10th International Symposium on Recent Developments in POPs Analysis: A Global Concern

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Recent Developments in Persistent Organic Pollutant Analysis

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Review presentations and articles from experts in environmental analysis and download for future reference. Join the experience and hear expert testimonials on current global POPs concerns. Check back for updated content including new articles written with The Analytical Scientist.

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Scientific Talks

Plenary Lecture: Human Biomonitoring: Parts per Quintillion— A New Frontier for Toxicologists and Risk Assessors

Donald G. Patterson Jr.
Exponent, Inc.
4921 Bill Cheek Rd., Auburn GA, 30011 USA

Abstract: Biomonitoring has become an indispensable tool for studying occupational and environmental exposure to chemicals, including persistent organic pollutants (POPs). POPs are lipophilic compounds and tend to accumulate in the lipid stores

of the body. Adipose tissue is mostly constituted by lipids (generally about 95% of the tissue) and therefore the POPs are generally found at the highest levels in this tissue. Because of the invasive nature of the surgical procedure required to obtain the adipose tissue sample, the analytical community then turned their attention to developing methods in blood which was a less invasive matrix, but the levels were much lower in the blood matrix due to the small amount of lipids (~0.6%) compared to adipose tissue (~95%). The methods required high resolution mass spectrometry (HRMS) in order to have the sensitivity required to measure normal background POPs levels in the picogram to femtogram range.

It is important to continue to try and develop more sensitive analytical methods for environmental chemicals: 1) to determine the normal human background levels of chemicals that we cannot detect with current methods; 2) to continue monitoring chemical levels that are decreasing in populations around the world (eg. dioxins, furans, PCBs, pesticides); 3) to provide better analytical CVs of chemicals that we can measure which will translate into lower measurement uncertainties; and 4) a lower analytical CV translates directly into higher statistical power in epidemiological studies.

Newer more sensitive analytical techniques are currently being developed using cryogenic zone compression and loop modulation coupled with HRMS to measure POPs. A modification of this technique called timed controlled cryogenic zone compression is being developed by Thermo Scientific. This technique allows targeted cryofocusing of certain peaks that might need enhanced sensitivity while allowing the remainder of the chromatographic separation to proceed unaltered.

The potential for these techniques to generate parts-per-quintillion detection will

be presented.

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Plenary Lecture: How Far Are We in the Chase after the Lowest Detectable Level for Dioxins?

Jean-François Focant, Ph.D.
Department of Chemistry, University of Liège, Belgium

Abstract: The chase for sensitive dioxin measurement methods has been going on for the last few decades. It opened when chickens were dying from Chick edema disease in the US in 1957. Today, one can surely say that we have reached a state-of-the-art in that field. Currently GC-MS methods can routinely work at the low picogram-high femtogram level, based on the use of high end MS sector instruments, but also the last generation of triple quadrupole analyzers. The latter being recently accepted as a tool for confirmatory measurement in food and feed under the maximum level (ML)-based EU legislation.

Once we consider human biomonitoring, the situation is somewhat different as there are no ML set for humans. The challenge there is to be able to measure the lowest possible quantity of target analytes, in order for the toxicologist working downstream of the analytical chemist to be able to properly analyze the situation in terms of global toxicity. Large volume injections (LVI), cryogenic compression (CZC), better ion production and transfer, longer ion accumulation times, … are the possible ways to enhance signals and improve instrument limits of quantification (iLOQs). Proper monitoring of blank levels is also of prime importance when considering method LOQs (mLOQs) as laboratory background levels are quite independent of sample sizes when chasing at sub femtogram levels and can easily exceed the levels to be measured in samples.

This presentation will focus on recent advances in the field of measurement at ultra-trace levels of dioxins and related compounds in the frame work of human biomonitoring.

