1ST GREEN ANALYTICAL CHEMISTRY WORKSHOP
5/6 FEBRUARY 2024 - NOVOTEL PARIS CHARENTON
09h30 - 09h55
Chemical transition in laboratories
Ismahane REMONNAY (Département des Expertises Scientifiques
et Techniques - Veolia, France).
09h55 - 10h20
Laboratories and environmental challenges: a paradigm shift
Christophe PÉRÈS (Le Labo Durable - LLD, France).
In October 2020, the European Commission published its sustainable development strategy, stating that 90% of Europeans are concerned about the impact of chemicals on the environment. Public distrust, concerns about dwindling supplies of raw materials and energy resources, and increasing environmental pollution have led the chemical community to redefine its thinking from a performance-driven approach to a broader vision geared towards sustainable development.
While the chemical synthesis community is well advanced in this direction thanks to the principles of green chemistry, analytical chemistry has received less attention on this subject. The multiplication of norms and quality standards has increased the need for precise measurements, and analytical chemistry is present at every stage of a product's development, potentially throughout its entire life cycle. The counterpart is the implementation of various intensive processes involving the consumption of resources, the production of data and the generation of high-impact waste. Today, the production figures for each analysis are rarely known, and the quantities of water, eluent, reagents, energy, special gases and consumables are often ignored. This makes it difficult to assess the impact of analytical processes, as there are no tools to quantify them.
10h20 - 10h45
Green Sample Preparation Solutions based on
Miniaturization and Automation
Frank DAVID (RIC technologies, Belgium).
Sample preparation is often considered as the step with the highest impact on the greenness of a complete analytical procedure. During the past years, numerous scientific papers have been published describing sample preparation techniques that reduce or even eliminate the use of solvents and toxic chemicals, that reduce the required sample size, and that are consuming less energy due to miniaturization. However, these new sample preparation methods do not always find their way to routine laboratories. Reasons for this can be based on the fact that no evidence is given that the new method delivers equivalent results (method not fully validated), but in a lot of cases the low implementation rate is also due to combinations of resistance to change, perceptions, and even biased assumptions. A traditional example of this is the assumption that stir bar sorptive extraction (SBSE) cannot be used on water samples containing some suspended organic material.
In this presentation, examples will be given how classical methods can be replaced by miniaturized and automated methods while analytical criteria such as limits of quantification, linearity and repeatability are maintained. Important bottlenecks will be discussed, and possible solutions presented. Applications include the analysis of priority pollutants in water samples, mineral oil analysis, profiling of aroma compounds and fatty acid methyl ester analysis in foods.
11h15 - 11h40
Development and transfer of methods with pure CO2 as
chromatographic solvent: considerations and advice
Stéphane DUBANT (Reach Separation, France).
When developing new analytical or preparative chromatographic methodologies, environmental issues are (rightly so) becoming increasingly important. Minimizing, or even eliminating, the use of traditional organic solvents is a “simple” way to reduce the impact of chromatography. We have therefore developed a multi-step approach, from the most virtuous to the least virtuous: 1-> pure CO2, 2-> CO2 + EtOH, 3-> H2O + EtOH, 4-> H20 + MeCN.
This approach is even more important since in our laboratory, we mainly develop methods intended for so-called “preparative” chromatography, which traditionally consume a lot of solvents and therefore generates large amount of waste that need treatment.
For this presentation, we will focus on methodologies in pure CO2, or in so-called "supercritical" chromatography. This chromatographic mode has advantages in terms of performance and selectivity (scientific point of view), cost (financial point of view) and waste (financial and environmental point of view). When using pure CO2 as the mobile phase, the density of the fluid is the key parameter allowing to alter the chromatographic profiles (through the pressure and the temperature applied to the column). We will see the effects of this parameter during method development but also during the transfer from one instrument to another: from one analytical system to another analytical system, but also from analytical to preparative or even to production, through examples made in the laboratory.
11h40 - 12h05
Automated extraction and analysis of drugs from biological fluids by Coated Blade Spray - Mass Spectrometry (CBS - MS)
Shane STEVENS (Restek Corporation, USA).
Many methods of ambient ionization mass spectrometry have been proposed as alternatives to LC-MS/MS offering advantages in analysis speed/turnaround time, minimal sample preparation and simplicity in workflow. Most; however, are very susceptible to matrix effects resulting in reduced sensitivity, reproducibility, and instrument contamination.
