Conference Schedule

Day1: March 12, 2018

Keynote Forum

Biography

Gary W Caldwell received his PhD in Physical/Organic Chemistry from Indiana University in 1982 under the direction of Dr. John E Bartmess using Ion Cyclotron Resonance Spectrometry to study gas-phase ion chemistry. Following his PhD, he performed Postdoctoral Research with Dr. Paul Kebarle at the University of Alberta in Canada using High-Pressure Mass Spectrometry to study gas-phase ion chemistry. He joined Janssen Pharmaceutical Research and Development, a subsidiary of Johnson & Johnson in 1985. During his 32-year career with Janssen Pharmaceutical R&D, he has managed a variety of functions within the drug discovery units. These functions include the NMR, GC/MS & LC/MS/MS pectroscopy group, the medicinal chemistry intermediates group, the large-scale separation group, the drug discovery in-vivo/in-vitro PK/ADME groups and the compound management group. His research interests primarily involve the use of advanced pectrometric and chromatographic techniques to chemically and biologically characterize new drug targets and drug entities. Presently, he is working on establishing targeted and untargeted metabolomic methods to understand “on” and “off” target effects to improve drug efficacy and reduce drug toxicity. He is the author of over 150 publications, two patents, over 50 poster presentations, and has given over 40 invited talks at universities, companies, and conferences. He is the Co-editor of Frontier in Drug Design & Discovery (volumes 1-4), and Editor of Optimization in Drug Discovery: In-vitro Methods (volumes 1-2). 

Abstract

Tracer metabolomics is a LC/MS/MS approach for simultaneous examination of multiple metabolic pathways by determination of deuterium enrichment in metabolites in the presence of deuterium oxide or by following the metabolism of isotopically stable molecules. By introducing a tracer into a biological system, pathways including gluconeogenesis, glyceroneogenesis, fatty acid/phospholipid/ triglyceride/cholesterol ester synthesis and many others can be followed for assessment of target engagement, mechanism of action and pharmacological effect. Several LC/MS/MS assays will be discussed that demonstrate the utility of tracer metabolomics in a drug discovery environment.

Biography

Magnus S Magnusson is a Research Professor of University of Iceland. He completed his PhD from the University of Copenhagen. He is the author of the T-pattern model and the corresponding detection algorithms in the THEMETM (PatternVision). He has focused on real-time organization of behavior, co-directed DNA analysis, published numerous papers and given invited talks and keynotes at international conferences in ethology, psychology, neuroscience, mathematical sciences, science of religion, proteomics and mass spectrometry, and at universities in Europe, USA and Japan. He has served as Associate Professor and Deputy Director (1983-1988), Anthropology Laboratory, Museum of Mankind, Museum of Natural History, Paris. He has been a repeatedly invited Professor in Psychology and Ethology (the biology of behavior) at the University of Paris, V, VIII and XIII. Since 1991, he has been the Founder and Director of the Human Behavior Laboratory (hbl.hi.is), University of Iceland. Since 1995, he is in collaboration between 24 universities on Methodology for the Analysis of Social Interaction (MASI) initiated at t

Abstract

With its roots in studies of social organization and communication in social insects and primates, this talk is about similarities and especially two kinds of self-similarity. The first concerns a type of pattern, named T-pattern, originating in ethological (biology of human behavior) work in the 70’s regarding children’s behavior and interactions when video recording and access to computers was becoming increasingly available; also, while behavioral and statistical methods of analysis were from earlier times of scarce computational power. The T-pattern and the corresponding dedicated T-pattern detection algorithm and software, THEMETM, was an answer to this situation around 1980s. It has since become clear that the T-pattern can be seen as a type of statistical pseudo or natural fractal. However, recently the whole universe out to its largest known structures has been shown to have a fractal and thus self-similar distribution. Applying T-pattern detection to many kinds of interactions in humans and animals and then between neurons in neuronal networks in rat brains showed an abundance of this kind of patterning at extremely different scales of size and time. When shifting to DNA analysis, the same kind of patterning is encountered and is the direct basis for, among other, motor proteins, behaving and working in “Cell Cities”, that is, proteins mass-societies, while human societies are the only, and biologically very recent, big-brain mass-societies. Could the detection and study of proteins thus provide keys to the understanding of human social phenomena, possibly including religion?
 

Tracks

  • New Advances and Development in Mass Spectrometry | Mass Spectrometry Applications | Mass Spectrometry in Pharmaceutical Industry | Spectroscopy | Mass Spectrometry Applications in Clinical Diagnostics | Capillary Electrophoresis | Tandem Mass Spectrometry | Environmental Analysis
Location: Armstrong
Speaker

Sherry A Tanumihardjo

University of Wisconsin, USA

Chair

Speaker

Lingzhi Gong

Guangzhou University of Chinese Medicine, China

Co Chair

Biography

Tanumihardjo studies vitamin a and carotenoid metabolism, serves as director of the undergraduate certificate in global health, and teaches at undergraduate and graduate levels including international field experiences.  she is on the executive board for the uw global health institute.  tanumihardjo has >160 publications and chapters.  she has presented at >280 domestic and international venues.  she has served as a reviewer for many journals.  awards: who’s expert advisory panel, g. malcolm trout visiting scholar at michigan state university, ruth pike lectureship at pennsylvania state university, alex malaspina ilsi future leader, dannon creative leadership institute, endowed chair and vilas associate at uw.

