Day 2 :
Keynote Forum
Steven J Soldin
National Institutes of Health, USA
Keynote: The role of mass spectrometry in improving the diagnosis and management of adrenal diseases
Time : 10:00-10:30
Biography:
Abstract:
After addressing several new findings which result in optimization of steroid profile measurement, the talk will focus on how mass spectrometry improves both diagnosis and treatment of patients with adrenal diseases (24 million people in the USA). The less than optimal use of serial cortisol measurements following ACTH stimulation to diagnose and treat adrenal disorders will be addressed. Replacement of cortisol by 11-DOC is preferable and in a fairly high percentage of cases (15–30%) disagrees with the diagnosis obtained using cortisol.
Keynote Forum
Nathan S. Jacobson
Keynote: Mass spectrometric sampling of high temperature inorganic vapors
Time : 10:30-11:00
Biography:
Abstract:
Mass spectrometry has long been applied to study inorganic vapors at high temperatures. Such measurements lead to a variety of useful information including thermochemical data on the both the vapor itself and condensed phase vapor source, identity of the vapor species above a particular condensed phase, and bond energies of the vapor species. However, sampling of high temperature vapors presents a series of unique challenges. Paricular care must be taken to have a clear, unobstructed path to the ionizer. In this paper, we describe the sampling system used at NASA Glenn and also a recently developed Monte Carlo simulation of this sampling system. The vapor source is a Knudsen cell, heated to temperatures up to 2000°C. An orifice in the cell of well-defined geometry forms a molecular beam, which is directed through a series of aperatures into the mass spectrometer ionizer. Following the method of Chatillon and colleagues, the first aperture has a smaller diameter than the Knudsen cell orifice so that the ionizer effectively ‘sees’ only into the cell, thus minimizing the effects of background gases. A second aperature further directs the molecular beam into the ionizing region. This molecular beam portion of this process can be modeled with a simple Monte Carlo simulation. Trajectories begin in the Knudsen cell and are traced through, till either the molecule leaves the sampling region or reaches the ionizer. This code allows optimization of aperature diameters and spacings as well as estimation of the efficiency of the sampling process. Future ideas are presented for high sensitivity measurements of inorganic vapors.
Keynote Forum
Wenjie Cao
SABIC Technology Center, Saudi Arabia
Keynote: Further investigation of the adduct ion complications on MRM at different LC-MS/ MS ionization techniques, ESI, APCI, MMI and APPI
Time : 11:15-11:45
Biography:
Wenjie Cao received a PhD from Professor John Calvin Giddings’ Group at the University of Utah, Salt Lake City, UT. He is a Contributor to the book, “Encyclopedia of Chromatography” and more than 20 publications and presentations in peer-reviewed scientific journals and international conferences. He has worked for Huntsman Polymers Corp, Sealed Air, and DuPont as a Research Investigator for 14 years in USA before joined SABIC in 2012. He is now the Technical Leader of chromatography and wet lab and a Staff Scientist of the Analytical Department of the SABIC Technology Center at Riyadh. He has filed six patents, delivered talks and made seminar presentations in the ISPAC Symposium, 2016 and in King Saud University since he joined SABIC Technology Center.
Abstract:
This presentation is going to extend the comparison of electrospray ionization (ESI) and atmospheric pressure chemical ionization (APCI) to multimode ionization (MMI) and atmospheric pressure photo ionization (APPI) per some of the audiences’ request during the Q&A section of my presentation during the 2015 International Summit on Current Trend of MS in New Orleans. For the triple quadrupole LC-MS/MS instrument, the primary purpose or the most significant feature is the highest sensitivity among almost all, if not all of the LC-MS/MS instruments by doing the multiple reaction monitoring (MRM) testing. Ionization efficiency, selectivity, adduct ion production are among the top parameters which affect the MRM testing and the sensitivity. From ESI to APCI to MMI and APPI, this presentation will show that the species and amount of adduct ions produced at each mode are quite different. Some type of the adduct ions may complicate the MRM testing by decreasing the sensitivities while some other adduct ions may prevent any reliable MRM tests being performed. Some examples will be presented to show how the typical adduct ions are produced in each mode from ESI to APCI and APPI, and how the typical adduct ions may complicate the MRM testing. The overall pros and cons, and the best ionization mode for some type of the targeted chemicals will be summarized for the different ionization techniques.
