Scientific Program

Conference Series Ltd invites all the participants across the globe to attend International Conference and Exhibition on Drug Safety & Pharmacovigilance Toronto, UK.

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Scientific Program Day 1
Scientific Program Day 2

Day 2 :

Keynote Forum

Brigitte Simons

Molecular Science Corp., Canada

Keynote: Differentiating cannabis products within the brands of the legalized adult use markets

Time : 09:00-09:40

Conference Series Drug Safety 2018 International Conference Keynote Speaker Brigitte Simons photo
Biography:

Brigitte Simons is a business development executive in support of leading-edge laboratory services and data management tools for the development of safe cannabis. Bridging expertise within analytical science, pharma drug development and environmental testing – Brigitte have a professional track record for laboratory testing instrumentation, software and sample contract design for the Canadian federal agencies, such as Canadian Food Inspection, Health Canada, Agriculture Canada and Environment Canada. She spent over 6 years working in the Drug Toxicology and Analysis Division at Health Canada in a mass spectrometry facility testing. She completed two post-doctoral fellowships at the Clinical Sciences Hospital of the National Heart, Blood & Lung Institute within the famous NIH campus in Maryland, USA. Continuing on in lab specialties, Brigitte then joined SCIEX, a global instrumentation vendor for hardware and software for mass spectrometry. With over 15 years experience with operating mass spectrometers, Brigitte managed Canadian federal and provincial government sales for full laboratory services, covering clinical, forensics to product health and environmental safety. Prior to working abroad, Brigitte received her Ph.D. in Chemical Biology at the University of Ottawa in a joint chemistry program with drug pharmacology at Health Canada. 

Abstract:

Within the framework of Bill C-45, Canada is positioned to become the global leader in the legal cannabis economy and global exporter. The enactment of this Canadian Cannabis Act provides legal access to marijuana and to control and regulate its production, distribution and sale. The primary objective of Health Canada’s regulatory policy bears stringencies with respect to public health and safety and strict requirements for quality assurance, record keeping and mandatory testing by 3rd party laboratories for product contamination. This opens an opportunity for advancing analytical development for cannabis metabolite profiling of active natural products and bleeds through to the accurate quantitative reporting of pesticides, mycotoxins and heavy metalloids that serve regulatory audit to clear products for sale. A complete LC-MS/MS workflow is described to quantitate 14 cannabinoids and screen for over 40 terpenoids to fingerprint various top cannabis dried flower brands from the large enterprise-producers in a method that is delivered in under 15 mins of analytical run time using a dual ESI and APCI ionization strategy. A wide linear dynamic range of 0.03 to 90% measurement (104 orders LDR) of cannabinoid per LC-MS injection can be reported to provide a more accurate view for product labeling and dosing recommendations. Terpene expression and metabolite measurement in plant cultivars are becoming less challenging with newly identified terpene synthases and availability of new mono-terpenes and sesquiterpene standards. It is of high interest for results of these metabolite profiling experiments to be correlated with plant cultivation parameters to achieve quality control and strengthen the consumer's experience with a brand of cannabis and differentiate products for retail. Furthermore, pesticide residue analysis in cannabis flower and oil formulations has been developed to meet the reporting requirements of Health Canada’s banned pest control ingredients list. With UHPLC linked tandem mass analysis covering all of the 96 banned pesticides except for 11 compounds best suited by GC separation, it is possible to achieve a validated cannabis product certificate of analysis for issuance to cannabis licensed producers in rapid turn-around. Analytical method details include LC separation using the Raptor Restek Column, Raptor Biphenyl and newly available mixtures of pesticide standards to meet the Canadian Pest Management Agency’s list of required pesticide maximum residual levels (down to 10 ppb in most cases). The addition of mycotoxins and other organo-contaminants can also be inserted into our methods with the use of optimized Scheduled MRM mass spec scanning techniques. The assembly of all the potency and ingredients data collection possible can provide information to consumers and track benefits to the cannabis producers stride to bring powerful brands to the global cannabis market.
 

 

Keynote Forum

Manuela G Neuman

University of Toronto, Canada

Keynote: Alcohol and drugs

Time : 09:45-10:25

Conference Series Drug Safety 2018 International Conference Keynote Speaker Manuela G Neuman photo
Biography:

Manuela Neuman is the CEO of In Vitro Drug Safety and Biotechnology, Toronto, ON., Canada. She is also teaching Pharmacology and Toxicology at the Faculty of Medicine, University of Toronto, Toronto, Canada. She is the Chair of Clinical Toxicology and Drug of Abuse Committee of the International Association of Therapeutic Drug Monitoring and Clinical Toxicology. She published 300 peer review articles. Her specialized laboratory provides personalized medicine and precision medicine results for Canada, USA and Europe. 

Abstract:

The interplay of alcohol with drugs includes multiple facets. These include the effects of alcohol on the effects of other hepatotoxicants and on the pharmacological effects of various drugs. Also relevant is the possible role of alcohol on the effects of carcinogenic agents. Less striking, but significant, are the effects of other drugs on the effects of ethanol. More difficult to identify but presumably significant, are the effects of alcohol-drug interplay on the development of an alcoholic liver disease. A common denominator of them is the role of ethanol-induced P-4502E1 (CYP2El) in affecting the toxicity of some hepatotoxicants and the effects of some drugs. Less prominent but also relevant is the effect of interplay with alcohol dehydrogenase and aldehyde dehydrogenase in the toxicity of some drugs. Alcohol has been shown to be responsible for cirrhosis in the 18th century and was labeled a hepatotoxin in the 19th century. During the second half of the 20th century alcohol has been recognize to enhance the toxic effect of other hepatotoxic agents such as acetaminophen, aflatoxin B1, allyl alcohol, bromobenzene, cocaine, enflurane, galactosamine, halothane, isoniazid, nitrosamines, thioacetamide, vinyl chloride and vitamin A. The toxicity of several hepatotoxicants is unaffected and of at least one, amanitine, is decreased by ethanol. The effect of ethanol on the toxicity of carbon tetrachloride and acetaminophen have been studied most extensively. The enhancement of toxicity by ethanol does not depend on an ethanol-induced hepatic injury but rather on the activity of the cytochrome P450 2E1 that converts the respective toxicants to their active metabolites. Nevertheless, inhibition by ethanol of regenerative response to injury may contribute to an enhancement of toxicity by ethanol. The toxicity produced by ethanol may have a bearing on the liver disease of alcoholism as well as on the toxicity and carcinogenicity of individual toxicants.
 

