This Technical Specification specifies a method for the determination of benzo[a]pyrene (BaP) plus benz[a<\/i>]anthracene (BaA), benzo[b<\/i>]fluoranthene (BbF) and chrysene (CHR) in several food matrices. The method is based on size exclusion chromatography (SEC) cleanup, followed by quantification with high performance liquid chromatography (HPLC) with programmable fluorescence detection. This method has been in-house validated via the analysis of spiked samples of edible olive oil, fresh mussels, smoked fish, smoked meat products, processed cereal-based foods for young children, infant formulae, chocolate and food supplements (isoflavones) at levels ranging from 0,25 \u03bcg\/kg to 1,00 \u03bcg\/kg and from 4,95 \u03bcg\/kg to 23,53 \u00b5g\/kg, depending on the Polycyclic Aromatic Hydrocarbon (PAH) or the matrix. This method complies with the performance characteristics specified for BaP, BaA, BbF and CHR in current legislation [3].<\/p>\n
The method has been shown to be applicable to a variety of additional matrices as meat products, fresh fish, paprika, roasted coffee, bread, herbs, breakfast cereals, beer, sunflower oil, olives and fried tomato, with a limit of quantification below 0,5 \u00b5g\/kg.<\/p>\n
In addition, the method was tested in-house and shown to be applicable also for the quantification of the other 12 PAHs of the 15+1 EU priority PAHs set (benzo[c<\/i>]fluorene (BcL), benzo[j<\/i>]fluoranthene (BjF), benzo[k<\/i>]fluoranthene (BkF), cyclopenta[cd<\/i>]pyrene (CPP), dibenz[a,h<\/i>]anthracene (DhA), dibenzo[a,e<\/i>]pyrene (DeP), benzo[ghi<\/i>]perylene (BgP), dibenzo[a,h<\/i>]pyrene (DhP), dibenzo[a,i<\/i>]pyrene (DiP), dibenzo[a,l<\/i>]pyrene (DlP), indeno[1,2,3-cd<\/i>]pyrene (IcP), 5-methylchrysene (5MC)) in all matrices listed above and at similar level ranges, except for CPP, where a UV detection had to be used with limits of quantification above 8 \u00b5g\/kg.<\/p>\n
For the determination of PAHs in edible fats and oils, two other standards are also available, EN ISO 22959 and EN ISO 15753 (see [1] and [2]).<\/p>\n
| PDF Pages<\/th>\n | PDF Title<\/th>\n<\/tr>\n | ||||||
|---|---|---|---|---|---|---|---|
| 5<\/td>\n | Foreword <\/td>\n<\/tr>\n | ||||||
| 6<\/td>\n | 1 Scope 2 Normative references 3 Principle 4 Reagents <\/td>\n<\/tr>\n | ||||||
| 9<\/td>\n | 5 Apparatus <\/td>\n<\/tr>\n | ||||||
| 10<\/td>\n | 6 Procedure 6.1 Sample preparation 6.2 Extraction <\/td>\n<\/tr>\n | ||||||
| 11<\/td>\n | 6.3 SEC cleanup 7 HPLC analysis 7.1 HPLC operation conditions Table 1 \u2014 Gradient conditions <\/td>\n<\/tr>\n | ||||||
| 12<\/td>\n | Table 2 \u2014 Fluorescent wavelength programme 7.2 Preparation of calibration solutions for HPLC <\/td>\n<\/tr>\n | ||||||
| 13<\/td>\n | Table 3 \u2014 Preparation of HPLC calibration solutions 7.3 Calibration curve 7.4 Determination of PAHs in test solutions <\/td>\n<\/tr>\n | ||||||
| 14<\/td>\n | 8 Calculation 9 Recovery 10 Test report <\/td>\n<\/tr>\n | ||||||
| 15<\/td>\n | 11 Precision data 11.1 General 11.2 Within-laboratory reproducibility <\/td>\n<\/tr>\n | ||||||
| 16<\/td>\n | Annex A (informative) Typical chromatograms Figure A.1 \u2014 Typical chromatogram of 15+1 PAHs from a sample of fresh mussels spiked at a level of 10 \u03bcg\/kg, each (CPP cannot be detected by fluorescence at that level) Figure A.2 \u2014 Typical chromatogram of 15+1 PAHs from a sample of extra virgin olive oil spiked at a level of 2 \u03bcg\/kg, each (CPP is cannot be detected by fluorescence at that level) <\/td>\n<\/tr>\n | ||||||
| 17<\/td>\n | Figure A.3 \u2014 Typical chromatogram of the PAH4 from a reference material of olive oil containing 2,06 \u03bcg\/kg of BaP, 3,17 \u03bcg\/kg of BaA, 2,40 \u03bcg\/kg of BbF and 9,52 \u03bcg\/kg of CHR, under the specific chromatographic conditions for the determination of the PAH4 Figure A.4 \u2014 Typical chromatogram from a sample of processed cereal-based foods for young children spiked with the 15+1 PAH at the level of 10 \u03bcg\/kg each, under the specific chromatographic conditions for the determination of the 15+1 PAHs. The chro… <\/td>\n<\/tr>\n | ||||||
| 18<\/td>\n | Annex B (informative) In-house validation data for the PAH4 in different matrices Table B.1 \u2014 In-house validation data for edible olive oil <\/td>\n<\/tr>\n | ||||||
| 19<\/td>\n | Table B.2 \u2014 In-house validation data for fresh mussels Table B.3 \u2014 In-house validation data for smoked fish <\/td>\n<\/tr>\n | ||||||
| 20<\/td>\n | Table B.4 \u2014 In-house validation data for smoked meat products Table B.5 \u2014 In-house validation data for processed cereal-based foods for young children <\/td>\n<\/tr>\n | ||||||
| 21<\/td>\n | Table B.6 \u2014 In-house validation data for infant formulae Table B.7 \u2014 In-house validation data for chocolate <\/td>\n<\/tr>\n | ||||||
| 22<\/td>\n | Table B.8 \u2014 In-house validation data for food supplements (isoflavones) <\/td>\n<\/tr>\n | ||||||
| 23<\/td>\n | Annex C (informative) In-house performance data with a mixture of cyclohexane and ethyl acetate as alternative extraction solvent Table C.1 \u2014 In-house performance data for olive oil Table C.2 \u2014 In-house performance data for fresh mussel Table C.3 \u2014 In-house performance data for smoked fish <\/td>\n<\/tr>\n | ||||||
| 24<\/td>\n | Table C.4 \u2014 In-house performance data for infant formulae <\/td>\n<\/tr>\n | ||||||
| 25<\/td>\n | Bibliography <\/td>\n<\/tr>\n<\/table>\n","protected":false},"excerpt":{"rendered":" Food analysis. Determination of benzo[a]pyrene, benz[a]anthracene, chrysene and benzo[b]fluoranthene in foodstuffs by high performance liquid chromatography with fluorescence detection (HPLC-FD)<\/b><\/p>\n |