4^th World Congress on Chromatography August 07-09, 2017 Rome, Italy

Biography:

Employed at Polymer Institute, Slovak Academy of Sciences in Bratislava. Served as elected member of the Presidium of the Slovak Academy of Sciences, President of the Slovak Chemical Society, Chairman of the Czecho-Slovak and Slovak National Committee of Chemistry for IUPAC. Corresponding member of the Central European Academy of Sciences and member of the Learned Society of the Slovak Academy of Sciences. Author or co-author of two monographs and 300+ scientific papers in extenso published in refereed periodicals, proceedings and chapters of books, as well as 60+ patents (four of them were licensed) - cited more than 3,000x. Presented over 120 invited plenary, key and main lectures, as well as over 900 regular lectures and poster contributions on symposia and conferences, as well as during lecturing tours to over fourty countries. Elected „Slovak scientist of the year 1999“ and „Slovak innovator of the year 2001“ 

Abstract:

The retention properties of high-performance liquid chromatographic (HPLC) columns belong to their most important characteristics. To test and compare the HPLC column retentivities, diff erent series of low molar mass probes bearing various functional groups are eluted in the appropriate mobile phase and the diff erences in their retention volumes, VR are evaluated. The nature of functional groups, overall polarity, basicity/acidity, etc., of the test substances are considered, as well as the size and shape of their molecules. However, the conclusions drawn from dissimilar sets of test substances may diff er remarkably. It seems that more unambiguous data can be obtained with series of chemical homologues as test probes, for example esters, which bear the same active group but their molecular size diff ers. Th e alternative approach considers application of oligomers, in which the active groups are repeated. However, the end-groups in oligomers can bias the results. Th e solution may bring polymeric test probes, in which the end-groups play negligible role. Polymer probes are available in diff erent both polarities and molecular sizes. Elution of series of polymers with distinct molar masses and sizes of their molecules in solution enables to independently evaluate both polar, adsorption and non-polar, partition properties of typical HPLC column packings such as bare and alkyl bonded silica gels. Plots of retention volume versus hydrodynamic volume of polymers Vh are constructed for the macromolecular probes with diff erent polarities. Such representation enables to compensate dependence of size of macromolecules in solution on their chemical composition and on eluent nature. The interactivity of columns is assessed from the courses of such dependences, which are denoted as universal calibrations. In the lecture, we will explain the principle of assessment of HPLC columns retentivity with the help of polymer probes and present typical examples of the procedure.

Publications

1. Berek D (2003) Evaluation of high-performance liquid chromatography columns retentivity using macromolecular probes, III partition properties of C18 phases traced by polymers. J Chromatography 1020:219–228.

2. Berek D (2002) Evaluation of high-performance liquid chromatography columns retentivity using macromolecular probes. J Chromatography 950:75–80

Biography:

Jorge Costa Pereira has completed his BsC, MsC and PhD in University of Coimbra in 2000 at the age of 35 years. Since than he was BsC fellow, Assitant Professor and is Full Proffesor in Department of Chemistry of Faculty of Science and Technology at the University of Coimbra.His main work is on Chemical Analytical Control and Environment Quality Control field in specially related with Statistics, Analytical Method Validation and Chemometrics. At the present moment have 30 published works in well reputed international journals and is member of the editorial board or Protuguese Electrochimica Acta

Abstract:

Chromatography, particularly High Performance Liquid Chromatography (HPLC), is probably the most used standard instrumental analytical technique, widespread over all analytical labs around the world, very convenient to perform fast separation and simultaneous quantifi cation of several analytes in several complex samples. Most convenient, reliable and roust detectors are usually related with UV-vis detection such as Diode Array Detectors (DAD). However, when analyzing real and complex samples, it is frequent to observe signal distortions related with analytes co-elution, spectral interference and column lack of selectivity. Is it possible to be aware of these situations and circumvent these problems? In this work we propose Unsupervised Contribution Analysis (UCA) to address these situations; all chromatographic data should be checked in order to fi nd out the real number of independent contributions that are consistent with dataset in study. In a recent study, we suggested how to retrieve independent component contributions in spectroscopic (synchronous fl uorescence) analysis of environmental samples. In this work, Independent Component Analysis (ICA) was used in order to retrieve spectral sources and respective component contributions. However, in order to preform ICA, it was necessary to previously determine the number of independent components responsible for data spectra. Using Principal Object Analysis (POA), we showed how it’s possible to fi nd out system independent contributions in a robust and reliable manner. With this same concept, we have proved the ability of UCA to determine independent contributions; their specifi c chromatogram and; respective contributions using same approach, especially when there are evidences for co-elution profi les in (UV/vis)-HPLC and other non-specific instrumental methods. Some examples, practical and theoretical considerations will be addressed in order to maximize the ability of Unsupervised Component Analysis (UCA) to evaluate chromatographic response in complex systems.