 

Keynote Forum

Abuzar Kabir

Florida International University, USA

Keynote: Fabric Phase Sorptive Extraction: A versatile sample preparation technology that meets the demands of twenty fi rst century modern analytical laboratories

Time : 10:45-11:25

Conference Series Drug Safety 2018 International Conference Keynote Speaker Abuzar Kabir photo
Biography:

Abuzar Kabir, a Research Assistant Professor at the International Forensic Research Institute (IFRI), Department of Chemistry and Biochemistry, Florida International University (FIU), Miami, Florida, USA, is a Separation Scientist and Materials Chemist. He has received his Ph.D. in analytical chemistry from University of South Florida (USF), Tampa, Florida, USA with specialization in sol-gel synthesis. He has invented 16-patented technologies in the area of chromatographic separation and analytical/bioanalytical sample preparation. He has also authored/co-authored 9 book chapters, 6 review articles, 46 research articles and 89 conference papers. 

Abstract:

 
Metabolomics plays an important role in discovering potential disease biomarkers from blood plasma or serum samples. Due to the distinctive complexity of whole blood as the sample matrix, either plasma or serum are used as the primary sample in metabolomics biomarker discovery research. During the transformation of whole blood into plasma or serum followed by extraction of targeted or non-targeted metabolites using conventional sample preparation techniques including solid phase extraction (SPE) and liquid-liquid extraction (LLE), a significant portion of the analytical information disappears, resulting in negligible success in discovering potential disease biomarkers. Fabric phase sorptive extraction (FPSE), a new generation sample preparation technology, has offered a paradigm shift approach in metabolomics sample preparation. FPSE innovatively combines the benefits of solid phase extraction (SPE) (works under exhaustive extraction principle) and solid phase microextraction (works under equilibrium extraction principle) into a single sample preparation technology platform. FPSE utilizes a flexible and permeable fabric substrate, coated with high-performance sol-gel sorbents as the extraction media. This uniquely designed extraction medium is capable of extracting target analyte(s) directly from whole blood. Due to the special geometry of FPSE medium (flexible, flat and permeable) and sponge-like porous architecture of sol-gel sorbents, rapid analyte mass transfer occurs between the bulk sample and the extraction medium, resulting in a near-exhaustive extraction within a fraction of time required for other comparable sample preparation techniques. FPSE is particularly suitable for analyzing target analytes e.g., metabolites, biomarkers directly from whole blood without requiring any protein precipitation or other pre-extraction sample cleaning/manipulation. After extracting the target analyte(s) directly from the whole blood sample, FPSE media is exposed to a small volume of organic/organo-aqueous solvent for eluting the extracted analyte(s). The low viscosity of the organic solvent, the capillary force of the fabric support and sponge-like porous sol-gel network allows fast diffusion of organic solvent into the FPSE medium for quick and complete recovery of the extracted analyte(s). As a result, FPSE completely eliminates time-consuming and error-prone solvent evaporation and sample reconstitution step often considered as an integral part of solid phase extraction/liquid-liquid e work-flow. During the solvent-mediated elution/back-extraction, any protein or matrix interferents adhered to the FPSE medium precipitates out and a final centrifugation of the resulting solution prior to injecting into the analytical instrument ensures clean particle-free highly concentrated target analyte(s). Fabric phase sorptive extraction has already developed a large number of sol-gel sorbents specifically suitable for polar metabolites/biomarkers such as sol-gel polyethylene glycol, sol-gel chitosan, sol-gel Carbowax 20M, sol-gel polycaprolactone-dimethylsiloxane-caprolactone to name a few. These high-efficiency sorbents have been found equally effective for analytes with a wide range of polarity. As a consequence, searching for a new disease biomarker from whole blood in presence of numerous endogenous and exogenous interferents is no longer a wishful thinking but an achievable reality. In the current talk, some new and fascinating data on metabolomics sample preparation using FPSE and a comparison between FPSE and conventional sample preparation techniques will be presented.
 

Keynote Forum

Peng Chen

Chemic Labs Inc., USA

Keynote: Applications of UPLC-MS QTOF in structural elucidation of small molecules

Time : 11:30-12:10

Conference Series Drug Safety 2018 International Conference Keynote Speaker Peng Chen photo
Biography:

Peng Chen received a Ph.D. in Analytical Chemistry from Indiana University in 1998 and a M.S. in Organic Chemistry from the University of Louisville in 1994. His graduate research includes the introduction of osazones as MALDI matrices for carbohydrate analysis and the structural elucidation of fluorescent aging markers. He has been working in various chemical industry sectors in the fields of chromatography and mass spectrometry. His work in recent years at Chemic Labs Inc. involves structural elucidation of small molecules in pharmaceuticals and medical devices by high-resolution QTOF mass spectrometry. 

Abstract:

The structural elucidation of small molecules by high-resolution mass spectrometry plays important roles in development and quality control of pharmaceuticals and medical devices. Trace amounts of small molecules can be present in forms of impurities, by-products or degradation products, etc. It is often difficult to separate and fractionate enough quantities of these analytes for conventional structural analysis by NMR and FTIR. Recent advances in instrumentation and software of UPLC-MS QTOF with MS/MS fragmentation capability can give structural insight into molecules of interest and in many cases offer structure candidates at high confidence. This presentation will use several practical examples in the analysis of synthetic compounds and identification of impurities associated with pharmaceuticals and medical devices to illustrate the convenience and power of UPLC-QTOF high-resolution mass spectrometry.
 

 

  • Analytical Chemistry and Engineering | Advances in Chromatography & HPLC Techniques | Pre- Clinical and Clinical Trials | Mass Spectrometry and Metabolomics | The key benefi ts of HPLC systems | Analytical Chemistry in Agricultural Research | Method Development and Validation | Analytical Chemistry In Veterinary Drug Analysis | Sensor Modifi cations
Location: Conference Hall: Frederick
Speaker

Chair

Milos Netopilik

Academy of Sciences of the Czech Republic, Czech Republic

Speaker

Co-Chair

Abuzar Kabir

International Forensic Research Institute, USA

Session Introduction

Marc Plante

Boston Analyticals, USA

Title: HPLC method development and troubleshooting

Time : 12:15-12:55

Speaker
Biography:

Marc Plante has been working in the field of analytical chemistry and chromatography for over 25 years, complete with a variety of publications, speeches, webinars and a wide range of experience. He first earned his BSc in Chemistry with two minors in engineering at Rensselaer Polytechnic Institute in Troy, New York and then a doctorate in Physical-Organic chemistry at Northeastern University. His first employment involved a novel semi-synthetic taxane process, for which he helped developed a now-patented chromatographic purification system. Over the course of his current career, he has largely worked with small-molecule chromatography, with much of his work involving the Corona Charged Aerosol Detector. At ESA, Dionex and Thermo Fisher Scientific, he created application notes and posters, gave many presentations and wrote two book chapters. He also provided some ideas for improving the detector's capabilities. 