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From Blue Sky to Field: Regulatory Drivers for the Development of New Methodology

Dr. David Mortimer, C Chem, MRSC
Food Standard Agency
United Kingdom

Abstract: Consumers are protected from health risks that arise from dietary exposure to POPs through a series of risk management measures that may include regulatory limits, action or guidance levels, codes of practice and precautionary advice. All stages from initial investigations into emerging risks, through surveys of levels in food, toxicology studies and investigations into contamination pathways, to in-process testing and the introduction of official controls to check compliance with regulatory limits, involve sometimes very challenging analytical method development and validation. The presentation will follow the route from the initial identification of an emerging POP to the stage when it is regulated in food. It will examine the types of challenges faced at different stages, whether these relate to incomplete knowledge of the determinant structures under investigation (for example, because the precise target compounds are unknown or because metabolites and degradation products may be significant); a need for very high sensitivity and/or selectivity; matrix complexity; good robustness; rapid turnaround; versatility; ease of use; and low cost. Real examples will be taken from chemicals or groups of chemicals that are already regulated under EU legislation; those that have been the subject of discussions at the European Commission’s Expert Committee on Persistent Organic Pollutants in Food; those that are the subject of calls for data through EC Recommendations based on advice from EFSA; and some that may arise in future discussions, for example because they are candidate compounds for listing under the Stockholm Convention.

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Assessment of Dioxin Exposure in the Communities Living Near the Da Nang Agent Orange Hotspot

Nguyen Hung Minh, Ph.D.
Vietnam Environment Administration
Hanoi, Vietnam

Abstract: Da Nang Airbase is one of the three major Agent Orange hotspots in Vietnam which showed extremely high level of dioxin in soil, sediment and biota. The elevated dioxin contamination was originated from uses of defoliants, especially Agent Orange, by the United States army during the wartime. The environmental contamination of dioxin has led to their bioaccumulation in the local food chains and consequently, the human exposure. Lack of knowledge and inappropriate awareness of the local people are key factors to increase the dioxin exposure and possible health consequences in the communities. This study explored dioxin exposure pathways in the local communities and as well as possible intervention measures to protect the people health.

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Integrated Solutions for GC-MS POPs Analysis

Dwain Cardona
Thermo Fisher Scientific
San Jose, CA, USA

Abstract: POPs sample analysis requires robust instrument performance with integrated solutions that streamline method development and standardize workflows.

While many tools exist that enable more efficient sample analysis, knowledge of

the various solutions is often lacking. In this informative presentation we will discuss specific features of the Thermo Scientific™ GC and GC/MS and sample preparation product portfolio focusing on tools and techniques useful for POPs analysis. The discussion will include sample preparation, mass spectrometry instrumentation and software platforms highlighting functionality and features with specific references to POP sample data.

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Dissemination Patterns and Risks Impersonated of Polycyclic Aromatic Hydrocarbons (PAHs) Contaminated in the Surface and Subsurface Soil

Mohamed H. EL-Saied
King Saud University
Riyadh, Kingdom of Saudi Arabia

Abstract: Pollution of agricultural land and urban areas in Riyadh is found affected with poly-cyclic aromatic hydrocarbons emitted through industrial processes, transport and local heating sources. Highest PAHs concentrations were found in soils near industry. The observed values are much higher than the reported concentrations of PAHs to non-industrial sites. oncentrations of PAHs in industrial land are higher than in urban soil, but the soil in relation to the other urban areas of the world. The highest observed concentrations of PAHs in soils of industry that have been collected near petroleum and charcoal industry. Elevated PAH concentrations were determined in places from the industrial area, but the oncentrations decreased compared to areas near the industry. Compared with the tonnage of PAH in other parts of the world, and shows that the soil in the area Riyadh. It is the main source of PAH pollution in coal tar refinery region. There are also contributions from other sources, district heating, transport and transport pollutants powerful. It is not possible to distinguish between the contributions of each source based on the proportions of the diagnosis.

Keywords: Polyaromatic Hydrocarbons, Soil, Environmental, Pollution, Dissemination patterns and risks impersonated

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Measurement of Ultra-Trace Levels of Dechloranes in Food and Feed. Assessment of Dietary Intake.

Chiara Calaprice
Department of Chemistry, University of Liège, Belgium

Abstract: Dechloranes, namely Dechlorane Plus (DP, C18H12Cl12), Dechlorane 602 (Dec 602, C14H4Cl12O), Dechlorane 603 (Dec 603, C17H8Cl12), Dechlorane 604 (Dec 604, C13H4Br4Cl6) and Chlordene Plus (CP, C15H6Cl12) are a family of emerging persistent organic pollutants (POPs) norbornene derivatives. They exhibit flame retardant and pesticide properties and are used as replacement of regulated compounds such as Mirex (also called Dechlorane). They and some of their by-products have been recently reported in dust from various environments [1, 2, 3, 4, 5]. Besides environmental issues, humans are exposed to these chemicals, and so far, barely any data about toxicity, metabolization, and prevalence in human body is available. Some very recent human biomonitoring studies have just highlighted levels at the ng/g lipid order in human serum from Norway [6] from France [7], and in breast milk from Canada [8].

