Finnish Custom Lab., Pekka Ravio Experiences on developing a screening method for Orbitrap mass spectrometer Esitys käsittelee kemiallista kvalitatiivista orgaanista analytiikkaa tänä päivänä nestekromatografia-massaspektrometrian näkökulmasta. 11. NPRW 1.-2.6.2015, Tallinn Finnish Custom Lab., Pekka Ravio

Finnish Custom Lab., Pekka Ravio Orbitrap Elite ETD Hankinta: olemassa olevasta 15 vuotta vanhasta ioniloukusta > ioniloukku-orbiloukkuun >> hinta >> FIMEAn mukaan tulo Laitteen osien kuvaus: neste – interface –ioniloukku – orbiloukku - ETD 11. NPRW 1.-2.6.2015, Tallinn Finnish Custom Lab., Pekka Ravio

Orbitrap Elite ETD: schematic Toimintaperiaatteen ero verrattuna moneen muuhun massaspektrometriin: analysoidaan ionipaketteja, ei jatkuvaa ionivirtaa Sähkösumutus interface – kaksiosainen ioniloukku – korkea paine loukussa erilaiset operaatiot – matalapaineloukusta skannataan – omat detektorit – C-trap - Orbitrap toimintaperiaate – HCD cell - ETD

Finnish Custom Lab., Pekka Ravio Pesticide residues screening with Orbitrap Elite Primary instrumental parameters to choose: 1. Mass resolution 15 000, 30 000, 60 000, 120 000 or 240 000 (at m/z 400) 2. Fragmention technique In-source CID, ion trap, HCD all ion fragmentation 3. Collision energy 11. NPRW 1.-2.6.2015, Tallinn Finnish Custom Lab., Pekka Ravio

Finnish Custom Lab., Pekka Ravio Measurement Alternating scanning Full scan, mass range 130 – 900 amu All ion fragmentation in-source CID, mass range 80-900 amu, collision energy 45 V Cycle time around 0.5 s (R = 60 000) > 15 or more scans over UPLC peak External mass calibration 11. NPRW 1.-2.6.2015, Tallinn Finnish Custom Lab., Pekka Ravio

Finnish Custom Lab., Pekka Ravio Alternating scan run, full scan – in source CID scan Zoom 11. NPRW 1.-2.6.2015, Tallinn Finnish Custom Lab., Pekka Ravio

Finnish Custom Lab., Pekka Ravio EUPT-SM06 2014, pepper + around 30 pestisides Furathiocarb, C18H26N2O5S, M+1 383.16352 383.16287 ∆ m =1,6 ppm Full scan C10H11O2S 195.0474, ∆ m = 4.1 ppm In-source CID 11. NPRW 1.-2.6.2015, Tallinn Finnish Custom Lab., Pekka Ravio

Finnish Custom Lab., Pekka Ravio Screening tests at the moment against a database and MS/MS library only, no validation with pesticide standards Database bought and collected from literature, 450 compounds Data processing software TraceFinder 3.2 Processing against database and MS/MS spectral library (bought) 11. NPRW 1.-2.6.2015, Tallinn Finnish Custom Lab., Pekka Ravio

Tullilaboratorio, Pekka Ravio Details in compound database 18.3.2015 Tullilaboratorio, Pekka Ravio

Tullilaboratorio, Pekka Ravio TraceFinder processing parameters 18.3.2015 Tullilaboratorio, Pekka Ravio

Finnish Custom Lab., Pekka Ravio Processing with TraceFinder software 11. NPRW 1.-2.6.2015, Tallinn Finnish Custom Lab., Pekka Ravio

Finnish Custom Lab., Pekka Ravio Finding correct fragments ” fragment soup” + library search in-source CID scan 0.019 mg/kg fenhexamid in raspberry Mass resolution 60 000 Fenhexamid MS/MS-library spectra 11. NPRW 1.-2.6.2015, Tallinn Finnish Custom Lab., Pekka Ravio

Tullilaboratorio, Pekka Ravio Evaluating isotopic pattern, red=meas., blue=theor. Fenhexamid 0,019 mg/kg 18.3.2015 Tullilaboratorio, Pekka Ravio

Tullilaboratorio, Pekka Ravio Screening results, EUPT-SM06 18.3.2015 Tullilaboratorio, Pekka Ravio

Tullilaboratorio, Pekka Ravio Screening results EUPT-SM07 18.3.2015 Tullilaboratorio, Pekka Ravio

Finnish Custom Lab., Pekka Ravio Conclusions Orbitrap Elite Easy to use Stable mass calibration Same mass calibration may be used 1- 3 days (error < 3 ppm) All ion fragmentation with HCD-cell would be nice Data processing with TraceFinder 3.2 Visual outlook very informative Should be able to use several collision energies 11. NPRW 1.-2.6.2015, Tallinn Finnish Custom Lab., Pekka Ravio

Finnish Custom Lab., Pekka Ravio Principle how the isotopic pattern is evaluated is unclear Overall conclusions Encouraging results All depends on the database A lot of work to build up the database, ”fine tune” it and ”fine tune” the processing parameters + MS/MS library The more compounds you have in database, the more false positives you get The software needs some upgrading and better manuals 11. NPRW 1.-2.6.2015, Tallinn Finnish Custom Lab., Pekka Ravio

Finnish Custom Lab., Pekka Ravio ”Our newest weapon” Customs moneydog Massi Thank You 11. NPRW 1.-2.6.2015, Tallinn Finnish Custom Lab., Pekka Ravio