MICROORGANIC POLLUTANTS: ACTION NEEDED TO SIMPLIFY THEIR ANALYSIS

MICROORGANIC POLLUTANTS: ACTION NEEDED TO SIMPLIFY THEIR ANALYSIS Mosca S., Guerriero E., Rotatori M.

The analysis of PCDD/Fs, PCBs and other related compounds requires complex analytical procedures using highly sensitive and selective instrumentation to meet very stringent data quality objectives.

At present, methods for analyses of dioxins and dioxin-like PCBs are expensive and slow. Therefore low-cost and fast methods have to be developed allowing to analyse in routinely manner a great number of samples and to provide quick, cheap, and reliable results on the presence of those compounds in the environment, feed and food

To speed up the time for analysis is important to many of today s GC analysts as they look for ways to improve sample throughput, without sacrificing the quality of the analysis. Benefits of increasing sample throughput include: Decreased costs: Less people and/or instruments are required to do the same amount of work. Increased profit: More samples can be processed in the same amount of time; Customers may be willing to pay more to get their results faster (surcharge for quick turnaround)

SCHEME OF ANALYSIS SAMPLING Chromatography column I Mixed column with differently treated silica CLEAN-UP EXTRACTION Chromatography column II Aluminium oxide B Super I AND FRACTIONATION PCBs fraction PCDD/Fs fraction HRGC/HRMS analysis HRGC/HRMS analysis ANALYSIS

SCHEME OF ANALYSIS SAMPLING Chromatography column I Mixed column with differently treated silica CLEAN-UP EXTRACTION Chromatography column II Aluminium oxide B Super I AND FRACTIONATION PCBs fraction HRGC/HRMS analysis PCDD/Fs fraction HRGC/HRMS analysis ANALYSIS

CNR-IIA SAMPLING CAMPAIGNS Industrial emission sampling Power plants Iron sintering ore plants Refineryies Medical waste incinerators Other plants Ambient air sampling

Sources of PCDD/Fs and PCBs in air - EU25 (5kg TEQ/year) Incinerators are no longer the main producers of dioxins due to regulatory restrictions and the subsequent compulsory technological evolution of plants (4.000 kg/year) From: European Commission. Identification, assessment and prioritisation of EU measures to reduce releases of unintentionally produced/released POPs. Study for implementation of the Regulation (CE) 850/2004 on Persistent Organic Pollutants (POPs) - June 2006

IRON ORE SINTERING PLANT - Trieste ng I-TEQ/Nm3 2005: Precautionary seizure from Law Court Company required to be within 0.4 ng I-TEQ/Nm 3 of PCCD/F Evaluation of influence of different charge typologies and additives on PCDD/Fs amount production and on congeneres profiles E. Guerriero et al., Environ. Eng. Sci. 26(1) (2009) 71-80 2,00 1,50 Studies about mechanisms of dioxin reduction 1,00 0,50 E. Guerriero et al. J. Hazard. Mater. 172 (2009) 1498-1504 0,00

POPs processes in the atmosphere A simultaneous monitoring of all affected environmental compounds is of great importance

AMBIENT AIR SAMPLING PCDD/Fs PCBs PAHs HCB Sintering plant

% I-TEQ Ambient air VS emission sampling % I-TEQ 60 Via Pitacco Ambient air sampling 50 40 30 20 10 0 media % febbraio media % luglio The congener pattern in ambient air samples is similar to the pattern in a stack emission sample 60 50 40 30 20 10 0 Flue gas stack emission emissioni forno di agglomerazione media % febbraio media % luglio

AMBIENT AIR REFERENCE VALUES European Directive 2004/107/CE (Italian Decree 152/2007) PAHs 1 ng/m 3 of B[a]P Air quality guidelines for Europe - WHO Regional Publications, 2000 PCDD/Fs PCBs 100 fg/nm 3 300 fg/m 3 indicate local sources to be identified and checked 3 pg/m 3 in non-industrial area 3000 pg/m 3 in industrial/urban area Average values Decision guidance document UNEP, 1996 HCB 300 pg/m 3

AMBIENT AIR STANDARD METHODS In Europe. ISO 12844 PAHs (2000) ISO 16000-13 PCDD/Fs and PCBs (2006) EN 15549 B[a]pyrene (2006) VDI 3498 PCDD/Fs (2008) ISO 16000-12 Sampling strategy for PCDD/Fs, PCBs and PAHs (2007) Common aspects: - Flow rate (15 m 3 /h) - Sampling time (24h) - Sampling system (High volume sampler equipped with a quartz fiber filter followed by an adsorbent)