Coated blade spray (CBS) is an easy-to-use solid phase microextraction (SPME)-based technology which combines sample extraction with direct ionization to a mass spectrometer. One coated blade is used to collect analytes of interest with subsequent introduction to MS by generating a Taylor cone.
The device comprises a small, thin, conductive substrate having a blade-shape with a flat surface terminating at a point. As a SPME device, the blade is coated at the terminal end with an extraction sorbent bed.
The CBS workflow reduces complexity, cost, and waste associated with other SPE methods. The SPME-based nature of CBS performs a selective extraction of desired analytes from the bulk sample matrix, resulting in very clean samples and improved instrument uptime. The Taylor cone is generated by applying a DC potential and adding a microliter-scale volume of solvent. CBS-MS eliminates the need for chromatographic separation, resulting in significant reductions in solvent consumption and analysis time.
In this work we demonstrate CBS-MS assays of drugs in biological fluids utilizing a modified laboratory pipetting robot to automate all functions associated with analyte collection as well as serving as the interface to a MS for analysis. We also present a modified pipettor capable of reorienting a blade from a vertical position required for extraction to a horizontal position required for MS analysis.
14h30 - 14h55
From the olfactory signature of hop cones to the aroma profile of beers: production of mini-beer by AMBERS (Automated Mini BEeR Screener) and analysis by SPME/GC/SCD
Damien STEYER (Twistaroma, France).
The choice of the hop variety is one of the key-factors that determines the final aroma of a beer. During beer production, a number of hop volatile compounds are chemically and biologically transformed to form new compounds that are not significantly present in the hop cones.
To understand the complexity of these modifications and their influence on the final beer aroma, 14 hop varieties were compared for their ability to provide aroma compounds to the beer. A new tool called AMBERS (Automated Mini BEeR Screener) was developed to mimic brewing (wort production) and fermentation in a small volume (5 mL). Here, this tool was used to compare the volatile profiles of 14 types of micro-beers. The hop cones and derived micro-beers from 14 hop varieties were analysed by SBSE-GC/ToF-MS.
14h55 - 15h20
Optimization of the extraction of the walnut oil volatiles using dynamic
headspace hyphenated to GC-MS: Adsorbent comparison.
Agnès CHARTIER (Institute of Organic and Analytical Chemistry
- University of Orleans, France).
Walnut oil is a highly valued vegetable oil due to its nutritional and medicinal properties. These properties are mainly attributed to the mono and polyunsaturated fatty acids, and to the presence of minor compounds such as antioxidants, amino acids and volatile compounds that are responsible for its organoleptic properties.
Unfortunately, this oil is not very stable and has a limited shelf life. Aging markers are correlated to the oxidation of the unsaturated fatty acids, which lead to the formation of various volatile compounds. These volatiles encompass five major chemical family such as acids, alcohols, aldehydes, ketones and furans.
The evolution over time of these different constituents induces a modification of the organoleptic properties of the walnut oil. Some of these compounds are difficult to trap before analysis. The use of experimental design made it possible to carry out the optimization parameters involved in a DHS-TDU-GC-MS method.
Different adsorbents such as Tenax, Carbopack, and mixed beds were tested and compared. This study has highlighted selective adsorbents with the aim of obtaining the profile of volatiles emanating from different nut oils from varied terroirs, and for some of them having an “organic” designation. From this study, markers of the aging walnut oil have been identified.
15h20 - 15h45
Automated and miniaturized method for the characterization of drugs in whole blood
Amélie LAGARDE (Laboratory of Scientific Police
- Saint Denis, France).
Forensic toxicology has to deal with many challenges. Among them, the quantity of sample available
to carry out a whole panel of analyzes and the increasingly low concentrations of new synthetic
substances. For example, in cases of chemical submissions, victims may have delayed filing a
complaint, thus extending the time between the facts and the samples and possibly leading to the
total elimination of the substances. The substances possibly ingested can be very different, so, a
certain quantity of blood and urine are necessary to be as exhaustive as possible. The new
psychoactive substances (NPS) are active with increasingly small quantities ingested, leading to very
low blood concentrations.
Scientific progress and innovation now make it possible to partially overcome these problems. SNPS’s Paris forensic laboratory acquired a new equipment : the CLAM-LCMSMS from Shimadzu. Based on their screening technique, we developed and validated a method for analyzing 22 substances belonging to the family of opiates, cocaine and amphetamines. The extraction is done on 50 μL of blood and the analysis is carried out in MRM mode. Lower limits of quantification obtained are between 2.5 and 5 ng/mL.
