The Analysis of Food Samples for the Presence of Genetically Modified Organisms

Transcription

1 The Analysis of Food Samples for the Presence of Genetically Modified Organisms Session 3 Samples Used during the Course M. Querci, N. Foti WORLD HEALTH ORGANIZATION REGIONAL OFFICE FOR EUROPE ORGANISATION MONDIALE DE LA SANTE BUREAU REGIONAL DE L'EUROPE WELTGESUNDHEITSORGANISATION REGIONALBÜRO FÜR EUROPA ВСЕМИРНАЯ ОРГАНИЗАЦИЯ ЗДРАВООХРАНЕНИЯ ЕВРОПЕЙСКОЕ РЕГИОНАЛЬНОЕ БЮРО

2 Samples Used during the Course 2 Table of Contents Session 3 Samples Used during the Course Samples used during the course 3 Certified Reference Material 3 Composition of raw and processed materials distributed to the participants 4 List of samples distributed during the course 6 Expected results by PCR 6 References 7

3 Samples Used during the Course 3 Samples used during the course During the course we will test different methods to detect the presence of MON810 maize and Roundup Ready soybean in different materials. For this purpose, we will use mixtures of non-gm and GM maize (MON810) and soybean (Roundup Ready soybean) respectively, at different concentrations. Two types of materials will be used: Certified Reference Materials Different raw and processed materials distributed to the participants Certified Reference Material Raw plant materials used during the course are Certified Reference Materials IRMM- 410S (Roundup Ready soybean) and IRMM-413 (MON810 maize). IRMM-410S and IRMM-413 consist of two sets of CRMs of dried soybean and maize powder, respectively, with different mass fractions (0, 0.1, 0.5, 1, 2, and 5%) of dried powder prepared from genetically modified Roundup Ready soybean and MON810 maize. The CRMs were produced by the Institute for Reference Materials and Measurements (IRMM - on behalf of Fluka Chemie AG (Buchs, Switzerland) in the frame of a collaboration with the Institute for Health and Consumer Protection (IHCP - of the Joint Research Centre of the European Commission (Ispra, Italy) (Trapmann et al., 2002 and 2001). They are intended for the validation of methods for the detection of genetically modified food. As DNA and/or protein quantification may depend on varieties, one has to exercise due care when drawing quantitative conclusions from measurements of unknown samples. The dried soybean powder containing GM Roundup Ready soybean has been produced from whole seeds of a non-modified soy line (Asgrow A1900) and the genetically modified event Roundup Ready soybean (Asgrow line AG5602 RR). The dried maize powder containing GM MON810 maize has been produced from whole kernels of the non-modified cultivar DK512 and MON810 cultivar DK513.

4 Samples Used during the Course 4 Composition of raw and processed materials distributed to the participants We will test different materials: biscuit, milk powder, snack food crumb and flour. Raw materials Mixed Flour. In order to obtain 0.5% of each GMO event (total dry weight) 1% of RR soya (IRMM-410S) was added to 1% MON810 (IRMM-413). 50 mg of both flours were weighed and added directly to reaction tubes ready for DNA extraction. Flour MON810 1% is a Certified Reference Materials IRMM-413 (1% MON810 maize). Snack food crumb This sample is derived from a GMO proficiency testing cheme (GeMMA Scheme, Round 06, test material GMO-06B). The material was prepared from commercially available GM free soya-based dried snack food and GM-containing soya snack food. Snack food crumb 1372 g GM-free soya snack food, 28 g GM soya snack food Before mixing, both materials were ground and sieved to give a homogenous crumb mix and then tumble-blended overnight. Finally, materials were mixed for approximately one hour using a rotary blender. Storage of the materials was at 20 C. Biscuit The material was produced at the JRC Institute for Health and Consumer Protection and was used to validate a PCR method for both Roundup Ready soybean and Maximizer maize (Bt-176) in processed food fractions (Lipp et al., 2001). Dry soybean and maize derived flour were weighed and mixed with the other ingredients in the proportion indicated below. Biscuits # g maize (0% GMO), 250 g soybean (0% GMO), 300 g wheat, 200 g sugar, 100 g butter, 10 g salt, 16 g vanilla baking powder, 2 eggs The ingredients were carefully mixed with 600 ml water and homogenised, spread out evenly on a baking plate and baked in a pre-heated oven at 180 C with

