18th International Conference on World HPLC & Separation Techniques August 29-30, 2018 Toronto, Ontario, Canada

Chair

Dusan Berek

Polymer Institute of the Slovak Academy of Science

Co-Chair

Brigitte Simons

Molecular Science Corp., Canada

Session Introduction

Milos Netopilik

Institute of Macromolecular Chemistry, Czech Republic

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

Time : 00:00-00:20

Biography:

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

Abstract:

The shape of the elution curves depends strongly on experimental conditions, in the first place on polymer molecular weight, upon this, on concentration and flow-rate. The effect of concentration is weak for polymers in theta solvents

up to the concentrations of overloading. On the other hand, in good solvents, the concentration effect is important. The effective hydrodynamic volume of dissolved macromolecules decreases with increasing concentration. The decrease in the hydrodynamic volume is of solvated molecules with increasing concentration is an established experimental factor which

theoretical explanation. The spatial distribution of the analyte with respect to the longitudinal axis of the separation system, developing in time, can be expressed by the binomial distribution. However, further treatments of this physical situation were approximative. The exact solution to the problem is obtained as the observation at a fixed point (the detector) of this binomial distribution developing in time after reaching the exclusion limit. This can be done numerically. The description of the concentration effect on SEC elution curves is possible on the basis of the displacement- equilibrium model. This is based on the concept of a theoretical plate on which the equilibrium is formed between molecules of the analyte moving together with mobile phase (MP) and those anchored on the or penetrated into the pores of the stationary phase (SP). The simulation of the concentration effect is possible with partition coefficient calculated numerically for each plate at each displacement.

Chair

Dusan Berek

Polymer Institute of the Slovak Academy of Science

Co-Chair

Brigitte Simons

Molecular Science Corp., Canada

Session Introduction

H W C Krishanthi Karunarathne

Government Analyst Department, Sri Lanka

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

Time : 00:00-00:20

Biography:

H W C Krishanthi Karunarathne has completed her MSc. in analytical chemistry, University of Peradeniya, Sri Lanka. She is the Assistant Government Analyst in

Government Analyst Department in Sri Lanka. She has published more than 10 papers in some reputed journals.

Abstract:

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

Chair

Dusan Berek

Polymer Institute of the Slovak Academy of Science

Co-Chair

Brigitte Simons

Molecular Science Corp., Canada

Session Introduction

Zahra Ramezani

Ahvaz Jundishapur University, Iran

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

Time : 00:00-00:20

Biography:

Zahra Ramezani is currently professor of analytical chemistry at Ahvaz Jundishapur University of Medical Sciences. She earns her BSc in pure chemistry, MSc and Ph.D. in analytical chemistry at Shiraz University, Iran. She is currently working on the synthesis of artificial antibodies for drug and diseases biomarker

determinations in complicated matrixes such as urine, saliva and plasma as well as drug delivery.

Abstract:

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

Chair

Dusan Berek

Polymer Institute of the Slovak Academy of Science

Co-Chair

Brigitte Simons

Molecular Science Corp., Canada

Session Introduction

Julio C Fernandez Travieso

National Centre for Scientifi c Research, Cuba

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

Time : 00:00-00:20

Biography:

Julio Cesar Fernandez Travieso is a Senior Investigator in the Clinical Trials Unit, National Centre for Scientific Research, Havana, Cuba. He has completed his BSc

in Pharmaceutical Sciences from Havana University, Cuba in 1996. He was awarded PhD in Pharmaceutical Sciences in 2003. He has published more than 130 publications and presented more than 100 papers in various scientific events. His research interest mainly focuses on clinical trials phase I-IV of different natural products: Policosanol, Abexol, Prevenox and Palmex.

Abstract:

Introduction: Stroke is one of the leading causes of mortality and disability. Clinical studies results show that policosanol (20 mg/day) + standard aspirin (AS) therapy had benefits versus placebo + AS given for 6 and 12 months to patients with recent ischemic stroke.

Objective: To analysis, the policosanol treatment effects in the hypertensive patients included in two ischemic stroke recovery trials.

Methods: This report was analyzed the records of all hypertensive patients included in two ischemic stroke recovery studies. Hypertensive patients with a modified Rankin Scale score (mRSs) 2 to 4 were randomized, within 30 days of onset, to policosanol/ AS or placebo/AS, for 6 months. The primary outcome was a mRSs reduction. Low-density Lipoprotein-cholesterol (LDL-C)

reduction and High-Density Lipoprotein-Cholesterol (HDL-C) increase were secondary outcomes.

