INSTITUTE OF CHEMICAL TECHNOLOGY Department of Food Chemistry and Analysis

1. INSTITUTE OFCHEMICAL TECHNOLOGY Department of Food Chemistryand Analysis Technicka 3, 16628 Prague, Czech Republic jana.hajslova@vscht.cz Rapid Control of Food / Environmental Safety: NEW CHALLENGES IN MASS SPECTROMETRY Jana Hajslova, Lukas Vaclavik, Tomas Cajka, Jan Poustka, Ondrej Lacina, Jakub Schurek

2. Ambient mass spectrometry

3. ….An awkward feature of mass spectrometry is that the sample must be introduced into vacuum or into an inaccessible region closely coupled to the vacuum system….. Ambient mass spectrometry provides solution!  the direct analysis of ordinary objects in the open atmosphere of the laboratory or in their natural environment is possible

4. DESI Purdue Univ. USA DART JEOL, USA

5. Recent review The ambient ionization methods retain the signature advantages of MS:  speed chemical specificity low detection limits moreover:no sample preparationis needed!!!

6. AMBIENT DESORPTION IONIZATION METHODS desorption production of ionizing chemical reagents a Techniques where ESI mechanisms are mainly responsible for ionization. b Methods where chemical ionization is responsible for ionization (photoionization - PI, ion evaporation - IE and electrical discharge)

8. DESI ► DART ▼

9. DESI ▼ Spray and fluid velocity illustration. Droplets are black sphereseight times their actual size. Background color indicates velocitymagnitude of surrounding fluid. The blue line shows the optimumcollection angle from experiments (~10°)

10. Because mass spectrometer orifice is continually bathed in hot inert gas, the DART source is remarkably resistant to contamination and sample carryover.

11. Comparison of the intensities of the molecular ion of warfarin, [M + H]+ = 309, dissolved in different solvents after DART ionization at 250 C.

12. Case study no. 1 DIRECT CHARACTERIZATION OF EDIBLE OILS Objectives of study: Optimization of conditions for triacylglycerols (TAG) dentification Identification of oil type Assessment of processing technology Assessment of

13. EXPERIMENTAL CONDITIONS • Neat oils were sampled with glass melting point tubes • A cotton swab dipped into diluted ammonium hydroxide was placed near the sampling gap to enhance formation of [M+NH4] ions from triACYLglycerides • AccuTOF-DARTTM system was operated under settings given below DART–TOFMS SETTINGS: Ion mode:POSITIVE Gas:HELIUM Gas temperature:300°C Needle voltage: 3000 V Electrode 1: 150 V Electrode 2:250 V Cone voltage:20 V Peaks voltage:1100 V MCP voltage: 2500 V Internal calib.: PEG 600

14. VIRGIN OLIVE OIL – DART positive mass spectrum, 300°C,m/z 100 - 1000 DAGs TAGs FREE FATTY ACIDS INTENSITY SQUALENE m/z

15. VIRGIN OLIVE OIL – DART positive mass spectrum, 300°C zoom m/z 250 - 310 OLEIC ACID [M+NH4]+ FFA are formed by heat-induced decomposition of TAGs OLEIC ACID [M+H]+ LINOLEIC ACID [M+NH4]+ LINOLEIC ACID [M+H]+ INTENSITY PALMITIC ACID [M+NH4]+ PALMITIC ACID [M+H]+ m/z

16. VIRGIN OLIVE OIL – DART positive mass spectrum OOO zoom m/z825 - 920 POO TAGs are detected as [M+NH4]+ SOO LOO POL PSO PPL LLO PLL PPL SSO m/z

17. AUTOOXIDATION OF RAPESEED OIL – DART POSITIVE SPECTRA, 300°C FRESH SUNFLOWER OIL (PEROXIDE VALUE = 0.17) SUNFLOWER OIL OXIDIZED IN MICROWAVE OVEN FOR 25´ (PEROXIDE VALUE = 27.31)

18. EXTRAVIRGIN (a) vs. POMACE OLIVE OIL (b), m/z 100 - 1000 (a) (b)

19. EXTRAVIRGIN (a) vs. POMACE OLIVE OIL (b), m/z 820 - 950 (a) (b)

20. DART positive mass spectra – various edible oils m/z 100-1000 OLIVE OIL SUNFLOWER OIL RAPESEED OIL PUMPKIN SEED OIL COCONUT FAT

21. DART+ various edible oils, m/z 820-920 - TRIACYLGLYCEROLS OLIVE OIL SUNFLOWER OIL RAPESEED OIL PUMPKIN SEED OIL COCONUT FAT

22. IS AUTHENTICATION BASED ON TGA PROFILE OBTAINED BY DART POSSIBLE? Principal Component Analysis (PCA) • STATISTICA v 7.0 software • PARAMETERS: relative intensities of [M+NH4]+ ions corresponding to TAGs • TAGs: PPL, PPS, LLL, POO

23. Principal Component Analysis (PCA) OLIVE OIL RAPESEED OIL PUMPKIN SEED OIL COCONUT FAT SUNFLOWER OIL Factor 1: PPL, PPS, LLL Factor 2: POO