As far as we know, at this time no data is available in Europe about Dechlorane levels in food and human exposure correlated to food consumption. The purpose of the study was to develop an analytical method for the analysis of 6 Dechloranes: Mirex, Dec 602, Dec 603, CP, DP syn, and DP anti isomers. The method is based on a sample preparation used for dioxins and on isotope dilution (ID) GC-QQQMS/MS for quantification. Maximization of signals was investigated for ultra-trace measurement by means of Experimental Design for large volume injection (LVI) operated in solvent vent mode. We have analyzed 104 different sample, including matrices such as milk, chicken, pork, eggs, pure animal fat, vegetable oil, salmon, feed additives, and corn usually received in our ISO17025 accredited routine dioxin laboratory and also bought from retail shops. We found low Dechlorane levels in feed (not reported) and assessed the average daily exposure dose of Dechloranes based on food habits of Belgian population for these products to be in the range 130-140 pg/day for the sum of all Dechloranes.

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Assessment of Polychlorinated Biphenyls (PCBs) in Soils and Water of an Old Transformer Oil Storage Site in Lagos, Nigeria

Babafemi A. Babalola
Department of Chemistry, Faculty of Science
University of Ibadan, Nigeria

Abstract: Polychlorinated biphenyls (PCBs) are toxic and persistent chemicals manufactured between 1929 and 1977 for insulating fluids in electrical equipments. They persist in the environment and accumulate in plants and animals. They induce reproductive disorders and endocrine disruption in humans. This study assessed the extent of polychlorinated biphenyl (PCB) contamination of soil and groundwater in a major storage site of old transformer oil in Lagos. 8 topsoil, 3 subsoil and 2 groundwater samples were collected from storage site of old transformer oil in Transmission Work Centre at Akangba, Lagos. Soil samples were extracted into hexane-acetone mixture (1:1) in an ultrasonic bath for three minutes and water samples were extracted with dichloromethane in separating funnel. The extracts were cleaned up by column chro-matography method using silica gel and analyzed by GC-MS. PCBs concentration in water (0.13-0.35 mg/L) and soil (1.00-39.0 mg/kg) were compared with AENV (2009) standard for industrial water (0.0094 mg/L) and soil (33 mg/kg). The source of con-tamination in soil and water samples can be traced to storage and use of PCB-con-taining transformer oil in the study site. Therefore, analysis of the groundwater in the vicinity of the study site is needed to be carried out.

Keywords: PCBs, endocrine disruption, old transformer oil, groundwater, GC-MS

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High sample Throughput in a Modern Dioxin Laboratory using Dual Acquisition

Wim Traag
Erichem, The Netherlands

Abstract: Feed business operators including traders and import operators placing oils or derived products intended for the use in animal feed on the market, must have these products analyzed in accredited laboratories for the sum of dioxins and dioxin-like PCBs. The operator placing the feed on the market is responsible for the sampling and analyses. Only if the analysis result is favorable, the feed or the ingredient can be delivered (together with the report of analysis of the concerned batch as a proof). These requirements are based on EU-legislation, as laid down in Reg. (EU) No. 225/2012 and Commission Regulation (EC) No 152/2009.

These “positive release” requirements puts great pressure on laboratories. Reports of analysis should not only be based on state of the art technology and fully accre-dited methods but more over they should be available, due to high cost before the ship can be unloaded (demurrage), as soon as possible. In order to be able to report results of analysis within 24 hours automation of the whole procedure is a must.

At Nofalab on request by the client a number of samples can be analyzed and reported within 8 hours. This has been achieved by introducing a new sample preparation system in combination with dual acquisition GC-HRMS.

The main advantages of the new sample preparation systems are:

• Extremely low solvent consumption (< 100ml="" per="">

• Rapid turnaround time of only 90 minutes

• Small final volume of fraction of interest 1-2 ml

• Columns easy to connect and no leakage

• No cross contamination

By using dual data acquisition the capacity of a GC-HRMS system is almost doubled. The Dual Data Acquisition option enables alternating injection series into two GCs both coupled to one high resolution mass spectrometer resulting in a comparable sample throughput of two single GC-HRMS systems.

