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CHROMATOGRAPHY AND SEPARATION SCIENCE

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SCIENTIFIC PROGRAM

Chromatography 2018

September 17-18, 2018 | Rome, Italy

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ICP-MS is a well-established analytical technique mainly applied to metal determinations both in inorganic chemistry and biological

samples (environmental and toxicological studies). The possibility of ICP-MS hyphenation to separation techniques, like HPLC and CZE, has shown interesting results in particular for the speciation of inorganic ions while applications in biological samples for pharmacological studies are rare. Along the years several pharmaceutical products containing metal, like iron, in high molecular weight complexes have been developed presenting difficult analytical problems especially in biological samples; indeed the determination of the metal itself is of modest values without concomitant information like molecular weight and polarity. In this context the combination with HPLC, permitting separations based on molecular weight, ionic strength and hydrophobicity, is an ideal tool to develop selective methods for such complex compounds. The possibility to correctly quantitate an element, irrespective of the chemical structure where is contained, is another interesting feature of ICP-MS and it offers interesting possibilities of metabolites quantitation and identification in biological samples of drugs containing not just metals but also sulfur and/or phosphor. A series of applications centered on these two classes of compounds will be presented showing the great potential of ICP-MS as a detector in combination with HPLC; as always nothing is perfect and therefore the limitations of this analytical approach will be discussed, in particular matrix effects and mobile phase compatibility.

BiographyDr. L Silvestro graduated in Medicine in Turin (Italy) in 1984 and specialized in Pharma-cology in 1988. From 1989 he is applying HPLC-MS in quantitation of bioanalytical samples as well as identification of drug metabolites. In 1990 he began to study the application of ICP-MS and other elemental analysis methods, hyphenated to HPLC, on bioanalytical samples. In 1996 he has co-founded 3S-Pharmacological Consul-tation & Research GmbH, a consultation company and CRO, in Germany and is still actively involved in the development of innovative analytical methods. From 1998 the company has expanded the activity in East Europe (Romania, Moldavia) cre-ating an analytical laboratory in Romania (Bucharest). In his scientific activity he has contributed to more then 60 articles in in-ternational scientific journals.

APPLICATIONS OF ICP-MS HYPHENATED TO HPLC ON BIOLOGICAL SAMPLES FOR PK STUDIES

SILVESTRO LGermany


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SCIENTIFIC TRACKS & ABSTRACTS

Principle of Chromatography | Application of chromatography | Advances in chromatography | Separation technique in Chromatography | Types of

chromatography techniques | Chromatography in analytical science

Title: Novel approach to prepare high quality photonic crystals for separation usageCHEN Yi, China

Title: High Performance Liquid ChromatographyWalter Jones, South Africa

Title: Applications of ICP-MS Hyphenated to HPLC on Biological Samples for PK StudiesSilvestro L, Germany

Title: Identifying, developing, and moving sustainable communities through application of bioenergy for energy Abdeen Mustafa Omer, UK

Title: Breadfruit (BUAH SUKUN) as nutraceutical Mukesh S. Sikarwar, Malaysia

Title: Nano chromatography and nano capillary electrophoresis: Need of this centuryImran Ali, Saudi Arabia

Title: Development of Non-contact Method for Autoclave Cure Monitoring of Carbon-Phenolic Composites by On-line Gas Chromatography TechniqueK.VeeraBrahmam, India

Title: Bioassay-Directed Isolation of Hypotensive Alkaloids from Holarrhena pubescensKhalid AFTAB, Pakistan

Title: Isolation of ulceroprotectivec cucurbitane type triterpenoids from cucumis melo seeds Gurpreet Singh Bal, India

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Online sample preparation method for complex sample analysis

Li Gongke

China

Professor Gongke Li obtained her PhD degree in analytical chemistry from Sun Yat-sen University of China in 1992. She then joined Sun Yat-sen University and became a professor since 2000. With expertise in chromatographic analysis and spectral analysis, her major research interests are focused on sample preparation techniques, analytical techniques for trace analysis of complex samples. She has published more than 350 research papers in reputed journals, 16 authorized invention patents and a book on sample preparation. She is currently the associate editor of Journal Separation Science and the member of editorial board of 15 academic journals including Journal of Chromatography A, Microchimica Acta and a member of the chemical teaching Advisory Committee for Colleges and Universities of the Ministry of Education. Science and Technology Awards and 2 honor prizes have been granted to her from the national and provincial governments of China. She was awarded the Chinese women's analytical chemist in 2015. She was also named as one of the 2016 Power List and the Top 50 most influential women in the analytical sciences in the world.

Sample pretreatment techniques have been regarded as the most important role of the whole analytical process. The online sample pretreatment mode is a promising way with the advantages of the automation, improvement of sensitivity and reduction of manual error. In this work, sample preparation media, including metal-organic frameworks, microporous organic polymers, graphene and molecularly imprinted polymers, were developed coupling to HPLC for online analysis of complicated samples. A NH2-MIL-53(Al)-polymer monolithic column was prepared and applied to in-tube SPME for online analysis of estrogens in urine. The enrichment factors were 180-304. A micro-solid-phase extraction (μ-SPE) column based on hydrazone linked covalent organic polymer was developed for online analysis of sudan dyes in chilli powder and sausage. The enrichment factors were 305-757.Acylhydrazone bond gel μ-SPE monolithic column was developed and applied to online enrichment and analysis of trace sulfonamides in weever and shrimp. The extraction capacity were 270-401 pmol. A monolithic column based on covalent cross-linked polymer gels for online extraction and analysis of trace aflatoxins in food sample was developed. The enrichment factors were 36-51. Acrylamide-modified graphene was applied to online μ-SPE for trace heterocyclic amines analysis in foods. The enrichment factors were 78-166. A molecularly imprinted monolithic column was developed for the online analysis of trace antimicrobials. The enrichment factors were 46-211. These online sample preparation methods were successfully applied to complex sample analysis.