24. TAG Sunflower oil

25. Sunflower seeds - direct sampling TAG Matrix components

26. Case studyno. 2 Traceability of honey origin

27. GCxGC/TOF MS

28. DART: comparison of two honey samples Different origin

29. Processing of measured data is not easy….. NameMeas.Calc.Diff(u)Abund. GBL 85.0294 85.0290 0.0005 23.1410 Pyruvic_acid 87.0129 87.0082 0.0047 28.4476 Lactic_acid 89.0241 89.0239 0.0002 12.2362 Cresol 107.0463 107.0497 -0.0034 3.4462 Uracil 111.0156 111.0195 -0.0038 19.2750 Creatinine 112.0506 112.0511 -0.0004 71.6850 Succinic/fumaric/maleic acid 115.0058 115.0031 0.0026 39.3193 Methylmalonic_acid 117.0225 117.0187 0.0038 13.1083 Computer analysis

30. Case study no.3 STROBILURIN FUNGICIDES IN GRAINS

31. WP6 Pesticides: target values for Strobilurin SPR bioassay • Extraction time: • Maximum of 3 hours for 20 samples • Working characteristics: • LODs<0.05 mg/kg (LOQs < 0.1 mg/kg) • Repeatability - %CV <40  Can be these parameters also achieved by DART?

32. CONCLUSIONS • The total time needed for analysis of one sample by novel validated approach (SLE extraction) is 8 min. • The screening of strobilurins direct in milled wheat grains is possible by analysis employing AccuTOF™DART™. • All target strobilurins can be determined by novel, simple and fast approach at concentration levels below LODs in

33. Case study no. 4

34. (1a) (1) Sample is placed in sampling hole, DIP-itTM sampler sticks are used for sample transfer (1b) (2) (2) Sampling stick is immersed into the samplefor 4s (3) (3)Sampling stick is transferred between the DART ion source and MS inlet, massspectra are acquired for 7s (4) (4) Used stick disposed DART AccuTOF LC-plus system with LEAP HTC PAL autosampler TOTAL RUN TIME: 35 s

35. DART-POSITIVE MASS SPECTRA OF COLA BEVERAGES GAS BEAM TEMPERATURE 250°C PEPSI-COLA PEPSI-COLA LIGHT caffeine caffeine COCA-COLA LIGHT COCA-COLA COCA marker? ? ?

36. SUGAR CONTENT IS VERY LOW (< 0.1g/L) HIGH SUGAR CONTENT (~100g/L) GAS BEAM TEMPERATURE 350°C HEAT-INDUCED SUGAR FRAGMENTATION IN NORMAL COLAS PEPSI-COLA PEPSI-COLA LIGHT COCA-COLA LIGHT COCA-COLA

37. GAS BEAM TEMPERATURE 350°C DETECTION AND IDENTIFICATION OF ASPARTAMEIN “LIGHT” PRODUCTS PEPSI-COLA LIGHT PEPSI-COLA COCA-COLA COCA-COLA LIGHT 277.11741

38. GAS BEAM TEMPERATURE 350°C DETECTION AND IDENTIFICATION OF ASPARTAMEIN “LIGHT” PRODUCTS PEPSI-COLA LIGHT PEPSI-COLA COCA-COLA COCA-COLA LIGHT 277.11741

39. 295.12984 277.11726 [M+H]+ ASPARTAME E951 meas. mass 295.1298 calc. mass 295.1294 difference+1.51ppm PEPSI-COLA LIGHT PEPSI-COLA [M-H2O+H]+ [M+H]+ 295.12968 COCA-COLA COCA-COLA LIGHT 277.11741

40. GAS BEAM TEMPERATURE 450°C PEPSI-COLA PEPSI-COLA LIGHT ASPARTAME COCA-COLA COCA-COLA LIGHT ASPARTAME

41. Example no 3 Coffee - quality, authenticity

42. Robusta Roasted (Douwe Egberts) Robusta Roasted reference match Arabica Roasted No match

43. SPME preconcentration – another challenge!!!

44. WHAT ABOUT USING SPME FOR SAMPLE FLAVOUR COMPONENTS COLLECTION / PRE.CONCENTRATION?? Blank SPME fiber (PDMS/CX/DVB, Supelco) Bleed: siloxans

45. Coffee (Nescafe Gold) 1g of matrix, HS-SPME (40°C, 60 min, PDMS/CX/DVB) coffee volatiles bleed

46. Coffee (Nescafe Gold) ZOOM (m/z 75-245) 1g of matrix, HS-SPME (40°C, 60 min, PDMS/CX/DVB) pyrazine 2-acetylpyrrole 2-(2-furfuryl)pyrazine methylpyrazine caffeine

47. Example no 5 BEERS PROFILING

48. DART- (350°C, 2400V, PV 600V) PRIMATOR: Light beer – porter (alc 9% vol.) PRIMATOR: Dark beer – porter (alc 10.5% vol.) m/z 250400 PRIMATOR: Light beer – lager (alc 5% vol.)

49. DART- (350°C, 2400V, PV 600V) PORTER (Dark) LAGER TRAPISTIC PORTER (Light)

50. Switching operation mode enables higher degree of confirmation. TRAPISTIC DART+ LAGER DART+ TRAPISTIC DART- LAGER DART-