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The role of Proficiency Tests in the Estimation of Measurement Uncertainty of Polychlorinated PCDD/PCDF and PCB Determination by Isotope Dilution Methods

Stefano Raccanelli
Environmental Ethical Scientist, Italy

Abstract: Uncertainty of POPs measurements is an increasingly important issue often neglected, in spite of the great importance that accuracy and precision have, for example, also in legal forums. Like other analytical measures, POPs concentration has several sources of uncertainty and proficiency tests, other than allowing participants evaluation, might provide a basis for defining uncertainty related to analytical method, to matrixes and to congeners. In this talk, we will evaluate how the European laboratories are moving toward estimating their uncertainty and how the continuous participation to proficiency tests can be the basis for a dynamic and updated estimation of laboratory uncertainty (Magnusson et al., 2003). In this framework, we present the results of Intercind, an international proficiency test for POPs resulting from merging of two long experiences (Intercal and CIND), for estimating the uncertainty of measurements of dioxins, furans and polychlorinated bypheniles.

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New and Emerging Techniques in POPs Analysis

Esteban Abad, Ph.D.
Barcelona, Spain

Abstract: Persistent organic pollutants (POPs) are environmental contaminants characterized by their persistence, long-range transport and biomagnification in food chains and bioaccumulation in humans. Polychlorinated dibenzo-p-dioxins (PCDDs), dibenzo-furans (PCDFs) and dioxin-like polychlorinated biphenyls (dl-PCBs) are commonly known as dioxins and furans and internationally recognized and the most important toxic members of POPs. The detection and confirmation of dioxins requires a high level of analytical performance. Until recently, EU legislation required the use of HRMS based on magnetic sector analyzers as a standard approach for confirmatory analysis of dioxins in feed and food. However, recent technological advances in GC-MS/MS instruments have enabled other alternatives (Commission Regulation (EU) No 589, 2014 and Commission Regulation (EU) No 709, 2014). These analytical techniques achieve the required sensitivity and selectivity for dioxin analysis. As result, GC-MS/MS is now permitted for use in the EU to control the maximum levels for dioxins in food and feed as a full confirmatory method. When using GC-MS/MS, specific performance criteria for dioxin confirmation must be met in addition to the criteria described by the European Commission regulations.

In this presentation the results of several experiments performed using triple quadrupole for dioxin analysis in food and feed samples is displayed. In addition, other GC- MS approaches which in the near future could be also suitable to be considered for same purposes will be noticed.

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The Organic Flame Retardant Story: Knowns and Unknowns

Stuart Harrad
School of Geography, Earth & Environmental Sciences
University of Birmingham, United Kingdom

Abstract: To meet flame retardancy regulations, a wide range of consumer goods are treated with chemical flame retardants. The discovery in the late 1990s that concentrations of one such class of chemicals (polybrominated diphenyl ethers – PBDEs) were increasing exponentially in human milk, triggered concerns about the environmental presence and effects of PBDEs and related brominated and organophosphate flame retardants (BFRs and PFRs). Because of their extensive indoor applications, the pathways of human exposure to these chemicals differ from those of organochlorine POPs like dioxins. Moreover, while manufacture and new use of PBDEs has ceased, there is a vast quantity of consumer goods containing them that have already or are yet to enter the waste stream, with concomitant potential for environmental releases. This presentation will summarize what is known about the sources, pathways, and extent of human exposure to BFRs and PFRs, covering emissions and exposure during both the in-use and end-of-life phases.

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From Emission to Immission: How POPs Stroll Around

Werner Tirler
Bolzano, Tirol

Abstract: Persistent organic pollutants (POPs) are toxic chemicals that negatively affects environment and human health. Since they can be emitted by different anthropogenic activities, like waste incineration and industrial activities, they are nowadays ubiquitously present in our environment. They are persistent and can be transported over long distances by wind and POPs generated in one country can even affect peoples health and wildlife far from where they were released. For this reason, it is considered that air is the key medium to be sampled in global monitoring programs.