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Novel approach to prepare high quality photonic crystals for separation usage

Chen YiChina

Chen Yi, received his Ph.D. degree from Institute of Chemistry, Chinese Academy of Sciences (CAS) in 1990, is presently PIs at the same institution and Beijing National Lab for Molecular Sciences, Chairing the Division Chemical Biology, and is Professor and Director of Teaching and Research Section at University of CAS. He has worked on capillary electrophoresis since 1984, later on chromatography and mass spectrometry, and surface plasmon resonance sensing and imaging since 1997, to have insights into the bio-functions of cells and biomolecules. Chen is presently Vice-presidents of several academic societies, Vice-directors of 4 Disciplinary/Specialized Committees at Chinese Chemical Society, a deputy editor of Electrophoresis and associate editors of Chinese Analytical Chemistry and Chinese Journal of Chromatography.

Photonic crystals (PCs) have ordered structure with optical band gap at the visible range or crystal lattice at the size of several hundreds of nano meters. PCs assembled from particles can offer pass-through nano-pores which, in theory, readily produce ultra-high performance of separation. Unfortunately, in practice their real applications were limited to the separation of a few compositions because they are hard to prepare unless highly mono-disperse nanopartciles are used that take time to prepare and cost high. In fact, mono-disperse particles do not necessarily uniformly order inside the assembled PCs but often separate into various domains. The domain borders can destroy the separation efficiency and easily cause clasps of PC-packed columns. Herein we present a novel approach to assemble PCs from polydisperse particles which are easily synthesized in a normal laboratory at a very low cost. The obtained PCs were measured to have sharp band gaps. They can be shaken in disarray but soon recover their ordered structure after settling for about decade seconds. Such a type of PCs can thus be made into various shapes so that they can be used to write iridescent letters, to paint colorful pictures, and of cause to fill capillary columns. The PC-filled capillary columns have been shown to yield fast (in seconds) and high separation efficiency in electro-chromatographic studies.

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High Performance Liquid Chromatography

Walter Jones

South Africa

Prof Jones studied Diploma in Military Medicine at the University of Moscow, Russia, continued at the University of Cambridge for Bachelor of Science in Medical Chemistry & Biology. He enrolled with Harvard University – Boston from the United States of America Completed Doctor of Medicine in Nuclear Medicine - MD. He further enrolled with the University of Johannesburg for extra-curricular and completed the course for One year Community Leadership before enrolling for Post-Doctoral, DSc in Energy Technology) from Anhalt University in German.

Abstract Liquid Chromatography and electrophoresis played a major role in the life science revolution, most strikingly in problem purification, peptide fractionation and sequencing, amino acid analysis, and DNA sequencing. The objective of this paper is to examine the potential role of separation systems in the continuing evolution of biochemistry and molecular biology, very small chip-based system may change how chemical analyst in biology, medical research and health care evolve over the next decade.

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Applications of ICP-MS Hyphenated to HPLC on Biological Samples for PK Studies

Silvestro L

Germany

Silvestro L graduated in Medicine in Turin (Italy) in 1984 and specialized in Pharmacology in 1988. From 1989 he is applying HPLC-MS in quantitation of bioanalytical samples as well as identification of drug metabolites. In 1990 he began to study the application of ICP-MS and other elemental analysis methods, hyphenated to HPLC, on bioanalytical samples. From 1998 the company has expanded the activity in East Europe (Romania, Moldavia) creating an analytical laboratory in Romania (Bucharest). In his scientific activity he has contributed to more then 60 articles in international scientific journals.

ICP-MS is a well-established analytical technique mainly applied to metal determinations both in inorganic chemistry and biological samples (environmental and toxicological studies). The possibility of ICP-MS hyphenation to separation techniques, like HPLC and CZE, has shown interesting results in particular for the speciation of in -organic ions while applications in biological samples for pharma -cological studies are rare. Along the years several pharmaceutical products containing metal, like iron, in high molecular weight comp -lexes have been developed presenting difficult analytical problems especially in biological samples; indeed the determination of the metal itself is of modest values without concomitant information like molecular weight and polarity. In this context the combination with HPLC, permitting separations based on molecular weight, ionic strength and hydrophobicity, is an ideal tool to develop selective methods for such complex compounds. The possibility to correctly quantitate an element, irrespective of the chemical structure where is contained, is another interesting feature of ICP-MS and it offers interesting possibilities of metabolites quantitation and identification in biological samples of drugs containing not just metals but also sulfur and/or phosphor. A series of applications centered on these two classes of compounds will be presented showing the great potential of ICP-MS as a detector in combination with HPLC; as alwa -ys nothing is perfect and therefore the limitations of this analytical approach will be discussed, in particular matrix effects and mobile phase compatibility.

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IDENTIFYING, DEVELOPING, AND MOVING SUSTAINABLE COMMUNITIES THROUGH APPLICATION OF BIOENERGY FOR ENERGY OR MATERIALS: FUTURE PERSPECTIVE THROUGH ENERGY EFFICIENCY

Abdeen Mustafa Omer

UK

Abdeen Mustafa Omer (BSc, MSc, PhD) is an Associate Researcher at Energy Research Institute (ERI). He obtained both his PhD degree in the Built Environment and Master of Philosophy degree in Renewable Energy Technologies from the University of Nottingham. He is qualified Mechanical Engineer with a proven track record within the water industry and renewable energy technologies. He has been graduated from University of El Menoufia, Egypt, BSc in Mechanical Engineering. His previous experience involved being a member of the research team at the National Council for Research/Energy Research Institute in Sudan and working director of research and development for National Water Equipment Manufacturing Co. Ltd., Sudan. He has been listed in the book WHO in the World 2005, 2006, 2007 and 2010.