Abstract:

Separation technology is a critical tool for the determination of many aspects for the goods and products that we use every day: from the multiple aspects of pharmaceuticals we take for our health, the researching and evaluation of new biosimilars, the products that we buy and use and to the food and water we consume every day. It's being an honor of being invited to speak on some of this technology, where I will have experience with the development and promotion of one of the more recent detector technologies for HPLC: the Corona charged aerosol detector. During my eight years as an applications chemist ESA Biosciences, Dionex and Thermo Fisher Scientifically its being able to create many novel applications encompassing lipids of all variety, biofuels, carbohydrates, foods, ions, pharmaceuticals, surfactants, proteins, industrial chemicals and pretty much everything else. Along with this work, it’s a unique opportunity to provide ideas for improving the detector, some of which were incorporated. This detector is still relatively new for HPLC and here at this conference, we will be hearing about the newer technologies that are in development and their uses, including new solid-phase extraction techniques, column materials and other separation technologies and methods.
 

 

Uchenna Samson Obi

University Hospital of the West Indies, Jamaica

Title: An assessment of nurses’ Knowledge Attitude and Practice (KAP) of pharmacovigilance

Time : 14:00-14:40

Speaker
Biography:

Uchenna Obi was born in Lagos Nigeria, with a migration to Jamaica in 2007 for educational pursuits, Uchenna Obi holds a Bachelor of Science degree in Nursing from the Northern Caribbean University in Mandeville Jamaica. While working as a Registered Nurse at the University Hospital of the West Indies Jamaica, he went on to obtain a Post-graduate certificate in Critical Care Nursing, together with a Master of Science degree in Applied Pharmacology from the University of the West Indies, Jamaica. Uchenna is a Registered/Critical Care Nurse who is committed to applying quality research to clinical practice for the improvement of patient outcomes and over-all health safety of the public. His research article is published in the Journal of Clinical Review & Case Reports. 

Abstract:

Purpose: The objective of the study is to assess nurses’ Knowledge, Attitude and Practice (KAP) of pharmacovigilance. Methods: This is a cross-sectional study that utilizes a questionnaire to evaluate nurses’ KAP of pharmacovigilance at the University Hospital of the West Indies, Mona. A sample size of 234 nurses was selected using a 95% confidence level with the raosoft online sample size calculator. Data obtained from filled questionnaires were analyzed using the SPSS 20 using descriptive and inferential measures. The chi-square test was used to test the association between two attributes at a P<0.05 significant level. Results: 209 responses were received from 260 distributed questionnaires, giving an 80% response rate. 13.5% of the nurses had heard of the term pharmacovigilance prior to the study. 58.4% correctly stated the functions of pharmacovigilance. Results for attitude towards pharmacovigilance revealed 93.7% of the nurses felt it was a professional obligation to report Adverse Drug Reactions (ADR). 55.3% of nurses indicated they had reported an ADR. A χ2 test of independence was performed to determine an association between nurses who noted ADRs in clinical practice and nurses who reported ADRs. Results showed a significant association between these two variables, χ2 (1) = 86.642, p<0.05. Conclusion: This study concludes that the registered nurses at UHWI had a good attitude towards pharmacovigilance, although their knowledge and practice was limited. Recommendation from the study includes instituting pharmacovigilance training programs that will improve nurses’ knowledge and hopefully impact their practice.
 

Dusan Berek

Polymer Institute of the Slovak Academy of Science, Slovakia

Title: Separation of parent homopolymers from block copolymers with advanced liquid chromatography methods

Time : 14:45-15:05

Speaker
Biography:

Dusan Berek is 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 130 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 2002".
 

Abstract:

Block copolymers present an important group of materials with numerous applications in science, medicine and technology. In a block copolymer, at least two chemically distinct polymer chains are connected with a chemical bond. Comprehensive molecular characterization of block copolymers is an analytical challenge. A special problem makes a determination of the amount and molar mass of parent homopolymers, which occur in most block copolymers and constitute highly undesired ballast. Gel permeation (size exclusion) chromatography GPC/SEC is commonly employed for the characterization of block copolymers. Molar mass of a precursor, the block polymerized as first, is determined by GPC/SEC and the same method serves for the approximate assessment of total molar mass of the block copolymer under study. Due to its low separation selectivity and detector sensitivity, GPC/SEC can hardly identify the presence of parent homopolymers and render their molar mass. We will discuss principles and applications of the new separation methods namely liquid chromatography under limiting conditions of enthalpic interactions, LC-LC and sequential two-dimensional polymer liquid chromatography, S2D LC to solve the above tasks. LC-LC methods are well robust and experimentally feasible. Their separation selectivity is very high and sample recovery is reasonable. LC-LC can efficiently separate both parent homopolymers from diblock copolymers in one single step. The separated sample constituents can be one-by-one forwarded into an on-line GPC/SEC column for determination of their molar mass average and distribution. Parent homopolymers present in the block copolymer at a very low concentration below 1% of can be tracked-down and characterized with help of the S2D LC.
 

Lounès Haroune

Pharmacology Institute of Sherbrooke, Canada

Title: (Not Yet Disclosed)

Time : 15:10-15:30

Speaker
Biography:

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).
 

Abstract:

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 (10µL–25µL) 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. 

Michael Anjello Jothi Rajan

Tamil Nadu State Council for Science & Technology, India

Title: Photoacoustics spectroscopy: The less explored non-destructive spectroscopy for multicharacterization

Time : 15:50-16:10

Speaker
Biography:

Michael Anjello Jothi Rajan, Ph.D., Doctorate in Physics, (Ph .D. Physics), now is Project Scientist in Tamil Nadu State Council for Science & Technology, Chennai – 600025, INDIA. He was an Associate professor of Physics, Director, Bio-Nano Research Laboratory and Dean of Research in Arul Anandar College (Autonomous), Karumathur – 625514, India.  He also headed the Department of Foundation Courses as an honorary member He is a popular science communicator and has started many science clubs in rural high schools in Tamil Nadu, India. He has specialized in Human Rights, Personality Development, Environmental Studies, Bioethics, Biocosmology and Peace education. He is a recipient of many awards in Physics as well as humanitarian.  He has written books on Human Rights, Bioethics, Personality Development and Environmental Studies. He has many projects in Biomedical engineering, Eco-Water, Sanitation and Hygiene and socio-economic development of Below Poverty Line families in rural Tamil Nadu. His team works seriously on Cancer Cells. This lab is short of much good instrumentations but his well-wishers in many countries had come forward to help his team. He and his team (8 Research Scholars) are open for any type of collaboration with teams of similar missions. 