Special active air samplers were used to measure selectively according to actual local wind conditions. This permits also to get information’s on how POPs once emitted are migrating by wind transport. By the use of a multi-stage impactor, cut points for the particulate fraction at aerodynamic diameter of 10 μm, 2.5 μm and 1 μm were obtained. The results indicate that POPs are mainly bound to the fine particulate fraction (< pm="" 1).="" in="" our="" study="" we="" found="" for="" urban="" areas="" a="" high="" contribution="" of="" small="" local="" emittents="" eg.="" domestic="" heating="" by="" household="" stoves)="" to="" the="" total="" pop="" concentration="" in="">

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Recent Advances in Lowering the Cost of Dioxin Analysis

David Hope
Pacific Rim Laboratories, Canada

Abstract: Conventional methods for PCDD/F and dlPCB analysis in food have been laborious and expensive, using soxhlet extraction, multi-column clean-ups (acid/base silica gel, Florisil, alumina, carbon) and ultimately high resolution mass spectrometry for analysis. Automated alternatives have been posed for the extraction and clean-up, however the costs can be prohibitive for smaller labs, especially in a competitive marketplace.

Lee et al1, and Yang et al2 have demonstrated the use of tandem acid silica / carbon columns in analyzing environmental samples for PCDD/F and dlPCB. We have taken previously analyzed food samples and developed a method for extraction by the Thermo Scientific™ Dionex™ Accelerated Solvent Extraction (ASE) system using Dioniun™ cells. Samples containing up to 5 g of lipid are cleaned up on acid silica gel columns and by coupling with a carbon column (Cape Technologies), can separate PCBs from PCDD/F.

Concentrating samples can be a laborious, time consuming step, however with the Rocket Evaporator, twelve samples coming off the Cape clean-up columns can be to the instrument in 1-1.5 hours. We will show how it is possible to prepare and analyze six samples in a day.

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Legacy Flame Retardants, Emerging Flame Retardants and Dioxins: Links and Tools for Risk Assessment

Martin D. Rose, Ph.D.
United Kingdom

Abstract: Some classes of brominated flame retardant (BFR) such as PBDEs and HBCDD

are now well established as legacy contaminants. More evidence and association between these classes of chemicals and negative impacts on the environment

and health are being reported, and as a result restrictions on their use are being imposed. As a result of these restrictions, new flame retardants are being placed

on the market. These are usually chemicals with similar properties and which are documented regarding production and use as FRs where they have been shown

to occur/ distribute to the environment and/or wildlife, humans or other biological matrices, are classed as ‘emerging’ contaminants. Chemicals that are documented as potential FRs and have been shown to be present in materials or products are termed ‘novel’ flame retardants, and the term ‘potential’ FRs has been used for chemicals reported to have applications as FRs (e.g. in patents).

Many of these organo-bromine compounds have the potential when they break down to form brominated dioxins and related compounds. These are potentially more toxic than the parent compounds but are rarely considered in risk assess-ments for BFRs. There is also a potential for mixed chloro- bromo- dioxins to be produced where there is also a source of chlorine present. Analysis for dioxins

is complex, and whilst more widely available now that a few years ago, is still usually confined to chlorinated congeners. Exposure estimates and risk assess-ments for dioxins do not usually consider brominated and mixed halogenated congeners.

Work done to estimate relative potencies for brominated and bromo- chloro-

dioxins and furans will be described and the importance of holistic risk assess-ments that covers both FRs and dioxins will be discussed.

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Use of Micro Scale Solid Phase Extraction and Automated Clean Up for POPs Analysis in Various Sample Matrices

Tom Hall
Toxic Reports Watertown, MA

Abstract: Laboratory analysis of human serum for Persistent Organic Compounds (POPs) has become increasingly important. The concept of biomonitoring in which levels of toxic chemicals are assessed in humans (typically in serum and urine) has received much attention in the last decade. Automation of the sample prep process can result in faster turnaround time of samples, lower costs, and improved quality of the data generated. As the amount of sample used in bio-monitoring as well as other markets with Solid Phase Extraction is typically lower than in, e.g., water analysis, a micro system is used.