In the U.S. AMBIENT AIR STANDARD METHODS EPA Compendium Methods TO-4A PCBs and pesticides in ambient air (1999) TO-9A PCDD/Fs in ambient air (1999) TO-13A PAHs in ambient air (1999) Common aspects: -Sampling system (High volume sampler equipped with a quartz fiber filter followed by an adsorbent) - Flow rate: 225 l/min (total sampled volume = 300 m 3 ) -Sampling time: 24h -

There is a lack of a standard method for simultaneous sampling of micro-organic pollutants (PAHs, PCDD/Fs and dl-pcbs). introduction of a single sampling method with increased sampling time, from 1 to 7 days, to enrich the samples and to determine different compounds simultaneously. Money saving: The method is less expensive than 24 h sampling, requires fewer samples to be treated and it permits weekly or monthly concentration trends to be obtained. Time saving: Finally, different classes of compounds can be analyzed simultaneously from only one sample.

SCHEME OF ANALYSIS SAMPLING Chromatography column I Mixed column with differently treated silica CLEAN-UP EXTRACTION Chromatography column II Aluminium oxide B Super I AND FRACTIONATION PCBs fraction PCDD/Fs fraction HRGC/HRMS analysis HRGC/HRMS analysis ANALYSIS

SCHEME OF ANALYSIS SAMPLING Chromatography column I Mixed column with differently treated silica CLEAN-UP EXTRACTION Chromatography column II Aluminium oxide B Super I AND FRACTIONATION PCBs fraction PCDD/Fs fraction HRGC/HRMS analysis HRGC/HRMS analysis ANALYSIS

AUTOMATED CLEAN-UP SYSTEM PrepLinc Platform Automated Sample Preparation System (J2 Scientific) A combination of: - GPC column (BioBead SX-3) - concentration module - SPE module - autosampler Mosca etal Organohalogen Compd 70 (2008) 501; Rossetti etal. OrganohalogenCompd 71 (2009) 2278

SPE column sample PCBs PCDD/F Concentration chamber GPC column AUTOMATED CLEAN-UP SYSTEM PrepLinc Platform Automated Sample Preparation System

GEL PERMEATION CHROMATOGRAPHY GPC is a highly effective cleanup method for removing high molecular weight interferences (i.e. lipids, polymers, proteins, natural resins, and cellular components) from sample extracts. This technique is ideal for post-extraction clean-up of pesticide residues, semi-volatiles, PAHs, PCBs, mycotoxins and antibiotic residues from animal tissue, fatty foods, plant tissue, soil, sediment, sludge and wastewater. Can GPC be suitable as a clean-up method of PCDD/Fs from environmental samples 22

Manual VS Automated Clean-up Total: 215 Ambient air 7% Incinerat ors 26% Kiln f or t he product ion of aluminium 1% Ref inery ( boilers) 2% Kiln f or t he product ion of clay expanded 13% RDF incinerator 19% Tyre incinerator 40% Medical waste incinerator 11% Cement plant 19% Pharmaceutical incinerator 7% Biomass incinerator 14% MSW incinerator 9% Sint ering plant 14% Cement plant ( co- burning) 18%

Manual VS Automated Clean-up 13 C CONGENER MANUAL CLEAN-UP AUTOMATED CLEAN-UP 2,3,7,8-TCDD 88 ± 4 80 ± 6 1,2,3,7,8-PeCDD 90 ± 3 81 ± 5 1,2,3,4,7,8-HxCDD 99 ± 3 85 ± 7 1,2,3,6,7,8-HxCDD 95 ± 3 85 ± 8 1,2,3,4,6,7,8-HpCDD 87 ± 6 70 ± 5 OCDD 67 ± 9 58 ± 11 2,3,7,8-TCDF 88 ± 6 75 ± 8 2,3,4,7,8-PeCDF 86 ± 4 70 ± 6 1,2,3,4,7,8-HxCDF 98 ± 3 91 ± 5 1,2,3,6,7,8-HxCDF 93 ± 4 91 ± 6 2,3,4,6,7,8-HxCDF 100 ± 3 90 ± 6 1,2,3,4,6,7,8-HpCDF 93 ± 6 71 ± 8 OCDF 68 ± 9 62 ± 9 mean (n=24) recoveries of EN-1948 ES in sintering plant samples OCDF 1,2,3,4,6,7,8-HpCDF 2,3,4,6,7,8-HxCDF 1,2,3,6,7,8-HxCDF 1,2,3,4,7,8-HxCDF 2,3,4,7,8-PeCDF 2,3,7,8-TCDF OCDD 1,2,3,4,6,7,8-HpCDD 1,2,3,6,7,8-HxCDD 1,2,3,4,7,8-HxCDD 1,2,3,7,8-PeCDD 2,3,7,8-TCDD AUTOMATED CLEAN-UP MANUAL CLEAN-UP 0 20 40 60 80 100 120