A second analysis method has been developed and includes around thirty NPS. This method is destined to evolve based on consumers' habits and new tendencies. The previous validated method required 2 mL of blood for the same research with a limit of quantification of 5 ng/mL. This method will be deployed in other laboratories of SNPS network.
16h15 - 16h40
Hydrogen as a carrier gas in GCMS, an upcoming
inevitability or an opportunity
David BENANOU (Scientific and Technical Expertise Department
- Veolia, France).
Helium is the second lightest and second most abundant element in the observable universe and is also the most widely used carrier gas for conventional gas chromatography and especially for gas chromatography-mass spectrometry. In recent years, the availability of helium has decreased and its cost has increased significantly. In 2021, the U.S. domestic helium reserve accounted for 30% of the world's helium and this reserve is expected to run out of helium by 2030.
This context of potential shortage and load increase, which until now seemed negligible, is prompting many chromatographers around the world to consider switching to the use of hydrogen.
We will present the chromatographic advantages inherent to the use of hydrogen in gas chromatography-mass spectrometry, its possible critical points and limitations and its "urban legends". An exhaustive economic assessment of the use of helium and hydrogen in different laboratories will be presented.
09h05 - 09h30
Blood Lipidomics and Metabolomics of Colorectal Cancer
Jeff FOCANT (ULiège University, Belgium).
Colorectal cancer (CRC) ranks as the third most frequently diagnosed cancer and the second leading cause of cancer-related deaths. The current endoscopic-based or stool-based diagnos-tic techniques are either highly invasive or lack sufficient sensitivity.
Thus, there is a need for less invasive and more sensitive screening approaches. We, therefore, conducted a study on 64 human serum samples representing three different groups (adenocarcinoma, adenoma, and control) using cutting-edge GC×GC–LR/HR-TOFMS techniques.
We analyzed samples with two different specifically tailored sample preparation approaches for lipidomics (fatty acids) (25 μL serum) and metabolomics (50 μL serum). In-depth chemometric screening with super-vised and unsupervised approaches and metabolic pathway analysis were applied to both datasets.
A Lipidomics study revealed that specific PUFA (ω-3) molecules are inversely asso-ciated with increased odds of CRC, while some PUFA (ω-6) analytes show a positive correla-tion. With the metabolomics approach, some proteogenic amino acids (Ala, Glu, Met, Thp, Tyr, Val), Myo-inositol, and 3- hydroxybutyric acid were found to be dysregulated in CRC.
This unique study provides a more comprehensive insight into molecular-level changes associated with CRC and allows for a comparison of the efficiency of two different analytical approaches for CRC screening using the same serum samples and single instrumentation.
09h30 - 09h55
Application of SBSE in analytics for ecotoxicological studies to meet requirements for next-generation product properties
Beate GRUBER (Analytical and Material Science at BASF SE Allemagne).
Information on acute or chronic effects of chemicals on aquatic organisms is an integral part of
environmental hazard assessment and risk assessment. In most regulatory studies, it is essential
to analytically verify an organism's exposure to a substance or its concentration in a test system.
Constantly increasing requirements for product registration as well as a higher complexity of next- generation products require analysis with sophisticated sample preparation. Commonly used liquid/liquid extraction for gas chromatographic analysis offers limited sensitivity due to dilution and consumes large amounts of hazardous solvents.
Therefore, Stir Bar Sorptive Extraction has been evaluated as an alternative to commonly used liquid/liquid extraction to improve sensitivity and reduce solvent consumption. The use of optimized method parameters resulted in a >100x improvement in sensitivity while improving safety and resource conservation by replacing solvents as extractants with a reusable adsorbent.
09h55 - 10h20
Evaluation of different head space techniques for flavor
trapping and profiling of meat alternative samples
Emilie USUREAU (Small Molecule Analysis at Center for Analytical Innovation
- DSM-Firmenich, Pays-Bas).
An important part of the acceptance of consumers in meat alternatives is their cooking experience. The smell released during baking or frying contributes to the overall perception of the product.
Often meat alternatives have a less appreciated smell during frying compared to meat burgers and knowing what kind of volatiles are responsible for that makes it possible to mitigate and work on improvements steps, increasing consumer acceptance.
Many headspace techniques are currently available to capture volatile organic compounds but there is so far no approach disclosed in literature or commercially available for fast profiling of volatile released during frying of the meat and meat alternative burgers.
In this lecture, the outcome of experiments where off-line air sampling for trapping the volatiles on adsorbent material, and on-line SPME sampling for volatiles profiling in fried meat and meat alternative burgers, followed by desorption and Gas Chromatography High Resolution Mass Spectrometry (GC-HRMS) analysis are presented.