The demand for energy continued to outstrip supply and necessitated the development of biomass option. Residues were the most popular forms of renewable energy and currently biofuel production became much promising. Agricultural wastes contained high moisture content and could be decomposed easily by microbes. Agricultural wastes were abundantly available globally and could be converted to energy and useful chemicals by a number of microorganisms. Compost or bio-fertiliser could be produced with the inoculation of appropriated thermophilic microbes which increased the decomposition rate, shortened the maturity period and improved the compost (or bio-fertiliser)quality. The objective of the present research was to promote the biomass technology and involved adaptive research, demonstration and dissemination of results. With a view to fulfill the objective, a massive field survey was conducted to assess the availability of raw materials as well as the present situation of biomass technologies. In the present communication, an attempt had also been made to present an overview of present and future use of biomass as an industrial feedstock for production of fuels, chemicals and other materials. We may conclude from the review paper that biomass technology must be encouraged, promoted, invested, implemented, and demonstrated, not only in urban areas but also in remote rural areas.

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BREADFRUIT (BUAH SUKUN) AS NUTRACEUTICAL

Mukesh S. Sikarwar

Malaysia

Mukesh S. Sikarwar is currently working as Senior lecturer in Faculty of Pharmacy, AIMST University, Malaysia. He received his PhD in Pharmacy from K.L.E. University, Belgaum India in year 2011. His current area of research is herbal antioxidants, antidiabetics and antihyperlipidemics, novel drug delivery in herbal drugs, formulation and evaluation of herbal drugs and food safety. He has more than 10 year of experience in mentoring students in Pharmacognosy and chemistry subjects. He has published 53 research and review papers in various peer reviewed internationals and national journals and also serving as reviewer for many international journals.

Breadfruit has potential for food and nutrition security and sustainable production in the tropics especially in Caribbean and ASEAN countries but it is under-utilised and commercial production is limited. It is rich source of nutrients and used as replacement for wheat and possess medicinal properties hence it has potential to be sold as nutraceutical. Over 130 phyto-compounds especially flavonoides and polypheols are identified in various parts of breadfruit plant. Nutritional compositions includes protein, carbohydrate, fat, calcium, phosphorus, K, iron, niacin, thiamine and vitamin C. Keeping above information in consideration nuratceutical product of soft gel capsules for breadfruit powder extract can be prepared and characterize for its marker compounds and antioxidant potential. It can be used as main source of antioxidant and will have application in treatment of oxidative stress caused by diabetes, inflammation, cancer and many such diseases. It can also be replacement for wheat flour, Gluten free, GMO free without trans-fat.

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Nano chromatography and nano capillary electrophoresis: Need of this century

Imran Ali

Saudi Arabia

Prof. Imran Ali, PhD, FRSC, C Chem, London is a world recognized academician and researcher. He completed his Ph.D from Indian Institute of Technology Roorkee, Roorkee, India. Prof. Ali is known globally due his great contribution in separation science, chiral drugs development and water treatment. He has published more than 380 papers in reputed journals including papers in Nature and Chemical Reviews of more than 41 impact factors. He has also written five books published by Marcel Dekker, Inc., USA; Taylor & Francis, USA; John Wiley & Sons, USA; John Wiley & Sons, UK; Elsevier, The Netherlands.

In spite of the curing properties of the drugs, these remain in our body tissues at very low concentrations for long time; leading to serious side effects. Similarly, some environmental pollutants exist at very low concentration in the earth’s ecosystem. The normal analytical instruments are not capable to detect them. Hence, we consider these species as absent, which is an illusion to the scientists and the society. The concentrations of some hormones, RNAs, DNAs, antibodies and other proteins are very low. Availability of infant plasma and cerebrospinal fluids is very poor. The proteomics and genomics researches are extremely difficult and need miniaturization of the separation techniques. Besides, the increasing economic pressure of the costly chemicals is a big issue globally nowadays. In view of these facts, there is big demand of microscale separations with low detection limits. Due these facts the micro-total-analysis system (µ-TAS) has been developed and is being used in few laboratories of the world. These are nano liquid chromatography (NLC) and nano capillary electrophopresis (NCE) techniques. The proposed lecture will highlight the importance of these techniques with special emphasis on the fabrication of microchip, instrumentation of NLC and NCE, detection, sample preparation, analyses and future perspectives

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Development of Non-contact Method for Autoclave Cure Monitoring of Carbon-Phenolic Composites by On-line Gas Chromatography Technique

K.VeeraBrahmam

India

K. Veera Brahmam, Indian engineer. Member of Indian Society Nondestructive Evaluation (life).Brahmam, K. Veera was born on May 15, 1975 in Hyderabad, India. Son of Kammari Seshaiah and Kammari Kasthuramma.Doctor of Philosophy, Sri Venkateswara University, Tirupati, 2003.Assistant professor Lenora College Engineering, Rajahmundry, India, 2000—2002. Scientist Advanced Systems Laboratory, Hyderabad, 2002—2009. First rank Sri Venkateswara University, Tirupati, Indonesia, 1997.With national cadet corps Indian Army, 1992-1995, Andhra Pradesh.

During aerodynamic loading of composite structures, experiences severe friction and high surface temperatures. In this context, carbon-epoxy shell is used as internal structural layer and carbon-phenolic shell is used as external layer for production of multi layered composite structure. Autoclave curing of phenolic composites involves a complex regime of time -temperature -pressure cycle where selection of pressure application point is the most important parameter, decides the quality of the component. Early pressure application with respect the gelation produces resin starved component, which degrade the thermal performance of the component whereas late pressure application produces more porosity in the component due to trapping of volatiles generated due to curing chemical reaction. Production of high resin content and minimum void distribution is most essential and which is achieved by analyzing the volatiles evolved during the process. During curing process, m-phenol and water are the one of the indicators and the concentrations are determined by suitable detectors as a function of temperature of the component. Based on the diminishing trend of the curves the gelation region for pressure application was determined. A new contact NDE method for cure monitoring of carbon-phenolic composites was developed right from development of concept, realization, development of proto type system for assessing the advancement of curing reaction and for determination of gelation region for pressure application. The system contains gas sample injection port, gas-chromatograph, detector unit, software for signal analysis. Finally the criteria for pressure application has been evolved.

