Day 3 :
- Chromatography as a Separation Technique|Hyphenated Techniques|Role of Spectroscopy as Separation Techniques
Location: Melia Meeting 1+2
Chair
Hayrunnisa Nadaroglu
Ataturk University, Turkey
Co-Chair
Snezana Maletic
University of Novi Sad, Republic of Serbia
Session Introduction
Nico Anders
RWTH Aachen University, Germany
Title: Anion exchange chromatography in lignocellulosics analysis
Time : 09:00-09:20
Biography:
Nico Anders has been working in the field of analysis and renewables since 2009. He has obtained his PhD from the TU Braunschweig in the group of Prof. Dr. Vorlop in Technical Chemistry. Since 2013, he is working as Junior Research Group Leader in the Aachener Verfahrenstechnik at the RWTH Aachen University. His research interests are analysis of lignocellulosic biomass, green analytical chemistry, conversion of lignocellulosic biomass and chromatographic separation.
Abstract:
Complex liquid samples such as lignocellulose hydrolyzates which contain both polar and nonpolar analytes as well as monomeric and oligomeric compounds are hard to analyze and require usually more than one analytical separation mechanism. In contrast, the reversed-phase mechanism anion exchange chromatography (AEC) allows for a separation of divergent analytes based on the acidity of the resulting anion. Thus, the aim of this study was to investigate the potential of AEC to analyze lignocellulosic biomass hydrolyzates completely using one analytical separation mechanism, as lignocellulose hydrolyzates contain a lot of potential anions with a variation in polarity and size. Therefore, the potential of generating anions from aldehydes, alcohols and phenols was investigated using an alkaline eluent. Additionally, the concentrations of aldehydes, alcohols and phenols derived from lignins were measured simultaneously with monosaccharides, oligosaccharides and uronic acids derived from cellulose, hemicellulose and pectin. Thus, parameters such as column temperature, eluent composition and chromatographic run time were examined. The final chromatographic methods was set to a column temperature of 40°C, an eluent flow of 1 mL/min and an eluent consisting of sodium acetate and sodium hydroxide as well as ultrapure water. This method allows for a complete characterization of lignocellulose hydrolyzates with limit of detections in the range of 0.014 mg/L for 2,6-dimethoxyphenol and 21.9 mg/L for 4-methoxybenzyl alcohol. Finally, this method was used to characterize 17 hydrolyzates from lignocellulosic biomass simultaneously for cellulose, hemicellulose, lignin and pectin derived degradation products.
Santerre Cyrille
ISIPCA, France
Title: SFC-MS as a tool for flower absolute composition analysis
Time : 09:20-09:40
Biography:
Santerre Cyrille has completed his Analytical Chemistry Engineering degree and Cosmetology Master’s degree from the Conservatoire National des Arts et Métiers (CNAM) and University of Chatenay Malabry Paris XI (France). He is Lab Manager and Member of a research team focusing on supercritical fluid chromatography (SFC) at ISIPCA and mass spectrometry at ICSN-CNRS (Gif sur Yvette-France). He has done several poster and oral communications about this subject.
Abstract:
Cosmetics and perfumes industries are very late for using supercritical fluids compared with pharmaceutical industry, for example. The protection and the environmental respect became a major subject of concern nowadays. Furthermore, with the increasing interest of the professionals for the natural raw materials with the complexity of their compositions, supercritical fluid chromatography (SFC) seems to be imperative to us as green chemistry tool. Hyphenation is very easy with mass spectrometry (MS) using various ionization sources (ESI, APPI, APCI). European legislation regulates 26 fragrance allergens at the moment with an exceeding stipulated cut-off level. In the near future, 32 other allergens will be newly added. In order to decrease analysis time and improve specificity, SFC coupled with diode array detector (DAD) has been tested. First step consists in stationary phase screening (Si, Hypercarb) using isocratic mode (95% CO2/5% Ethanol as modifier). Hypercarb was selected as the most retentive stationary phase for this type of analysis. Then chromatographic resolution has been improved by checking the effect of column temperature, CO2 back pressure, nature of modifier (methanol, ethanol, isopropanol) and gradient mode. First results indicate that at least 20 compounds can be efficiently separated in less than 12 minutes. Subsequently two methods have been developed: One, SFC-UV to quantify major components in essential oils (e.g. Eucalyptus and clove); and as second one based on SFC-MS to study flower absolutes composition. Indeed volatile compounds are already well known and described in the literature using gas chromatography (GC) coupled with MS but unvolatile fraction remains incompletely described.