Abstract:

Optical spectroscopy remains a widely used and most important tool for investigating and characterizing the properties of matter. The energy used in optical spectroscopy exists in the form of optical photons or quanta, with a wavelength ranging from less than 1AËš in the x-ray region to more than 10 6AËš in the far-infrared. It is highly versatile, widely ranged and nondestructive in nature. Optical Spectroscopy has been a scientific tool for over a century and a half and it has proven invaluable in studies on reasonably clear media, such as solutions and crystals and on specularly reflective surfaces. There are, however, several instances where conventional transmission spectroscopy is inadequate even for the case of clear, transparent materials. Such a situation arises when one is attempting to measure a very weak absorption, which in turn involves the measurement of a very small change in the intensity of a strong, essentially unattenuated, transmitted signal. Although this problem occurs for all forms of matters, it has received particular attention in the case of transparent gas mixtures containing minute quantities of an absorbing species or pollutant. Various techniques develop to overcome this difficulty, such as derivative spectroscopic, have proven to generally inadequate. In addition to weakly absorbing materials, there are a great many nongaseous substances, both organic and inorganic, that are not readily amenable to the conventional transmission or reflection modes of optical spectroscopy. These are usually highly light-scattering materials, such as powders, amorphous solids, gels, smears and suspensions. Other difficult materials are those that are optically opaque and have dimensions that far exceed the penetration depth of the photons. Over the years, several techniques have been developed to permit optical investigation of highly light – scattering and opaque substances. The most common of these are diffuse reflectance, attenuated total reflection (ATR) and internal reflection spectroscopy and Raman scattering. All these techniques have proven to be very useful, yet each suffers from serious limitations. In particular, each method is applicable to only a relatively small category of materials, each is useful over a small wavelength range and the data obtained are often difficult to interpret. The modern scanning and tunneling microscopic techniques in spite of their versatility are having inherent inadequacies and economically very costly. The photoacoustic spectroscopy strikes a balance between the optical spectroscopy and the modern microscopic techniques in that it is relatively cheaper, highly efficient over a wide range of wavelengths, applicable for any type of material. The newly developed electronics technology is highly assisting the versatility of the photoacoustic spectroscopy (PAS), Ultrasonic photoacoustic microscopy and Piezoelectric Photoacoustic microscopy (PPAM) to study the thermal and optical characteristics of any type of materials in the micro and nanoscales. In this work, we present the thermal diffusivity measurement of Poly (methyl methacrylate) (PMMA) - montmorillonite (MMT) clay nanocomposite by PPAM and compare it with the X-ray diffraction studies.

Alina Vasilescu

International Centre of Biodynamics, Romania

Title: Poster: Electrochemical sensors coated with prGO coated enable to capture different aggregation behaviors of proteins and peptides
Speaker
Biography:

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.

Abstract:

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 analysis.
 

Nives Galic

University of Zagreb, Croatia

Title: Poster: Structural investigation of Fe(III) and Ga(III) complexes with aromatic hydrazones by ESI MS/MS
Speaker
Biography:

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 (IP2014-09-4841).
 

Abstract:

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.
 

Lounès haroune

Pharmacology Institute of Sherbrooke, Canada

Title: Poster: What about dried blood spot for cannabinoid quantification?
Speaker
Biography:

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).
 

 

Abstract:

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 (10µL–25µL) 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. 

Sabrina Saibi

Université de Sherbrooke, Canada

Title: Poster: A simple analytical method for the detection and quantifi cation of a pharmaceuticals and pesticides in complex environmental matrices
Speaker
Biography:

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).
 

 

Abstract:

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.

 

Isil Yasa

Bristol-Myers Squibb, USA

Title: Poster: Platform Size Exclusion Chromatography (SEC) method development for a broad range of monoclonal antibodies
Biography:

Abstract:

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

Imtakt Corporation, Japan

Title: Poster: LC-MS analysis of intact amino acids on a novel mixed-mode HPLC column
Speaker
Biography:

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.
 

Abstract:

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.
 

Fekadu Desta

Addis Ababa University, Ethiopia

Title: Poster: Comparison of immune cells subsets, ex-vivo and in-vivo expression of T cell activation and memory marker between LNC and corresponding PBMC from calves exposed to natural mycobacterium bovis infection
Speaker
Biography:

Fekadu Desta is a veterinarian with a dream and responsibility to control and prevent the existing endemic and emerging new diseases of livestock and companion animals, Ethiopia. For the last 6 years, he has been doing his PhD in Tropical Infectious Diseases with a PhD dissertation entitled “Comparison of Immune Cell Subsets, ex-Vivo and in-Vitro Expression of Activation and Memory Marker Between LNC and the Corresponding PBMC from Calves Exposed to Natural Mycobacterium bovis Infection in BCG Efficacy Trial” which is one of a research priority of the country. Currently, he is on data analysis, interpretation and result dissemination stage. He has published one paper on molecular epidemiology of M. bovis and preparing 3 more manuscript on bovine immunology.
 

Abstract:

Cell-mediated immunity and development of necrotic granulomas in Mycobacterium bovis (M. bovis) infected lymph node (LN) is pathognomonic for bovine tuberculosis (BTB). This delayed hypersensitive host response involves a complex interaction of cellular and immune mediators within systemic circulation and LN. Hence, tuberculosis immunological response should be independently investigated at the peripheral blood and LN tissue level. The objective of this study was, therefore, to compare the cell surface and cytokine expression between immune cell from peripheral blood and lymph node cells (LNC) from calves on BCG efficacy trial. Twenty pairs of peripheral blood mononuclear cells (PBMC) and) LNC from M. bovis naturally infected calves during BCG vaccine experiment trial were isolated and investigated in two phases of the flowcytometry experiment. In the first phase of a flow-cytometry experiment the proportion of ex-vivo CD25+ expressing cells was significantly higher (P<0.05) in CD4+ and CD8+ T node than that of peripheral blood. However, such difference in CD25+ expression was not observed in WC1 γδ T cells. Contrary to CD25+ ex-vivo expression, in-vitro IFN-γ and TNF-α producing cells were greater (P<0.05) in T cells of the peripheral blood than T cells of lymph node after PMA + ionomycin stimulation. This difference in IFNγ and TNFα responses was also statistically significant between a vaccinated and non-vaccinated group. An IL-4 producing cell was not evident in PBMC and LNC. During the second phase of flow-cytometry experiment additional surface marker; CD2, CD21, CD205, CD335 and CD1W2 were included to add more panels for immune cell subset. The second experiment revealed that PBMC CD4–WC1+ and CD8–WC1+ γδ T cells and CD205+D1W2+ DC subset exhibited lower percentage than γδ T cells and DC of LNC respectively (p=0.0001, p=0.0061). However, PBMC CD335+CD2+ NKT cells subset exhibited a higher percentage than NKT cells of LNC (p=0.0129). No difference was observed between groups in the percentage of the rest of T-cell and B-cell (p>0.05). Findings of this study suggest the existence of phenotypic immune compartmentalization between the two tissue compartments.
 