Abstract

Vitamin A is a micronutrient essential in vision, reproduction, immune function, cellular differentiation, and bone health.  Assessment of vitamin A status is not straightforward.  Concentrations of retinol circulating in blood are homeostatically controlled and therefore are not useful indicators of status until liver reserves are dangerously low.  The retinol isotope dilution (RID) tests have been validated against liver reserves of vitamin A from deficiency through hypervitaminosis A, have a working range that far exceeds all other currently available indirect biomarkers of vitamin A status, and are very conducive to assessing long-term vitamin A status as it relates to vitamin A exposure.  RID was very good at determining vitamin A status of rats over a very broad range of liver vitamin A reserves (2-10 fold difference), while serum retinol did not detect a difference.  RID has been used to estimate total body reserves of retinol in groups of humans.  Both deuterium and 13C have been introduced into the retinol moiety with various enrichments.  Depending upon the enrichment and mass spectrometer sensitivity, traditional gas chromatography (GC) mass spectrometry (MS), GC-combustion isotope ratio-MS, and liquid chromatography tandem MS have been used for analysis.  The most sensitive method, GCCIRMS, allows researchers to use small doses of tracer to not perturb normal vitamin A metabolism.  Eleven African countries were involved in an International Atomic Energy Agency project to assess the vitamin A status of groups of 3 to 5 year old children.  The dose level of 13C2-retinyl acetate used in the African children was 1 µmol/child.  The RID test using either D4 or D8-retinyl acetate used 6 mmole/child, which is less than prior doses.  Although vitamin A deficiency has been the most concerning over the past few decades, widespread fortification and the continued use of high-dose supplements cause new concerns for hypervitaminosis A demanding more sensitive methodology. 

Biography

Zhaohua peng is a full professor in the department of biochemistry, molecular biology, entomology, and plant pathology at mississippi state university. He received his bs and ms degrees in biophysics at china agriculture university and his Ph.D degree in plant molecular biology at ohio state university. From 1997 to 2001, he received training at yale university as postdoctoral fellow. He joined mississippi state university faculty as an assistant professor in 2002. He was promoted to associate professor in 2008 and full professor in 2013. His research focuses on the regulation of rice seed development and seed storage nutrient production with emphasis on proteomics and epigenetic studies. 

Abstract

Protein lysine malonylation, succinylation, crotonylation, and 2-hydroxyisobutyrylation have been recognized as new post-translational modifications (PTMs) in recent years. It is plausible that these modifications may have a greater functional impact than lysine acetylation due to bulkier structural changes and larger charge differences introduced on the modified lysine residues. However, the identity of proteins harboring these modifications and their corresponding functions in cereal plants remain largely unknown. Using antibody-based affinity enrichment of modified peptides followed by nano-HPLC/MS/MS analyses, we identified from a few hundreds to over nine thousands modification sites for these modifications in developing rice (Oryza sativa) seeds, respectively. Distinct sequence motifs at the modification sites were identified for each of the modification.  Proteins with different sequence motifs were shown to be favorably associated with unique metabolic pathways or protein function domains. Many of the modified proteins and the corresponding modification sites were conserved from E. coli, human, to plants.  Remarkably, heavy modifications were detected on major seed storage proteins together with the key enzymes participating in central carbon metabolism and storage starch biosynthetic pathways, which are essential for rice seed nutrition reservoir development. Rice proteins with co-modifications of succinylation, malonylation, crotonylation, 2-hydroxyisobutyrylation, acetylation, ubiquitination, and phosphorylation were studied through a comprehensive comparison analysis. In addition, the impact of lysine modification on lysine bioavailability in rice proteins is also analyzed. Our study delivers a platform for expansive investigation of the molecular networks administrating cereal seed development via post-translational modifications. 

Biography

Gary w. Caldwell is a working in johnson & johnson, usa. His recharge skills on chromatography ,mass spectrometry,liquid chromatography, sample preparation,analytical chemistry instrumentation spectrometrychromatographic method development.

 

Abstract

Rheumatoid arthritis (RA) is an autoimmune disease that causes chronic inflammation of the synovial lining of the joints. It affects females more frequently than males and is typically diagnosed between the ages of 40-60 years with symptoms developing gradually.  If untreated, RA can lead to permanent joint damage and an increased risk of premature mortality. Synthetic DMARDS (sDMARDs) such as methotrexate (MTX), and biological DMARDS (bDMARDs) such as TNF-α antagonists (e.g., Etanercept, Adalimumab, or Infliximab), and antibodies that deplete B cells (e.g., Rituximab) are widely used in inhibiting synovial inflammation, and retarding bone erosion. It has been estimated that 30-40% of patients have poor or no response to these treatments. Metabolomics is a rapidly developing approach in biomarker research which involves the measurement of the set of final products and by products of metabolic pathways using NMR and mass spectrometry methods. Since RA disrupts metabolite (products) pathological processes, metabolomics can measure the alterations in these metabolite profiles long before overt signs and symptoms of the disease appears. Metabolomics should provide a more accurate representation of the RA phenotype than that accounted for by genetic, epigenetic, gastrointestinal microbiome and environmental factors. Many metabolomics studies have been published identifying potential RA biomarkers for use in diagnosis, prognosis or prediction of drug treatment. Here we discuss the advantages and disadvantages of metabolomics RA biomarker research using mass spectrometry techniques.

