Alina Vasilescu is an analytical chemist with expertise in the development and validation of novel analytical methods. Her experience encompasses both academic research and analytical research in the pharmaceutical industry. She currently works as a Senior Researcher at the International Centre of Biodynamics in Bucharest, where she coordinates several research projects, focussing on the development of (bio)sensors for practical applications. Study of protein aggregation is a primary research area and she collaborates with other groups for including nanomaterials in the development of novel sensors.

Statement of the Problem: Protein instability due to misfolding and aggregation is of big concern for protein-based therapeutics because it impacts the bioavailability and immunogenicity of such drugs. Simple and cost-effective analytical methods, indicating the presence or absence of protein aggregates, are consequently of high importance. Methodology & Theoretical Orientation: Porous reduced graphene oxide (prGO) coated electrodes have been investigated for the early and sensitive identification of protein aggregation. The detection principle lies in following the change in the oxidative current of the proteins. The sensor architectures studied included glassy carbon electrodes with drop cast prGO and disposable, screen printed electrodes modified with prGO using the layer-by-layer deposition technique. The studies focused on the protein lysozyme and the pharmaceutical polypeptide calcitonin having the ability to form aggregates in different conditions of pH and temperature. Parallel experiments were performed by fluorescence with thioflavin T, size exclusion chromatography, and Atomic Force Microscopy Imaging. Findings: Comparing the oxidation peak of lysozyme by differential pulse voltammetry for different electrode architectures allowed validating the higher sensitivity of the prGO-coated interfaces versus bare ones. Moreover, the modified electrodes allowed detecting in a fast and reliable manner the changes in the protein structure occurring at pH 2 and pH 7.4, as per processes leading to the formation of amyloid and amorphous aggregates, respectively (Fig.1). Screen printed electrodes modified with prGO enabled to differentiate between the amyloid-type aggregation of calcitonin (2 mg mL-1) in citrate buffer and no amyloid formation in acetate buffer. These electrodes were also applied to the analysis of a pharmaceutical drug product of low potency, Miacalcic (8.3 µg mL-1 calcitonin), where no aggregation was observed. Conclusion & Significance: Electrochemical sensors coated with prGO coated enable to capture different aggregation behaviors of proteins and peptides and represent a complementary tool for biopharmaceutical

Nives Galic was born in Zagreb, Croatia. She received a Ph.D. degree in Analytical Chemistry (1999). In 2016 she was elected to the position of Full Professor. During the period 2011-2017 she was the Head of the Division of Analytical Chemistry. She is a leader of the project funded by Croatian Science Foundation (IP-2014-09-4841).

Aroylhydrazones can act as neutral, monoanionic or dianionic ONO tridentate ligands [1]. The coordination abilities of aromatic hydrazones derived from nicotinic acid hydrazide and differently substituted 2-hydroxybenzaldehydes towards Fe3+ and Ga3+ will be discussed. Different techniques, like UV-Vis, vibrational spectroscopy, and mass spectrometry were used for structural investigation of the complexes in solution and in the solid state. In this work, the ESI MS and MS/MS spectra, including fragmentation pathways of Fe(III) and Ga(III) complexes with aroylhydrazones are presented.

Caijun Zhang has built up his expertise in analytical chemistry support to pharmaceutical industrial for more than 20 years. He recently brings his knowledge to Terramera Inc, an agriculture biotech company and support the product development from prove- concept stage to the commercialization of finished product.

Statement of the Problem: Neem oil contains more than 100 biologically active compounds (limonoids). Azadirachtin as a marker for neem oil is well adopted in the biopesticide industry. However, this marker is insufficient to distinguish whether the neem oil is contaminated with other botanical oil because azadirachtin (A and B) represent only the polar and hydrophilic limonoids, very small portion of bioactive ingredients in neem oil. Methodology & Theoretical Orientation: Terramera develop a finger-print HPLC method which can detect most bioactive ingredients in neem oil. According to the HPLC profile (figure 1), a new marker system was developed in Terramera. This new marker system includes a wider spectrum of active ingredients and can protect from the adulterated neem oil because any foreign peaks can be easily detected under other limonoids region Conclusion & Significance: The new marker system for neem oil provides a much better representation of the quality of neem oil than measuring azadirachtin alone

Lounès Haroune is manager of the bioanalytical platform of the pharmacology institute of Sherbrooke University. After 5 years as analytical development manager in an analytical laboratory and graduated in analytical chemistry, he is working on the development of analytical methodology and sample preparations for the detection and quantification of biomolecules in biological matrices. He also works on the metabolomics and peptidomics methodologies in complex matrices. He also focuses on the development and implementation of new analytical workflow for molecular characterizations (biocatalysis, physico-chemistry reaction, etc).