Aleksandra Tubic
University of Novi Sad, Republic of Serbia
Title: Evaluation of solid-phase chromatography as a method for natural organic matter characterization
Time : 09:40-10:00
Biography:
Aleksandra Tubić is Assistant Professor at the Chair of Chemical Technology and Environmental Protection, University of Novi Sad, Faculty of Sciences. She completed PhD in Chemistry from University of Novi Sad Faculty of Sciences, Department of Chemistry, Biochemistry and Environmental Protection in 2010, and BSc in Chemistry from University of Novi Sad, Faculty of Sciences, Department of Chemistry in 2003. Her research interests include: Environmental protection, chemical technology, ambient and drinking water treatment. She also worked as Quality Manager of the Laboratory for the Analysis of Environmental Samples, accredited according to ISO 17025 protocols.
Abstract:
Natural organic matter (NOM) is a ubiquitous constituent of surface and ground waters worldwide. It is well known that various inorganic and organic species interact with NOM in water matrices and these interactions can have a great impact on their mobility and behaviour, as well as the expression of toxic effects. NOM, if present in source water, can have a great negative influence on drinking water treatment, through disinfection by-products formation, requirements for increasing amounts of chemicals during treatment and deterioration of water quality in the distribution system. NOM is generally site specific, and the approximation of its influence is rather difficult to make based on simulations or previous knowledge gained from other sites. Thus, the characterization of NOM at the specific site of interest is of great help in understanding the processes in ambient waters as well as during treatment. There are various possibilities for NOM characterization, including solid-phase fractionation using XAD resins. The aim of this paper is to evaluate solid-phase chromatography as a method for NOM fractionation through a review of the results obtained by fractionation of 3 ground waters and 1 surface water, which was repeated 2-5 times for each water. The method is based on fractionation of the dissolved organic matter into four fractions: The humic acid, fulvic acid, hydrophilic acid and hydrophilic non-acid fractions; and measuring the DOC value after the fractionation. The fractionation method showed great recovery values for DOC (> 98%) compared to the DOC measured in the bulk water samples. The relative standard deviations (RSD) of the DOC values measured in the same fraction in each water sample range from 0.71-9.9%. An additional benefit of the method is that it gives satisfactory recovery results even after resin regeneration, with recoveries of 99%, 97% and 88%, for the resins used before and after the first and second regeneration cycles.
Jong-Sup Jeon
Gyeonggi Province Institute of Health and Environment, Republic of Korea
Title: Robustness test of the chromatographic method for the quantification of chlorogenic acid in coffee brew
Time : 10:00-10:20
Biography:
Jong Sup Jeon has completed his PhD from College of Veterinary Medicine, Konkuk University (Republic of Korea). He is DVM and Research Scientist for the Public Health in Gyeonggi Province Institute of Health and Environment, Republic of Korea. He has published variable domestic and international papers (subjects: Antibiotics, hazardous materials in food and functional ingredients in cosmetics) in reputed journals and served as a co-worker in various field.