Biography

Master in organic chemistry from Université de Sherbrooke, he worked for more than 10 years in the pharmaceutical industry in Canada and the United States. For the last 8 years he has pursued his career at the Centre de Toxicologie du Québec (CTQ) where he is involved in the development of new and specialized analytical methods, to assess exposure to heavy metals, solvents and their metabolites, pesticides and other environmental pollutants.With the help of his colleagues at the CTQ he recently published a paper on the matrix effects subject.  

Abstract

The effects of the matrix on the analysis may have numerous impacts on the quantification of an analyte. This may lead to confusion on what is actually measure, due to a non-universal terminology used in the scientific literature. In the hope bringing more clarity to the MEs we introduced new terms, and showed the relationship between them. We design a series of experiments to visualise how well the MEs are controlled, and to gain knowledge of where the perceived variations are coming from. The links between the different MEs and descriptors are presented, providing a pathway to follow when doing troubleshooting work. The results obtained with the test provided information on both the analytes and the internal standards facilitating the analyte/internal standard association for a correct quantification in the sample matrices. The technique presented is applicable to matrices containing a base quantity of the analyte, and to different types of instruments like LC, GC and ICP. This methodology facilitates the decision-making process while lowering the required costs and time.
 

Biography

Aneta Wojnicz has completed her Master Degree in Environmental Biotechnology at Warminsko-Mazurski University (Poland)/Leibniz Hannover University (Germany) and her PhD in Clinical Pharmacology at Autonomous University Madrid (Spain). Currently, Aneta Wojnicz is working as Analytical and Pharmacokinetic Unit Responsible at Clinical Pharmacology Department, Hospital Universitario de la Princesa‟ (Madrid, Spain). The aim of the Unit is to develop and validate analytical LC-MS/MS methods for drug quantification in biological fluids. She has published more than 10 papers in reputed journals.

Abstract

Liquid chromatography-tandem mass spectrometry (LC–MS/MS) method has been developed and validated for simultaneous quantification of five tirosine kinase inhibitors (imatinib, dasatinib, nilotinib, bosutinib, ponatinib) and one Bruton's tyrosine kinase inhibitor (ibrutinib) in human plasma. Stable isotope-labeled internal standards have been applied for each compound to minimize matrix effect which commonly occurs in MS analysis. The analytes and their internal standards were extracted from only 200 µL of human plasma using solid phase extraction (Versaplate-SCX). The compounds were eluted under gradient conditions using a Poroshell 120 EC-C18 column (2.1 x 75 mm, 2.7 mm) at flow rate of 0.5 mL/min and 60ºC. Analytical column was protected by a 0.2- μm on-line filter. The mobile phase consisted of 0.1% formic acid in MilliQ water, pH=2 (solution A) and 0.1% formic acid in acetonitrile (solution B) (80:20; v/v). Total run time was 9 minutes, including acquisition time of 6 minutes followed by a re-equilibration time of 3 minutes. Five μL of the sample was injected into the chromatographic system. All analytes were detected using the mode multiple reaction monitoring in the positive ionization mode. The method was validated according to the recommendations of regulatory agencies through tests of precision, accuracy, recovery, matrix effect, stability, sensitivity, and selectivity. Sample preparation method applied in the present assay removes more than 95% of main plasma phospholipids compared to protein precipitation. The method enables selective quantification of six compounds. Present assay is currently being applied to therapeutic drug monitoring of imatinib, dasatinib, nilotinib and ponatinib in clinical practice in order to individualize dose adjustment and manage adverse effect.

Biography

Yajie Wang has completed her PhD of Organic Chemistry in University of Chinese Academy of Sciences(UCAS). She has worked as a specialist associate staff scientist of Center for High Pressure Science and Technology Advanced Research (HPSTAR) since July, 2017. Her research interest is focused on high pressure chemistry and the development of chemical analysis method using high resolution mass spectrometry.

Abstract

Pressure is one of the most important physical factor and a powerful thermodynamic variable. As the recent great advancements in high pressure technologies, numerous unique structures and behaviors have been discovered under high pressures, which is very significant for  underlying the evolution of physics and chemistry of the Earth and the planetary system.  The pressure effect on organic chemicals involves promoting intra/inter-molecular interaction, inducing crystalline phase transitions or even triggering chemical reactions, which allows producing new dense materials with peculiar structure and properties.  However, as the reaction paths are complex and the corresponding products are in mixed phases, most of reaction mechanisms in terms of high pressure is not clear so far. The mass spectra are distinctive to every molecule that will be pre-separated by chromographic columns, and thus lead to a ready identification of the compounds. Applying high resolution mass spectrometry with ultra-high sensitivity is greatly helpful for the detection of small amounts of polymeric products synthesized by high pressure methods. 