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Bioassay-Directed Isolation of Hypotensive Alkaloids from Holarrhena pubescens

K Aftab

Pakistan

K Aftab, PhD male, Pharmacologist, graduated from department of Pharmacology, Faculty of Pharmacy, University of Karachi, Pakistan in 1995. He worked for Pharmaceuticals industry as Quality Control & Quality Assurance professional and was actively involved in Research & Development of Pharmaceutical preparations. He has worked in few Medical & Dental Colleges & Universities as Assistant, Associate and became full Professor Pharmacology in 2006 and worked as visiting Professor in different Universities & research institutions. From 2009-2011, he has worked in Kingdom Saudi Arabia as a full Professor Pharmacology and currently working as Professor & HOD Pharmacology in SMC, University of Health Sciences.

Holarrhena pubescens belongs to the family Apocynacea, commonly known as “kurchi” is highly reputed in traditional medicine as a remedy for amoebic dysentery and other intestinal ailment. Bioassay-directed fractionation by chromatographic methods the ethanolic extract of Holarrhena pubescens resulted in the isolation of steroidal alkaloids i.e. Holamide and Pubscinine. Holamide showed a three proton doublet at 1.45 (J=6.56 Hz) and two AB doubles at 3.17 and 3.00 each for on proton (J=12.06 Hz) in the 1H NMR spectrum suggested that it belongs to conanine series of alkaloid (A class of compound with the steroid nucleus and a five members heterocyclic ring with nitrogen). In contrast Pubscinine showed one methyl at 1.28 while the doublet is missing a three proton singlet was observed at 2.28 due to a vinylic methyl indicated a double bond in the 18,20 – epimino ring of the conanine series of alkaloids. In anaesthetized rats, the Holamide and Pubscinine caused a fall in blood pressure in a dose-dependent manner. Pretreatment of animals Atropine completely abolished the hypotensive response of Acetycholine; whereas hypotensive effect of Holamide and Pubscinine were not modified by Atropine. Similarly Acetylcholine produced contractile effect in guinea-pig ileum, which was antagonized by atropine, however both (Holamide and Pubscinine) failed to produced any stimulant response on guinea-pig ileum. These data indicate that the steroidal alkaloids i.e. Holamide and Pubscinine from Holarrhena pubescens mediated hypotensive response through a mechanism different to that of Acetycholine.

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ISOLATION OF ULCEROPROTECTIVEC CUCURBITANE TYPE TRITERPENOIDS FROM CUCUMIS MELO SEEDS

Gurpreet Singh Bal

India

Gurpreet Singh Bal completed his post graduation in April 2003 with Pharmaceutical Chemistry from Punjab Technical University, Jallandhar. Now working as an Assistant Professor in Rayat Institue of Pharmacy, India since Jan 2007 to till date.

Medicinal plants are the richest bio-resources of drugs in traditional medicinal systems, modern medicines, folk medicines, intermediate and chemicals entitled for synthetic drugs. Plants provide a source of inspiration for novel drug development as they contain a vast array of substances that treat chronic diseases. Cucumis melo seeds have been traditionally used for treating various health ailments. The main aim of our current study is to isolate Cucurbitanetype triterpenoids from Cucumis melo seed extract and conduct antiulcerogenic activity of the isolated compound. Phytochemical investigations of methanolic seed extract of Cucumis melo was carried out which showed the presence of various important phytoconstituents. The main active constituents of Cucumis melo have shown a number of potent pharmacological activities. The isolation of Cucurbitane-type triterpenoids was carried out by column chromatography using methanolic seed extract of Cucumis melo. Mobile phase hexane and hexane-ethyl acetate (98:2) was used to run the column. TLC profiling was done simultaneously in an appropriate solvent system (hexane: ethyl acetate, 97:3). Various fractions were collected. The fractions with similar Rf value were pooled together. Fractions giving single spot in the TLC were regarded as pure. The isolated compound showed positive result for Liebermann-buchard test from which we can conclude that the isolated compound might be triterpenoid. The structure of the isolated compound was determined by IR, 1HNMR, 13CNMR techniques. The spectral analysis of the isolated compound showed following results: IR: It showed the peaks at 3383, 2976, 2814, 1721, 1465, 1123 cm-1 indicated the presence of alcoholic group. 1H NMR (400 MHz, CDCl3): δ 0.66-1.29 (m, 24H, -CH3), δ 1.32-1.38 (m, 4H, H7, H8, H9, H10), δ 1.40-1.51 (m, 4H, H10,H19, H20, H21), δ 1.52-1.59 (m, 3H, H11, H6, H22), δ 1.61-2.38 (m, 2H, H4, H3), δ 3.16-3.20 (m, 6H, H1, H2, H12, H13, H15, H17). 13C NMR (400 MHz, CDCl3): δ 15.99, 16.13, 18.01, 18.33, 19.32, 20.94, 25.16, 27.43, 27.46, 28.00, 29.71, 29.86, 34.30, 35.60, 37.18, 38.07, 38.73, 38.87, 40.02, 40.85, 42.84, 43.01, 47.99, 48.32, 50.45, 55.32, 79.00, 109.34, 109.67 (C=C), 150.96 (C=O). From the above result, the isolated compound was elucidated to be tetracyclic triterpenoid. As triterpenoids are mostly responsible for anti-ulcerogenic activity so the isolated compound was further evaluated for antiulcer activity by pyloric ligation induced gastric ulcer, water immersion stress ulcer and indomethacin induced ulcer models in wistar albino rats. In the pyloric ligation induced gastric ulcer model, the isolated compound at the dose of 300 mg kg-1 showed significant reduction in gastric volume, free acidity and total acidity i.e., 1.79±0.12, 31.58±0.31 and 72.95±0.11 respectively. The percentage inhibition was found to be maximum at the dose of 300 mg kg-1 in all the three animal models. The percentage inhibition was 56.6, 66.3 and 61.2 in pyloric ligation induced gastric ulcer, water immersion stress ulcer and indomethacin induced ulcer models respectively. All the above gathered results the isolated compound i.e., Cucurbitane-type triterpenoids was found to be potent against gastric lesions and therefore can be used as future natural antiulcerogenic agent.