Abstract:
Coffee has been for decades the most commercialized and widely consumed beverage in the world. Coffee beans contain a large variety of substances, which in many cases are biologically active, such as caffeine and chlorogenic acids. Chlorogenic acids are water-soluble phenolic components of coffee and other plants formed by the esterification of certain trans cinnamic acids, such as caffeic, ferulic and p-coumaric acids,with(-)-quinic acid. The main subgroups of chlorogenic acid isomers in coffee are the caffeoylquinic acids, feruloylquinic acids, dicaffeoylquinic acids and, in smaller amounts, p-coumaroylquinic acids. In this study, we carried out robustness evaluation for method validation of quantification of chlorogenic acid in coffee brew, using Youden’s test. It was possible to determine the effect of each analytical parameter in the final analysis results. Seven analytical factors were selected and small variations were induced in the nominal values of the method. Then, 8 runs were performed aiming to determine the influence of each parameter in the final result. The 7 factors are: Concentration of KH2PO4 in mobile phase, column temperarture, flow rate, wavelength of detector, column supplier and initial mobile phase composition. Standard deviation of the differences Di (SDi) was calculated and that value was not larger than the standard deviation of the method carried out under within-laboratory reproducibility conditions. The experimental t values resulted below the 2-tailed t-critical value for all 7 factors. The tested procedure proved to be fairly robust since minor fluctuations in the operative parameter that can occur during the routine application of the method do not significantly affect its performance characteristics.
Hayrunnisa Nadaroglu
Ataturk University, Turkey
Title: Purification, characterization of lipase enzyme from lactobacillus brevis and immobilization onto magnetic florosil NPs
Time : 10:20-10:40
Biography:
Hayrunnisa Nadaroglu has completed her PhD and MSc from Ataturk University, Graduate Institute of Sciences, Department of Biohemistry (Erzurum, Turkey) in Bioorganic Reactions using Purified Carbonic Anhydrase Isoenzymes. She is Scientific Expert in process development on bioremediation of waste water, some nano-biotechnological applications onto environmental pollution and some industrial enzyme applications onto food technology (clarification of fruit juice, hydrolyzation of phytate ext.). She has published more than 120 papers in the field of biochemistry, food technology and nano-biotechnology in journals and conferences.
Abstract:
Lipases (E.C.3.1.1.3; triglycerol acylhydrolases) are enzymes catalyzing reversible hydrolysis of animal and vegetable oils under normal conditions. Besides, they also catalyze reactions such as esterification and transesterification. In this study, a new lipase enzyme was isolated from Lactobacillus brevis and immobilized onto modified florisil with iron NPs and the usability of free and immobilized lipases as a detergent additive material was investigated. Lipase enzyme was purified using ammonium sulphate precipitation, DEAE-Sephadex ion-exchange chromatography and sephacry S200 gel filtration chromatography techniques. Its molecular mass was determined to be 57 kDa by SDS-PAGE and gel filtration chromatography. Purified lipase was immobilized onto magnetic florisil NPs and determined immobilization conditions. Also immobilized lipase characterization was done using SEM, FTIR and XRD techniques. Immobilized lipase showed good thermo-stability and retained its activity at 80%, than free lipase enzyme at 60°C. The free and immobilized lipase enzymes were most stable in the alkaline pH. Also, immobilized lipase had more stability towards metal ions than free lipase enzyme. Washing performances of some detergents formulation were done and maximum percentage of olive oil was removed by the immobilized lipase than commercial detergents. The study on oil stain removal from cotton cloth indicated that oil removal was superior in the presence of immobilized lipase and immobilized lipase with detergent than the detergent alone.
Miguel Angel Lopez Zavala
Tecnologico de Monterrey, Mexico
Title: Ultrahigh pressure liquid chromatography as a separation technique for the simultaneous extraction and determination of four different groups of pharmaceuticals in compost
Time : 10:55-11:15
Biography:
Miguel Ángel López Zavala has completed his PhD and Post-doctoral studies at the Hokkaido University, Japan, in the field of Urban and Environmental Engineering. He is Professor at the Tecnológico de Monterrey, Mexico. He is Member and Evaluator of the National Research System of the National Council of Science and Technology. He was Researcher of the Japan Science and Technology Agency at Hokkaido University, Japan. He is author of more than 55 scientific papers published in international journals and proceedings of international conferences and congresses. He is active Member of the International Water Association. He is Treasurer of la IWA-Mexico from 2010 to 2011.