Biography

Seitaro Kamiya has completed his PhD in Pharmaceutical Technology from the University of Shizuoka. He has worked as Senior Assistant Professor of Pharmaceutical Technology at Nagasaki International University. He has published more than 12 papers as a first author in reputed journals. He has succeeded in commercializing vitamin C related products through collaborative research with a health food company. 

Abstract

Powder X-ray diffractometry (PXRD) and differential scanning calorimetry (DSC) are employed to investigate the property of crystalline powders. In the case of PXRD, powders with crystal phase produce diffractive peaks and powders with amorphous phase produce hallow pattern. In the case of DSC, the powders with crystal phase has endothermic peak because they have melting point. Meanwhile, powder with amorphous phase has no endothermic peak because they have melting point. In the present study, we investigated about a relationship between various saccharides and nanoparticles during freezedrying or normal drying. Because nanoparticles suspension is thermodynamically labile, we focus on freeze-drying and normal-drying method. Trisaccharides, tetrasaccharides, or pentasaccharides were added to the nanoparticle suspensions, followed by rehydration of the samples, which had been either dried normally or freeze-dried. The particle size after rehydration each sample at that time was then measured. The particle size of each freeze-dried saccharide was maintained. Meanwhile, the particle size of each normal dried saccharide indicated various particle size. We studied the association between the nanoparticles aggregation and the crystal form of saccharides and their mechanisms by using the obtained results of the data of particle size, powder X-ray pattern, and DSC curves. The relationship between degrees of the nanoparticles aggregation and degree of crystallization are gradually figured out. By continuing this research, the saccharide additives map for preservation nanoparticles will be drawn. 

Biography

Yahdiana Harahap is a Professor in the field of Pharmaceutical Chemistry especially Bioanaysis related to bioequivalence study and DNA Adduct. She received her Master Degree in 1994 and Doctoral degree in 2003 from Department of Pharmacy, Faculty of Mathematics and Natural Sciences, Institute Teknologi Bandung. She has been The Head of Bioavailability-Bioequivalence Laboratory Faculty of Pharmacy Universitas Indonesia since 2008. She serves as member of Expert Council Indonesian Pharmacist Association, reviewer in several international journals, President elected on Asian Federation of Pharmaceutical Science, Head of Sub Collegium Indonesia Pharmaceutical Industry and as The Head of Pharmacy Division in Indonesian Accreditation Agency for Higher Education in Health (LAM PT-Kes). She has generated more than 80 scientific works published in international and national journals, thus presented them in national and international conferences.       

Abstract

Methamphetamine is a very strong central nervous system stimulant of the amphetamine group. Methamphetamine concentration in the body is usually determined in the blood and urine. Saliva as a biological matrix is simpler and is more efficient for methamphetamine tests in the body although it is rarely used. The aims of this study is to develop analytical methods for methamphetamine in saliva using gas chromatography tandem mass spectrometry. This technique is suitable to be applied for forensic drug abuse test using small sample and short analysis time. The chromatography conditions were DB MS-5 capillary columns with a length of 30 m; inner diameter of 0.25 mm; mobile phase Helium gas of 99.999%; flow rate of 0.8 mL/min; detection of MS at m/z values of 58.00 and 91.00, thus ephedrine HCl was used as an internal standard. Sample preparation using liquid-liquid microextraction with cyclohexane solvent and the residue was dried and reconstituted with 100 µL of methanol. The method was valid in term of selectivity, accuracy and precision, carry over, matrix effect, stability test and was linear in the concentration range of 15.0 to 300.0 ng / mL. The method was successfully applied to the saliva sample of methamphetamine users. Prior to sample analysis saliva was collected from the users less than 24 hours after the consumption of methamphetamine. The levels of methamphetamine in saliva was 106.99 ng/mL and 177.14 ng/mL.

Biography

Lingzhi Gong general research interests include mass spectrometry based analysis of small molecules and biomolecular molecules (nucleic acids, peptides, proteins) through hyphenated to a chromatographic method (mainly liquid chromatography), and research into chromatographic retention mechanisms. His current work focuses on characterizing single- and double-stranded DNA/RNA, and protein/peptide - DNA crosslinking complex using hyphenated liquid chromatography and electrospray mass spectrometry (LC-ESIMS). Lingzhi currently runs Core Mass Spectrometry Research Facility at Queen Mary University of London

Abstract

Synthetic oligonucleotides are widely used in the polymerase chain reaction (PCR) as DNA primers or in molecular biology as probes to screen for diseases, viral infections, and to identify genes. Sensitive and selective methods have always been demanded for the characterization of oligonucleotides, especially, when the oligonucleotides are applied as therapeutics. Ion-pair reversed-phase (IP-RP) liquid chromatography has been commonly used for the analysis of oligonucleotides, but ion suppression is a major problem when coupling with electrospray mass spectrometry (ESI-MS). Although the introduction of hexafluoro isopropanol (HFIP) in the mobile phase has improved MS detection sensitivity of oligonucleotides, this mobile phase system results in a severe problem with adducts formation particularly if large oligonucleotides are analysed. An alternative chromatographic approach, hydrophilic interaction liquid chromatography (HILIC), was recently employed for the analysis of oligonucleotides. It provided enhanced MS sensitivity with fewer adducts but lacked chromatographic resolution for omeoligonucleotides. Here we improve chromatographic resolution whilst maintaining MS sensitivity by adding an ion-pairing reagent, triethylammonium acetate (TEAA), into the HILIC mobile phase. The IP-HILIC approach produces lower retention capacity and has the added benefit of providing simpler MS spectra, with fewer charge states, when comparing with HILIC. 

