complex samples.

The ACQUITY UPLC System has low-dispersion characteristics, precise flow delivery characteristics, and uses a sensitive detector to realize the chromatographic potential of sub-2 µm porous chromatographic material. The rugged design of the instrument facilitates the use of high operating backpressures, permitting the use of longer columns for high-resolution separations.

The use of elevated temperature in liquid chromatography does not increase the maximum obtainable performance in either isocratic or gradient mode for a given column. It does, however, allow the same performance to be obtained in less time. Increasing the column temperature allows the use of longer columns, generating gradient peak capacities in excess of 1000 in just 60 minutes.

The chromatograms produced using these sub-2 µm particles are significantly superior to those produced by 3 to 3.5 µm particles, which generate separations of peak capacities in the region of 200 to 300 per hour.

The combination of sub-2 µm particles used at elevated operating temperatures, with optimized flow rates, and in long column formats allows for complex samples, including natural products (i.e. ginseng), biological fluids (i.e. bile, urine), and pharmaceutical compounds, to be investigated in much greater detail than before – all in a time frame of one hour or less.

This improves the ability of scientists to understand more about their samples and maximize return on large capital investments such as mass spectrometers.

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