10h20 - 10h45
Development, comparison and applications of Vaccum Assisted Sorbent Extraction (VASE): new perspectives on headspace analysis.
Jérôme COTTON (Vittel research laboratory, France).
A significant proportion of customer complaints reported each year for Nestlé Waters are related to
off-taste or off-odor in water. These organoleptic defects are caused by chemical molecules such as
pyridazines, halophenols or haloanisoles, which have very low human perception thresholds at ng/L
Analytical tools, in particular gas chromatography coupled to mass spectrometry, do not allow these detection limits to be reached without appropriate sample preparation. The VASE (Vacuum Assisted Sorbent Extraction) is a new technology that proposes, in a new way, to apply a vacuum in an analysis vial containing the sample in order to increase extraction yields to meet this type of analytical challenge.
Nestlé Vittel’s laboratory has developed a VASE-GC-MS method for analyzing liquid matrices and optimized extraction parameters such as contact and water management time, extraction and cooling temperature, sample/headspace ratio, and sorbent pen temperatures and desorption times. The VASE was also compared in terms of sensitivity, comprehensiveness, and usability with other techniques such as SBSE, Purge&Trap or liquid/liquid extraction for the analysis of undesirable organic compounds in water or beverages.
11h15 - 11h40
New Developments in Sorptive Extraction
Christophe DEVOS (RIC technologies, Belgium).
11h40 - 12h05
Artificial intelligence at the service of laboratories
Yvon GERVAISE, founder of E.S.C. ( ExpertScience- Consulting and Leader of the GT #LaboratoireduFutur at SECF ).
While A.I. at the service of the Laboratory is, surprisingly, a long story that has had its
winters and its summers, the recent boom in the use of artificial neural networks with
machine learning associated with deep-Learning raises the urgency of thinking about
the Laboratory of the future.
As early as 1965, mass spectrometry laboratories were using the first Expert system, called Dendral, which enabled chemists to automatically identify molecules from their simple mass spectrum.
In the 2020s, after the days of symbolic AI, connectionist AI based on artificial neural networks, with the power of its Deep-Learning algorithms, has accelerated A.I. applications in the laboratory world.
Google DeepMind's AlphaFold illustrates this success with its ability to automatically predict the 3D structure of proteins by reading their amino acid sequence. We will shed some light on the different types of A.I. (symbolic, connectionist, generative) and explain the concepts and mechanisms that govern them.
We will explain examples of AI applications in the laboratory (quantification of non- targeted pollutants, interpretation of results and exploitation of laboratory data (Big Data), automatic prediction of physico-chemical properties and toxicity (QSPR), automation in organic synthesis with the orchestrated laboratory, etc.).
We conclude by addressing these 3 questions:
- will AI lead to the replacement of the laboratory technician or, on the contrary, turn him or her into an augmented researcher?
- Will AI become the Laboratory Matrix of the future?
- Is A.I. an opportunity to be seized by laboratory players?
12h05 - 12h30
Determination of haloanisoles and halophenols in wine and in materials likely to carry contamination.
Rémy FULCHIC (Château Leoville las cases, France).
Wine is a matrix that can easily pick up all kinds of contaminants present in the materials with which it comes into
contact, or in the environments in which it is stored. Numerous molecules are known to be responsible for organoleptic
deviations, notably haloanisoles, which irreversibly cause musty odors and tastes at very low concentrations, just a few
This contamination can occur throughout the winemaking and ageing process, and even after bottling via the cork, which is sometimes responsible for the famous corky taste when it releases 2.4.6-Trichloroanisole into the bottle.
At Château Léoville Las Cases, a Gironde vineyard located in the Médoc in the Saint-Julien appellation, the in-house laboratory regularly analyzes new materials used in buildings, corks and mains water for haloanisoles (2.4.6- trichloroanisole, 22.214.171.124-tetrachloroaisole, 2.4.6-tribromoanisole and pentachloroanisole) and their corresponding halophenol precursors.
The latter do not present a direct olfactory risk, but they can be biomethylated into anisoles by certain molds, and therefore present a real risk if they are present, bearing in mind that vats or cellars are humid environments highly conducive to mold growth.
Their simultaneous determination is carried out by SBSE-GC-TOF, either directly for water and wine, or after extraction with ASE (Accelerated Solvent Extraction) for solid matrices. Phenols are acetylated during the SBSE phase, to improve extraction yield and thus the sensitivity of the method, as well as the chromatographic behavior of the compounds.
