LC-UHPLC Hybrid 2D Platform For Biological Samples LC/MS Analysis: A New Paradigm | 18661
Journal of Analytical & Bioanalytical Techniques
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Modern UHPLC systems have important advantages over previous generation HPLC systems. These are: a) higher pressure
limits, and b) considerably smaller delay and post column volumes, which are essential for the development of fast
gradient applications. Fast gradient applications by UHPLC/UV systems are easily adopted in QC/QA applications. Analysis
of complex samples was found to be more complicated and costly, if a UHPLC system was used. Clinical sample analysis
usually utilizes a large volume injection of a diluted sample, which is suboptimal for a UHPLC autosampler. Extracted samples
obtained after sample preparation steps, drying and reconstitution may have small particulates that shorten UHPLC column
We developed a cost-efficient hybrid LC platform that�s very efficient for biological sample analysis by LC/MS. This
platform is based on Agilent 1100 and Agilent 1290 series LC devises. The sample was injected by a standard (not UHPLC)
autosampler into pre-analytical column, where it was desalted and partially purified. A standard pressure range binary HPLC
pump was used.
The fraction containing analytes of interest was transferred through a UHPLC valve to a fused core column. A fast gradient
was performed by using a UHPLC 1290 pump. The addition of a UHPLC valve and pump to a standard 1100 series LC
system (autosampler, pump and column compartment) greatly extends operational flexibility including column selection,
while standard LC - which is already available in the lab, performs the initial steps of sample loading and clean-up.
This platform for LC/MS analysis was successfully used. Instead of retiring an entire functioning Agilent 1100 LC system,
it was added just one UHPLC pump to achieve much greater overall performance, functionality and lower cost, compared to a
single pump UHPLC system purchase.
Eduard Rogatsky completed his MSc in physical chemistry from Belarus State University, PhD in Bioanalytical chemistry from Bar-Ilan University (Israel) in 1999, and postdoctoral studies at Albert Einstein College of Medicine, NY. He joined the faculty there in 2001, and is currently a Senior Associate Scientist and Director of Mass Spectrometry in the Biomarker Analytical Resource Core Laboratory, Einstein-Montefiore Institute for Clinical and Translational Research, Bronx, NY, USA.
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