Improving mycotoxins measurements by combining rapid analysis with dust sampling

Transcription

1 Improving mycotoxins measurements by combining rapid analysis with dust sampling Project presentation Dr. Scarlett Biselli, Mareike Reichel Mycotoxin Forum , Brussel

2 Dust Sampling WEJ Contaminants 2

3 Principles and Hypothesis Dust of cereal kernels & corn Are distributed all over the truck load are easy to separated from the grains Fast & representative sampling Are homogeneous particles no grinding necessary Show accumulation of mycotoxins Fast & simple sample preparation Fast, sensitive & cost-efficient analysis Show less matrix effects Correlation between the mycotoxin Fast results for whole grain lots contaminations of dust and grains

4 Project considerations so far o Development of an applicable workflow in practice o Development of reliable and robust detection method either in the Lab or on-site: LC-MS/MS (Multitoxin Screening) Lateral-flow devices (single toxin) o Development of reliable Data Model for each species and each toxin o Evaluation of representativeness of the dust : through parallel suction experiments of truckloads Comparison of results from legal volume of aggregate sample with dust results Repeatability trials in dust samples o Field application, prove of concept

5 Applicable workflow in practice 1. Suction of dust 2. Perform Rapid Test (adapted) 3. Read out contamination of the grain Lot

6 Place of Sampling- Process chain Suction of the relevant dust is everywhere possible where grain is moved or treated unloading loading In Process At the back or at the side Feed inlets, chain or belt conveyors Conveyed length, Elevators, Cleaning facilities for cereals 6

7 Method Validation: Suction system Dust sampler Currently manual sampling via a lance Installation at any unloading / loading station Filter-free technology (Cyclon) Adjustment of air speed to separate particles of specific size Possible to retain reference sample Prove of concept currently only for truckloads

8 Validation of the method Data model for data models > 5-10 kg grain samples were mobilised using a high volume blender on-site during loading during unloading of trucks Specific data for each toxin and cereal species By Investigation of : Wheat / Durum Rye Barley Corn 8

9 Method of analysis Dust Sample Kernel Sample On-site (Lateral Flow) Multi-toxin Method Reference Method Sample preparation by dry milling Extraction & Analytical Preparation 2 min, 0.5 g + 4 ml (MeOH)/H2O Depending on Test 2 min, 0.5 g + 4 ml ACN/H 2 O 80:20 addition of IS Extraction & IAC e.g. CEN calibrated Test Reader for dipsticks Measurement LC-MS/MS: or; LC-HR-MS HPLC-FD (Cobracell reaction)

10 In Comparison.. Homogenous sample (variance < 5%) No homogenisation step needed Small particles < 1 mm No grinding needed Mycotoxin concentrations is up to 20 fold higher in dust than in grain Dust Sample Extraction Only dilution and no concentration steps are needed Reduced matrix effects Reduced effects and interferences in immunochemical tests Rapid test 10

11 Results Wheat Deoxynivalenol & Zearalenon 1/4 Conversion from contamination in the dust to contamination in the grains Higher amounts in dust, thus. Results at low legal limits possible Rapid tests for every toxin will be applicable Data models for DON, ZON, Fumonisines, Aflatoxins, Ochratoxin A Correlation >0.8 No false negative results at MRL over all trials observed Due to the higher enrichment, toxins could be always detected. Over estimation is possible.

12 concentration dust [µg/kg] Results Corn ,0 concentration dust [µg/kg] 0 600,0 R² = 0,89 R² = 0,83 Fumonisin B1 Fumonisin B concentration grain [µg/kg] R² = 0,84 Aflatoxin B1 0,0 0,0 150,0 concentration grain [µg/kg] concentration dust [µg/kg] concentration dust [µg/kg] R² = 0,80 Ochratoxin A 0,0 0,0 100,0 concentration grain [µg/kg] 5000 R² = 0,77 Deoxynivalenol concentration grain [µg/kg]

