The gmp.galis project: Four years survey on PCDDs/Fs and dl-pcbs in raw materials for animal feed Dra. Verónica Fernández-Villarrenaga Martín Unit of Chromatographic Techniques Scientific Research Support Services 1 University of A Coruña
A Coruña is a medium size city in the NW coast of Spain 246.056 hb (year 2009) Galician Autonomy 2
Since 2009 the lighthouse at he entrance of the harbour, the Tower of Hercules is inscribed ib on UNESCO s World Heritage List It was built by Romans in late 1 st century A.D and was restored in the 18 th century which is unique as it is the only lighthouse of Greco-Roman antiquity to have retained a measure of structural integrity and functional continuity. (http://whc.unesco.org/en/list/1312) 3
Unit of Chromatographic Techniques Part of the Scientific Research Support Services of University of A Coruña. Services work for scientist from our and other universities, public organisations and private customers. Instruments for GC, HPLC, GC/MS, LC/MS and sample preparation 2 Thermo Polaris Q ion traps 1 AB API-3200 3Q 1 Thermo Orbitrap Discovery mass spectrometer 1 Thermo MAT 95 XP magnetic sector mass spectrometer coupled to two Trace GC analysis of PCDDs/Fs and dl-pcbs Since May 2009 we are accredited as test laboratory according to ISO/UNE/EN 17025 for the determination of PCDDs/Fs in food and feed. We are the 4th laboratory in Spain achieving this accreditation. 4
What is gmp.galis? FOOD SAFETY PROGRAM FOR THE ANIMAL FEED SECTOR IN GALICIA Asociación Gallega de Fabricantes de Alimentos Compuestos (AGAFAC) g m p galis Seguridad Alimentaria del Noroeste S.L. (SANOR) 5
The Galician sector in Spain Total production 2,700,000 tons 2008 15% Spain 5 th producer of Spain gmp.galis borns to give an answer to Pressure of administration that is applying more strictly controls due too new legislations. Pressure of livestock sector to increase levels on food safety. Pressure of public opinion that is increasingly sensitized by the recent crises. Pressure of surrounding countries with high controls, that increase the risk of arrival of raw materials with poorer quality. 6
gmp.galis AREAS gmp.galis Ser services to producers gmp.galis Lab coordinate analysis SAI-UDC PCDDs/Fs and dl-pcbs Pesticides PAH Mycotoxins Trace elements LIGAL Nutrient value CIAM Aflatoxins MOURISCADE Nutrient value Microbiological Other pathogens Minerals 7
PCDDs/Fs and dl-pcbs: working scheme Sample selection Administrator SANOR SL Sampling Specific personnel SANOR SL Homogenisation i Division in portions Coding Distribution Mouriscade Laboratory Sample preparation and HRGC-HRMS analysis SAI -UDC Results Congener concentration (ng/kg or pg/g) Concentration as WHO-TEQ (ng/kg or pg/g) Individual congeners Total WHO-TEQ 8
PCDDs/Fs and dl-pcbs: analysis Samples Vegetables < 5% fat Inorganic samples Vegetables > 5% fat Fat samples LCS HUMIDITY LCS HUMIDITY LCS HUMIDITY Soxhlet extraction Soxhlet extraction Solution in hexane Aid Acid attack Aid Acid attack Clean-up (Power Prep) Clean-up (Power Prep) Clean-up (Power Prep) Concentration Concentration Concentration ISS HRGC-HRMS ISS HRGC-HRMS ISS HRGC-HRMS 9