14h00 - 14h25
Presentation on Cognac and GCxGC-ToF MS
Xavier POITOU (Hennessy, France).
14h25 - 14h50
Automatic Robot for Preparation of Organic Samples:
ARPOS - TD-GC-MS/MS
Noemi SANTIAGO SANCHEZ (Labaqua, Espagne).
Pesticides, PAHs, PCBs and other contaminants coming from the human activity are transported and heavily presented in soil. Usually the extraction techniques for this type of compounds in solid samples are tedious and use large amounts of organic solvents in order to reach the quantification limits required by the regulations. This entails a high economic cost of material and time.
For this reason, there is a need to automate this process due to it is a completely manual extraction and currently there are a high number of soil samples that require to analyse organic compounds. Labaqua, in collaboration with GERSTEL, has developed a robot that get to automate this extraction, eliminating in this way most of the manual process for organic compounds analysis.
In addition this automation increases considerably the productivity of the analysis. ARPOS is able to add all the extraction reagents, as well as homogenizing, concentration and reconstitution of the sample in a fully automatic way, so that the sample is ready for direct analysis by TDU-GC-MS/MS. Furthermore, the chromatography analysis of the extracted sample by TDU-GC-MSMS allows the injection of a large volume of extract, achieving in this way a high sensibility and a low limit of detection of organic compounds.
By means of a validation of the process carried out in Labaqua, its repeatability and reproducibility has been demonstrated, achieving a high profitability of the process. Therefore, ARPOS robot is presented as a bet on the future for the automation of large-scale sample analysis.
14h50 - 15h15
Migration testing from plastic materials: analysis of trace levels in water by automated high-capacity sorptive extraction and GC-TOFMS
Catherine BRASSEUR (Certech, Belgium).
The migration of NIAS (Non-Intentionally Added Substances) into water is a recurring topic for the plastics industry. For example, antioxidants used in polyolefins to stabilize polymers against processing and thermal stress can generate fragments able to migrate into water. The identity of these migrating compounds was published in the 2000s and they have since been known as Arvin #1 through #10 substances. Other NIAS are also present in polymer materials. The importance of using sensitive and screening methods for migration testing analysis is reinforced by the emergence of recycled plastics.
Standard methods for monitoring the migration of small molecules into water by GC-MS analysis were initially based on large volume solvent extraction. Over the last decades, a lot of methods were developed based on sorptive extraction for water analysis and should be now better considered as a standard. From SPME coupled to GC-MS SIM, to high capacity sorptive extraction such as SBSE or Hisorb coupled to GC-TOFMS, allowing to improve sensitivity and screening. The methods can also be combined to high resolution mass spectrometry for identification of unknowns. It is important to convince and help industries to refer to this type of methods to reduce the amount of solvent used, increase automation and improve the characterization of migration tests.
15h45 - 16h10
Of mice and wine, an approach using SBSE-GC-MS
Céline FRANC (Institut des Sciences de la Vigne et du Vin (ISVV)
- Villenave d'Ornon, France).
Mousy off-flavour is a wine defect of modern concern among wine producers. Olfactory descriptors that characterise this particularly unpleasant defect include rodent urine (“dirty mouse cage”) and grilled foods such as popcorn, rice, crackers, and bread crust. These past years, wines spoiled by mousiness are increasing in frequency. This is often associated with rising pHs as well as certain oenological practices which significantly decrease the use of sulphur dioxide, thus favouring the development of microorganisms.
Three major molecules, 2-acetyl-1-pyrroline (APY), 2-acetyltetrahydropyridine (ATHP) and 2-ethyltetrahydropyridine (ETHP), were identified as responsible for mousiness in wines. However, to date, quantification data reported in the literature are limited due to analytical issues related to the nature of these compounds. Indeed, a simple and effective analytical method to determine trace levels of these three mousy N-heterocycles simultaneously was unavailable. To fill the gap and later understand the parameters influencing mousiness, a method using stir bar sorptive extraction (SBSE) followed by GC-MS analysis was developed.
After optimisation and performance evaluation, the quantification method was applied to analyse 6 control wines and 68 wines produced without added sulphites. This method was simultaneously adapted to microbiological media and used to monitor the production of the three mousy N-heterocycles based on the microorganism studied.
The SBSE-GC-MS analysis method proved its efficiency and paved the way to further studies aimed at understanding the conditions influencing the occurrence of mousiness in wine and the oenological parameters that modulate its expression.
16h10 - 16h35