Aims The method for the confirmation of drugs of abuse for addiction testing within King’s College Hospital prior to 2008 was a labour intensive thin layer chromatography method. To replace this with a faster method more suited to future requirements, the laboratory bought a liquid chromatography system with ion trap mass spectrometric detection. The development of the routine analytical method and the implementation of this method within the laboratory using on-line solid phase extraction and Turboflow® sample preparation will allow the laboratory to operate successfully in the field of clinical drugs of abuse testing in the future. Method Analyses are performed on an ion trap mass spectrometer with an electrospray ion source following reversed phase liquid chromatography, initially using on-line solid phase extraction with a Jasco XLC® series autosampler and pump and later a Thermo Turboflow® on-line extraction method with a CTC Combi-Pal® autosampler and Agilent 1100 series liquid chromatography system. Elution of drugs and metabolites is performed with a multi-step gradient of ammonium formate buffer and acetonitrile, followed by regeneration of the extraction and analytical columns to starting conditions. Detection is achieved with a Thermo LCQ Fleet ion trap mass spectrometer with a combination of full spectrum survey scans, dedicated product ion scans, neutral loss scans and data dependent product ion scans in two analysis segments. Total run time is only 20 minutes, allowing a throughput of around 65 samples per day. Results The methods include the novel combination of the elimination of any hydrolysis step, on-line SPE or Turboflow extraction, detection of multiple drug groups, full spectrum analysis and library matching, the use of data dependent scans and ion trap mass spectrometry using MS3 and neutral loss scans. The methods developed were validated using a departmental method validation protocol and accepted for routine use. Simultaneous detection of over fifty analytes has been found possible in a range of clinically relevant drug groups, including opiates, amphetamines, methadone, propoxyphene, cocaine, ketamine and their metabolites. The use of neutral loss scans and product ion scans of phase 2 drug metabolites permits the addition of previously unidentified drugs and metabolites to the method, allowing the laboratory’s services to develop in line with requirements of the service. Quality is maintained through the use of standard operating procedures, staff training, quality control samples and external quality assessment. Conclusion Drugs of abuse testing is key for treatment and monitoring of drug addiction. The introduction of modern mass spectrometry techniques has reduced the turnaround time of routine analysis for a range of drugs and metabolites and increased the range of drugs that can be analysed. The methods introduced have revolutionised testing at King’s College Hospital and produced a method which is capable of evolving with the needs of the service to keep abreast of future requirements of the service.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:655160 |
| Date | January 2014 |
| Creators | Evers, Frank Richard |
| Contributors | Mills, Graham |
| Publisher | University of Portsmouth |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Source | https://researchportal.port.ac.uk/portal/en/theses/development-of-a-liquid-chromatography-ion-trap-mass-spectrometer-method-for-clinical-drugs-of-abuse-testing-with-automated-online-extraction-using-turbulent-flow-chromatography(c047da7d-eb8a-41d0-ad1c-830deca531dc).html |
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