Abstract:
Extracting and separating pharmaceuticals from complex environmental matrices such as compost is particularly challenging because of the complex nature of the samples, the different chemical characteristics of the compounds and the low detection limits required. In this study, ultrahigh pressure liquid chromatography (UHPLC) was used as a separation technique for the simultaneous extraction and determination of 4 different groups of pharmaceuticals in compost obtained from the thermophilic aerobic treatment of placenta. The pharmaceuticals were 2 non-steroidal anti-inflammatory drugs, ketorolac and naproxen, usually administered to humans; two fluoroquinolones- ofloxacin and ciprofloxacin (which are among the most commonly prescribed class of antibiotics in Mexico); 2 anti-cancer (antineoplastic or cytotoxic) chemotherapy drugs- ifosfamide and cyclophosphamide; and 2 b-blockers- atenolol and propranolol, also called b-adrenergic blocking agents, which treat a variety of conditions, such as high blood pressure, glaucoma and migraines. The pharmaceuticals of each group were selected because they are commonly used in Mexico and environmental and health impacts have been reported. The clustering was based on the use of the drug and not on the similarity of the structure. The use of UHPLC allowed better detection and quantification of all pharmaceuticals; furthermore, shorter analysis time was required and lower costs were involved. Recovery values of the ultrasonic extraction for all compounds were on the range of 87% and 113%. The limits of detection and quantification for the eight pharmaceuticals were on the order of 0.66 ng g−1 and 2 ng g−1 respectively for all the pharmaceuticals analyzed. These values are lower than those values reported in the literature.
Aneta Iwona Wojnicz
Autonomous University of Madrid, Spain
Title: Simple HPLC–MS/MS method for simultaneous determination of aripiprazole and dehydroaripiprazole in human plasma using microelution solid phase extraction
Time : 11:15-11:35
Biography:
Wojnicz A is doing her PhD from Autonomous University of Madrid, Spain. She is Bioanalyst Scientist, working at Analytical and Pharmacokinetic Unit of Clinical Pharmacology Service of ‘Hospital Universitario de la Princesa’ (Madrid–Spain). The unit develops and validates analytical methods to determine drug levels in biological fluids by LC-MS/MS. She has spent a 3-months period at Department of Pharmacy & Pharmaceutical Science and Biochemistry of University of California San Diego (UCSD) to improve her knowledge with experts in mass spectrometry. She has published more than 7 papers in reputed journals.
Abstract:
A selective and accurate high pressure liquid chromatography-tandem mass spectrometry (HPLC–MS/MS) method has been developed and validated for simultaneous monitoring of Aripiprazole and its active metabolite Dehydroaripiprazole in human plasma using aripiprazole-d8 as the internal standard (IS). The analytes and IS were extracted from 200 µL of human plasma by solid-phase extraction using Oasis PRiME HLB 96-well µElution Plate, 3 mg sorbent per well (Waters, Madrid, Spain). Separations were carried out at 25°C in an ACE C18-PFP column (4.6 mm×100 mm and 3-μm particle size (SYMTA, Madrid, Spain) protected by a 0.2-μm on-line filter. The mobile phase consisted of a combination of 0.2% formic acid and 0.3% ammonia in MilliQ water pH=4.0 (solution A) and ACN (solution B) (65:35, v/v). The chromatogram was run under gradient conditions at a flow rate of 0.6 mL/min. Run time was 5 min followed by a re-equilibration time of 3 min, to give a total run time of 8 min. The volume injected into the chromatographic system was 5 μL. The analytes were detected using the mode multiple reaction monitoring in the positive ionization mode. The linearity of the method was established in the concentration range 0.15–110 ng/mL and 0.35–100 ng/mL for Aripiprazole and Dehydroaripiprazole, respectively. We validated the analytical method according to the recommendations of regulatory agencies through tests of precision, accuracy, recovery, matrix effect, stability, sensitivity and selectivity. The method was applied to 6 different bioequivalence studies of 10 mg aripiprazole formulation in 40 healthy Caucasian subjects.