- Fundamentals of Mass Spectrometry | Mass spectrometry imaging | New Approaches in Mass Spectrometry | Recent Advances and Development in Mass Spectrometry | Mass Spectrometry in Proteome Research
Session Introduction
Eduard Rogatsky,
Albert Einstein College of Medicine, USA
Title: Impact of mass spectrometer-friendly mobile phases on reverse-phase columns selectivity
Biography:
Abstract:
Despite all C18 reverse-phase (RP) columns being based on the same general principle and even with the same non-polar ligand as stationary phase, many RP columns exhibit different selectivity. Due to strong variability in chromatographic behavior, RP columns are extensively tested and compared for LC/UV applications in conjunction with mobile phases containing 0.1% TFA (trifluoroacetic acid) or phosphate, and therefore are not optimal for MS analysis. In the current investigation, we tested 28 different brands of RP columns at different temperatures, and compared their performance and selectivity using MS-compatible mobile phases versus a “standard” mobile phase containing 0.1% TFA. Analytes tested were human insulin (~5,808 m.w.), glucagon (~3483 m.w.), C-peptide (~3020 m.w.) and angiotensin 1 (~1,296 mw). To conduct the current study, we built a simple and inexpensive system for automated column testing based on a column switching system from Valco Instruments. While all the tested columns demonstrated low bleed, stable background and also excellent analyte peak shape in the presence of 0.1% TFA, we found that only a fraction of the columns tested could provide good peak shape without TFA for all tested peptide analytes. For some columns, low concentrations of TFA in the mobile phase (which is compatible with LC/MS analysis) may improve peak shape. In many cases however, this did not completely eliminate peak broadening and tailing. We also noticed that silica pore size had only a minor influence on peak shape and utilization of wide pore silica (300A) could not be considered an efficient strategy for improvement of the peak shape for peptide analytes tested. The C18 columns tested demonstrated almost identical selectivity when using 0.1% TFA and 0.015% TFA in the mobile phase, while utilization of MS–compatible mobile phases for peptide separations with formic acid instead of TFA demonstrated strong, unpredictable variability in column selectivity with changes in the retention order of the peptides used in our study. These results emphasize the necessity of column testing if maximization of LC/MS assay performance (by enhancing of peak shape and maximizing column separation efficiency) is the goal.
Josef Havel
Masaryk University, Czech Republic
Title: MS for characterization and analysis of inorganic and nano materials - mass spectrometer as a kind of synthesizer
Biography:
Abstract:
Mass spectrometry (MS) is one of the most important techniques to fulfil demand for chemical/structural information frombiological systems. Structural analysis of organic compounds, biomolecules, etc. iswell known and widely used. Inorganic materials extensively penetrate to medicine as drugs, nano-carriers for drug delivery and also to MS as matrices (nano-gold, nano-diamonds) or as SALDI (Surface Assisted Laser Desorption Ionisation) or SELDI (Surface Enhanced LDI) matrices - to
enhance ionisation. However, MS of inorganic materials requires different approach: (i) mostly, usual MALDI matrices cannotbe used,
(ii) instead of MALDI just LDI is to be applied
(iii) fragmentation of materials (mostly unknown) accompanies LDI
and should be taken into account
(iv) interaction of the ions in plasma plume should be taken into consideration. In spite of
the problems it is demonstrated that MS can be applied for characterization/analysis of inorganic materials:
(a) nano-clusters
(or graphene oxide, etc.) can increase ionisation of biomolecules in MALDI
(b) in situ generated nano-clusters of monoisotopic
elements can be used as internal/external calibration standards
(c) structures of amorphous chalcogenides (atomic
switch memory Ge2Sb2Te5 thin films; As2Ch3S3 chalcogenides; As-S-Se or AgAsS2 glasses; complex glasses (GeSe2)(100-x)
(Sb2Se3)(x), etc. can be elucidated or determined (d) MS can be used for laser ablation synthesis (carbides, selenides, etc.) of
novel compounds. Concluding, even if the fragmentation accompanying often MS of inorganics is not completely known, MS is useful for materials analysis and in addition the instrumentation can be as a kind of a synthesizer applied for generation of new compounds.