While there has been a growing interest in understanding the pharmacological and physiological properties of cannabinoids in the last decades, analytical methodologies including sample preparations, remain one of the most challenging topics for their quantification in biological matrices. Moreover, the low sample weight or volume coupled to the complexity of biological samples (i.e. whole blood, plasma, etc.) could overwhelm the analyst expectations. In this study, we explored different possibilities to quantify a mixture of 8 natural phytocannabinoids present in biological samples (cannabinol, cannabigerolic acid, cannabinochromene, cannabigerol, cannbidiolic acid, tetrahydrocanninol, tetratracannabinolic acid and cannabidiol). The evaluation was carried-out using plasma and whole blood samples using different usual extraction protocols (solid phase extraction, liquid-liquid extraction, protein precipitation and blood spot sampling). Stability of tested molecules was also evaluated in several matrices (plasma, serum, ex vivo and pharmacokinetic profiles). The results showed a moderate matrix effect resulting by signal suppression (≤ 30 %) and acceptable recoveries (≥ 60 %) for most of the different tested extractions and matrices, except for whole blood when using acetonitrile for protein precipitation, which appears to be the less efficient approach for cannabinoid extraction, with a recovery lower than ≤ 40 %. The applicability of tested methodologies was also applied for the determination of pharmacokinetic profiles and showed that dried blood spot sampling (DBS) could become an interesting alternative for in vivo studies. DBS is a rapid, acute and minimally invasive technic based on a single blood drop (10 µL – 25 µL) that reduces handling and quantity of blood to be sampled, which consequently also reduces the cost of analysis. According to these aspects, DBS could become a reference methodology for in vivo pharmacokinetic experiments.

During the last years, the use of sewage sludge as soil amendments for crops has gain in interest. It limits the addition of fertilizers, it reduces the land occupation and it promotes waste valorization. However, the presence of organic compounds such as pharmaceuticals and pesticides (PhPCs) may cause the transfer of these contaminants to the soil, and to the groundwaters. In this work, an analytical method for the simultaneous extraction of 70 compounds from complex matrices was developed and validated using an experimental design plan. Firstly, the targeted compounds were extracted by an optimized QuEChERS approach using ethyl acetate/water (4/1, v/v) as the extraction solvent and a dispersive solid phase extraction (dSPE) with C18/Na2SO4. Then, the analytes were quantified using a LC-MS/MS methodology. The method was validated in terms of accuracy and precision. The results obtained showed a strong matrix effect resulting by signal suppression. Therefore, the solvent matched calibration approach was chosen for the quantification. The applicability of the method for different matrices was demonstrated through the analysis of biosolids samples from Magog (Qc) waste water treatment plant, sediment samples from Massawippi (Qc) and Montjoie (Qc) Lakes and benthic organisms (cheronomedae and oligochaete). The recoveries were higher than 50 % for most of the targeted compounds in all tested matrices. 10 compounds (acetaminophen, caffeine, carbendazim, naproxen, carbamazepine, atrazine, ibuprofen, fenofibrate and ketoprofen metolachlor) were quantified in the samples at concentration ranging from ≈ 5 ng.g-1 to ≈ 40 ng.g-1

Sabrina Saibi is PhD student at université de Sherbrooke. She is working on the development of analytical methodology and samples preparations for the detection and quantification of contaminants of emerging concern (CEC) in complexes matrices (biological, environmental). She also works on the monitoring and occurrence of the CEC in the environment. She also focuses on the development and implementation of new biotechnologies process for the removal of organic contaminants (fungi, bacteria, enzyme catalysis).

Understanding of size heterogeneity in biotherapeutic proteins is essential since it is one of the Critical Quality Attributes (CQA) due to its impact on safety and efficacy. The size heterogeneity covers the product related species/impurities that includes fragments, monomers, and aggregates. Size exclusion chromatography (SEC) has been widely used to separate aggregates, monomer, and fragments of monoclonal antibodies (mAbs) that might form during manufacturing, storage and shipping. The progress in biologics pipeline and the urgency to reach first-in-human (FIH) has provided an opportunity to develop a generic method that can serve as a platform method for monoclonal antibodies. This study describes platform SEC method development for monoclonal antibodies using commercially available highperformance liquid chromatography (HPLC) and ultra-performance liquid chromatography (UPLC) SEC columns. For this purpose, several antibodies covering a broad range of isoelectric points (pI) and hydrophobicity were analyzed. Initial comparison was performed using six different mAbs to understand the impact of mobile phase and organics on separation of aggregates and fragments.

Itaru Yazawa has co-founded a company, Imtakt Corporation in 1999 at Kyoto Japan to focus on separation technology providing his own designed and manufactured HPLC columns to the global market. He used to work for Shimadzu Corporation (instrument development) and YMC Co.Ltd (column development), and then he wanted to supply his own unique column products such as RP+AX+CX muli-mode ODS column "Scherzo C18 Family" and 2um Non-porous ODS column "Presto FF-C18" etc. which are based on his own technical idea. Now he will introduce a next-generation novel amino acid analysis column for LC-MS "Intrada Amino Acid" for this conference

There are four established methods for analyzing amino acids: pre-labeled, post-labeled, ion-pairing reversed phase and normal-phase, but each of these methods has disadvantages. The pre-labeled method has problems with derivitization efficiency and cost, while the post-labeled method is usually not compatible with LC-MS due to non-volatile mobile phases. The ion-pairing reversed-phase method has difficulty separating polar amino acids; on the other hand, the normal-phase mode has problems separating all the compounds, especially the Leu and Ile isomers. We have developed a novel amino acid separation column for LC-MS (MS) which can separate all 20 amino acids in protein using a mixed-mode stationary phase structure. We have also estimated separation and detection characteristics using LC-MS instruments. We found two methods to successfully analyze the complete array of 20 amino acids: 1) high throughput separation with Leu/Ile separation in 5min, and 2) simple gradient separation. We also found that detection can occur not only in single MS mode but also in triple MS mode. In addition, no derivitization is required, and a standard LC-MS (MS) system is sufficient for the analysis. This novel HPLC method will be a powerful tool for amino acid LC-MS(MS) analysis in many different biochemistry applications.