Martin Topiar
University of Chemistry and Technology Prague, Czech Republic
Title: Supercritical CO 2 in its multiple roles: solvent for fractionation of complex mixtures and blowing agent in polymer foaming
Time : 11:35-11:55
Biography:
Martin Topiar has completed his Master of Science in Synthesis and Production of Drugs from University of Chemistry and Technology Prague. Now he is a PhD student at Institute of Chemical Process Fundamentals of the CAS, v.v.i. focusing on the SFE from plants with particular interest in a study of several types of fractionation techniques. He has published 3 papers in reputed journals and presented his work in many international conferences dealing with supercritical fluids.
Abstract:
Carbon dioxide in its supercritical state (sc-CO2) can find application in formation of microcellular polystyrene foams (PSF) with improved thermal properties in comparison with PSF on the market. A most important application of sc-CO2 on industrial scale is supercritical fluid extraction (SFE) from plants. Both processes have been studied in our laboratory. Although the SFE is more selective than common extraction methods, CO2 extracts still contain a mixture of chemical compounds. If a high concentrated isolate is required, suitable fractionation methods have to be involved into the process. Supercritical adsorption, a novel promising method combining SFE from plants with selective adsorption of extracted compounds, could be a solution when high purity isolates free of any traces of organic solvents are required. We used this method for fractionation of turmeric (Curcuma longa L.) isolate obtained by SFE. The major compounds analyzed by GC were turmerone (22.7 wt.%), ar-turmerone (13.2 wt.%) and curlone (13.8 wt%). Turmerones belong to widely studied substances with potential use in the treatment of neurodegenerative diseases. The efficiency of fractionation was studied in terms of adsorption conditions, adsorbent type and sorbent-to-feed ratio. The concentration of turmerones in isolate increased from the initial 49.7 wt.% up to 93.8 wt% with using a particular type of silica gel. Moreover, the concentration of β-sesquiphellandrene, a compound with a potential anticancer activity, in volatile fraction obtained from supercritical adsorption process was almost 16 times higher than in the initial sample. These are promising results for a more detailed subsequent research on this method.
Lukasz Antoni Sterczewski
Princeton University, USA
Title: Resolving terahertz spectral mixtures using the blind source separation approach – a method to study the dehydration kinetics
Time : 11:55-12:15
Biography:
Lukasz A Sterczewski received MSc degree in Electronics from Wroclaw University of Technology, Poland in 2014, working on terahertz time-domain spectroscopy. He is currently working towards a PhD degree from WrUT. In September 2015 he joined PULSE (Princeton University Laser Sensing Laboratory) as a Visiting Student Research Collaborator. His research is focused on development on new spectroscopic techniques and advanced signal processing tools.
Abstract:
Terahertz spectroscopy has gained popularity as a promising non-invasive investigation tool in recent decades. In studies of solid-state pharmaceuticals, its usefulness is enhanced, since it enables one to distinguish different polymorphic and pseudo-polymorphic forms. Terahertz absorption spectra are additive, meaning that the resulting spectrum of two or more compounds in the sample is an algebraic sum, in the linear region of the Beer’s law. For this reason, the evolving-in-time linear mixture of unknown pure components can be resolved using the blind source separation approach, where both spectral sources and their concentrations are to be estimated. Such an evolution occurs naturally in temperature-induced (pseudo)polymorphic transitions, i.e. dehydration, where the terahertz spectrum is the hydrate and dehydrates’s response with time-varying proportions. Conventionally, to estimate the kinetics of a reaction–an important physical-chemical parameter–the area of the unique spectral peak is integrated and normalized to evaluate the abundance, but a difficulty of strongly overlapping peaks unable its application. To overcome this problem, we show, that by employing the blind source separation procedure, we can resolve a complex, multi-compound spectral mixture with significant cross-bands, where the peak area method failed. We evaluate the performance of our approach in studies on dehydration of a well-known polycrystalline hydrate–α-D-glucose monohydrate. Seeing that the polymorphic transitions can completely change the properties of a pharmaceutical, our approach can find application in a drug development process, where their careful characterization is of utmost importance.
