AUTOMATION OF SAMPLE PRAPARATION IN BIOANALYTICS FOR HIGH-THROUGHPUT, ACCURATE LC-MS/MS ANALYSIS AND LABORATORY INFORMATION MANAGEMENT SYSTEM

Abstract

Developing high-capacity sample preparation systems and strategies are of key importance in providing breakthrough in the time required to develop a drug by increasing the number/time of analyzed samples. This objective must be achieved without loosing quality within the obtained data and it is of paramount importance in Pharmacokinetic/dynamic studies. The aim of this PhD project in Analytical Chemistry is to automize the manual sample preparation processes for LC/MS analysis using high throughput techniques, and to insert the developed process in the frame of a pharmaceutical bioanalytical process managed by a LIMS system. Sample preparation by protein precipitation is routinely used for removal of matrix component from biological fluids (typically plasma) prior to analysis. It is a generic sample preparation technique, applicable to a broad range of analyte types. Conventional protein precipitation is carried out in vials or collections microplates, with subsequent centrifugation and supernatant removal. Recently, filterplate precipitation has increased in acceptance, because this approach significantly reduces manual liquid handling steps and is readily automated. Automate an assay is important to increase process reproducibility and often the throughput in the laboratory make free from time-consuming repetitive tasks and gave more time for creative thinking. Automated systems will not automatically improve the quality of pipetting and yield. Only in the case of a well-defined assay with a protocol well adapted to the automated process, yield can be expected to be equal or better than in the manual process. Some assays require manual steps that require close eye-hand interaction: e.g. moving the tip of a droplet and aspirate following the droplet. In other words: mimicking complex manual processes can be a challenge even with a robotic workstation’s technology that is using advanced monitoring features to imitate such manual operations. Most liquid handling workstations have a similar hardware architecture. They consist of a deck that is the workbench, where labware with reagents and samples are placed, a pipetting arm that holds the actual pipetting units. The arm moves across the deck so that the pipetting units can reach the labware. There are individual pipetting units as well as pipetting “blocks” in 8-well format, 96-well format or 384-well format. The main and basic parameters that make Hamilton automation so efficient are Liquid Level Detection (LLD), Monitored Air Displacement (MAD), CO-RE, Total Aspirate and Dispense Monitoring (TADM) and Anti Droplet Control (ADC) Technologies. With this preliminary remarks and with a good skill of Microlab Star Vector Software it could be possible to adapt and to perform laboratory methods to increase processes reproducibility and the throughput in the laboratory. The first aim of my project, after having learnt how to use the software and how to adapt the methods to the robot ability, has been to perform a Protein Precipitation Method with a high level of…