HPLC INTERFACE FOR ELECTROSPRAY MASS SPECTROMETRY

THE GOAL OF THIS RESEARCH IS TO DEVELOP AND ADAPT VESTEC CORPORATION'S "UNIVERSAL INTERFACE" TO COUPLE CONVENTIONAL HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC) TO ELECTROSPRAY MASS SPECTROMETRY. THE INTERFACE, WHICH HAS BEEN SUCCESSFULLY APPLIED TO COUPLING REVERSED-PHASE HPLC AT FLOW RATES AS HIGH AS 1.5 ML/MIN OF AQUEOUS EFFLUENT TO ELECTRON IMPACT MASS SPECTROMETRY, PRODUCES AN AEROSOL OF DROPLETS CONTAINING THE SAMPLE AND ALLOWS REMOVAL OF AS MUCHSOLVENT AS DESIRED WITHOUT LOSING THE SAMPLE OR DESTROYING THE CHROMATOGRAPHIC FIDELITY. ELECTROSPRAY MASS SPECTROMETRY IS AN EXCITING NEW TECHNIQUE WHEREBY MOLECULAR-WEIGHT INFORMATION CAN BE OBTAINED ON PROTEINS ANDOTHER BIOPOLYMERS WITH NO APPARENT LIMIT ON SIZE. THIS PROJECT WILL DEVELOP A TECHNIQUE FOR CHARGING THE DROPLETS EMERGING FROM THE UNIVERSAL INTERFACE AND TRANSMITTING THE RESULTING CHARGED DROPLETS AND/OR MULTIPLY-CHARGED MOLECULAR IONS TO A QUADRUPOLE MASS SPECTROMETER. IN THIS WAY, SPECTRA ON PROTEINS AND OTHER LARGE BIOMOLECULES WILL BE OBTAINED SIMILAR TO THOSE ALREADYDEMONSTRATED FOR ELECTROSPRAY, BUT WITHOUT THE RESTRICTIONS ON LIQUID FLOW AND COMPOSITION INHERENT IN THE CONVENTIONAL ELECTROSPRAY TECHNIQUE. THE ULTIMATE GOAL IS AN INSTRUMENT FOR DETERMINING THE MOLECULAR WEIGHTS ON SUBPICOMOLAR COMPONENTS OF COMPLEX BIOLOGICAL EXTRACTS ONLINE WITH HPLC SEPARATION.