Julio C Fernandez Travieso

National Centre for Scientific Research, Cuba

Title: Poster: Effects of policosanol in older patients consuming nitrates vasodilators
Speaker
Biography:

Julio Cesar Fernandez Travieso is a Senior Investigator in Clinical Trials Unit, National Centre for Scientific Research, Havana, Cuba. He has completed his BSc in Pharmaceutical Sciences from Havana University, Cuba in 1996. He was awarded with PhD in Pharmaceutical Sciences in 2003. He has published more than 130 publications and presented more than 100 papers in various scientific events. His research interest mainly focuses on clinical trials phase I-IV of different natural products: Policosanol, Abexol, Prevenox and Palmex.
 

Abstract:

Introduction: Policosanol is a cholesterol-lowering drug with concomitant antiplatelet effects. The efficacy and safety of policosanol have been investigated in clinical studies and post-marketing surveillance. Policosanol is very safe and no drugrelated Adverse Events (AE) have been demonstrated, even in population subsets with high consumption of concomitant therapy, indicating that the potential risk of Drug-Drug Interaction (DDI) for policosanol is low. Vasodilators are used in geriatric populations mainly to treat congestive heart failure and acute decompensating of heart failure, although associated with other anti-hypertensive are also used to manage arterial hypertension. Vasodilators, however, have a considerable risk of drugrelated toxicity, the most frequent symptoms being those derived from excessive vasodilation and hypotension, such as nausea, vomiting, loss of consciousness and reflex tachycardia. Vasodilators show important DDI derive from pharmacodynamic interactions with several drugs, those associated with the concomitant use of other vasodilators and diuretics being the most relevant. Considering such facts, the interest to study putative DDI between policosanol and vasodilators is supported. Objective: To investigate whether policosanol administered to older patients consuming vasodilators induces any specific disturbance on safety indicators and/or increase the frequency or severity of AE in such patients. Methods: This report was based in the analysis of the records of all patients (185) taking nitrates vasodilators included in a prevention study in the elderly randomized to policosanol 5 mg/d or placebo for 3 years. The analysis was by Intention-to-treat. Results: Baseline characteristics were well balanced in both groups. After one year on treatment, policosanol lowered significantly Low-Density Lipoprotein-Cholesterol (LDL-C) (20.9%), Total Cholesterol (TC) (15.9%) and triglycerides (19.3%), whereas raised High-Density Lipoprotein-Cholesterol (HDL-C) (8.3%). Policosanol effects persisted, even increased, during the 3 years treatment. At the end of the study, policosanol reduced LDL-C (35.0%), TC (25.0%), triglycerides (19.3%) and raised HDL-C (16.7%). Of 185 randomized patients taking vasodilators, 44 (23.8%) withdrew from the trial. The frequency of withdrawals in placebo (31/95; 32.6%) was greater (p<0.01) than in the policosanol group (13/90; 14.4%). Overall, 26/185 (14.1%) patients discontinued due to some AE: 23 placebo (24.2%) and 3 policosanol patients (3.0%) (p<0.01). Policosanol did not impair safety indicators compared with placebo but induced additional decreases in systolic pressure compared with placebo. The frequency of policosanol patients experiencing Serious Adverse Events (SAE) (3/90; 3.3%) was lower (p<0.01) than in the respective placebo (23/95; 24.2%). Likewise, the frequency of policosanol patients who experienced some mild or moderate AE during the study (10/90; 11.1%) was lower (p<0.05) than in matched placebo (28/95; 29.5%). Conclusions: Policosanol was well tolerated in older subjects with high coronary risk-taking vasodilators, not impairing safety indicators or increasing any AE respect to placebo. Policosanol, however, produced additional decreases of arterial pressure and reduced the frequency of SAE compared with placebo. Cholesterol-lowering efficacy of policosanol was persistent and consistent with that expected. These results indicate that policosanol can be administered to older patients taking vasodilators without risk of relevant adverse DDI.
 

Julio C Fernandez Travieso

National Centre for Scientific Research, Cuba

Title: Poster: Concomitant use of policosanol and antiplatelet drugs in older patients
Speaker
Biography:

Julio Cesar Fernandez Travieso is a Senior Investigator in Clinical Trials Unit, National Centre for Scientific Research, Havana, Cuba. He has completed his BSc in Pharmaceutical Sciences from Havana University, Cuba in 1996. He was awarded with PhD in Pharmaceutical Sciences in 2003. He has published more than 130 publications and presented more than 100 papers in various scientific events. His research interest mainly focuses on clinical trials phase I-IV of different natural products: Policosanol, Abexol, Prevenox and Palmex.
 

Abstract:

Introduction: Policosanol is a cholesterol-lowering drug with antiplatelet action. Policosanol effects have been investigated in clinical trials and post-marketing surveillance, its efficacy and safety are demonstrated. Policosanol has resulted very safely, even when administered to special populations with high consumption of concomitant drugs, without demonstration of drugrelated Adverse Events (AE). Accordingly, the potential risk of Drug-Drug Interaction (DDI) with policosanol appears to be low. DDI generally comes from pharmacokinetic or/and pharmacodynamic actions. Experimental data show that DDI with policosanol derived from pharmacokinetic interactions is not very probable. Nevertheless, DDI based on pharmacological interactions needs to be investigated. Antiplatelet drugs are widely used in middle-aged and geriatric populations mainly to prevent recurrent coronary or cerebrovascular events. Experimental and small clinical studies have shown that policosanol enhances the antiplatelet effects of aspirin in an additive manner. Hence, the interest to study putative DDI between policosanol and antiplatelet drugs in a population sensitive to drug-related effects, as the elderly, is supported. Objective: The objective of the present analysis as a part of a prevention study, we investigated whether policosanol administered to older individuals taking antiplatelet drugs supposes concern regarding a potential risk for adverse drug-drug interactions. Methods: We randomized 1470 elderly patients at high coronary risk to policosanol 5 mg/day or placebo for 3 years. For this analysis, the records of all patients (334) taking antiplatelet drugs were included. The analysis was by intention-to-treat. Results: After one year, policosanol decreased significantly Low-Density Lipoprotein-Cholesterol (LDL-C) (21.0%), total cholesterol (16.9%) and triglycerides (19.6%), while raised High-Density Lipoprotein-Cholesterol (HDL-C) (6.2%). Policosanol effects were maintained, even improved, during the follow-up. At study completion policosanol lowered LDL-C (34.3 %), total cholesterol (23.9%), triglycerides (22.2%) and raised HDL-C (14.5%). Sixty patients (41 placebo, 19 policosanol, p<0.01) withdrew from the study, 33 (23 placebo, 10 policosanol) (p<0.01) due to some adverse event, all serious. Policosanol did not impair safety indicators and did not increase any adverse event with respect to placebo. Conclusions: The policosanol can be administered to older patients taking antiplatelet drugs without risk of relevant adverse drug-drug interactions.
 

