Robotic systems preparation

In the last several years, on-line extraction systems have become a popular way to deal with the analysis of large numbers of water samples. Vacuum manifolds and computerized SPE stations were all considered to be off-line systems, i.e., the tubes had to be placed in the system rack and the sample eluate collected in a test-tube or other appropriate vessel. Then, the eluted sample had to be collected and the extract concentrated and eventually transferred to an autosampler vial for instrumental analyses. Robotics systems were designed to aid in these steps of sample preparation, but some manual sample manipulation was still required. Operation and programming of the robotic system could be cumbersome and time consuming when changing methods. 

Typical protein precipitation procedures use one volume of plasma plus three to six volumes of acetonitrile or methanol (or a mixture) with the internal standard at an appropriate concentration for the assay. Poison et al.102 reported that protein precipitation using acetonitrile eliminates at least 95% of the proteins after filtration or centrifugation, the supernatant can often be directly injected into the HPLC/MS/MS system. Usually this step is performed using 96-well plates that are ideal for semi-automation of sample preparation. Briem et al.103 reported on a robotic sample preparation system for plasma based on a protein precipitation step and a robotic liquid handling system that increased throughput by a factor of four compared to a manual system. 

MS has recently been used to measure compounds with significant levels of impurities and solubilities below the quantitation limits of other methods. Guo et al.46 described the use of LC/MS for solubility measurements in buffer solutions in a 96-well plate. Fligge et al.47 discussed an automated high-throughput method for classification of compound solubility. They integrated a Tecan robotic system for sample preparation in 384-well plates and fast LC/MS for concentration measurement. This approach is limited by LC/MS throughput. 

Lester et al. [24] have described a robotic system for the analysis of arsenic and selenium in human urine samples which demonstrates how robotics has been used to integrate sample preparations and instrument analysis of a biological matrix for trace elements. The robot is used to control the ashing, digestion, sample injection and operation of a hydride system and atomic absorption instrument, including the instrument calibration. The system, which routinely analyses both As and Se at ppb levels, is estimated to require only... 

The automated robotic system was used to prepare and analyse more than 200 spiked water samples in order to evaluate the stabifity of the system. The system was very stable and no major problems were encountered during the stabifity studies. 

The apphcations described here illustrate the wide range of uses for robotic systems. This chapter is not intended to he exhaustive there are many other examples of successful applications, some of which are referenced below. For instance, Brodach et al. [34] have described the use of a single robot to automate the production of several positron-emitting radiopharmaceuticals and TTiompson et al. [3S] have reported on a robotic sampler in operation in a radiochemical laboratory. Both of these apphcations have safety imphcations. CHnical apphcations are also important, and Castellani et al. [36] have described the use of robotic sample preparation for the immunochemical determination of cardiac isoenzymes. Lochmuller et al. [37], on the other hand, have used a robotic system to study reaction kinetics of esterification. 

A list of the 64 analytes and their method performance is shown in Table 6.6. The increased number of analytes is possible because of improvements to the collision region of the MS/MS system that provide increased sensitivity and reduced memory effects. In addition, robotic systems for sample handling and on-line solid-phase extraction (SPE) of plasma samples were integrated with the LC/MS/MS system (Figure 6.22). An isocratic reversed-phase HPLC method provided a cycle time of 4.5 min per sample. The on-line sample preparation and short analysis resulted in an increased sample throughput that required less time from the scientist. The... 

When considering automating the sample preparation steps and interfacing with chromatographic systems, laboratory robotics has been the method of choice. A laboratory robotics system has a robotic arm and controller, a computer linked to a controller or connected directly to the robotic arm, and application peripherals for performing specific functions in the application process. 

In 1996, workers at GlaxoWellcome disclosed the solution-phase preparation of 2-aminothiazole combinatorial libraries [99], Utilizing 20 glass vials and a DPC liquid dispensing robot, they prepared a library of 2500 compounds by this procedure. Also in 1996, Argonaut reported the use of their computer-controlled fluid delivery system for biaryl synthesis via a Suzuki coupling reaction [61],... 

This robotic sample preparation and counting technology, together with mechanical improvements in the chemical separation system, has resulted in an automated column chromatography system that can run almost autonomously, whereas several people were required to operate the ARCA II system for a transactinide chemistry experiment. 

Compound dilutions should be performed from 10 mM compound stock solutions in DMSO at the day of experiment. Compounds should be taken from micro well plates prepared by the different robot systems dilution is prepared with buffer or intermediate steps (containing DMSO) to avoid precipitation of compounds with low solubility. Final DMSO concentration should not exceed 0.5%. Alternatively, compound dilutions are prepared manually to result in maximal final concentration of DMSO of 0.5 %. 

For sample preparation, a Packkard MultiProbe II liquid handling robotic system was used (Packard Instruments, Meride, CT, USA) using the WinPrep software. 

The operator s attention is still necessary during continuous screening modes, and he or she can be alerted via email or telephone of problems arising during off hours. Cooperation between assay development scientists and robotic engineers is required for reagent preparation and trouble shooting. Some of the operational issues of fully automated robotic systems include ... 

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