Biography:

Dina Shokry completed her Bachelor degree in Pharmacy in 2009 at Ain Shams University then Master’s degree in Analytical Chemistry at Cairo University in 2013. Now, she is about to complete her PhD as a member of Dr Waters group for fi nding alternatives to animal testing at Huddersfi eld University. She worked as a Teaching Assistant then as an Assistant Lecturer of Analytical Chemistry at Future University. She produced high quality research that was published in anumber of reputed peer reviewed journals and presented her work in nine conferences. Her work is focused on developing models for prediction of human intestinal absorption through in vitro-in vivo correlation studies which has economic impact in the pharmaceutical industry fi eld. She developed prediction models from MLC,solubilization and permeation studies where the obtained in vitro data correlated well with the in vivo absorption data and resulted in two recently published papers.

Abstract:

Most of the new released drug compounds are formulated as orally administered drugs due to the convenience of the oral administration route. However, the properties of some compounds could be incompatible with oral administration. In fact, major fi nancial losses were suff ered by pharmaceutical industry because some new drugs were discovered to have poor bioavailability aft er their oral administration when tested in later clinical stage of development. Th erefore, drugs with poor aqueous solubility and oral bioavailability that are considered poor candidates should be spotted as soon as possible before reaching fi nal clinical stages of development where the costs spent on research carried out in such stages for studying the
biopharmaceutical properties of the drug is signifi cantly high, it is even better to discover these properties before the drug is synthesized to save time and money. Over the past three decades, there has been growing interest in the prediction of the biopharmaceutical properties as aqueous solubility and intestinal permeability of new drug entities (NDE) that resulted in the development of a large number of experimental (in vitro and in situ) and mathematical models. In addition to being cost
effective and time saving, some of these models help in the determination of best drug candidates during drug discovery and
development stage. Drug intestinal permeability is one of the most important biopharmaceutical properties that are worth investigating and predicting using the previously mentioned models. In the spectroscopic and permeation methods, we
developed mathematical models generated for prediction of human intestinal absorption (HIA) through the determination
of the micelle/water partition coeffi cients (logKxm/a) for a series of 20 compounds using UV spectroscopy and also through
determination of the permeation constants (log Kp) of a number of drugs through gels made from bile salt saturated with
infi nite dose of these drugs. Prediction models with good predictability were developed using the obtained data from both
methods along with the reference absorption data and other physicochemical properties to develop prediction equations
through simple and multiple linear regression respectively. In another work, we developed a model using MLC method which
was proved successful for prediction of HIA.

Biography:

Melissa Graewert is a Structural Biologist. Currently, she works at EMBL's Outstation in Hamburg, Germany located at PETRA III, one of the most brilliant storagering-based X-ray radiation sources in the world. Her expertise includes “Biophysical and structural characterization of proteins”. Her main research focus is on
“The implementation and constant development of small angle X-ray scattering as an emerging technique for the characterization of biological therapeutics such as monoclonal antibodies”.