Results: One hundred forty-two hypertensive patients (mean age: 66 years) were included in the analysis. Policosanol/AS decreased significantly the mRSs mean from the first interim check-up (3 months) (p<0.0001 vs. placebo/AS). The policosanol treatment effect did not wear off, on the contrary, even improved after 6 months therapy (p<0.0001 versus placebo/AS). Moreover, policosanol/AS (57/71; 80.3%) treatment achieved significant results (mRSs ≤1; p<0.0001). Whereas the placebo/ AS did not (6/71; 8.5%). Treatments were well tolerated. Two patients discontinued prematurely and four patients (2 from policosanol/AS group and 2 from placebo/AS) referred mild AE.

Conclusions: Six months administration of policosanol/AS given to hypertensive patients after suffering ischemic stroke demonstrated to be better than placebo/AS in improving functional outcomes at 3 and 6 months when used among hypertensive patients with ischemic stroke.

Chair

Dusan Berek

Polymer Institute of the Slovak Academy of Science

Co-Chair

Brigitte Simons

Molecular Science Corp., Canada

Session Introduction

Ishiaq Omotosho

University of Ibadan, Nigeria

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

Time : 00:00-00:20

Biography:

Abstract:

The incidence of kidney failure is on the increase, unfortunately, traditional renal function markers are equivocal especially at the early stage until end-stage renal disease when kidney transplant becomes inevitable. Hence, the need for an early and more sensitive marker of renal damage indicating the presence of covert renal damage in occupational lead toxicity is imperative. This work is proposing diagnostic methods that could predict the development of Chronic Renal Failure (CRF) especially in occupational lead-exposed subjects combining results of conventional and new biomarkers of kidney damage using

a mathematical model based on Area under the Receiver Operating Characteristics (AUROC). Traditional Renal Function markers (TRF) (plasma creatinine, urea and uric acid) were determined in one hundred each of Lead-Exposed Subjects (LES) and non-exposed, non-nephrotic adults (control) along with sixty Chronic Renal Failure patients (CRF) (all age-matched) using standard spectrophotometric methods. Blood lead level (Pb) was determined in all participants using Atomic Absorption Spectrophotometry (AAS) while levels of novel urinary renal enzymes - Glutathione-S-transferase (GST) and N-acetyl-β-Dglucosaminidase (NAG)- activities were also evaluated using ELISA techniques. Pb was used as True Positive Indices (TPI) and TRF along with NAG and GST were used as False Negative Indices (FNI). Ratios of mean, Creatinine : GST (A) (0.01, 0.02 and 0.09), Creatinine:NAG (B) (0.03, 0.08 and 0.6), Uric acid : GST (C) (0.05, 0.08 and 0.08), Uric acid : NAG (D) (0.29, 0.3 and 0.55), Urea : GST (E) (0.17, 0.55 and 0.93), Lead : GST (F)(0.42, 0.59 and 0.88), Lead : NAG (G) (2.56, 2.28 and 6.09), Lead : Creatinine (H) (80.62, 30.37 and 10..22), Lead : Urea (I) (2.46, 1.07 and 0.95) and Lead : Uric acid (J) (8.66, 7.61 and 11.12) for LES, control and CRF groups respectively were computed and used to plot an ROC curve using the FNI values as the abscissa and the TPI values as the ordinate while their AUC were calculated. The AUC values for Lead : Creatinine, Lead Urea and Lead

: Uric acid were 1.00, 0.917 and 0.833 respectively. We suggest that application of this model after proper standardization may be useful in early identification of covert kidney damage especially in occupationally vulnerable group.

Alina Vasilescu

International Centre of Biodynamics, Romania

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

Time : 00:00-00:20

Biography:

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

Abstract:

Lysozyme is used as a model to study protein function and enzyme catalysis, is suggested as a biomarker in various diseases and also used as an antimicrobial agent in the food industry. Various methods have been reported for lysozyme detection based on its physicochemical properties, enzymatic activity or affinity for biological receptors. The aptasensors with detection by Surface Plasmon Resonance (SPR) developed by our group are versatile tools for the detection of residual lysozyme in wines or of lysozyme dimer in aggregated solutions. Advancing from these concepts relying on thiol coated plasmonic interfaces, we report the development of graphene oxide (GO) coated plasmonic interfaces via the layer-by-layer method, as robust and sensitive platforms with controlled thickness. Furthermore, the GO-coated interfaces were easily modified with whole cells of Micrococcus lysodeikticus- an enzymatic substrate for lysozyme. Detection of lysozyme in spiked serum samples was achieved on the principle of lysozyme’s lytic action causing desorption of bacteria from the interfaces and consequently changes in the

SPR signal. The analysis time was 3 minutes and the detection limit was 3.5 nM. A second sensing concept exploited the affinity of lysozyme for an aptamer, fixed covalently to the GO-coated interfaces. In this case, a detection limit of 0.71 nM and a linear range of 2-21 nM were observed. The two analytical strategies are based on different sensing mechanisms, nonetheless, both are sensitive and easy to implement with GO-coated interfaces suggesting a high potential and versatility of these interfaces for bioanalytical purposes.

