Supply, Installation, Validation and Verification of Tandem Mass Spectrometer – Liquid Chromatography System

The CUH Blood Sciences Endocrine Laboratory is looking to replace a high sensitivity Triple Quadrupole Tandem Mass Spectrometer (TMS) coupled to an ultra-high pressure liquid chromatography (HPLC) system. This system runs 10 methods that were developed in-house.

The Blood Sciences labs currently have four Sciex triple quadrupole tandem mass spectrometers (TMS) linked to Shimadzu or Sciex (Exion) liquid chromatography (HPLC) systems. This includes the system that we are replacing. The combined laboratories currently process around 120,000 samples a year through the four LC-Tandem Mass Spectrometer systems. Results generated from the systems are from technically/clinically validated methods and are used for diagnosis of endocrine and metabolic disorders, as well as for neonatal screening tests.

The replacement system will be capable of quantitation of a range of hormones or metabolites in human serum, urine and saliva. Some of the analytes are at pmol/L concentrations in human serum e.g. aldosterone/metanephrines, so a high sensitivity system is essential. The TMS – HPLC system must have the following critical elements:

TMS

• The TMS will have multiple modes of operation including multiple reaction monitoring and scan modes. Scan modes must include precursor ion scan, product ion scan, neutral loss scan.
• The TMS must be capable of Atmospheric Pressure Chemical Ionisation (APCI) and electrospray (ESI) ionisation modes with proven use of APCI for clinical hormone analyses.
• The TMS must be capable of positive and negative modes.
• The TMS must be capable of polarity switching within a sample acquisition run with a minimum settling time of 5msec between polarities.
• The TMS must accept a fluid flow rate from 5µL/min to 3000µL/min, without flow splitting.
• To allow for future development needs the TMS must have an option to be upgraded to a linear ion trap system that will provide additional scan modes such as MRM3, Enhanced Product Ion Scan.
• The MS must have an integral infusion pump under full control of the system software to allow infusion rates up to 30µL/min to allow compound tuning and for ion suppression testing.
• The MS control software must be compatible with multi-vendor HPLC systems e.g. Shimadzu, Agilent, Waters.
• The system must be capable of the self-generation of all gases required for routine use (i.e. no requirement for a gas cylinder in the lab).
• A suitable roughing pump must be supplied. This must be an oil-free pump.
• Results must be able to be exported in .txt format to allow uploading to our current hospital information system.
• Current methods in use in the Endocrine Lab must be directly transferrable without the need for re-development.

HPLC

• The HPLC system must have a dual binary pump arrangement allowing pressures up to 10,000PSI. This setup is to allow online solid phase extraction using 2-position valves.
• There must be a minimum of 4 solvent lines per pump with inline degassers. If binary pumps are proposed, there must be 4 solvent lines for each side of the binary pump. The LC must have the ability to switch to different solvent input lines, analytical columns without operator intervention to allow running of multiple assays to be processed consecutively.
• The HPLC autosampler must be capable of injecting from 1 to 50µL.
• The HPLC autosampler must have at least six positions for plate/vials and must be able to inject from 96 shallow well plates, 96 deep well plates and vials. It must be capable of using these in combination without changing hardware configuration.
• The column oven must have an operating temperature range of 5 to 85°C.
• The column oven and must be able to accommodate six analytical columns of 25cm length.
• The column oven must be able to accommodate and drive 2, 7-port 6-position valves to allow switching between columns and therefore multiplexing of analytical methods.
• The system must be able to drive 3 2-position 6-port valves independently to allow sequential running of different methods using online solid phase extraction.

The Trust believes that only Sciex 6500+ Tandem Mass Spectrometer, in conjunction with Shimadzu dual binary HPLC, can fulfil all of the essential criteria stated, and that only this equipment will allow rapid direct transfer of our current methods without the need to re-develop them.