Biography

Mahmoud Rafea is working in Central Lab of Agriculture Expert Systems, Egypt. Initially, he is a Medical Doctor. He has a double qualification in Clinical Pathology and Computer Science. He worked as a Researcher in Artificial Intelligence (AI). He was promoted by his institution (ARC) to a Professor of Computer Science. Currently, he is working on medical R&D using his AI and medical knowledge. 

Abstract

Erythrocytes dynamic antigens store (EDAS) is a new discovery. In patients with infectious diseases, malignancies, Alzheimer’s disease, and idiopathic disorders, EDAS contains antigens related to those disorders. Storing EDAS of patients in a database has, at least, two benefits. First, it can be queried with EDAS of a patient presenting with the unknown disorder to diagnose his condition. Second, it can be mined to determine biomarkers, which can be used in developing a new line of clinical laboratory kits. We show how this approach can be implemented using hypothetically created EDAS. The mathematical model of hypothetical EDAS together with software tools for biomarker discovery and direct diagnosis are described. The different possibilities that may occur in reality are experimented. We found that the tool can identify biomarkers’ proteins for pathogens and malignancies. Also, the tool can take EDAS of a case, as an input, to reach to a diagnosis. In effect, the tool can be used to diagnose quickly conditions that may be difficult or take longer time and effort to diagnose by conventional methods. The presented tool can be used in clinics to diagnose disease disorders. This work is done as an initial step to study different possibilities and situations to be prepared for using real big data. 

Biography

Rahul Hajare has completed his PhD from Vinayaka Missions University. He was a Post-Doctoral Fellow ((2013)) at the Indian Council of Medical Research New Delhi under the guidance and supervision of Dr. Ramesh Paranjape, Retd Director and Scientist ‘G’, National AIDS Research Institute, India.

Abstract

Although pharmaceutical impurity has not yet been accepted as monograph facility in pharmaceutical products, it has shown extraordinary diversity. The US FDA has raised concerns on the quality, purity and efficacy of medicines being sold in India, throwing the issue back into spotlight. Over the last few years, many domestic majors, including multinational and listed pharmaceuticals in stock exchange have faced regulatory issues over quality and purity of medicines exported from India and sold in US and other overseas markets. We report here, the non listed impurity profile study of Ciprofloxacin Hydrochloride from API industry. Of these, pharma impurity (A1) and API impurity (B) are most prevalent in India, while raw material impurity (C) and cleaning impurity (D) have occasionally been reported. Apart from these A1/C (1, 2) and B/C (3) intersubtype recombination impurity have also occasionally been reported. The presence of these have listed, so specified impurity and other detectable impurity in ciprofloxacin hydrochloride, these substances present at a sufficient level, be detected by one or other of the test in the monograph European pharmacopeia. They are limited by the general acceptance criteria for other/ unspecified impurities and/or by the general monograph substance for pharmaceutical use. In this study, we characterize 
inconsistent unspecified impurity that has been associated with specified impurity and adjacent to other detected impurity in pharmaceutical ciprofloxacin hydrochloride. Here, we report for the first time in India, a unique impurity recombination between subtype C (raw material acrylic acid) and environmental impurity (C1) and novel recombination between subtype A1 and C1. Unique recombination formation impurity between crude and pure products have been reported many a times from pharmaceutical industries (Database form patent literature and market analytical survey).The ciprofloxacin hydrochloride was amplified using Aarti Drugs Limited, Mumbai store database and stored co-cultured 84 hrs and drug ciprofloxacin hydrochloride by UPLC describe (Applied waters system acquity UPLC column). 

Biography

Abhishak C Gupta is working in Indian Institute of Technology, Delhi. He is a Project Scientist in Department of Textile Technology, IIT, Delhi. 