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The Role of Programmed Cell Death 'Apoptosis' in the Development of Inner Sulcus in the Cochlea

Tarfa M. Peter

Nigeria

Tarfa M. Peter has completed his M.Sc. from University of Maiduguri, Borno State, Nigeria. He is the Histology lecturer, in the Department Human Anatomy, Gombe State University. He has published more than 10 papers in reputed journals and is presently undergoing his Ph. D in Ahmadu Bello University Zaria.

Hearing loss is one of the most common chronic diseases that affect both young and old but it is most prevalent in old people. This condition is generally irreversible in humans and can be due to the loss of hair cells, which are unable to regenerate. However, recent evidence of some regenerative ability reported in a number of non-mammal vertebrates have given us hope that, in the future a solution may be discovered. Although several advances have been recorded in this field in recent times and ere are still challenges ahead. This study tried to investigate he formation of the inner sulcus located in the cochlea, as it is thought that, the processes involved during the development of this important region are most likely due to apoptosis or another type of programmed cell death, although this has not yet been confirmed. Mouse expressing an EGFP (green fluorescent protein) reporter at the Tecta locus was used. Specimens were stained with phalloidin as a general cell stain of f-actin and this was combined with (Terminal deoxynucleotidyl transferase dUTP Nick End) TUNEL staining in order to observe whether dying cells are the result of programmed cell death. Very little TUNEL staining was observed in the developing sulcal region, although some were seen in the associated mesenchymal cells in the cochlea. In some of the sections, Blebbing as well as extrusion of some cells that are thought to be undergoing programmed cell death were evident during the formation of the sulcus. The formation of the sulcus occurs earlier in the basal region of the cochlea than in the apical part following the regression of the greater epithelial ridge (GER) cells. Counting of nuclei in the sulcal region during the formation suggest that cells are being lost.It is not easy to establish whether these cells that are being removed could be due to apoptosis or another type of programmed cell death.

High Performance Liquid Chromatography | Chromatography in separation science | Ion exchange chromatography | Multidimensional chromatography Advances in gas chromatography | Market growth of chromatography | Biomedical chromatography | Extraction chromatography |

Partition chromatography | Adsorption chromatography

Title: The Role of Programmed Cell Death 'Apoptosis' in the Development of Inner Sulcus in the CochleaTarfa M. Peter, Nigeria

Title: Development of Dispersive liquid-liquid microextraction procedure for trace determination of pesticide malathion in urine samplesSeyed Jamaleddin Shahtaheri, Iran

Title: Dispersive liquid-liquid microextraction in chromatographic analysis of drug of abuseRajeev Jain, India

Title: Characterizations of the Effect of alkaline earth cations on Amlodipine Besylate and Croscarmellose sodium drug-excipient interactionPrasad Panzade, Canada

Title: Toxicity of organometallic compounds with medical extractedNagham Mahmood Aljamali, Iraq

Title: Chromatographic methods developed for purity characterization natural compounds with applications in analytical research of biotechnologyBUTNARIU Monica, Romania

Title: Abuse Drugs by Gas Chromatography Mass SpectrumRaafat Abdeldayem, Egypt

Title: RegenerAge System: Therapeutic effects of combinatorial biologics (mRNA and allogenic MSCs) with a spinal cord stimulation system on a patient with spinal cord section. Joel I. Osorio, Mexico

Title: Efficiency of Ionic-MOFs in ion-exchange applicationAli Morsali, Iran

Title: Comparative pharmacokinetics of trandolapril, its active metabolite, and verapamil in human plasma of Egyptian population using HPLC–MS/MS Ahmed Hemdan, Canada

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Development of Dispersive liquid-liquid microextraction procedure for trace determination of pesticide malathion in urine samples

Seyed Jamaleddin Shahtaheri

Iran

Professor Seyed Jamaleddin Shahtaheri has completed his PhD at the age of 38 years from Surrey University, Guildford, Surrey, England in 1996. He is an academic member of Department of Occupational Health Engineering, Tehran University of Medical Sciences, Iran, acting as the Dean Research Deputy at Institute for Environmental Research the same University too. Dr Shahtaheri is the Persistent Organic Pollutant Review Committee (POPRC) Member under the Stockholm Convention, UNEP, UN. He has published more than 150 papers in reputed journals and has been serving as an editorial board member of 7 national and International Journals.

Background: Measurement of pesticides in biological matrices is a serious challenge for researches because of their very low concentration in different matrices. The aim of this study was to develop a new sample preparation method with high accuracy and validity, simplicity as well as a short retention time for chromatographic determination of pesticide malathion. Methods: Dispersive liquid-liquid micro-extraction (DLLME) technique coupled with high performance liquid chromatography equipped with ultra violet detector (HPLC-UV) was developed for trace extraction and determination of pesticide malathion in human urine samples. One variable at a time (OVAT) method was used to optimize parameters affecting the malathion extraction. Different parameters such as extraction solvent, disperser solvent, and volume of the extraction solvent, volume of the disperser solvent, centrifugation time and speed, salt addition, and sample pH were studied and optimized. Results: Under the optimized conditions, the limit of detection and enrichment factor of the developed procedure were 0.5 µg L-1 and 200, respectively. The calibration curve was linear in the concentration range of 2-250 µgL-1. The relative standard deviation for six replicate experiments at 200 µgL-1 concentration was less than 3%. The relative recoveries of spiked urine samples were 96.3%, 101.7% and 97.3% at three different concentration levels of 50, 200, and 1000 µgL-1, respectively. Conclusion: According to the obtained results, DLLME procedure was successfully developed for the extraction of malathion from human urine samples. Compared to other extraction techniques, the proposed procedure had some advantages such as shorter extraction time, better reproducibility, and the higher enrichment factor. Keywords: Malathion, dispersive liquid–liquid microextraction, one variable at a time, urine, HPLC.

