A new gold standard for Dioxin analysis A combined approach ; automated efficient clean-up together with a new Triple Quadrupole GC-MS/MS
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1 A new gold standard for Dioxin analysis A combined approach ; automated efficient clean-up together with a new Triple Quadrupole GC-MS/MS Haruhiko Miyagawa, GCMS Product Manager Analytical &Measuring Instruments Division SHIMADZU Cooperation DIOXIN 2016
2 2 Recent demands for Dioxin analysis in food/feed Reliable results Easier and faster operation Automated procedure Lower running cost Dioxins S 3, Smart Solution Systems Analysis
3 Benefits of the system Fat Fatty Acid Sample Preparation Extraction Purification Concentration Measurement 1 day > 1 day > 2 days 0.5 day Analysis of 12 samples >2hr >3hrs 3
4 BUCHI SpeedExtractor E-914 / E-916 >10x Faster extraction Extracts the samples at elevated temperature and pressure High throughput Parallel extraction of up to 6 samples Less solvent use Sample volumes ml, no limitation for large sample size Reliable and efficient Direct sample transfer between extraction and concentration Extract collection glassware is compatible to evaporation instruments from BUCHI 4
5 MIURA GO-HT series Less solvent No DCM use Small fraction Automated & shorter time process Consumables Total Solvents required Solvents Silver Nitrate Si column Sulfuric Acid Si column Carbon column AlOx column 110mL Hex, Tol Cost Sol( 120/euro) 221 ( 2) Final fraction volume 1 ~ 1.5 ml Max. Sample Weight 3 g fat Max. Capacity 6 samples Basic Time 94 mins No Cross-contamination Reliable Results! Washing the system is not necessary! 5
6 Shimadzu GCMS-TQ8050 A new GC-MSMS, our latest model! Ultra-high sensitivity for the fg quantitative analysis Durable hardware for routine work Easy to use, simple operation and maintenance 6
7 7 Samples Foods : Pork meat, Egg, Salmon, Olive oil from the market, 3g fat collected. Standard Reference Material :Cod liver oil (EDF-5462, EDF-5463 from CIL), 1g fat collected. Sample were extracted by SpeedExtractor (BUCHI) and purified by GO-HT (MIURA) Obtain 1.3mL Dioxin fraction and 1.0mL PCB fraction, concentrated to 20uL and 50uL.
8 8 GC-MS/MS conditions GC Column: Glass Insert: Injection Volume: Injection Mode: Injection Temp: 280 Column Oven Temp. program: DB-5MS (60 m, 0.25 mm I.D., df =0.25 μm) Custom SkyLiner, Splitless (Restek) 2 μl Splitless Food New Regulation Commission Regulation No 589/ (1 min) (20 /min) 220 (2 /min) 260 (3 min) (5 /min) 320 (8.5min) Flow Control Mode: Constant Linear Velocity 29.4 cm/sec MS Ionization Mode: Acq.Mode: Ionization Volt.: Emission current: Transision, CE: Loop Time: Pressure of CID gas: MS resolution: Detector volt.: Calibration range: EI MRM mode 70 ev 150 μa refer to Smart Env. Database 1 sec (Target: 0.8 sec., ISTD: 0.2 sec.) 150 kpa Q1 Unit, Q3 Unit 2.1 kv conc ppb ~ 2 ppb (x10,000) > > ,2,3,4,7,8- HxCDF ,2,3,6,7,8- HxCDF