5 Samples Used during the Course 5 recirculating air for 10 min. The material was removed from the oven, covered to avoid contamination and allowed to cool to room temperature. Storage was at 20 C. Soya milk powder This sample is derived from Round 05 GeMMA proficiency testing scheme g of US soybean milk powder were tumble blended overnight with 300 g of Roundup Ready soybean protein isolate. Individual sub-samples (10 g) were dispensed into screw topped plastic containers and stored at ambient temperature prior to distribution. Biscuits MON810 This material was produced at the JRC Biotechnology and GMOs Unit. Dry maize derived flour was weighed and mixed with the other ingredients in the proportions indicated below. Biscuits MON g wheat flour, 100 g maize flour* (2 * % GMO), 150 g sugar, 100 g butter, 1 egg *2% MON810 maize flour was obtained by adding wild type maize flour to 100% MON810 flour and mixing for 30 minutes. The ingredients were carefully mixed, spread out evenly on a baking tray and baked in a pre-heated oven at 180 C with recirculating air for 10 min. The material was removed from the oven, covered to avoid contamination and allowed to cool to room temperature. Storage was at 4 C until required.

6 Samples Used during the Course 6 List of samples distributed during the course Sample Characteristics % GMO (specific ingredient) RR soybean MON810 maize Biscuits #1 0% 0% Mixed flour 1% 1% Flour MON810-1% Snack food crumb 2.2% - Soya milk powder 8.9% - Biscuits MON810-2% Expected results by PCR Sample zein lectin 35S nos E35S/hsp70(b) CTP/EPSPS IRMM- 410S-0 (0%) IRMM- 410S-1 (0.1%) IRMM- 410S-2 (0.5%) IRMM- 410S-3 (1%) IRMM- 410S-4 (2%) IRMM (0%) IRMM (0.1%) IRMM (0.5%) IRMM (1%) IRMM (2%) Biscuits # Mixed flour Flour MON Snack food crumb Soya milk powder Biscuits MON

7 Samples Used during the Course 7 References Lipp, M., Bluth, A., Eyquem, F., Kruse, L., Schimmel, H., Van den Eede, G. and Anklam, E. (2001). Validation of a method based on polymerase chain reaction for the detection of genetically modified organisms in various processed foodstuffs. European Food Research Technology 212, Lipp, M., Brodmann, P., Pietsch, K., Pauwels, J. and Anklam, E. (1999). IUPAC collaborative trial study of a method to detect genetically modified soy beans and maize in dried powder. Journal of AOAC International 82, Trapmann. S., Catalani, P., Conneely, P., Corbisier, P., Gancberg, D., Hannes, E., Le Guern, L., Kramer, G. N., Prokisch, J., Robouch, P., Schimmel, H., Zeleny, R., Pauwels, J., Van den Eede, G., Weighardt, F., Mazzara, M. and Anklam, E. (2002). The certification of reference materials of dry-mixed soya powder with different mass fractions of Roundup Ready soya. Certified Reference Materials IRMM-410S. ( chements/erm-bf410_report.pdf) (accessed January 2010) Trapmann, S., Le Guern, L., Prokisch, J., Robouch, P., Kramer, G. N., Schimmel, H., Pauwels, J., Querci, M., Van den Eede, G. and Anklam, E. (2001). The certification of reference materials of dry-mixed maize powder with different mass fractions of MON810 maize. Certified Reference Materials IRMM-413. ( chements/erm-bf413_report.pdf) (accessed January 2010) GeMMA Scheme-GMO analysis Round 06. GMO Proficiency Testing, (October 2001). Report N. GMO 06. GeMMA Scheme-GMO analysis Round 05. GMO Proficiency Testing, (October 2001). Report N. GMO 05.