Abstract

Backgrounds & Aim: In the absence of any symptomatic clinical features, liver biopsy remains the gold standard to assess disease progression in nonalcoholic fatty liver disease (NAFLD) patients. Oxidative stress is postulated to play an important role in liver disease progression. Albumin is the most abundant plasma protein with antioxidant activity. The degree of oxidized cysteine 34 (Cys34) in human serum albumin (HSA) is correlated with oxidative stress related pathological conditions and modulates its physiological function, as well as serves as a biomarker for oxidative stress. The aim of the present study was to develop a noninvasive diagnostic plasma marker for NAFLD by studying the differential modification pattern of plasma albumin in NAFLD patients.
Patients & Methods: We analyzed purified plasma albumin from 46 biopsy-proven NAFLD patients (17 with benign fatty liver and 29 with NASH) and 21 healthy non-smoker/ non-alcoholic blood donors matched with age and BMI. The structural modifications of albumin were analyzed by direct measurement of plasma albumin using liquid chromatography coupled with electrospray ionization time-of-flight mass spectrometer (ESI-TOF/MS). Relative percent abundance of unmodified (intact) and modified isoforms of albumin was compared between patients and controls.
Results: Three most prominent isoforms of albumin were observed in the deconvoluted ESI spectrum with molecular masses of 66.438±2.8, 66.559±4.8 and 66.603±6 Da in controls and NAFLD patients represents intact, cysteinylated and glycated isoforms of albumin respectively. Unmodified albumin was the predominant peak with 100% relative abundance in healthy subjects in perfect agreement with calculated theoretical mass (66.438 Da, 542aa). In contrast, the relative abundance of modified form with addition of +119Da (cysteinylation) of albumin was predominant (100%) in NAFLD patients. Cysteinylated isoform of albumin (cys-Alb) was significantly higher in NAFLD patients than controls [100% vs. 52% (p<0.01)]. Although NAFLD showed 100% relative abundance of cys-Alb isoform, further fatty liver and NASH patients differ on the basis of unmodified albumin isoforms [82% vs. 60% (p<0.05)] suggesting varied oxidative stress.
Conclusion: Our results showed that sustained oxidative stress is reflected by high levels of cysteinylated albumin in NAFLD patients. The measurement of cysteinylated albumin in suspected NAFLD patients might prove to be a useful marker to assess the degree of oxidative damage, inflammation and severity of the disease.
 

Day2: March 13, 2018

Keynote Forum

Biography

Sherry A Tanumihardjo studies vitamin A and carotenoid metabolism, serves as Director of the Undergraduate Certificate in Global Health, and teaches at undergraduate and graduate levels including international field experiences. She is on the Executive Board for the UW Global Health Institute. She has more than 160 publications and chapters published. She has presented at more than 280 domestic and international venues. She has served as a reviewer for many journals and is the recipient of the following awards: Who’s Expert Advisory Panel, G Malcolm Trout Visiting Scholar at Michigan State University, Ruth Pike Lectureship at Pennsylvania State University, Alex Malaspina ILSI Future Leader, Dannon Creative Leadership Institute, Endowed Chair and Vilas Associate at UW

Abstract

Maize is a staple crop in many parts of the world and has been targeted for biofortification with provitamin A carotenoids to provide more vitamin A (VA) to consumers. VA assessment can be challenging because static serum retinol concentrations do not always reflect total body stores. Maize is a C4 plant, meaning that it is naturally enriched with 13C. For effectiveness studies with provitamin A carotenoids, changes in 13C natural enrichment of serum retinol can be determined as a measure of biofortified (orange) maize consumption. To show proof-of-concept, a study in Mongolian gerbils (n=55) investigated changes in the 13C/12C (δ13C) of serum retinol from feeding orange maize (C4) and carrots (C3) for 62 d. The design was a 2x2x2 maize (orange vs. white) by carrot (orange vs. white) by VA fortificant (VA+ vs. VA-). Liver and serum VA δ13C was analyzed by gas chromatography-combustionisotope ratio mass spectrometry. Serum and liver VA δ13C were significantly correlated (R2=0.92, P<0.0001). Serum retinol δ13C differentiated controls consuming white maize and white carrots (-27.1±1.2 δ13C‰) from groups consuming orange maize and white carrots (-21.6±1.4 δ13C‰, P<0.0001) and white maize and orange carrots (-30.6±0.7 δ13C‰, P<0.0001). Natural abundance measurements were applied to an efficacy study in Zambian children, in which orange or white maize was fed for 90 days. In these children, 13C natural abundance was higher 
after two months in the orange maize group compared with the white maize group (P<0.05). Predictions made from equations developed in the gerbil study estimate that maize provided 11% (2-21%, 95% CI) of recent dietary VA in this group. Shifts in serum retinol δ13C can be used for maize and other biofortified C4 crop effectiveness studies. Advantages of this method in effectiveness trials include no extrinsic tracers, one blood sample, and high sensitivity compared with measuring serum retinol alone.
 

Biography

Aneta Wojnicz has completed her Master’s Degree in Environmental Biotechnology at Warminsko-Mazurski University (Poland)/Leibniz Hannover University (Germany) and her PhD in Clinical Pharmacology at Autonomous University Madrid (Spain). Currently, she is working at the Analytical and Pharmacokinetic Unit in the Clinical Pharmacology Department, Hospital Universitario de La Princesa, Spain. The aim of the unit is to develop and validate analytical LC-MS/MS methods for drug quantification in biological fluids. She has published more than 10 papers in reputed journals.
 

 

Abstract

Mass spectrometry (MS) is one of the most important analytical techniques in the twenty-first century. It has been widely applied in several fields of medicine, chemistry, and physics and also in human habitat studies. MS can be implemented as a targeted or an untargeted approach. Here we present three different examples of liquid chromatographytandem mass spectrometry (LC-MS/MS) applications. Two of them focus on targeted method development and validation and have been successfully validated according to the recommendations of the regulatory agencies. The third approach aims to evaluate the chemical background of modern human habitat designed as untargeted analysis based on three-dimensional (3D) cartography MS. The first example of targeted LC-MS/MS method applicability has been shown in clinical practice. Different methods validated in our laboratory have been applied in pharmacokinetic studies and therapeutic drug monitoring. The second type of applications enforced the importance of MS in basic science studies, such as monitoring neurotransmitters in tissue and cell cultures. We have been able to simultaneously determine four different groups of neurotransmitters, including monoamines, amino acids, nucleotides and neuropeptides in bovine chromaffin cell cultures, rat brain extracts and human plasma samples, where altered neurotransmitter levels could be detected due to stress. The untargeted approach, 3D cartography MS, examines the chemical background associated with modern human habitat in order to detect the chemicals distributed in humans and their environment. Consequently, a great utility of MS has been shown as it has been successfully applied to different research purposes.
 