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Dispersive liquid-liquid microextraction in chromatographic analysis of drug of abuse

Rajeev Jain

India

Rajeev Jain has completed his PhD at the age of 28 years from CSIR-Indian Head of the Department of Toxicology Division at Central Forensic Science Laboratory, Guwahati (India), a premier Forensic Laboratory under Directorate of Forensic Science Services, Ministry of Home Affairs, Govt. of India. His research work focuses on development of green and cost-effective analytical methods for the analysis of various drugs and toxicants. He has published more than 20 papers in reputed journals and has been serving as a referee of various journals of repute.

Analysis of drugs of abuse in biological matrices is always been a challenging task for forensic chemists due to availability of limited amount of sample and unknown nature of analyte. Generally, traditional sample preparation methods such as liquid-liquid extraction (LLE) and solid-phase extraction (SPE) are employed for the extraction of drugs of abuse from biological matrices which are time consuming, labor intensive, costly and consumes large volumes of toxic organic solvents. In recent years, dispersive liquid-liquid microextraction (DLLME) has attracted the interest of researchers because of being simple, fast, inexpensive and green with high enrichment factors and extraction efficiencies. Beside of this, DLLME can be coupled with majority of chromatographic instrument such as GC-MS, GC-FID, GC-ECD and HPLC. Due to aforesaid advantages, DLLME has been widely applied for the determination of drugs of abuse such as amphetamines, barbiturates, benzodiazepines, cannabinoids, cocaine, opiates, hallucinogens, anti-depressants etc. Moreover, the capability of DLLME of simultaneous derivatization and extraction of polar drugs and coupling with injector port silylation (IPS) makes it a method of choice of sample preparation for GC-MS analysis of drugs of abuse. The present talk will be focused on applications, operating procedures and future trends of DLLME for chromatographic analysis of various classes of drugs of abuse.

Chromatography 2018

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Rome, Italy



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World Congress on


Characterizations of the Effect of alkaline earth cations on Amlodipine Besylate and Croscarmellose sodium drug-excipient interaction

Prasad Panzade

Canada

Prasad Panzade is an executive with a broad vision and strategic entrepreneurial mind set with over 23 years of experience in the multidimensional chemical industries including the Pharmaceutical and biopharmaceuticals. Dr. Panzade holds a Ph.D. degree in Pharmaceutical sciences with specialization in the field of Pharmaceutical Analytical Sciences and Technology. Dr. Panzade is a speaker who provides active insights on the latest in the fields of analytical sciences. He has several publication and patients to his credits.

Making a stable drug formulation is a challenging task in many instances with respect to compatibility studies. The compatibility study between Croscarmellose (an excipient) and Amlodipine Besylate is studied during the stable formulation drug development of Amlodipine Besylate and is discussed in this presentation. Drug-excipient interaction between primary amine drug substances and Croscarmellose sodium is described in different literature sources and confirmed experimentally in our laboratory. The effect of alkaline earth cation that added to the sample solvent in the form of Calcium acetate salt was discovered and studied in our laboratory to reduce such interaction and improve the recovery of the drug substance during sample preparation for assay determination by reversed phase HPLC. The interaction mechanism and determination of optimal concentration of Calcium Acetate in the sample solvent was determined. The determination was based on Amlodipine Besylate assay sample preparation, in the presence of Croscarmellose Na for achieving the complete recovery of Amlodipine Besylate from the sample matrixes.

Chromatography 2018

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Rome, Italy



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World Congress on


Toxicity of organometallic compounds with medical extracted

Nagham Mahmood Aljamali

Iraq

Dr.Nagham is a member of more than 35 scientific world journals, has published two books in Germany house of publication. Nagham Mahmood has two printed books in Chemistry field. Dr.Nagham Mahmood has PhD – certification in Organic Chemistry. She is working as an Assist. Professor, Department of Chemistry, IRAQ.

Many bio-chemical compounds were prepared for bio-experiments experiments for biochemistry studying like anti fungi, antibacte rial, tissue of animals. The structure of all formatted compounds were characterized by using many techniques((FT.IR,1H.NMR, Mass Spectra, Chromatography Measurement-Thermal-Measurements)) then studying of physical and chemical characterization and other chemical studies.

Chromatography 2018

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Name & Country

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Rome, Italy



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World Congress on


Quick HPLC-PDA Method for Simultaneous Determination of Five Selected Quinolones in Commercial Pharmaceutical Formulations

Muhammad S. Tahir

Pakistan

Muhammad S.Tahir is highly qualified quality assurance manager with more than sixteen years of experience in the field. He has experience in quality control, quality improvement and quality management.He Possess good communication and problem solving skills. He Practice and knowledge of specialized laboratory equipments and techniques: Spectroscopy; FT-IR, UV-Vis, Atomic (Absorption and Emission) SpectroscopyChromatography; GC, HPLC, UPLC, GCMS, LCMS, TLC etc.NMR; (1H and 13C)Electroanalytical techniques; Potentiometry, Conductometry etc.CHNS Elemental Analyzer Single Crystal X-Ray Diffractometer Gravimetric determinationKarl Fischer water determination Particle Size Analysis Polarimeters Titrations Heavy Metal Analysis

A rapid and simple isocratic liquid chromatographic method has been developed, optimized and subsequently validated for the simultaneous separation and unified quantification of five frequently prescribed quinolones, namely, pipemidic acid, ciprofloxacin hydrochloride, gemifloxacin mesylate, levofloxacin hemihydrate and moxifloxacin hydrochloride in commercial pharmaceutical formulations. The chromatographic resolution of selected quinolones was done employing Purospher Star RP-18 (4.6 mm × 25 cm, 5 μm) column maintained at 40 °C and a detector set at preferential detection wavelength of 278 nm. The optimized mobile phase was comprised of tri-sodium phosphate buffer (0.02 M), methanol, acetonitrile and methane sulfonic acid pumped at a ratio of 50:25: 25:0.1 (v/v). The mobile phase flow rate employed was 1 mL/min and run time of analysis was 8 min. The calibration curve was linear from 24-60 μg/mL for pipemidic acid and moxifloxacin, 15.0-40.0 μg/mL for ciprofloxacin and levofloxacin, and 19.20-51.20 μg/mL for gatifloxacin, with a coefficient of determination ≥ 0.9996, while repeatability and intermediate precision (expressed as relative standard deviation) were ≤ 1.91 and 1.65 %, respectively. The proposed HPLC method was rapid and simple for the determination of selected quinolones in commercially available tablet formulations providing recoveries ≥ 99.31 %. Validation studies proved the precision and accuracy of the method in the established linearity range. Complete separation of analytes in the presence of placebo provided the justification of method specificity.