% Recovery AVERAGE AUTOMATED CLEAN-UP RECOVERIES 120 110 100 90 80 70 60 50 dl-pcbs PCDD/Fs It avoids the double analysis of dl- PCBs both in PCBs and in PCDD/Fs fraction: it means less standard solution to use and just two chromatographic runs 100 90 80 70 60 50 40 30 20 10 0

ADVANTAGES of the GPC/alumina automated purification system (1/2) Fast: in just over 1h it fully processes the sample from the extraction to the GC injection. The manual procedure, however, requires about 6h tomake the same process Time-saving: the automated system may work at night, standalone and it can processes up to 27 samples Money-saving: the GPC column is the same for hundreds of samples and the alumina microcolumn is one for each sample, whereas in the manual clean-up the operator prepares the adsorbents and then packs the column, one for each sample.

ADVANTAGES of the GPC/alumina automated purification system (2/2) Safer: the automated evaporation step is on-line: this allows a minor exposure of the operator to toxic solvents and is never exposed to chemical risk from agents like silica dust, acidic or AgNO 3 silica Green: it achieves the separation with a lower consumption of solvents (only DCM and Hex) Non-distructive : it is possible to analyze several classes of compounds within the considered mass range

SCHEME OF ANALYSIS SAMPLING Chromatography column I Mixed column with differently treated silica CLEAN-UP EXTRACTION Chromatography column II Aluminium oxide B Super I AND FRACTIONATION PCBs fraction PCDD/Fs fraction HRGC/HRMS analysis HRGC/HRMS analysis ANALYSIS

SCHEME OF ANALYSIS SAMPLING Chromatography column I Mixed column with differently treated silica CLEAN-UP EXTRACTION Chromatography column II Aluminium oxide B Super I AND FRACTIONATION PCBs fraction PCDD/Fs fraction HRGC/HRMS analysis HRGC/HRMS analysis ANALYSIS

Our goal: Separate and detect PCDD/Fs in the shortest amount of time possible HOW TO ACHIEVE OUR GOAL? Use different columns stationary phase type stationary phase thickness column length Use different temperature (or pressure) programs for the column Use different carrier gas (inert gas) Helium Hydrogen

ANALYTICAL INSTRUMENTATION GC/MS Thermo ITQ Thermo TSQ Quantum Autospec Ultima We are comparing the performances of LowRes MS (Ion Trap and Triple Quadrupole) vs HiRes MS in POPs analysis in different matrices

HRMS vs MS/MS: selectivity Precursor ion: [M] +, [M+2] + and [M+4] + (Resolution > 8200) CID (Precursor ion) (Product Ion) Product ions: [M-CO 35 Cl] +, [M+2-CO 35 Cl] +, [M+2-CO 37 Cl] +, [M+4-CO 37 Cl] +

Low Resolution Mass Spectrometer new method for the trace-level screening of PCDD/Fs, following the analytical strategy of U.S. EPA Method 1613 using TSQ Quantum XLS triple quadrupole gas chromatography tandem mass spectrometry (GC/MS/MS) instrument Using highly selective precursor ion selection and high-precision hyperbolic quadrupole technology available with the TSQ Quantum XLS, superior and uniform selectivity for screening for PCDD/Fs and PCBs at imposed MRLs in complex matrices is possible.

CONCLUSIONS In order to simplify POPs analysis, we suggest: SAMPLING Ambient air: simultaneous sampling, longer sampling time Industrial emission: Long-term sampling CLEAN-UP Automated clean-up (i.e. GPC+SPE based system) ANALYSIS Use of LowRes MS Use of a versatile GC column

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