Abstract:

Small angle X-ray scattering (SAXS) is a universal and powerful method to analyze proteins and other macromolecules in
solution, in a broad range of sizes and conditions. Th e synergistic improvement in hardware as well as soft ware over the
last decade has transformed SAXS into a high-through put technique, which became highly attractive for the pharmaceutical
industry. SAXS provides direct insights in the quaternary state; however, it is oft en hampered by inherent sample polydispersity.
At EMBL’s P12 beamline (@Petra III, DESY, Hamburg, Germany), we are developing novel in-line purifi cation systems such as
the implementation of an extended size exclusion chromatography set-up for the parallel biophysical and SAXS characterization
on separated components. Th e eluting protein is suffi ciently concentrated and pure, so that SAXS data can be directly collected
and used for structural and biophysical studies. Modes of access to this set-up (including European funded translational
activities such as iNEXT and industrial service provision) are discussed.

Biography:

Giacomo Russo is a Post-doctoral Scientist in Pharmaceutical Sciences. His research fi eld is intended to elucidate the mechanisms of drug interactions with biological membranes involved in bioavailability and distribution processes. His additional interest is in the development and validation of analytical methods aimed at determining endocrine disrupting agents in food/beverage and biological matrices.

Abstract:

The present study proposes a method for an in silico calculation of phospholipophilicity. Phospholipophilicity is intended as the measure of analyte affi nity for phospholipids; it is currently assessed by HPLC measures of analyte retention on phosphatidylcholine-like stationary phases (IAM-Immobilized Artifi cial Membrane) resulting in log kWIAM values. Due to the amphipathic and electrically charged nature of phospholipids, retention on these stationary phases results from complex mechanisms, being aff ected not only by lipophilicity (as measured by n-octanol/aqueous phase partition coeffi cients, log P) but
also by the occurrence of polar and/or electrostatic intermolecular interaction forces. Diff erently from log P, to date no method
has been proposed for in silico calculation of log kWIAM. Th e study is aimed both at shedding new light into the retention
mechanism on IAM stationary phases and at off ering a high-throughput method to achieve such values. A wide set of physicochemical and topological properties were taken into account, yielding a robust fi nal model including four in silico calculated
parameters (lipophilicity, hydrophilic/lipophilic balance, molecular size, and molecule fl exibility). Th e presented model was
based on the analysis of 205 experimentally determined values, taken from the literature and measured by a single research
group to minimize the inter laboratory variability; such model is able to predict phospholipophilicity values on both the two IAM stationary phases to date marketed, i.e., IAM.PC.MG and IAM.PC.DD2, with a fairly good degree (r2=0.85) of accuracy.
The present work allowed the development of a free on-line service aimed at calculating log kWIAM values of any molecule included in the PubChem database, which is freely available at http://nova.disfarm.unimi.it/logkwiam.html

Biography:

Yadira S Prieto Curbelo completed her Graduation in Biochemistry in 2005 and MSc in 2010 at University of Havana, Cuba. She has expertise in different areas,such as: purifi cation and development of analytical techniques and; in proteomic studies of expression stability in recombinant NS0 cell lines. She developed and
validated chromatographic techniques for the mAbs characterization and quantifi cation. She has expertise in mass spectrometric for characterization of proteins and oligosaccharide residues. She has been serving as Reviewer of Cytotechnology journal and member of Cuban Society of Pharmacy.

Abstract:

Techniques based on high-resolution liquid chromatography are currently used to quantify recombinant proteins from culture supernatants, as well as their characterization. Such assays can be easily and rapidly developed, this is the case of reverse phase chromatography. In this study, we describe the development and validation of an analytical technique for quantifying HER1 extracellular domain (HER1 ECD) in bioreactor supernatant using reversed-phase chromatography with a C8 column. On the other hand, we validate the methodology for the peptide mapping of monoclonal antibody using C4 column. For both study cases, the proteins were analyzed by monitoring the absorbance of the sample at 214 nm. Th e resulting analytical methodology was found to provide precise and accurate results for a wide range of concentrations (10–120 μg/mL) of HER1 ECD. Th e accuracy of the method varied from 86 to 109%, while the repeatability and the day-to-day intermediate precision were less than 7.25 and 7.85%, respectively. In the case of peptide mapping of mAb, the methodology provides a range of 35-40 well resolved peaks. As a criterion is set RT ≤0.5 min and the percentage peak height relative to the reference material must be 70% to 130%. These methodologies constitute a useful tool that can be applied during the production of the HER1 ECD vaccine and in the identifi cation of modifi cations on the primary structure of the mAb due to changes in biomanufacturing process.