Yongtaek Lee
Chungnam National University, Korea
Title: Numerical analysis on purification of methane through the hollow fiber membrane system
Time : 12:15-12:35
Biography:
Yongtaek Lee has completed his PhD from the state univrsity of New York at Buffalo. He is currently the head of the department of Chemical Engineering, Chungnam National University. He has published more than 100 papers in reputed journals .
Abstract:
The polysulfone membrane was found to be applicable to the selective removal of CO2 from a biogas, resulting in preparation of the relatively concentrated methane. The purified methane might be utilized as a vehicle fuel if the methane concentration is kept higher than the 95 mole %. Generally speaking, the concentration of methane in the biogas is around 20 mole %, which should be upgraded using a cascade operationg system. Even if there are several conventional technologies, the membrane system is known to be not only a new technology, but also an economial method. The cascade membrane system may be constituted of either two membrane modules or three membrane modules, depending on the input biogas conditions and the degree of purification of methane. In this study, a theoritical analysis on the cascade membrane system is proposed and a proper numerical technique is applied to solve the obtained differential equations where the permeate flow rate and the concentration of the each component will be described in terms of the permeability and the partial pressure difference across the membrane. As the operating conditions are changed, the obtained flow rate and the concentration of methane as a product are found to be severely affected. As a design tool, this numerial analysis is strongly recommended since the proper operating condition will be suggested together with the suitable recycle flow rate for the target goal such as the recovery ratio and the purity of methane in the final product. The cascade schemes and the analysis results will be presented.
Mariana Campos Assuncao
PSL Research University, France
Title: Purification of phosphoric acid by liquid-liquid
Time : 12:35-12:55
Biography:
Mariana Campos Assuncao is currently a PhD student at Chimie ParisTech conducting a research project on the purification of phosporic acid in collaboration with Prayon. Before this experience, she has obtained a Master’s degree in Nuclear Energy at Chimie ParisTech and an Engineering degree in Chemical Engineering at INSA de Rouen.
Abstract:
Phosphoric acid is a weak oxyacid with many industrial applications depending on its degree of purification, including surface treatments or fertilizer production (merchant grade Phosphoric Acid, MPA), salt production for animal feeding or acidification of food and drinks for human consumption (Food grade Phosphoric Acid, FPA), and pharmaceutical industry (Pharmaceutical grade Phosphoric Acid, PPA). In order to produce these different grades, various purification technologies can be implemented including hydrometallurgical processes as it is the case in the Prayon’s process based on the selective liquid-liquid extraction of phosphoric acid. The current solvent used by Prayon’s process for purifying phosphoric acid is a mixture of 90 wt% di-iso-propylether (DIPE) and 10 wt% tri-n-butylphosphate (TBP). There is very few information about the physicochemistry involved in the purification of wet phosphoric acid by liquid-liquid extraction because wet phosphoric acid is a very complex medium (highly concentrated medium which can reach 14 M, high complexing power, only few data on metal speciation in phosphoric acid, etc.). Furthermore, liquid-liquid extraction of phosphoric acid involves very complex phenomena such as phase splitting and third phase formation which can be explained by the presence of supramolecular species in solution as well as coextraction of large amount of water. In the present paper, the physicochemistry involved in phosphoric acid and water extraction by new extraction solvents is presented with a focus on decrypting the role of the supramolecular organization. Inedit data on the transitions from triphasic systems towards biphasic systems are presented and a focus has been placed on the description of forces playing a role in these transitions.