Tracks

  • Protein Mass Spectrometry | Ionization techniques Mass Spectrometry | Forensic Analysis | Mass Spectrometry in Medicine | Imaging Mass Spectrometry | Analytical Chemistry
Location: Armstrong
Speaker

Magnus S Magnusson

University of Iceland, Iceland

Chair

Speaker

Lingzhi Gong Guangzhou University of Chinese Medicine, China

Co Chair

Biography

Semra G Tuncel is working in Middle East Technical University, Turkey.

Abstract

Seventeen organochlorine pesticides (OCPs) were evaluated in 14 surface sediment samples from a dam lake in Northwestern Turkey. As analytical tool GC-Mass system; HP (Hewlett Packard) 6890 series gas chromatograph coupled with HP 5973 mass spectrometer was used. The HP 5 MS capillary column had 30 m length with 0.32 mm internal diameter. A 0.25 mm film thickness cross-linked with stationary phase of 5% phenyl methyl siloxane and ultra-pure helium gas was used as mobile phase. Ultrasonic bath extraction method was applied and cleaned up process were carried up with anhydrous Na2SO4 and Florisil column. Total pesticides concentrations were ranged from 0.237-2.39 mg/kg for dry weight. Percent total organic carbon (TOC) were observed between 1 and 3%. Average total OCP concentrations was 58.00±45.44 mg/kg. The total concentrations of OCPs in sediment samples ranged from 12.9 to 169.9 mg/kg, with a mean value of 58.00 mg/kg. Although organochlorine pesticides have been banned in Turkey, the residues can still be seen in sediment samples indicating the use of prohibited pesticides in the country. Comparison of organochlorine pesticide concentrations in sediment samples with other lakes in Turkey implies higher concentration and therefore higher usage of synthetic chemicals

Biography

Rahul Hajare has completed his PhD in Pharmacy Medicinal Chemistry from Vinayaka Mission’s Research Foundation and Postdoctoral Studies from ICMR-National AIDS Research Institute (NARI), ICMR PDF grant of 7th Batch (2013). He has worked as Associate Professor of Pharmaceutics at BSPM College of Pharmacy-Dr. Babasaheb Ambedkar Marathwada University Aurangabad. He has published more than 58 papers in reputed journals

Abstract

There has been much recent talk about how to target the rising tide of ART resistance across the world, one of the biggest threats to global health today. While there is no doubt about the problems faced by scientists, healthcare professionals and the pharmaceutical industry, there are innovative ways we can target ART resistance in the short term. With only a few ART in development and a long drug development process (often 1015 years), there is concern that what is being done to combat ART resistance may be ‘too little, too late’. “If virus continues developing resistance to multiple ART at the present rate, at the same time as the ART pipeline continues to dry up, there could be catastrophic costs to healthcare and society globally”, reported in reputed articles. While any ART resistance is concerned, the increasing incidence of ART-resistant mutant virus has become a particular problem as strains resistant to multiple ART are becoming common and no new drugs to treat these infections (e.g., New entrance-resistant Enterobacteriaceae) will be available in the near future. These mutant viruses are considered the most critical priority in the list of the 12 families of virus that pose the greatest threat to human health that was just released by the World Health Organization. The reasons for high levels of ART resistance observed in these critical Gram-negative organisms are explained in another research article in the same issue and extended literature. After a close monitoring, one of the main contributing factors to the increased resistance observed in Gram-negative bacteria is the permeability barrier caused by their additional outer membrane. An innovative strategy that is gaining momentum is the synergistic use of antibiotics with FDA-approved non-antibiotics. Using this novel approach, an FDA-approved nonantibiotic drug is combined with a specific antibiotic that enables it to breach the outer membrane barrier and so restore the activity of an antibiotic. 

Biography

Rashid Mahmood has a Master’s Degree in Analytical Chemistry and MS in Total Quality Management. He has 14 years of experience in pharmaceutical quality operations and has participated in many international conferences as a Keynote Speaker. He has presented various talks in USA, Canada and China on cleaning validation, cgmp guidelines and quality risk management. Currently, he is working as a Senior Executive Manager, Quality Operations for Surge Labs, Pakistan. He is engaged in the manufacturing of microencapsulated APIs, liquid and dry powder parenterals.