Biography:

Hermes Licea Perez is a Senior Scientifi c Advisor and Technology Leader in Department of Bioanalysis, Immunogenicity & Biomarkers at GlaxoSmithKline, USA. He has been recently selected as GSK Fellow for his scientifi c contribution to the analytical community at GSK. He has completed his Master of Science degree in Chemistry at Moscow State University and a PhD degree at Stockholm University. His PhD research was focused on “Quantifi cation of haemoglobin adducts of industrial chemicals under the supervision of Prof. Siv Osterman-Golkar”. His interests at GlaxoSmithKline include method development and validation of
pharmaceutical drugs and metabolites in biological matrices using techniques such as protein precipitation, Solid Phase Extraction (SPE), Liquid Liquid Extraction (LLE), and chemical derivatization (chiral and achiral) for LC (or SFC)-MS/MS detection.

Abstract:

In recent years, interest has expanded to perform chiral separations by supercritical fl uid chromatography (SFC) which is proven to be superior to conventional liquid chromatography in separating structurally related compounds, such as diastereoisomers and enantiomers. Several examples will be described for separation of multiple stereoisomers in biological samples, confi rming SFC to be a powerful tool for stereoisomeric resolution for drug metabolism and pharmacokinetics (DMPK) applications. Two of these examples are summarized below: Gradient UPLC methodologies have previously been applied to separate a drug development compound and its six polyoxygenated metabolites (M2-M6 and M13), supporting numerous non-clinical and clinical PK studies. However, each of these metabolites exists in diff erent stereoisomeric forms,
resulting in 14 separate species. Initial attempts at developing UPLC methodologies were not capable of adequately separating these complex species; separation was unsuccessful using chemical derivatization, chiral and conventional reversed-phase liquid chromatography. Th e application of SFC is described herein to separate this complex mixture of 14 stereoisomeric metabolites; these data provided important data on which species circulate in human. SFC in combination with chemical derivatization was proven superior for separation of four diastereomeric species of another drug development compound.Th is method was fully validated and applied to evaluate potential in vivo chiral conversion in pooled clinical and preclinical samples.

Biography:

Field experiments usually give ecologically relevant results contributing to understand the ecosystem deeply. In marine research, aquacultures are one of the most popular procedures used to carry out fi eld experiments (fi gure a). In this study, the aquaculture of the green macroalga Ulva mutabilis was inoculated for the fi rst time with freshly induced gametes (7-day old germlings) to culture large volume (200 L) with small axenic gametes. Two sets of aquaculture: defi ned community (inoculated with axenic gametes of U. mutabilis and two associated bacteria: Roseovarius sp. strain MS2. and Maribacter sp. strain MS6), and undefi ned community (inoculated of U. mutabilis axenic gametes only). In defi ned community U. mutabilis showed a healthy growth and development (fi gure b) whereas in undefi ned community U. mutabilis lost its ability for growing and developing and formed only callus-like colonies (fi gure c). Multivariate statistics of the GC/MS and LC/MS analyses along with acquisition of biological metadata revealed that the waterborne metabolites in defi ned community were aff ected qualitatively and quantitatively by the growth phases of U. mutabilis as was proven in previous study of bioreactor cultures.

Abstract:

Taghreed Alsufyani is an Assistance Professor of Bioorganic Chemistry at Taif University. She received her Bachelor and Master degrees at King Abdul Aziz University, Jeddah, KSA. After that, she joined Chemistry department at Taif University where she got a scholarship to join PhD program at Friedrich Schiller University Jena, Germany, under the supervision of Dr. Thomas Wichard. By the end of 2014, she completed her PhD. In 2015, she was promoted to Assistant Professor at Taif University. Since 2015, she has established Algal Research Laboratory and began her investigation of algal chemical ecology as well as the applications of algae in biotechnology processes such as water treatment and bioenergy production.