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Recent Activities of the EURL/NRL Network for Dioxins and PCBs in Feed and Food

Alexander Schächtele
EURL for Dioxins and PCBs in Feed and Food
State Institute for Chemical and Veterinary Analysis of Food (CVUA)
Freiburg, Germany

Abstract: The organization of proficiency tests is one of the important tasks of the European Union Reference Laboratory (EURL) for Dioxins and PCBs in Feed and Food. In 2014 a proficiency test on determination of PCDD/Fs and PCBs in cod liver and

fish liver oil was performed. Additionally participants were asked to analyze selected brominated flame retardants described in Commission Recommendation 2014/118/EU. 93 participants reported results for PCDD/Fs and/or PCBs and 27 participants reported results for at least one of the listed brominated flame retardants.

Another important topic of the network of the European Union Reference Laboratory (EURL) and the National Reference Laboratories (NRLs) of the European member states is the development of guidance document for the estimation of measurement uncertainty for persistent organic pollutants (POPs) quantified by isotope dilution-mass spectrometry based techniques, in particular PCDD/Fs and PCBs. Within this working group approaches based on the bottom-up and top-down approach are discussed and developed. The measurement uncertainty is also closely linked with the estimation of the limit of quantification. In cooperation with the three other EURLs working in the field of contaminants (EURL for heavy metals in feed and food, EURL for mycotoxins and EURL for polycyclic aromatic hydrocarbons) a guidance document is developed.

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10 Years Dioxin Gold Standard: DFS GC-HRMS System

Dirk Krumwiede, Ph.D.
Thermo Fisher Scientific, Bremen, Germany

Abstract: The DFS magnetic sector GC-HRMS system celebrates its 10th birthday in 2015. Introduced on the Dioxin conference in 2005 in Toronto it soon became the market leading solution in Dioxin and POPs analysis – the “gold standard for Dioxins” - in following years. The DFS system has been specifically designed for Dioxins & POPs analysis based on 20 years of experience in this field from its predecessor the MAT95 GC-HRMS. If offers a number of important and several unique features:

- Worldwide compliance with official Dioxin & POPs methods

(e.g. EPA 1613, 1668, 1614, etc.)

- The best Dioxin installation specification: 20 fg 2378 TCDD with SN > 200:1

(4 Sigma noise definition) – Demonstrated with every installation of a DFS

- A large volume ion source specifically designed for highest sensitivity in combi-nation with robustness for routine analysis - “robustness -by-size”. Tested over 10 years by now in Dioxin & POPs routine analysis in many Dioxin & POPs expert labs all over the world.

- Unique dual GC configuration

This presentation intends to have a look back into DFS beginnings, discuss where we are now and have an outlook into the future. Where are we now? Where could we go? How close are we to detect attogram amounts of 2378-TCDD and/ or other POPs?

Analysis data of 2 fg 2378-TCDD on a TR5ms type 60 m column (i.D. 0.25 mm, 0.25 um film) will be shown and data from human serum extracts at lowest levels.

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Things You Can Do to Make GC Systems Stay Up Longer to Analyze More Samples

Jack Cochran
Restek, USA

Abstract: The so called “dilute-and-shoot” method is an approach to avoiding “detector” issues such as matrix-suppressed/enhanced compound response in LC-MS/MS. The sample is diluted in mobile phase prior to injection so matrix no longer negatively influences LC-MS/MS analysis of trace level components.

In GC-MS, most problems occur on the front end, at the GC inlet, where compounds can degrade during hot splitless injection, active compounds can be irreversibly adsorbed to inlet liner surfaces, and nonvolatile material from dirty samples can compromise the transfer of less volatile compounds of interest from the inlet to the GC column. These issues are magnified due to the very slow inlet flow during splitless injection, which is typically less than 2 mL/min. One way to mitigate them is to instead use split injection, what we term “shoot-and-dilute”, where the much higher flow rate through the inlet results in a substantially reduced residence time and a proportionately higher transfer for difficult compounds of interest. This technique is especially appropriate with ultra-sensitive detectors like the electron capture detector (ECD), magnetic sector high resolution mass spectrometers, and newly introduced GC-MS/MS systems.

Yet another way to suppress chromatography problems, especially in splitless injection GC, is through the use of “analyte protectants”, which are essentially masking agents such as sugars, diols, etc. that are co-injected with samples and standards to tie up active sites in GC inlet liners and columns. While this technique is somewhat common in the GC analysis of pesticides in food, its use is underexplored in environmental analytical work.

This paper will demonstrate the possibilities of using split injection and analyte protectants to keep GC systems up longer to analyze more samples.

Better chromatographic peak shapes and more stable responses are the goals.

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