Abstract

Mass spectrometry is an analytical technique to measure the molecular or atomic weight of samples. It is an instrument that produces charged molecular species in vacuum, separates them by means of electric and magnetic fields and measures the mass-to-charge ratios and relative abundances of the ions thus produced. It is being increasingly used for detection and analysis of proteins from complex samples. The emergence of mass spectrometry as a powerful analytical tool in the health, life sciences and pharmaceutical sector associated with drug discovery and development process is rich and varied. Many of the initial efforts were associated with online high performance liquidchromatography-mass spectrometry in drug metabolism, pharmacokinetic and pharmacodynamic studies. Pharmacokinetic studies with mass spectrometry can provide quantitative information about a compounds half life in the body and how quickly it is metabolized or excreted. the increase in sensitivity and resolution of the mass spectrometer has opened new dimensions in analysis of pharmaceuticals and complex metabolites of biological systems. Compared to others, mass spectroscopy is only the technique for molecular weight determination, which can further help in prediction of molecular formula. It is also used as a sensitive detector for chromatographic techniques like LC-MS and GC-MS. 

Biography

Ania Cabrales Rico is working as a Research Assistant at the Center for Genetic Engineering and Biotechnology (CIGB), Cuba. She graduated from University of Havana and got her PhD in Pharmaceutical Sciences in 2015 when she was promoted to Head of Purification and Analytics group, from the Physicochemical Characterization Department, in Biomedical Research. She has been working with synthetic peptides for many years, but particularly in the field of bioanalytical methods based on mass spectrometry (MS), since 2008. This multidisciplinary work allowed her to gain expertise in: (1) isotope labeled internal standards design, (2) sample processing previous to LC-MS analysis, (3) peptide quantitation by MS, (4) bioanalytical method validation and (5) pharmacokinetic analysis. Peptides are quite diverse regarding to their physicochemical properties, so the bioanalytical methods needs to be tailored, becoming each experience in unique opportunity to overcome an analytical challenge in a successful outcome. 

Abstract

Mass spectrometry (MS) has impressive capabilities in terms of sensitivity, resolving power, mass accuracy and different scan-modes versatility. Either alone or in combination with liquid chromatography, it is the analytical tool of choice for synthetic therapeutic peptide characterization. Nevertheless, for peptide quantitation in human plasma or other biological samples, the design of the internal standard (IS) and the optimization of the sample processing and LC-MS analysis are also key elements for a successful outcome. In consequence, all strategies involving the peptide quantitation in biological fluids are still a challenge and need to be tailored. We present here our recent experiences in the development and validation of customized bioanalytical methods applied to pharmacokinetic studies included in phase I clinical trials. For the absolute quantitation of these three therapeutic peptides, alternatives to the AQUA® methodology were used. However, the design of the IS, sample processing and mass spectrometry techniques were optimized case by case for CIGB-500, CIGB-300 and CIGB-814 candidates. IS for CIGB500 and CIGB-814 were synthetic peptides labeled with stable isotopes (13C and/or 15N) in specific residues within the amino acids sequence, instead of IS for CIGB-300 that was a N-terminus acetylated peptide. Sample processing, mainly based on plasma proteins organic or acid precipitation was adapted according to the peptide recovery. In the particular case of CIGB-300, no liquid hromatography separation was needed before MS analysis by MALDI-TOF MS. For CIGB-500 it was applied LC-MS analysis with simultaneous ion monitoring (SIM) in full scan mode. For CIGB814 it was used LC-MS analysis in single reaction monitoring mode (SRM). The three bioanalytical methods were fully validated and applied to pharmacokinetic (PK) analysis in a phase I clinical trials. It was possible to obtain PK profiles and main PK parameters for all of the assessed candidates. 

Biography

Rashid Mahmood has a Master’s Degree in Analytical Chemistry and MS in Total Quality Management. He has 14 years of experience in pharmaceutical quality operations and has participated in many international conferences as a Keynote Speaker. He has presented various talks in USA, Canada and China on cleaning validation, cgmp guidelines and quality risk management. Currently, he is working as a Senior Executive Manager, Quality Operations for Surge Labs, Pakistan. He is engaged in the manufacturing of microencapsulated APIs, liquid and dry powder parenterals. 

Abstract

In the pharmaceutical industry every product and every process associated with risks. To maintain product quality throughout the product life cycle, too much time and resources are allocated. Risk is described in - recent guidance as a combination of the probability of occurrence of harm and the severity of that harm. The Quality Risk Management (QRM) approach initiated by regulatory agencies with recognized management tools along with support of statistic al tools in combination allows for a risk-based approach to quality management, thus ensuring that resources are deployed in a timely and expeditious manner to areas that need them most. QRM improves risk awareness and accelerates detection of potential issues by analyzing and comparing existing data from a quality perspective to manage product quality, manufacturing processes, validation and compliance within a risk based Quality Management System. In addition quality risk management improves decision making if a quality problem arises. It should include systemic processes designated to coordinate, facilitate and improve science-based decision-making with respect to risk. Quality Risk Management can be applied not only in the manufacturing environment, but also in connection with pharmaceutical development and preparation of the quality part of marketing authorization dossiers. The guideline applies also to the regulatory authorities in the fields of pharmaceutical assessment of the quality part of the marketing authorization dossier, GMP inspections and the handling of suspected quality defects. ICH Q9 - Quality Risk Management provides an excellent high-level framework for the use of risk management in pharmaceutical product development and manufacturing quality decision making applications. It is a landmark document in acknowledging risk management as a standard and acceptable quality system practice to facilitate good decision-making wit h regard to risk identification, resource prioritization and risk mitigation / elimination, as appropriate.