9 Instrument performance check Standards compound RRF at conc. (ppb) LOQ LOD σ 10σ 2,3,7,8-TeCDD ,2,3,7,8-PeCDD ,2,3,4,7,8-HxCDD ,2,3,6,7,8-HxCDD ,2,3,7,8,9-HxCDD ,2,3,4,6,7,8-HpCDD OCDD ,3,7,8-TeCDF ,2,3,7,8-PeCDF ,3,4,7,8-PeCDF ,2,3,4,7,8-HxCDF ,2,3,6,7,8-HxCDF ,3,4,6,7,8-HxCDF ,2,3,7,8,9-HxCDF ,2,3,4,6,7,8-HpCDF ,2,3,4,7,8,9-HpCDF OCDF ,3,7,8-TeCDD > > ,2,3,7,8-PeCDD > > ,2,3,4,7,8-HxCDD > > ppb 9
10 Stress test : Stability check Standards Pig fat Olive Egg Salmon EDF-5462 EDF-5463 Compound Name Injection number Mean Conc value (ppb) %RSD 2,3,7,8-TeCDD ,2,3,7,8-PeCDD ,2,3,4,7,8-HxCDD ,2,3,6,7,8-HxCDD ,2,3,7,8,9-HxCDD ,2,3,4,6,7,8-HpCDD OCDD ,3,7,8-TeCDF ,2,3,7,8-PeCDF ,3,4,7,8-PeCDF ,2,3,4,7,8-HxCDF ,2,3,6,7,8-HxCDF ,3,4,6,7,8-HxCDF ,2,3,7,8,9-HxCDF ,2,3,4,6,7,8-HpCDF ,2,3,4,7,8,9-HpCDF OCDF
11 Evaluation of quantitative values Analysis of the food samples (pg/g) Pig fat Olive Egg Salmon 3.5 (x10,000) > Compound Name Pig fat > Oliveoil Egg fat Salmon 3.0 GC-MS/MS GCHRMS 2.5 GC-MS/MS GCHRMS GC-MS/MS GCHRMS GC-MS/MS GCHRMS 2,3,7,8-TeCDD ND ND2.0 ND ND ND ND ND ND 1,2,3,7,8-PeCDD ND 1.5 ND ND ND ND ND ND ND 1.0 1,2,3,4,7,8-HxCDD ND ND ND ND ND ND ND ND 0.5 1,2,3,6,7,8-HxCDD ND ND0.0 ND ND ND ND ND ND 1,2,3,7,8,9-HxCDD ND ND ND ND ND (0.3) ND ND 1,2,3,4,6,7,8-HpCDD ND ND OCDD ND ND 0.6 (0.5) ND (0.2) OCDD 1.0 (1.1) (in ND Egg fat) ND ND (0.5) 2,3,7,8-TeCDF ND ND ND ND (0.2) (0.3) (0.3) (0.4) (x1,000) 1,2,3,7,8-PeCDF ND ND >240.95ND ND ND (0.2) ND ND > ,3,4,7,8-PeCDF ND ND ND ND (0.1) (0.2) (0.2) (0.3) 1,2,3,4,7,8-HxCDF ND ND5.0 ND ND ND ND ND ND 1,2,3,6,7,8-HxCDF ND ND ND ND ND ND ND ND 1,2,3,7,8,9-HxCDF ND 2.5 ND ND ND ND ND ND ND 2,3,4,6,7,8-HxCDF ND ND ND ND ND ND ND ND 1,2,3,4,6,7,8-HpCDF ND 0.0 ND 20.0ND 21.0 ND ND ND ND ND 1,2,3,4,7,8,9-HpCDF ND ND 2,3,7,8-TeCDF ND ND ND ND ND ND OCDF ND ND (in ND Egg fat) ND ND ND ND ND ( ) : LOD < value < LOQ 11
12 Evaluation of quantitative values Quantitative values of EDF-5462 Comparison of the individual dioxin concentrations using GC-HRMS and GC-MS/MS. Compounds Name Mean value %RSD(n=3) GC-MS/MS GCHRMS GC-MS/MS GCHRMS 2,3,7,8-TeCDD ,2,3,7,8-PeCDD ,2,3,4,7,8-HxCDD ,2,3,6,7,8-HxCDD ,2,3,7,8,9-HxCDD ,2,3,4,6,7,8-HpCDD OCDD ,3,7,8-TeCDF * ,2,3,7,8-PeCDF ,3,4,7,8-PeCDF ,2,3,4,7,8-HxCDF ,2,3,6,7,8-HxCDF ,3,4,6,7,8-HxCDF ,2,3,7,8,9-HxCDF ,2,3,4,6,7,8-HpCDF ,2,3,4,7,8,9-HpCDF OCDF *Peak was saturated EDF-5462 EDF
13 Summary Dioxins S 3, Smart Solution Systems contribute Speed of the sample preparation Reduction of running cost Reliability of the results Food New Regulation Commission Regulation No 589/2014 Shimadzu Europe 13
14 Future work More evaluations Faster GC analysis Food Feed New Regulation Commission Regulation No 589/2014 Commission Regulation (EC) No 709/ min. (Column:DB-5MS 60 m)) > > min. (Column:DB-5MS 30 m) Peak separation of 1,2,3,4,7,8- HxCDF to 1,2,3,6,7,8-HxCDF is satisfactory to the criteria
15 Thank you for your attention 15
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