PCDDs/Fs and dl-pcbs: results Samples since late 2005 PCDDs/Fs Samples since mid 2006 PCDDs/Fs and dl-pcbs 137 samples of 47 suppliers 30 animal fat samples 51 vegetable fat samples 41 vegetable samples 15 mineral samples 10
PCDDs/Fs and dl-pcbs: results 040 0.40 Animal fat max=0.985 0985 max=0.449 30 samples TOTAL WHO O-TEQ (pg/g) 0.30 020 0.20 0.10 0.00 max=0.748 Directive 2006/13/EC 3.0 pg/g WHO-TEQ PCDD/F + dl-pcb All samples were below limit LARD ANIMAL FAT (BLEND) POULTR Y FAT 1.00 Vegetable fat max=1.552 51 samples TOTAL WHO-TEQ (pg/g) 0.80 0.60 040 0.40 0.20 0.00 max=0.227 SOYA OIL max=0.512 PALM OIL HYDR. FAT >90% max=1.089 PA ALM SO OAP Directive 2006/13/EC 1.5 pg/g WHO-TEQ PCDD/F + dl-pcb Several hydrogenated fat samples were above limit 11
030 0.30 PCDDs/Fs and dl-pcbs: results max=0.510 max= =0.280 max= =0.249 max=0.090 max=0.465 max=0.322 max=0.268 0.25 0.20 0.15 0.10 0.05 0.00 Vegetables 41 samples Directive 2006/13/EC 1.25 pg/g WHO-TEQ PCDD/F + dl-pcb All samples were below limit TOTAL WHO-TEQ (pg/g) ALFALFAA DDG GLUTEN FEED COTTON WHOLE SEEDSS SOYA RAPESEED BEET PULPP SUGARCAN MOLASSESS E WHEAT MIDDLINGS PALMKERNE MEAL SOLVENTS L 0.60 0.50 0.40 030 0.30 0.20 0.10 0.00 max=0.846 15 samples Directive 2006/13/EC max=0.314 max=0.180 max=0.166 1.5 pg/g WHO-TEQ PCDD/F + dl-pcb All samples were below limit 12 TOTAL WHO-TEQ (pg/g) SEPIOLITE CALCIUM CARBONATE DICALCIUM PHOSPHATE ZINC OXIDE
PCDDs/Fs and dl-pcbs: results At the end of 2009 we had a food safety alert Hydrogenated fat WHO-TEQ PCDDs/Fs 0.916 pg/g >> 0.75 pg/g (2006/13/EC) 1.6 1.4 WHO-TEQ PCB WHO-TEQ PCDD/F 1.2 Administrator sent other 8 samples from the same producer All of them had a WHO-TEQ PCDDs/s > 0.75 pg/g WHO-TEQ (pg/g) Total 1.0 0.8 0.6 0.4 But only 2 had a WHO-TEQ PCDDs/Fs + dl-pcbs > 1.5 pg/g (2006/13/EC) 0.2 0.0 H.F. 1 H.F. 2 H.F. 3 H.F. 4 H.F. 5 H.F. 6 H.F. 7 H.F. 8 H.F. 9 WHO-TEQ PCB 0.219 0.098 0.059 0.219 0.193 0.088 0.052 0.145 0.039 WHO-TEQ PCDD/F 0.916 0.930 1.493 0.916 0.808 0.820 1.316 1.261 1.469 13
PCDDs/Fs and dl-pcbs: results RASFF notification https://webgate.ec.europa.eu/rasff-window/portal/index.cfm?event=notificationdetail¬if_reference=2010.0087 Notification is only for PCDDs/Fs because Local Government subtracted uncertainty to all results According with this criterion: 1. Sample H.F. 3 was the only one that exceeded the legal limit for WHO-TEQ PCDDs/Fs 2. TOTAL WHO-TEQ was always under legal limit 14
PCDDs/Fs and dl-pcbs: results No analysis of the original palm oil Analysis of a palm soap produced with the same oil Analysis of 2 feeds containing the hydrogenated fat 1.40 Both feeds had very low levels of WHO-TEQ Total WHO-TE EQ (pg/g) 1.20 1.00 0.80 0.60 0.40 0.20 WHO-TEQ PCB WHO-TEQ PCDD/F Palm soap levels l were low and lower than directive limit Palm oil was not contaminated PCDDs/Fs can be formed during hydrogenation 0.00 H.F. MEAN PALM SOAP FEED 1 FEED 2 WHO-TEQ PCB 0.124 0.061 0.042 0.009 process WHO-TEQ PCDD/F 1.103103 0.270 0.080080 0039 0.039 15