Maria Ramos Payan

University of Seville, Spain

Title: Video: New trends in sample miniaturization and its applications: On-chip devices

Time : 16:50-17:10

Speaker
Biography:

Maria Ramos Payan has completed her Ph.D. from University of Seville, Spain and postdoctoral studies from University of Copenhagen (Denmark), University of
North Carolina (USA) and Microelectronic National Center of Barcelona (Spain). She is the leader of the microfluidic research line. She has published more than
30 papers in reputed journals and has been serving as an editorial board member of repute.

Abstract:

Sample preparation miniaturization is one of the latest trends in analytical chemistry. The development of new sample
preparation procedures is closely linked to the new on-chip microfluidic devices. These microchips have proven to offer new
advantages over traditional methodologies, such as a decreasement of the reagent volume, organic solvent and sample volume.
It also reduces the analysis time and offers very high extraction efficiencies when working under double flow conditions or
good enrichment factors when working under stationary conditions. These microchips systems have been employed using two
different extraction techniques: liquid phase microextraction and electromembrane. The parameters that affect both extractions
are the composition of the sample and the acceptor phase, the flow of the acceptor and donor phase, the organic solvent used as
a liquid membrane supported and the extraction time. The voltage is one of the extra parameters to be determined in the case
of the electromembrane. However, one of the most influential variables is related to the geometry of these microchips systems,
since depending on their length, width and depth, different extraction and enrichment efficiencies will be obtained. In this
work, new trends in the geometric study of microfluidic devices and their application to both environmental and biological
samples are presented.

Maria Ramos Payan

University of Seville, Spain

Title: Video: New trends in sample miniaturization and its applications: On-chip devices
Speaker
Biography:

Maria Ramos Payan has completed her Ph.D. from University of Seville, Spain and postdoctoral studies from University of Copenhagen (Denmark), University of North Carolina (USA) and Microelectronic National Center of Barcelona (Spain). She is the leader of the microfluidic research line. She has published more than 30 papers in reputed journals and has been serving as an editorial board member of repute.
 

Abstract:

Sample preparation miniaturization is one of the latest trends in analytical chemistry. The development of new sample preparation procedures is closely linked to the new on-chip microfluidic devices. These microchips have proven to offer new advantages over traditional methodologies, such as a decreasement of the reagent volume, organic solvent and sample volume. It also reduces the analysis time and offers very high extraction efficiencies when working under double flow conditions or good enrichment factors when working under stationary conditions. These microchips systems have been employed using two different extraction techniques: liquid phase microextraction and electromembrane. The parameters that affect both extractions are the composition of the sample and the acceptor phase, the flow of the acceptor and donor phase, the organic solvent used as a liquid membrane supported and the extraction time. The voltage is one of the extra parameters to be determined in the case of the electromembrane. However, one of the most influential variables is related to the geometry of these microchips systems, since depending on their length, width and depth, different extraction and enrichment efficiencies will be obtained. In this work, new trends in the geometric study of microfluidic devices and their application to both environmental and biological samples are presented.
 

  • Polymer and Material Chemistry | Disease Detection and Formulation Development | Good Pharmacovigilance Practice | Mass spectroscopy and Gas chromatography | Hyphenated HPLC Methods | Electrophoresis | Principles and Applications of HPLC | Chromatography | Analytical Chemistry | Nuclear Magnetic Resonance
Location: Conference Hall : Frederick
Speaker

Chair

David M Parish

Sherwin-Williams Company, USA

Speaker

Co-Chair

Brigitte Simons

Molecular Science Corp., Canada

Session Introduction

Dusan Berek

Polymer Institute of the Slovak Academy of Science, Slovakia

Title: Retention mechanisms in liquid chromatography of synthetic polymers
Speaker
Biography:

Dusan Berek is 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 130 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 2002". 

Abstract:

At present, methods of liquid chromatography, (LC) provide the most important information on both average values and distributions of molecular characteristics – molar mass, chemical structure (composition) and physical architecture (topology) of synthetic macromolecules. Gel permeation (size exclusion) chromatography, GPC/SEC, is commonly employed for the determination of the molar mass of macromolecules. Its basic retention mechanism is steric exclusion based on the changes of conformational entropy of coiled macromolecules entering the pores of the column packing. However, GPC/SEC cannot give information about polymer molar mass in presence of the changing second or even third molecular characteristic. To simultaneously determine two molecular characteristics of a complex polymer such as copolymers or polymer blends, the entropy controlled retention mechanism is combined, coupled with the interaction retention mechanisms. The most common interaction retention mechanism employed in coupled LC methods is adsorption i.e. the distribution of a solute between a solid surface and a volume of its solution in a mobile phase. Adsorption usually results from the polar interactions among active sites on macromolecules and on the column packing surface, which are controlled by eluent polarity and less often by temperature. The appropriate stationary phase is bare silica gel. Another LC retention mechanism is absorption, (enthapic partition), the distribution of a solute between the volumes of mobile and stationary phases. The practically applicable volume of LC stationary phase is produced by the chemical attachment, bonding of appropriate groups usually C18 alkyl groups onto a carrier, mainly silica gel. In practice, both adsorption and enthalpic partition retention mechanisms are applied either isocratically or applying mobile phase with the gradually changing composition. All enthaphy based processes in the LC columns are accompanied with the large changes of conformational entropy of macromolecules so that all coupled polymer LC procedures present a combination of enthalpy and entropy-based processes.
 

Yintang Zhang

Shangqiu Normal University, China

Title: Advances in determination of Alzheimer’s -amyloid peptide
Speaker
Biography:

Yintang Zhang has completed his PhD at the age of 29 years from Central South University. He is the director of Henan Joint International Research Laboratory of Chemo/Biosensing and Early Diagnosis of Major Diseases. He has published more than 25 papers in reputed journals. 

