FEATURE

Chromatography in action

Warren Clark on acquisition applications

The Institute of Chemical Technology, Prague

The Institute of Chemical Technology, Prague (ICT) is the largest educational institution of its kind in Central Europe. There are four faculties and many departments with different laboratories at the school. More than 50 independent chromatography stations, provided by DataApex, are in place at ICT to aid students and researchers in a variety of educational and research needs.

All students at ICT must pass a course in core laboratory practices, so that they become acquainted with the basics of chromatography. Students who are being exposed to chromatography for the first time need to know how to acquire and evaluate chromatographic data. Because these courses are mandatory for students, it is vital to ensure that the software used is equipped with an intuitive interface, and that it is simple to use. ICT selected Clarity Chromatography Software precisely because it meets all of these requirements. Also, Clarity EVAL, a supplementary offline version of the same software, is available so that work with data can be continued after the measurement session has ended.

Researchers enjoy the intuitive interface and its versatility. Advanced calculations and the ability to compose one’s own calculation columns are necessary for research. Modular design allows Clarity to be used in special applications such as gel permeation chromatography (GPC/SEC), spectral processing for photo diode array detectors (PDA) or capillary electrophoresis (CE). It can also be used for data acquisition in non-chromatographic applications like thermogravimetry, porosimetry or cyclic voltametry.

Associate professor Jana âopíková, at the department of Carbohydrate Chemistry and Technology, uses the Clarity GPC module for size exclusion chromatography while researching the immobilisation of metals to acid polysaccharides.

ICT Prague is a typical educational user of Clarity. Clarity can also be found in other laboratories, including environmental, pharmaceutical, food control, chemical industry and so on.

Institute for Research in Immunology and Cancer, Montreal

The Institute for Research in Immunology and Cancer (IRIC), based at the Université de Montréal, Canada, brings together 17 research teams and 10 core facilities led by Dr Pierre Thibault. His lab focuses on the development and application of proteomics and bioanalytical mass spectrometry to identify trace-level proteins in complex cell extracts. His lab utilises a multidisciplinary team approach, using methods and techniques from bioanalytical chemistry, protein chemistry, biochemistry and cell biology.

Essentially he is trying to find needles in haystacks, using a combination of highly sensitive data acquisition tools, to collect the relevant information from large milieus of proteins. He then applies powerful profiling techniques to eliminate most of the haystack to hone in on the key target proteins associated with immune and cancer diseases. These proteins may be of very low abundance and only change slightly – and therefore peak capacity and resolution power need to be maximised, without losing the ability to see the ‘bigger picture’. Such a process requires LC/MS products and carefully designed profiling tools.

Dr Thibault spent three years developing the software for analysis and profiling and has relied on products from Agilent Technologies, such as the HPLC Chip and 6210 TOF LC/MS system with ESI ion source. Using these, he can identify all proteins that change in abundance, at whatever level that is. One of the keys to the research is the Agilent HPLC Chip’s ability to fraction the complex protein mix ready for use on the ESI-TOF LC/MS system or an ion-trap MS. This is important, since the TOF-based MS is excellent at identifying target proteins, whereas the ion-trap MS is better for quantification and qualification of known target proteins. By having the same fractionation method, both techniques will have the same input. This then removes further complex calculations from the analyses and profiling stages making the complex task more streamlined.

The Dow Chemical Company, Philadelphia, USA

Many chromatography laboratories, particularly in petrochemical and chemical facilities, require 24/7 operation. Unfortunately, routine operating system updates and reboots that are required to ensure network security can cause unnecessary downtime. The impact is reduced productivity and possible delays in product delivery. The Dow Chemical Company took a unique approach to this problem through the clever application of firewalls and system isolation, to ensure the security of its labs without disrupting productivity.

Previously, Dow’s laboratories were on the main corporate network and there was no difference in the management of a desktop PC or a PC acquiring chromatography data. Dow realised that its mission critical systems, such as Laboratory Information Management Systems (LIMS) and Chromatography Data Systems (CDS), could not undergo an update every time Microsoft issued an emergency security patch. Labs need to be able to plan downtime so that it does not delay delivery or interfere with important analytical work.

Dow decided to isolate its labs and those PCs from the network to ensure system security without compromising the needs of the business.

Dow uses the Atlas Chromatography Data System (CDS) from Thermo Electron as well as Thermo’s headless PC, the DataServer, to acquire data. The DataServer can be locked down to prevent access from external sources. The DataServers were locked down to only allow access to specific protocols and ports. Dow was able to establish that only the application server can access the DataServer, thus ensuring that viruses cannot make it into the DataServer.

Dow deployed this feature, but took the extra precaution of isolating the lab as a whole. All DataServers and instruments are now isolated into what they call a ‘Zone’ and these systems are maintained by the lab. To ensure that the Zone was truly isolated from the rest of the network and the outside world, all of the PCs in the Zone were only able to communicate with each other and a limited set-up of safe PCs on the Dow network. These safe PCs were defined through static IP addresses, but Active Directory or LDAP could also have been used. This strategy allowed the application server and clients to remain outside of the Zone. This was possible because Atlas does not require a connection with the server or the clients to acquire data. When the server is rebooted to install security patches or company operating system images, the lab continues to run.

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