Chair

Milos Netopilik

Institute of Macromolecular Chemistry, Czech Republic

Co-Chair

Abuzar Kabir

International Forensic Research Institute, USA

Session Introduction

Dusan Berek

Polymer Institute of the Slovak Academy of Science, Slovakia

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

Time : 00:00-00:20

Biography:

Dusan Berek is employed at Polymer Institute, Slovak Academy of Sciences in Bratislava. Served as elected member of the Presidium of the Slovak Academy of Sciences, President of the Slovak Chemical Society, Chairman of the Czecho-Slovak and Slovak National Committee of Chemistry for IUPAC. Corresponding member of the Central European Academy of Sciences and member of the Learned Society of the Slovak Academy of Sciences. Author or co-author of two monographs and 300+ scientific papers in extenso published in refereed periodicals, proceedings and chapters of books, as well as 60+ patents (four of them were licensed) - cited more than 3,000x. Presented over 130 invited plenary, key and main lectures, as well as over 900 regular lectures and poster contributions on symposia and conferences, as well as during lecturing tours to over fourty countries. Elected "Slovak scientist of the year 1999" and "Slovak innovator of the year 2002".

Abstract:

Block copolymers present an important group of materials with numerous applications in science, medicine and technology. In a block copolymer, at least two chemically distinct polymer chains are connected with a chemical bond. Comprehensive molecular characterization of block copolymers is an analytical challenge. A special problem makes a determination of the amount and molar mass of parent homopolymers, which occur in most block copolymers and constitute highly undesired ballast. Gel permeation (size exclusion) chromatography GPC/SEC is commonly employed for the characterization of block copolymers. Molar mass of a precursor, the block polymerized as first, is determined by GPC/SEC and the same method serves for the approximate assessment of total molar mass of the block copolymer under study. Due to its low separation selectivity and detector sensitivity, GPC/SEC can hardly identify the presence of parent homopolymers and render their molar mass.

We will discuss principles and applications of the new separation methods namely liquid chromatography under limiting conditions of enthalpic interactions, LC-LC and sequential two-dimensional polymer liquid chromatography, S2D LC to solve the above tasks. LC-LC methods are well robust and experimentally feasible. Their separation selectivity is very high and sample recovery is reasonable. LC-LC can efficiently separate both parent homopolymers from diblock copolymers in one single step. The separated sample constituents can be one-by-one forwarded into an on-line GPC/SEC column for determination of their molar mass average and distribution. Parent homopolymers present in the block copolymer at a very low concentration below 1% of can be tracked-down and characterized with help of the S2D LC.

Michael M Shapi

Mangosuthu University of Technology, South Africa

Title: Occurrence and levels of potentially harmful elements (PHEs) in natural waters of the gold mining areas of the Kette-Batouri Region of Eastern Cameroon

Time : 00:00-00:20

Biography:

Abstract:

This study was designed to determine the extent to which alluvial and bedrock gold mining activities in the Kette-Batouri region of Eastern Cameroon is responsible for the discharge of potentially harmful elements (PHEs) into natural water bodies. This is considered to be of great public concern, since elevated levels of PHEs in these water bodies, which are widely used for domestic purposes, could have adverse health and environmental effects on the population and nearby ecosystems. The investigation into the levels of PHEs in natural water bodies of this area was conducted to ascertain the toxicity posed by mine waste stockpiles as data from the literature on this subject are scarce. Fourty two water samples from the region were analyzed for some 60 PHEs by ICPOES, of which 22 that were not below the detection limit were considered for further data analysis. Maximum total concentrations in water of As, Cr, Pb and V and Zn are above the World Health Organization (WHO) maximum allowable concentrations (MAC) levels and are as follows (μg l−1): As (21.90-50.9); Cr (1.80-57.30); Pb (0.50-34.70) V (24.70-77.20) and Zn (3.10-481.70). This information is consistent with our recent research efforts which have indicated moderate pollution by heavy metals in the soils with a slight deterioration of site quality in this region. The data generated from this investigation is important in the formulation of water management strategies and recommendations for remediation of water bodies at abandoned mine sites for meeting water quality standards.