13 Homogeneity Distribution of toxins in large Lots Especially storage toxins Aflatoxins and Ochratoxin A are well known to be uneven distributed in the Lots: Study: Parallel repeated analysis of corn samples and dust derived from a Lot > 100t during loading Results of this sampling study show that the standard deviation between the single results of multiple aggregate samples is much smaller for the dust procedure. Corn results CV %: 39 % Samples of a Lot > 100 t Concentration [µg/kg] Dust dust of results corn CV %: 14 % Samples ofa Lot > 100 t

14 Aflatoxins parallel sampling results of one large Lot (100t)

15 Varieties of toxins detectable via dust - corn- Fusarium toxins Aspergillus & Penicillium toxins Alternaria toxins Deoxynivalenol 3-Acetyl DON 15-Acetyl DON Nivalenol Fusarenon X T2 toxin HT2 toxin Diacetoxyscirpenol Monoacetoxyscirp. Neosolaniol Zearalenone Fumonisin B1 Fumonisin B2 Fumonisin B3 Moniliformin Beauvericin Enniatine A Enniatine A2 Enniatine B Enniatine B2 Aflatoxin B1 Aflatoxin B2 Aflatoxin G1 Aflatoxin G2 Sterigmatocystin Citrinin Cyclopiazonic acid Ochratoxin A Roquefortine C Mycophenolic acid Alternariol AOH methylether Tenuasonic acid Altenuene Tentoxin AAL Toxin C01 F C02 F C03 D C04 UK C05 UK C06 D C07 UK C08 UK C09 D C10 D C11 D C12 D C13 D C14 D C15 D C16 D C17 SB C18 RO C19 RO C20 RO C21 RO C22 RO C23 RO C24 RO C25 RO

16 -Wheat- Deoxynivalenol 3-Acetyl DON 15-Acetyl DON Nivalenol Fusarenon X T2 toxin HT2 toxin Diacetoxyscirpenol Monoacetoxyscirp. Neosolaniol Zearalenone Moniliformin Beauvericin Enniatine A Enniatine A2 Enniatine B Enniatine B2 Aflatoxin B1 Aflatoxin B2 Aflatoxin G1 Aflatoxin G2 Sterigmatocystin Citrinin Cyclopiazonic acid Ochratoxin A Roquefortine C Mycophenolic acid Alternariol AOHmethylether Tenuasonic acid Altenuene Tentoxin AAL Toxin Ergosine Ergocristine Ergocryptine Ergocornine Ergotamine Ergometrine W01 W02 W03 W04 W05 W06 W07 W08 W09 W10 W11 W12 W13 W14 W15 W16 W17 W18 W19 W20 W21 W22 W23 W24 W25 W26 W27 W28 W29 W30 Fusarium toxins Aspergillus & Penicillium toxins Alternaria toxins Ergot alkaloids

17 Possible Fields of Application Within the trade of chain and during processing of cereals Farm [Cutter] Loading / harvest transport Aquisition Reception control Re-arrangement Loading Trade Reception Re-arrangement Loading Food Feed Acception In-Process Control 17

18 Estimation of risk within the Trade 100% Risk Seller Rejection of a Lot Propability of Acceptance Acceptance of a Lot Decision Level Sampling~1 kg pertruck Sampling~10 kg per Truck Sampling of Dusts Full analysis of a Lot Risk byer 0% 0 MRL Content of mycotoxins in the Lot Due to faster and more representative testing, dust procedure decrease the risk level and increase knowledge and opportunities 18

19 To summarize Dust sampling and mycotoxin analysis of grain dusts is a promising field of application for rapid test systems as it allows: Fast & more representative sampling Fast & simple sample preparation without any grinding step Fast, sensitive & cost-efficient analysis using simple methods Application of the procedure enabled possible detection of mycotoxins which are regulated in the lower ppb range Thanks to the accumulation of toxins on the surface, determination of a broad variety of toxins is possible within laboratory analysis.

20 Thanks for your attention! For detailed information please contact us! Mareike Reichel Dr. Scarlett Biselli Eurofins WEJ Contaminants GmbH Neuländer Kamp 1a D Hamburg