Abstract:

Alzheimer’s disease (AD), the most common type of dementia in the elderly, is a progressive and devastating neurodegenerative disease causing memory loss, impaired thinking and other symptoms. β-amyloid peptide (Aβ) indicates a biomarker for an AD in cerebrospinal fluid, blood, plasma and serum. Therefore, it is important to determine the Aβ concentration for early diagnosis and treatment of AD. In this study, micellar electrokinetic capillary chromatography (MEKC) combined with an online preconcentration method sweeping was established to determine Aβ1-42. Electrophoresis was carried out at a constant voltage of 5 kV in an uncoated fused-silica capillary. The 15 mmol/L borate buffer containing 20 mmol/L sodium dodecyl sulfate (SDS) with pH 9.3 was used as the running buffer. The samples were injected into the capillary by applying a pressure of 50 mbar for 60s. Under the optimal conditions, the detection limit of Aβ1-42 was as low as 0.08 μmol/L. The recoveries of the normal addition method in real human serum sample analysis were found to be between 89.2% and 98.5%. The relative standard deviation (RSD) of the determination was less than 6%. 

Milos Netopilik

Institute of Macromolecular Chemistry, Czech Republic

Title: Combinatorial model of chromatography applied on optimizing operational conditions in SEC
Speaker
Biography:

Milos Netopilik has completed his Ph.D. at the age of 30 years from Institute of Macromolecular Chemistry and postdoctoral studies from Virginia Polytechnic Institute and Technical University. He has published more than 65 papers in reputed journals.
 

Abstract:

The shape of the elution curves depends strongly on experimental conditions, in the first place on polymer molecular weight, upon this, on concentration and flow-rate. The effect of concentration is weak for polymers in theta solvents up to the concentrations of overloading. On the other hand, in good solvents, the concentration effect is important. The effective hydrodynamic volume of dissolved macromolecules decreases with increasing concentration. The decrease in the hydrodynamic volume is of solvated molecules with increasing concentration is an established experimental factor which theoretical explanation. The spatial distribution of the analyte with respect to the longitudinal axis of the separation system, developing in time, can be expressed by the binomial distribution. However, further treatments of this physical situation were approximative. The exact solution to the problem is obtained as the observation at a fixed point (the detector) of this binomial distribution developing in time after reaching the exclusion limit. This can be done numerically. The description of the concentration effect on SEC elution curves is possible on the basis of the displacement- equilibrium model. This is based on the concept of a theoretical plate on which the equilibrium is formed between molecules of the analyte moving together with mobile phase (MP) and those anchored on the or penetrated into the pores of the stationary phase (SP). The simulation of the concentration effect is possible with partition coefficient calculated numerically for each plate at each displacement.
 

H W C Krishanthi Karunarathne

Government Analyst Department, Sri Lanka

Title: Determination of the concentration of benzoic acid and sorbic acid in ready to serve products using High Performance Liquid Chromatography (HPLC)
Speaker
Biography:

H W C Krishanthi Karunarathne has completed her MSc. in analytical chemistry, University of Peradeniya, Sri Lanka. She is the Assistant Government Analyst in Government Analyst Department in Sri Lanka. She has published more than 10 papers in some reputed journals.
 

Abstract:

Sodium benzoate and potassium sorbate are two major chemical preservatives which are used in ready to serve products. In this study, a total of 50 commercial brands of highly consumed ready to serve products were analyzed. The HPLC determination of the preservatives was performed using a reversed – phase C18 column and UV detection at 235 nm. Flow rate approximately 1.2 ml/min. Eluent for HPLC, mix 50 volume parts of ammonium acetate solution with 40 volume parts of methanol for HPLC and adjust to a pH of 4.5 to 4.6 with acetic acid. The preservative concentration in samples was using authentic external standard sodium benzoate and potassium sorbate. Among 50 samples, the minimum and maximum concentration of benzoate content in various brands were 80ppm and 874ppm and for sorbate was 60ppm to 562ppm respectively, 22% of samples do not compliance with standard regulations in Sri Lanka. Exposure to these chemical preservatives could be a risk factor for the human health, especially when their intake was being occurred by various foodstuffs simultaneously.

Zahra Ramezani

Ahvaz Jundishapur University, Iran

Title: Fabrication of a magnetic molecularly imprinted polymer for pre-concentration and cleanup of sarcosine, a prostate cancer metabolic biomarker, in urine prior to capillary electrophoresis determination
Speaker
Biography:

Zahra Ramezani is currently professor of analytical chemistry at Ahvaz Jundishapur University of Medical Sciences. She earns her BSc in pure chemistry, MSc and Ph.D. in analytical chemistry at Shiraz University, Iran. She is currently working on the synthesis of artificial antibodies for drug and diseases biomarker determinations in complicated matrixes such as urine, saliva and plasma as well as drug delivery.
 

Abstract:

Sarcosine, a non-proteinogenic amino acid, is an intermediate product in the synthesis and degradation of amino acid glycine. Recently, it has been investigated as a prostate cancer (CaP) biomarker, a most common type of tumor disease in men. It is a candidate for diagnosis of early stages of CaP in the body fluid such as urine instead of prostate-specific antigen (PSA). PSA can only be detected in plasma when the disease progresses. Scientists have different opinion about its suitability as a biomarker in early diagnosis of CaP. This is due to false positive results because of interference from amino acid, aniline in the quantitation methods. Therefore, liquid chromatography or gas chromatography equipped with tandem mass detection is applied for its quantitation. For the accurate determination in biological matrixes such as urine, a good cleanup and pre-concentration technique is also required. Molecularly imprinted polymer as a synthetic antibody is a good approach. In the present survey, a new magnetic molecularly imprinted polymer (MMIP) using a chelate-Cu-sarcosine as the template, methacrylic amide as the monomer, ethylene glycol dimethacrylate as crosslinker and 2,2-azobis boutirnitril as an initiator is introduced. Synthesis of the MMIP was optimized by two heating methods, microwave irradiation and conventional heating. On column derivitization capillary electrophoresis analysis was used to determine 13 amino acids including sarcosine. Fig. 1 shows electrophoregram for the mixture of amino acids determined by the proposed method. In the electrophoregram, aniline and sarcosine are appeared at different times. So, the interference of aniline is eliminated.
 

