Doping Evaluation with NuGenesis SDMS and TargetLynx
News Nov 16, 2010
The Doping Control Lab of the Karolinska University Hospital in Stockholm, Sweden is one of 33 doping control labs worldwide accredited by the World Anti-Doping Agency (WADA). The lab’s nine chemists and lab technicians analyze approximately 7,500 samples annually, including 4,717 athlete samples (compared to 183,337 samples worldwide). 87 positive samples were found at the Karolinska lab in 2006 (3,909 worldwide), using hyphenated chromatographic techniques such as GC/MS, GC/NPD, and LC/MS/MS.
Karolinska’s Doping Control Lab receives samples from a variety of customers, including the Swedish Sports Confederation, WADA, and other International sports confederations, as well as from correctional institutions and Public Health services.
The Doping Laboratory is primarily financed by payments for analyses performed, therefore the more samples that are run and results secured, the more income is generated to finance research and extended testing.
The WADA’s World Anti-Doping Code provides an internationally standardized list of prohibited substances for athletes both in and out of competition, as well as some specific regulations for particular sports disciplines. A laboratory’s work is assessed by WADA on the basis of a “proficiency testing program”, which must be successfully completed if the laboratory is to retain its status as an official doping control laboratory.
The Karolinska lab focuses specifically on testing for the 44 diuretics and other masking agents, as well as the 58 stimulants listed in the WADA code. In the future, they are planning to add tests for narcotics and beta-blockers to their spectrum.
Tests for diuretics and narcotics have been conducted using GC/MS, involving complex sample preparation with hydrolyzation, extraction, and derivatization. Stimulant tests using GC/NPD involve similarly complex preparation steps.
Increasing regulatory pressure and introduction of new technologies have driven up the number of samples analyzed and increased the quality of data produced.
Managing information and controlling the flow of data were major challenges. Enhancing the lab’s information and knowledge management processes is an area where Waters Laboratory Informatics has played a crucial role.
New analytical methods have been introduced using LC/MS/MS. The sample preparation for this method is significantly easier, since the diluted urine sample can be directly injected into the instrument with an internal standard added. This allows for more samples to be prepared and analyzed.
The solution chosen by the Karolinska Doping Control Lab is the Waters® ACQUITY UPLC® System with a BEH shield RP18 column, 50 x 2 mm, 1.7 μm, with a flow rate of 0.4 mL/min (MeOH/NH4OAc 10 mM gradient, 6 minutes). A QC-sample is run at every 12th injection; water and urine are used as blanks. For detection, Waters’ Quattro Premier™ XE tandem quadrupole Mass Spectrometer is operated in MRM mode. The instruments are controlled by Waters’ MassLynx™ Software, with processing and reporting done using the TargetLynx™ Application Manager. TargetLynx is specifically designed for quantitation and specific analyte confirmation and it streamlines and automates the analysis of these samples.
All raw instrument data are automatically and securely captured and stored within Waters’ NuGenesis® Scientific Data Management System (SDMS).
The Karolinska lab has found that with the UPLC® System, they can now screen up to 105 analytes in eight minutes, injection to injection.
The hospital uses TargetLynx Application Manager to find analytes that comply with three criteria:
• Correct retention time
• Above signal-to-noise
• Above concentration threshold
If a positive hit of an illegal drug is detected, TargetLynx will flag this analyte.
Having automated the analysis and post analysis calculations, they were still faced with the need to manually review 140 compounds per sample and determine the positive hit patterns. Today, there are more than 100 samples in each batch. This manual process is tedious and time-consuming, and mistakes can easily be made. Most samples are negative (only 2.5% of samples are positive).
The next step was to automate and streamline the TargetLynx report utilizing the print capture capabilities within SDMS.
The analyst now prints all reports into SDMS with a TargetLynx format, that include the stimulants, ephedrines, and diuretics as individual reports. These reports are normally several hundred pages long.
When the analyst runs the custom application provided by the Waters Enterprise Software Services Group, they have the choice of defining the batch number by parameters that include standards, controls, and blanks. Then all reports can be extracted from a particular batch, excluding all standards, QC’s, and internal control analytes. QC results are automatically exported to their LIMS.
The end result is an SDMS report of analytes that were found, detailed by batch number and name of compound. Within this “hit list”, the user can click a link and SDMS will output the printed TargetLynx report that is stored within SDMS. Analysts can review the report just as they always have, but now they only have to concentrate on verifying the true positive hits (approximately 187 per year) instead of reviewing all 7,500 reports.
The Waters solution is providing a number of benefits to the Karolinska University Hospital’s Doping Control Laboratory operations including:
• Increased revenue potential – greater sample throughput with UPLC/MS/MS.
• Greater productivity – approximately two hours are saved per batch in reviewing results for illegal compounds with SDMS and TargetLynx reports.
• Reducing risk of data loss – SDMS automatically manages raw data backups from all LC/MS instruments.
• Better data accessibility – all reports and raw data are fully text searchable from within SDMS, reducing the time to locate critical information.
• Connecting science to the business of the hospital – TargetLynx and SDMS have automated and streamlined results calculations, review, and consolidated reporting.
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