Biography:

Yuliya E Silina completed her Doctorate degree in Analytical Chemistry. She is a Principal Investigator in the analytical team at Leibniz Institute for New Materials, Germany, focusing on innovations in chromatography, modern mass-spectrometry and environmental sensing. She has published more than 30 papers in reputed journals.

Abstract:

Recently, a link between changes in the cellular state refl ected by nucleotide and lipid profi les has been established. The analytical determination of nucleotides is not trivial, however, because of their high polarity and hydrophilic nature. Liquid chromatography (LC) has been widely implemented for determination of nucleotides from biological samples. Most of these LC-based techniques usually require ion-pairing reagents, thus making them unsuitable for LC-mass spectrometry (MS), timeconsuming, however, and also exhibited limited chromatographic resolving and strong matrix background power for biological samples. On the other hand, because of complexity of lipids, lipids analysis is still full of challenges. Meanwhile, due to the vital roles of the changes in human physiological and pathological process, lipidomics is attracting more and more attention. Hydrophilic interaction chromatography (HILIC) has become a powerful tool for the retention of polar analytes, because of its excellent mobile phase compatibility and complementary selectivity to RP chromatography. Herein, we implemented HILIC-MS for separation and quantifi cation of low molecular weight nucleotides and phospholipids in a nanoparticle-treated lung cells and cells under at diff erent stages of hypoxia. Th e challenges, namely necessity of liquid-liquid and solid phase extraction, samples stability, dilution re-assay and matrix effect in tissues are resolved and discussed. Th e elution conditions were subsequently optimized by evaluating organic content, pH and salt concentration in the mobile phase allowing a short simple isocratic run of only 20 min for nucleotides and 9 min for phospholipids, respectively.

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Ng Mei Han is a Principal Research Offi cer at Malaysian Palm Oil Board (MPOB). MPOB focuses on the research and development for the wellbeing of the oil palm industry. She primarily focuses on the green processing of the palm oil downstream products and developing methods for the analyses of palm oil minor components.

Abstract:

Natural tocols, which is a powerful antioxidant, are classifi ed into two major homologues series, the tocopherols and tocotrienols. Each of these homologue consists of α-, β-,
- and δ- isoforms. While tocopherols are more commonly found in natural plants, the availability of tocotrienols is more limited. Studies have found that both tocopherols and tocotrienols are present in palm oil. In fact, palm oil is known to contain highest amount of natural tocotrienols. Earlier studies documented that palm tocols consist of α-tocopherol (α-T), α-tocotrienol (α-T3), γ- tocopherol (γ-T), γ-tocotrienol (γ-T3)
and δ-tocotrienol (δ-T3). Studies in later years reported the presence of α-tocomonoenol (α-T1) in palm oil. Accuracy and reliability of analyses for tocols have always been a cause for concern. Th is is due to the fact that the tocols are highly potent antioxidative compounds and thus, are easily susceptible to oxidation or degradation. Th e most debated question remained the use of individual standards for the analyses of α-, β-, γ- and δ-tocotrienols. Th e question on the concentration of the standards were oft en raised as the tocotrienols, especially δ-tocotrienol is easily susceptible to oxidation. Th e scarcity of tocotrienols and
tocomononeol standards also contributed to the diffi culty in analyses and calibrations. Offi cial AOCS method recommended
the use of α-tocopherol as reference in the absence of the tocotrienols standards, which is a common practice. However, this is
oft en not accepted by the industry players mainly due to the question on the diff erent response of the individual tocopherols and
tocotrienols in UV spectroscopic analyses. In addition, the concentration of α-tocomonoenol is oft en not reported although
its presence is quite signifi cant in palm oil. Moreover, to date, there is no reliable offi cial method and authentic standards for
such analyses. Th e American Oil Chemists’ Society (AOCS) has documented a method for the analyses of tocopherols and tocotrienols, however, the method is deemed useful only when the standards for the individual tocopherols and tocotrienols are available for calibration. Th e present paper reports on a comprehensive and reliable method for the HPLC analyses of
tocopherols, tocomonoenol and tocotrienols in palm oil, taking into consideration the limit of detection, limit of quantitation, linear range of analyses as well as the calibration of the tocols in the absence of authentic standards.