MSACL 2016 EU Abstract

Evaluation and Integration of Accurate Mass LC-MS into the Routine Clinical Laboratory

Joanne Adaway (Presenter)
University Hospital South Manchester

Bio: • Graduated with a First Class (Hons) BSc in Biochemistry from UMIST in 2000 • Started training in Clinical Biochemistry at Wythenshawe Hospital in 2003 whilst writing up a PhD thesis entitled ‘Proteomic Analysis of Stem Cell commitment’. • Awarded PhD from the University of Manchester in 2005. • Graduated with an MSc in Clinical Biochemistry from the University of Manchester also in 2005. • Finished Clinical Scientist training in 2006 and started work in the Clinical Biochemistry department at Christie Hospital. • Returned to Wythenshawe Hospital in 2009 as a Senior Clinical Scientist and was promoted to Principal Clinical Scientist in 2010. • Awarded FRCPath in January 2011. • Honorary Lecturer at the University of Manchester since April 2011. • Current research interests: Biogenic amines and drug monitoring by LC-MS/MS.

Authorship: Joanne E Adaway, Brian G Keevil, Laura J Owen
Department of Clinical Biochemistry, Wythenshawe Hospital, Southmoor Rd, Wythenshawe, Manchester, M23 9LT and Manchester Academic Health Science Centre, University Hospital South Manchester, The University of Manchester, Manchester, UK.

Short Abstract We evaluated a Waters Xevo G2-XS QToF for 6 months in an NHS Clinical Biochemistry laboratory. We assessed small molecule quantification, identification and quantification of proteins, and also drug screening. Small molecule quantification was analyte dependent; an aldosterone assay compared well to a triple quadrupole method, whereas the sensitivity of the instrument was insufficient to quantify oestradiol. Drug screening was much simpler and faster to develop than on a triple quadrupole instrument. A wealth of information was produced on peptides and proteins to enable their accurate quantification. The Xevo G2-XS was easily integrated into the workflow of our laboratory.

Long Abstract

The linear measurement range of accurate mass instruments such as Time of Flight analysers has improved over the past few years and the cost has decreased to the extent that their introduction into the routine clinical laboratory is becoming feasible. We evaluated a Waters Xevo G2-XS QToF for 6 months in an NHS Clinical Biochemistry laboratory to determine how it would fit into the routine lab and the different ways in which it could add to the repertoire of the laboratory.

Small molecule quantitation is routinely carried out using triple quadruple instruments. We developed methods for oestradiol, aldosterone and plasma renin activity on the G2-XS and compared the results to our previously published triple quadrupole methods. Screening of samples for a range of analytes such as drugs is carried out on a variety of instrumentation, including accurate mass instruments and nominal mass LC or GC-MS. We evaluated the ease of use of the instrument for screening of a range of drugs, and also determined the sensitivity of the instrument for these drugs compared to a Waters Xevo TQS Micro. Finally, accurate mass instruments are invaluable in the discovery and quantitation of peptides and proteins, and we developed methods for a range of proteins and peptides to determine whether this functionality could be useful in an NHS laboratory.

The integration of the instrument into the routine laboratory proved simpler than expected, with instrument set up and calibration required only once a week. Infusion of a reference solution into the mass spectrometer at specified time intervals during analysis compensated for any fluctuations in detector conditions or calibration during the run, which aided the accurate mass measurement and assured data integrity between calibrations.

We discovered that the small molecule quantitation is very analyte dependent. The performance of the aldosterone and plasma renin activity assays compared favourably with the performance on a triple quadrupole instrument, however the sensitivity of the ToF assay for oestradiol was poor and would not be acceptable for routine use.

Screening of samples for drugs using the Xevo G2-XS was much simpler than an equivalent assay on a triple quadrupole instrument. The assay development was much faster on the QToF, as ready-made libraries were available containing many of the drugs of interest, and it was quick to add extra analytes to the library. In contrast, screening for the sample compounds using a triple quad was much more laborious and reporting of the results was also much more involved and time consuming than using the software on the Xevo G2-XS.

The QToF was able to provide a wealth of information on the peptides and proteins we analysed, and this data could be used to develop quantification methods on the G2-XS as well as MRM methods on a triple quad.

The Xevo G2-XS was easily integrated into the workflow of our NHS laboratory. It could be used to carry out quantification of some small molecules, as well as to provide information and quantification on larger molecules and to simplify drug screening methods.

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