Deepa Bisht
National JALMA Institute for Leprosy & Other Mycobacterial Diseases (ICMR), India
Title: Exploring aminoglycosides resistance in Mycobacterium tuberculosis isolates by proteomics and mass spectrometry
Biography:
Abstract:
Tuberculosis (TB) caused by the pathogen Mycobacterium tuberculosis (MTB) still remains a major global public health issue. India is a country burdened with high TB. Increasing incidences of multidrug-resistant strains and of co-infection with HIV, urgently demands better control measures. Aminoglycosides are broad spectrum antibiotics and are an important component of any antituberculosis therapy regimen and drugs of choice especially for category II patients. Streptomycin (SM), kanamycin (KM) and amikacin (AK) are the key aminoglycosides drugs against TB and resistance to these severely affects the options for treatment. They inhibit protein synthesis by interacting with steps of translation. Several explanations have been put forward for aminoglycosides resistance but still our understanding is fragmentary. As proteins manifest most of the biological processes, these are attractive targets for developing drugs, immunodiagnostics or therapeutics. Two-dimensional gel electrophoresis is an extremely powerful tool to dissect multiprotein complexes. Whole proteome analysis of aminoglycosides susceptible and resistant isolates by two-dimensional gel electrophoresis and their identification by mass spectrometry and bioinformatics tools have been carried out. Some proteins were found to be overexpressed in resistant isolates. Few of these were identified as hypothetical proteins. In silico docking analysis showed significant interactions of these drugs with hypothetical proteins. Characterization of hypothetical proteins may provide information about new targets against resistant TB or some new more effective vaccine candidates.
Jianke Li
Chinese Academy of Agricultural Science, China
Title: Proteomics reveals the molecular underpinnings of stronger learning and memory in eastern compared to western bees
Biography:
Jianke Li is form Chinese Academy of Agricultural Science, China
Abstract:
The characterized and compared the proteomes of mushroom bodies (MBs), antennal lobes (ALs) and optical lobes (OLs) in the brain of both species, and biologically validated the functions related to learning and memory. Acc and Aml have evolved similar proteome signatures in MBs and OLs to drive the domain-specific neural activities. In MBs of both species, commonly enriched and enhanced functional groups related to protein metabolism and Ca2+ transport relative to ALs and OLs, suggests that proteins and Ca2+ are vital for consolidatin g learnin g and me ory via modulati n of s nap tic struc ture an d signal tra nsduction. F urthe more, in O L sof both specie ,the main ly enriche d ribonucleosid metabo lis sugge ts its vita l role as secon d messenger i npromoting ph oto transd uction. Nota bly , in A sof both s pec es,distinct pr oteo me setting hav e shaped to p rim e olfactor y learni g and me ory. In AL of Acc this i s sup ported by the enr iched c yto kel ton or gan zati n to susta i nol actory signal ng through modu lation of pl asticity i ng lomeruli a nd in tracellu lar t rans port In AL of Aml, h owever, t he enriche d functional gro ups i pli ate d in hydrog e nio n transport are in dicativ e of their impo rta nce i su pport ing olfact ory proces ses by regu lati n of synap tic transmis sion. The biologic al c nfirmati n of enhance d act iviti sof protei metaboli s ma nd si nal trans duction in A Lsan d MBs of Acc rela tive t o in A ml demo nstr tes that a stron ger sense o f olf ctory l arn ng and memor y ha s evolved i Acc. The rep orte d first in depth proteome data of honeybe e br in s ub- organ s provide a nove l insigh t into the m olecular basis of neurobiolog y, and is po tent ially u seful for further neurologic al s tudies in ho neybees and other inse cts.
A P Sarycheva
National Research Nuclear University, Russia
Title: Mass-analyzer for micro and gaseous impurities analysis in solids
Biography:
Abstract:
The report describes a novel scheme of mass-analyzer. Such an analyzer allows performing an analysis of bothmicroimpurities and gaseous impurities with ppb level detection limit. The analyzer is based on ion gate between two wedge-shaped reflectors. Tandem laser is used for solids analysis. Such a laser provides with removing of adsorbed particlesfrom the surface, evaporation of solid sample and laser plasma generation. Ion gate allows intensive matrix ions package cut-off during ions’ transit, thus detection limits can be improved due to exclusion of scattered matrix ions’ influence on detection of microimpurities’ ions. Suggested evaporation and plasma generation regimes from solid surfaces allow standardless analysis of both microimpurities and gaseous impurities. Achievable resolution is over 7000 within ion drift of 37 cm. Proposed ion-optical scheme of mass-analyzer allows one to build the mass-spectrometer with compact design, featuring standardless analysis.