Ishiaq Omotosho

University of Ibadan, Nigeria

Title: Area under the receiver–operating characteristics as a model for evaluating and predicting biomarkers of early renal tubular damage in subjects occupationally exposed to lead
Biography:

Abstract:

The incidence of kidney failure is on the increase, unfortunately, traditional renal function markers are equivocal especially at the early stage until end-stage renal disease when kidney transplant becomes inevitable. Hence, the need for an early and more sensitive marker of renal damage indicating the presence of covert renal damage in occupational lead toxicity is imperative. This work is proposing diagnostic methods that could predict the development of Chronic Renal Failure (CRF) especially in occupational lead-exposed subjects combining results of conventional and new biomarkers of kidney damage using a mathematical model based on Area under the Receiver Operating Characteristics (AUROC). Traditional Renal Function markers (TRF) (plasma creatinine, urea and uric acid) were determined in one hundred each of Lead-Exposed Subjects (LES) and non-exposed, non-nephrotic adults (control) along with sixty Chronic Renal Failure patients (CRF) (all age-matched) using standard spectrophotometric methods. Blood lead level (Pb) was determined in all participants using Atomic Absorption Spectrophotometry (AAS) while levels of novel urinary renal enzymes - Glutathione-S-transferase (GST) and N-acetyl-β-Dglucosaminidase (NAG)- activities were also evaluated using ELISA techniques. Pb was used as True Positive Indices (TPI) and TRF along with NAG and GST were used as False Negative Indices (FNI). Ratios of mean, Creatinine : GST (A) (0.01, 0.02 and 0.09), Creatinine:NAG (B) (0.03, 0.08 and 0.6), Uric acid : GST (C) (0.05, 0.08 and 0.08), Uric acid : NAG (D) (0.29, 0.3 and 0.55), Urea : GST (E) (0.17, 0.55 and 0.93), Lead : GST (F) (0.42, 0.59 and 0.88), Lead : NAG (G) (2.56, 2.28 and 6.09), Lead : Creatinine (H) (80.62, 30.37 and 10..22), Lead : Urea (I) (2.46, 1.07 and 0.95) and Lead : Uric acid (J) (8.66, 7.61 and 11.12) for LES, control and CRF groups respectively were computed and used to plot an ROC curve using the FNI values as the abscissa and the TPI values as the ordinate while their AUC were calculated. The AUC values for Lead : Creatinine, Lead Urea and Lead : Uric acid were 1.00, 0.917 and 0.833 respectively. We suggest that application of this model after proper standardization may be useful in early identification of covert kidney damage especially in occupationally vulnerable group.
 

Julio C Fernandez Travieso

National Centre for Scientifi c Research, Cuba

Title: Effects of policosanol in the functional recovery of ischemic stroke hypertensive patients
Speaker
Biography:

Julio Cesar Fernandez Travieso is a Senior Investigator in the Clinical Trials Unit, National Centre for Scientific Research, Havana, Cuba. He has completed his BSc in Pharmaceutical Sciences from Havana University, Cuba in 1996. He was awarded PhD in Pharmaceutical Sciences in 2003. He has published more than 130 publications and presented more than 100 papers in various scientific events. His research interest mainly focuses on clinical trials phase I-IV of different natural products: Policosanol, Abexol, Prevenox and Palmex.
 

Abstract:

Introduction: Stroke is one of the leading causes of mortality and disability. Clinical studies results show that policosanol (20 mg/day) + standard aspirin (AS) therapy had benefits versus placebo + AS given for 6 and 12 months to patients with recent ischemic stroke. Objective: To analysis, the policosanol treatment effects in the hypertensive patients included in two ischemic stroke recovery trials. Methods: This report was analyzed the records of all hypertensive patients included in two ischemic stroke recovery studies. Hypertensive patients with a modified Rankin Scale score (mRSs) 2 to 4 were randomized, within 30 days of onset, to policosanol/ AS or placebo/AS, for 6 months. The primary outcome was a mRSs reduction. Low-density Lipoprotein-cholesterol (LDL-C) reduction and High-Density Lipoprotein-Cholesterol (HDL-C) increase were secondary outcomes. Results: One hundred forty-two hypertensive patients (mean age: 66 years) were included in the analysis. Policosanol/AS decreased significantly the mRSs mean from the first interim check-up (3 months) (p<0.0001 vs. placebo/AS). The policosanol treatment effect did not wear off, on the contrary, even improved after 6 months therapy (p<0.0001 versus placebo/AS). Moreover, policosanol/AS (57/71; 80.3%) treatment achieved significant results (mRSs ≤1; p<0.0001). Whereas the placebo/ AS did not (6/71; 8.5%). Treatments were well tolerated. Two patients discontinued prematurely and four patients (2 from policosanol/AS group and 2 from placebo/AS) referred mild AE. Conclusions: Six months administration of policosanol/AS given to hypertensive patients after suffering ischemic stroke demonstrated to be better than placebo/AS in improving functional outcomes at 3 and 6 months when used among hypertensive patients with ischemic stroke.

Alina Vasilescu

International Centre of Biodynamics, Romania

Title: Biosensing approaches for lysozyme detection with graphene oxide-coated plasmonic interfaces
Speaker
Biography:

Alina Vasilescu has completed joint Ph.D. studies from the University of Bucharest, Romania and University of Perpignan, France and postdoctoral studies from University of Toronto, Canada. She has worked in analytical development in the pharmaceutical industry and is currently a researcher at the International Centre of Biodynamics in Bucharest, Romania working on practical applications of biosensors. She has published more than 30 papers in the field of biosensors.

Abstract:

Lysozyme is used as a model to study protein function and enzyme catalysis, is suggested as a biomarker in various diseases and also used as an antimicrobial agent in the food industry. Various methods have been reported for lysozyme detection based on its physicochemical properties, enzymatic activity or affinity for biological receptors. The aptasensors with detection by Surface Plasmon Resonance (SPR) developed by our group are versatile tools for the detection of residual lysozyme in wines or of lysozyme dimer in aggregated solutions. Advancing from these concepts relying on thiol coated plasmonic interfaces, we report the development of graphene oxide (GO) coated plasmonic interfaces via the layer-by-layer method, as robust and sensitive platforms with controlled thickness. Furthermore, the GO-coated interfaces were easily modified with whole cells of Micrococcus lysodeikticus- an enzymatic substrate for lysozyme. Detection of lysozyme in spiked serum samples was achieved on the principle of lysozyme’s lytic action causing desorption of bacteria from the interfaces and consequently changes in the SPR signal. The analysis time was 3 minutes and the detection limit was 3.5 nM. A second sensing concept exploited the affinity of lysozyme for an aptamer, fixed covalently to the GO-coated interfaces. In this case, a detection limit of 0.71 nM and a linear range of 2-21 nM were observed. The two analytical strategies are based on different sensing mechanisms, nonetheless, both are sensitive and easy to implement with GO-coated interfaces suggesting a high potential and versatility of these interfaces for bioanalytical purposes.