Chromatography 2018

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Rome, Italy



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World Congress on


Chromatographic methods developed for purity characterization natural compounds with applications in analytical research of biotechnology

BUTNARIU Monica

Romania

Monica BUTNARIU has completed her PhD in 2005 (Banat University, RO), since 2013 is habilitated doctor from Cluj Napoca University, RO. She is the professor of University of Agricultural Sciences and Veterinary Medicine of Banat "King Mihai I of Romania" in Timisoara, 300645, Romania (Chemistry & Biochemistry Discipline). She has numerous articles published in the Science Citation Index (SCI) / International Scientific Indexing (ISI) / Web of Science Core Collection-Clarivate Analytics that have been cited over 1200 times, and her publication H-index is 25 (Citation Metrics) and she has been serving as an editorial board member of reputed Journals and she has peer review activity. So far, he has written several books and book chapters in different areas of chemistry. It should be noted that currently is head of the Head–CONEM Romania Biotechnology and Environmental Sciences Group and also National Correspondent of the International Association for Plant Biotechnology.

Chromatographic methods could solve the problem of purity, natural compounds but also the stability of organic molecules, which has an important role in predicting the stability of the active substance, which will provide an important response to analytical research in biotechnology. All this demonstrates that the properties that matter, are considered from the perspective of the role in predicting the efficacy of the active substance and, also, in establishing the optimal biotechnological form. Other considerations on purity assessment, are determined by methods such as HPLC / GC-MS, to determine the specified and non-specified impurities. In this context, the amount and number of methods that can be developed, is very high due to the complexity of the studies involved. One consideration is based on the ability of the active substance and related substances to produce a characteristic signal. The physicochemical properties of the active substance play an important role in the development and specificity of chromatographic methods.

Chromatography 2018

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Name & Country

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Rome, Italy



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World Congress on


Abuse Drugs by Gas Chromatography Mass Spectrum

Raafat Abdeldayem

Egypt

Dr. Rafaat is an associate Professor of Toxicology at the Emergency Hospital in the Faculty of Medicine at Mansoura University, Egypt. He obtained a PhD of hydro geochemistry in 2004, at Mansoura University and previously completed an MSc in hydro geochemistry in 2001 at the same institution. He also has a Diploma of toxicology and forensic chemistry (1996) from the Faculty of Medicine and a Diploma of applied chemistry (1993). Dr. Rafaat specializes in medical analysis, toxicology & forensic chemistry as well as water pollution. He has published papers over 10 local paper and 13 in international publications. He has spoken at 55 local conferences and workshops and 15 international meetings. He is currently a member of eight international scientific societies.

Background: Drugs of abuse are any drug or substance which if taken by any route (oral-snuffing-injection) will lead to mood change, psychological disturbance and can affect brain functions and level of perception. The aim of this work is to evaluate the usefulness of using urine immunoassay kits for detection of some drugs of abuse, and study the correlation between the results obtained by EMIT and GC / MS techniques. Materials & Methods: Drug toxicology tests are most commonly performed on urine, since most drugs and their breakdown products are excreted in the urine at higher concentration. A total number of 449 inhabitant urine samples were collected from patients admitted to emergency hospital, Mansoura University. 449 urine samples were analyzed by EMIT and GC/MS for benzodiazepine, barbiturate, opiate and cannabinoid. Results: Revealed that urine immunoassay kit is useful for rapid preliminary screening of abuse drug. GC/MS results confirm that 245 samples (54.56 %) are positive of the total number of samples. These positive samples by GC/MS were as follows; benzodiazepines; 159 (clonazepam, oxazepam, temazepam), barbiturates; 58 (thiobarbiturate, butabarbital, seconal) and opiates; 28 (methadone metabolite). Conclusions & Recommendations: GC/MS analysis must be done for accurate identification and confirmation of EMIT results. In addition, it is recommended as the most suitable technique for obtaining optimum analytical results.

Chromatography 2018

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Rome, Italy



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World Congress on


Chemical characterization of polyphenols from Daucus muricatus growing in Algeria by RP-UHPLCESI-QTOFMS/MS

Salah Akkal

Algeria

Salah Akkal received his Ph.D. in Organic Chemistry from University of Constantine, Algeria in 2001. He was supervised on 4 M.Sc. Theses and 8 Ph.Ds. He is professor in natural product chemistry since 2007 in Chemistry Department, Faculty of science. He has published more than 90 papers in reputed journals and has been serving as an editorial board member of repute and has been serving as a reviewer for many articles in her specialization. He attended more than 30 International Conferences He teaching all the organic chemistry and natural product courses for the students of Faculty of Science and Faculty of Technology in University of Constantine, Algeria.

In the present work, reversed phase (RP) ultra-high-performance liquid chromatography (UHPLC) coupled to quadrupole-time-of flight (QTOF) mass spectrometry in tandem has been used for the identification of the main phenolic compounds in the aerial parts of Daucus muricatus. The characterization of the compounds was carried out taking into account retention time, mass accurate measurements, the generated molecular formulae and fragmentation pattern. These data were contrasted with literature and databases, as well as with those of authentic standards when possible. The proposed method provided tentative identification of 30 phenolic and other polar compounds, including hydroxycinnamic acid derivatives, flavonoids and hydroxybenzoic acid derivatives. As a note, hydroxycinnamic acid derivatives were found to be the most diverse phenolic class of Daucus muricatus.

Chromatography 2018

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Name & Country

Abstract

Rome, Italy



Y E A R S


World Congress on


RegenerAge System: Therapeutic effects of combinatorial biologics (mRNA and allogenic MSCs) with a spinal cord stimulation system on a patient with spinal cord section.