Hala Dadi
Université Paris-Sud-CNRS, France.
Title: Growth hormone: Chromatographic separations coupled with mass spectrometry towards a novel method for anti-doping tests
Biography:
Abstract:
Statement of the Problem: The recombinant human growth hormone (rhGH) is now widely manufactured by biotechcompanies. The rhGH is used to treat growth hormone deficiency diseases. On the other side, the availability of rhGH in the black-market has continuously increased because of doping in sports. To date, the detection of GH doping is still verychallenging. In fact, the natural and the biosynthetic hGH have identical peptide sequences. So far, the valid human growth hormone anti-doping tests are based on immunological recognition. However, immunoassays have their own limitations. Thehuman growth hormone concentration in blood depends on multiple factors. The secretion of GH is increased by physical activity and thus athletes produce more GH. In addition, the secretion of GH is pulsatile and influenced by nictemeral variations. These variations may lead to non-representative GH measurements. Therefore, the next generation analysis of GH has to be more specific and accurate. Mass spectrometry coupled with separation methods such as electrophoresis and chromatography could represent a more advanced instrumental set-up to find GH doping practices.
Methodology & Theoretical Orientation: Mass spectrometry coupled to reversed phase chromatography was used to find sachemical differences between the pituitary hGH and the rhGH. Intact GH proteins were separated by C8 or C18 columns prior to mass analysis.
Findings: The pituitary extracted hGH is glycosylated, whereas the biotech product is sugar free. Moreover, by chance thisglycosylation is bounded to a tryptic fragment that is proteospecific of the GH protein. Thus, we expect to be able to measure the concentration ratio between the wild type and the synthetic one.
Conclusion & Significance: The present work represents the first building blocks towards a novel methodology for a novel hGH anti-doping test. In addition, the collected data from the analysis of different hGH preparations should lead to other practical analytical application for quality control. The availability of a test that quantifies the natural and the rhGH will also be beneficial for GH deficiency diagnosis and treatment follows
C.J. Chukwu
Federal University, Nigeria
Title: Isolation, purification and characterization of flavor compounds in the fruit pulp of Synsepalum dulcificum
Biography:
Abstract:
Biography:
Abstract:
The family Cucurbitaceae includes pumpkins, squash, musk melon, watermelons and cucumbers which are known as vine crops. Netted muskmelon or cantaloupe is one of the most widely cultivated cucurbits. Cucumis melo var. cantalupensis and Cucumis melo var. reticulatus known in Egypt as Baladi and French cantaloupe, respectively are the most famous varieties in Egypt. Previous studies have shown that cantaloupe pulp extract possesses high antioxidant and anti-inflammatory properties but nothing was traced about its peels. The objective of the present study is to evaluate the in vivo anti-inflammatory activity of the peels and pulps of the two Egyptian varieties of cantaloupe, together with UPLC-MS/MS analysis of their constituents. Pretreatment of rats with the 95% ethanol extract of the pulps and peels of the two varieties at the two dose levels of 25 and 50 mg/kg, significantly inhibit the carrageenan-induced increase in the edema volume of rat paws after three hours, except the low dose levels of the French pulps. Injection of carrageenan into the rat hind paw induced a significant increase in the hind paw prostaglandins (PEG-2), tumour necrosis factor (TNF-α) and interleukins (IL-6 and IL-1β) concentration, three hours after injection, compared with those found after saline injection. Treatment of rats with the tested extracts at both dose levels caused a significant reduction of increased PEG-2, TNF-α IL-6 and IL-1β generation by carrageenan (P<0.05). UPLC-MS/MS allowed the identification of 46 phenolic compounds including phenolic acids and flavonoids. It is worthy to note that this is the first report for the chemical and biological study of the peels of Cucumis melo var. cantalupensis and Cucumis melo var. reticulatus.
Tehami Wafaa
Université Djillali Liabes, Algeria
Title: Contribution to the phytochemical and antimicrobial study of the leaves and roots of Salvia argentea from western Algeria
