BioTrove's ultra high throughput mass spectrometry system consists of a proprietary computer-controlled fluidic robot interfaced with a triple-quadrupole mass spectrometer. Robotics replace conventional HPLC (high performance liquid chromatography) as the sample injection and purification system used in conventional LC-MS applications.
The RapidFire™ system can quantify multiple analytes simultaneously, so substrate and product of an assay can be quantified simultaneously, along with one or more internal standards as necessary.
High quality data with coefficients of variability under 10% are routinely achieved in high-throughput screening applications. The RapidFire™ system can routinely analyze a 384-well plate in around 32 minutes (5 seconds per sample). As many as fifteen 384-well plates (5760 individual samples) can be analyzed by a single instrument in an 8-hour shift.
With RapidFire™, a 1 to 6 µl aliquot of sample is withdrawn from each well of a standard 96 or 394-well microtiter plate. A microfluidic module cleans and purifies the sample by removing components in the reaction mixture that are incompatible with API (atmospheric pressure ionization) mass spectrometry. Such components typically include salts, buffers, detergents, proteins, and microsomes or sub-cellular material. These components are required for the biochemical assay but cause mass spectrometry ion suppression and tend to precipitate in the mass spectrometry source.
The purification and mass spectrometry analysis steps described above are performed in a serial manner, eliminating the need for up-front sample preparation or purification. A wide range of buffers, salts, detergents, and compounds such as ATP or NADPH are fully compatible the RapidFire™ system. The purified samples can be analyzed by electrospray ionization (ESI) or atmospheric pressure chemical ionization (APCI), as required by the specific analytes in the assay.