9.0 8.0 7.0 6.0 PCDDs/Fs and dl-pcbs: results Animal fat samples LARD ANIMAL FAT (BLEND) POULTRY FAT 140 120 100 80 60 40 LARD ANIMAL FAT (BLEND) POULTRY FAT (pg/g) Concentration (pg/g) 5.0 4.0 3.0 2.0 1.0 00 0.0 20 0 Concentration 2378-TCDD 12378-PeCDD 123478-HxCDD 123678-HxCDD 123789-HxCDD 1234678-HpCDD OCDD 2378-TCDF 12378-PeCDF 23478-PeCDF 123478-HxCDF 123678-HxCDF 234678-HxCDF 123789-HxCDF 1234678-HpCDF 1234789-HpCDF OCDF PCB81 PCB77 PCB123 PCB118 PCB114 PCB105 PCB126 PCB167 PCB156 PCB157 PCB169 PCB189 120 100 80 Vegetable fat samples SOYA OIL PALM OIL HYDROGENATED FAT >90% PALM SOAP 80 70 60 50 40 30 20 SOYA OIL PALM OIL HYDROGENATED FAT >90% PALM SOAP n (pg/g) Concentration (pg/g) 60 40 20 0 10 0 16 Concentration 2378-TCDD 12378-PeCDD 123478-HxCDD 123678-HxCDD 123789-HxCDD 1234678-HpCDD OCDD 2378-TCDF 12378-PeCDF 23478-PeCDF 123478-HxCDF 123678-HxCDF 234678-HxCDF 123789-HxCDF 1234678-HpCDF 1234789-HpCDF OCDF PCB81 PCB77 PCB123 PCB118 PCB114 PCB105 PCB126 PCB167 PCB156 PCB157 PCB169 PCB189
9.0 8.0 7.0 6.0 PCDDs/Fs and dl-pcbs: results Vegetable samples 40 ALFALFA DDG 35 GLUTEN FEED COTTON WHOLE SEEDS SOYA 30 ALFALFA DDG GLUTEN FEED COTTON WHOLE SEEDS SOYA n (pg/g) Concentration (pg/g) 5.0 4.0 3.0 2.0 1.0 0.0 25 20 15 10 5 0 Concentration 2378-TCDD 12378-PeCDD 123478-HxCDD 123678-HxCDD 123789-HxCDD 1234678-HpCDD OCDD 2378-TCDF 12378-PeCDF 23478-PeCDF 123478-HxCDF 123678-HxCDF 234678-HxCDF 123789-HxCDF 1234678-HpCDF 1234789-HpCDF OCDF PCB81 PCB77 PCB123 PCB118 PCB114 PCB105 PCB126 PCB167 PCB156 PCB157 PCB169 PCB189 2.0 1.8 1.6 1.4 1.2 RAPESEED BEET PULP SUGARCANE MOLASSES WHEAT MIDDLINGS PALMKERNEL MEAL SOLVENTS 50 70 60 RAPESEED BEET PULP SUGARCANE MOLASSES WHEAT MIDDLINGS PALMKERNEL MEAL SOLVENTS on (pg/g) 2378-TCDD 12378-PeCDD 123478-HxCDD D 123789-HxCDD 1234678-HpCDD OCDD 2378-TCDF 12378-PeCDF 23478-PeCDF 123478-HxCDF 123678-HxCDF 234678-HxCDF 123789-HxCDF 1234678-HpCDF 1234789-HpCDF OCDF Concentrationn (pg/g) 1.0 0.8 0.6 0.4 0.2 0.0 40 30 20 10 0 17 Concentratio 123678-HxCD PCB81 PCB77 PCB123 PCB118 PCB114 PCB105 PCB126 PCB167 PCB156 PCB157 PCB169 PCB189
PCDDs/Fs and dl-pcbs: results Mineral samples 7 6 5 SEPIOLITE CALCIUM CARBONATE DICALCIUM PHOSPHATE ZINC OXIDE tion (pg/g) 4 3 Concentrat 2 1 0 2378-TCDD 12378-PeCDD 123478-HxCDD 123678-HxCDD 123789-HxCDD 1234678-HpCDD OCDD 2378-TCDF 12378-PeCDF 23478-PeCDF 123478-HxCDF 123678-HxCDF 234678-HxCDF 123789-HxCDF 1234678-HpCDF 1234789-HpCDF OCDF 140 120 100 80 60 40 SEPIOLITE CALCIUM CARBONATE DICALCIUM PHOSPHATE ZINC OXIDE Concentration (pg/g) 20 0 18 PCB81 PCB77 PCB123 PCB118 PCB114 PCB105 PCB126 PCB167 PCB156 PCB157 PCB169 PCB189
ACKNOWLEDGEMENTS UTC PERSONNEL CONSUELO LÓPEZ BOLAÑO PAULA MARTÍNEZ TOJEIRO CRISTINA MONTOIORO PEREIRO GERARDO FERNÁNDEZ MARTÍNEZ THERMO ENGINEERS ANDREW BALDWIN MIGUEL ÁNGEL SÁNCHEZ JAVIER FERNÁNDEZ 19