Joel I. Osorio

Mexico

He is CEO and Co- Founder of Biotechnology and Regenerative Medicine at RegenerAge International. Vice President of International Clinical Development for Bioquark, Inc. and Chief Clinical Officer at Re-Anima Advanced Biosciences. Advance Fellow by the American Board of Anti-Aging and Regenerative Medicine (A4M), Visiting Scholar at University of North Carolina at Chapel Hill. Fellow in Stem Cell Medicine by the American Academy of Anti-Aging Medicine and University of South Florida.

As it has been previously demonstrated that coelectroporation of Xenopus laevis frog oocytes with normal cells and cancerous cell lines induces the expression of pluripotency markers and in experimental murine model studies that mRNA extract (Bioquantine) purified from intra and extra-oocyte liquid phases of electroporated oocytes) showed potential as a treatment for a wide range of conditions, including Spinal Cord Injury (SCI) among others. The current study observed beneficial changes with Bioquantine administration in a patient with a severe SCI. Pluripotent stem cells have therapeutic and regenerative potential in clinical situations CNS disorders even cancer. One method of reprogramming somatic cells into pluripotent stem cells is to expose them to extracts prepared from Xenopus laevis oocytes. The positive human findings for spinal cord injury with the results from previous animal studies with experimental models of traumatic brain injury and SCI respectively as our evidence and due to ethical reasons, legal restrictions and a limited number of patients, we were able to treat only a very small number of patients, deciding to include in our protocol the RestoreSensor SureScan to complete it. Based on the electrical stimulation for rehabilitation and regeneration after spinal cord injury published by Hamid and MacEwan, we designed an improved delivery method for the in-situ application of MSCs and Bioquantine in combination with the RestoreSensor SureScan. To the present day the patient who suffered a complete section of spinal cord at T12-L1 shows an improvement in sensitivity, strength in striated muscle and smooth muscle connection, 13 months after the first treatment and 6 months after the placement of RestoreSensor at the level of the lesion, showing an evident improvement on his therapy of physical rehabilitation (legs movement) on crawling forward and backwards and standing on his feet for the first time and showing a progressively important functionality on both limbs.

Chromatography 2018

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Rome, Italy



Y E A R S


World Congress on


Efficiency of Ionic-MOFs in ion-exchange application

Ali Morsali

Iran

Ali Morsali has completed his PhD in Inorganic Chemistry with Dr. Ali Reza Mahjoub in 1999-2003,Tarbiat Modares University (TMU), Tehran, Iran. He is the professor of Tarbiat Modares University, IRAN. He has over 500 publications that have been cited over 10000 times, and his publication H-index is 50 and has been serving as an editorial board member of some journals.

Ionic porous metal–organic frameworks (MOFs) materials consisting of porous anionic or cationic frameworks and cationic or anionic guests. The structure and performance of cationic and anionic MOFs are influenced by several factors, such as organic ligands, metal ions, and charge-balancing anions. These types of anionic and cationic materials have distinct advantages in comparison with neutral MOFs in the design of functional materials, and their synthesis affords several distinct advantages over the routine neutral frameworks by virtue of the isolated charged species in confined nano-spaces. The nanosized and charged pores in these ionic structures make a strong interaction between the host and guest molecules including enhanced adsorption towards small gases and solvent molecules. Via ion exchange, Ionic-MOFs can be easily modified and accommodate other charged guest molecules, making them an ideal platform for different applications such as functional materials. Ion-exchange chromatography and ion-exchange solid phase extraction based on ion-exchange resins are known as the first generation of powerful tools for the separation of charged molecules, so the researchers are interested to find new materials with higher performance in their ability for the ions separation. Another separation method, size-exclusion chromatography, is based on the size or molecular weight of the analytes. Although these methods are especially useful, but they are used only for separation of very large species, such as proteins and polymers. But, by mixing the unique structural of MOFs, especially their porous structures with the efficiency of ion chromatography, it is possible to develop a novel separation system, in which charged MOFs replace the conventional ion-exchange resins. The structural characteristics of porous MOFs such as precisely defined pores can result in useful properties including size-exclusion effect for guest molecules in a size regime that cannot be achieved by conventional ion-exchange resins

Chromatography 2018

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Name & Country

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Rome, Italy



Y E A R S


World Congress on


Comparative pharmacokinetics of trandolapril, its active metabolite, and verapamil in human plasma of Egyptian population using HPLC–MS/MS

A. Hemdan

Canada

Dr. Ahmed Hemdan has completed his PhD at the age of 30 years from Ain Shams University, Egypt. He is now a post-doc researcher in University Medical Center Hamburg-Eppendorf, Germany. He has over 20 publications that have been cited over 60 times, and his publication H-index is 4 and has been serving as an editorial board member of reputable Journals.

Trandolapril and verapamil are commonly used antihypertensive drugs. However, there is a lack of available data on the change of their pharmacokinetics in patients with liver or kidney impairment and hence the need for dose adjustment. In this article, a high performance liquid chromatography–tandem mass spectrometry (HPLC–MS/MS) method was developed and validated for the monitoring of trandolapril, its active metabolite trandolaprilat, and verapamil in human plasma of patients with renal impairment and/or liver insufficiency. The chromato -graphic separation was achieved on a Gemini C18 reversed phase column using a gradient elution mode with a run time of 10 minutes. The mobile phase consisted of a mixture of methanol and 2% acetic acid. The electrospray ionization MS/MS analysis was performed in multiple reaction monitoring (MRM) mode. The assay was validated as per Food and Drug Administration (FDA) guidelines for bioanalytical method validation and proved to be suitable for the determination of therapeutic drug levels in plasma. The inter‐group changes in pharmacokinetic data were compared to that of healthy volunteers. The comparison showed a significant difference in the pharmacokinetic parameters between the studied groups. The presented results exhibit the benefits of the proposed assay as a validated analytical tool for the continuous drug monitoring.

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