Journal of Analytical & Bioanalytical Techniques
Open Access

Abstract

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 lifetimes. 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.

Biography

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.