Day 2 :
Keynote Forum
Brigitte Simons
Molecular Science Corp., Canada
Keynote: Differentiating cannabis products within the brands of the legalized adult use markets
Time : 00:00-00:40
Biography:
Brigitte Simons is a business development executive in support of leading-edge laboratory services and data management tools for the development of safe cannabis. Bridging expertise within analytical science, pharma drug development and environmental testing – Brigitte have a professional track record for laboratory testing instrumentation, software and sample contract design for the Canadian federal agencies, such as Canadian Food Inspection, Health Canada, Agriculture Canada and Environment Canada. She spent over 6 years working in the Drug Toxicology and Analysis Division at Health Canada in a mass spectrometry facility testing. She completed two post-doctoral fellowships at the Clinical Sciences Hospital of the National Heart, Blood & Lung Institute within the famous NIH campus in Maryland, USA. Continuing on in lab specialties, Brigitte then joined SCIEX, a global instrumentation vendor for hardware and software for mass spectrometry. With over 15 years experience with operating mass spectrometers, Brigitte managed Canadian federal and provincial government sales for full laboratory services, covering clinical, forensics to product health and environmental safety. Prior to working abroad, Brigitte received her Ph.D. in Chemical Biology at the University of Ottawa in a joint chemistry program with drug pharmacology at Health Canada.
Abstract:
Within the framework of Bill C-45, Canada is positioned to become the global leader in the legal cannabis economy and global exporter. The enactment of this Canadian Cannabis Act provides legal access to marijuana and to control and regulate its production, distribution and sale. The primary objective of Health Canada’s regulatory policy bears stringencies with respect to public health and safety and strict requirements for quality assurance, record keeping and mandatory testing by 3rd party laboratories for product contamination. This opens an opportunity for advancing analytical development for cannabis metabolite profiling of active natural products and bleeds through to the accurate quantitative reporting of pesticides, mycotoxins and heavy metalloids that serve regulatory audit to clear products for sale. A complete LC-MS/MS workflow is described to quantitate 14 cannabinoids and screen for over 40 terpenoids to fingerprint various top cannabis dried flower brands from the large enterprise-producers in a method that is delivered in under 15 mins of analytical run time using a dual ESI and APCI ionization strategy. A wide linear dynamic range of 0.03 to 90% measurement (104 orders LDR) of cannabinoid per LC-MS injection can be reported to provide a more accurate view for product labeling and dosing recommendations. Terpene expression and metabolite measurement in plant cultivars are becoming less challenging with newly identified terpene synthases and availability of new mono-terpenes and sesquiterpene standards. It is of high interest for results of these metabolite profiling experiments to be correlated with plant cultivation parameters to achieve quality control and strengthen the consumer's experience with a brand of cannabis and differentiate products for retail. Furthermore, pesticide residue analysis in cannabis flower and oil formulations has been developed to meet the reporting requirements of Health Canada’s banned pest control ingredients list. With UHPLC linked tandem mass analysis covering all of the 96 banned pesticides except for 11 compounds best suited by GC separation, it is possible to achieve a validated cannabis product certificate of analysis for issuance to cannabis licensed producers in rapid turn-around. Analytical method details include LC separation using the Raptor Restek Column, Raptor Biphenyl and newly available mixtures of pesticide standards to meet the Canadian Pest Management Agency’s list of required pesticide maximum residual levels (down to 10 ppb in most cases). The addition of mycotoxins and other organo-contaminants can also be inserted into our methods with the use of optimized Scheduled MRM mass spec scanning techniques. The assembly of all the potency and ingredients data collection possible can provide information to consumers and track benefits to the cannabis producers stride to bring powerful brands to the global cannabis market.
Keynote Forum
Manuela G Neuman
University of Toronto, Canada
Keynote: Alcohol and drugs
Time : 00:00-00:40
Biography:
Manuela Neuman is the CEO of In Vitro Drug Safety and Biotechnology, Toronto, ON., Canada. She is also teaching Pharmacology and Toxicology at the Faculty of Medicine, University of Toronto, Toronto, Canada. She is the Chair of Clinical Toxicology and Drug of Abuse Committee of the International Association ofa Therapeutic Drug Monitoring and Clinical Toxicology. She published 300 peer review articles. Her specialized laboratory provides personalized medicine and precision medicine results for Canada, USA and Europe.
Abstract:
The interplay of alcohol with drugs includes multiple facets. These include the effects of alcohol on the effects of other hepatotoxicants and on the pharmacological effects of various drugs. Also relevant is the possible role of alcohol on the effects of carcinogenic agents. Less striking, but significant, are the effects of other drugs on the effects of ethanol. More difficult to identify but presumably significant, are the effects of alcohol-drug interplay on the development of an alcoholic liver disease. A common denominator of them is the role of ethanol-induced P-4502E1 (CYP2El) in affecting the toxicity ofsome hepatotoxicants and the effects of some drugs. Less prominent but also relevant is the effect of interplay with alcohol dehydrogenase and aldehyde dehydrogenase in the toxicity of some drugs. Alcohol has been shown to be responsible for cirrhosis in the 18th century and was labeled a hepatotoxin in the 19th century. During the second half of the 20th century alcohol has been recognize to enhance the toxic effect of other hepatotoxic agents such as acetaminophen, aflatoxin B1, allyl alcohol,bromobenzene, cocaine, enflurane, galactosamine, halothane, isoniazid, nitrosamines, thioacetamide, vinyl chloride and vitamin A. The toxicity of several hepatotoxicants is unaffected and of at least one, amanitine, is decreased by ethanol. The effect of ethanol on the toxicity of carbon tetrachloride and acetaminophen have been studied most extensively. The enhancement of toxicity by ethanol does not depend on an ethanol-induced hepatic injury but rather on the activity of the cytochrome P450 2E1 that converts the respective toxicants to their active metabolites. Nevertheless, inhibition by ethanol of regenerative response to injury may contribute to an enhancement of toxicity by ethanol. The toxicity produced by ethanol may have a bearing on the liver disease of alcoholism as well as on the toxicity and carcinogenicity of individual toxicants.
Keynote Forum
Abuzar Kabir
Florida International University, USA
Keynote: Fabric Phase Sorptive Extraction (FPSE): A versatile sample preparation technology that meets the demands of twenty first century modern analytical laboratories
Time : 00:00-00:40
Biography:
Abuzar Kabir is a Research Assistant Professor in the Department of Chemistry and Biochemistry, Florida International University (FIU), Miami, Florida, USA. His research interest primarily focusses on synthesis and applications of novel sol-gel derived advanced material systems (chromatographic stationary phases, surface coatings of high-efficiency microextraction sorbents, nanoparticles, microporous and mesoporous functionalized sorbents) for analyzing polar, medium polar, nonpolar, ionic analytes, heavy metals and organometallic pollutants from biological/pharmaceutical/clinical/environmental sample matrices. He is an ardent advocate of Green Analytical Chemistry (GAC). His recent inventions, fabric phase sorptive extraction (FPSE), dynamic fabric phase sorptive extraction (DFPSE), Capsule Phase Microextraction (CPME), substrate-free liquid chromatographic stationary phases and extraction sorbents, organic polymeric liquid chromatographic stationary phases and extraction sorbents and universal molecular imprinting technology have drawn tremendous interests among the researchers. He has published more than 50 peer-reviewed journal articles, 9 book chapters and 90 conference proceedings. Dr. Kabir has invented numerous chromatographic stationary phases and sample preparation technologies, resulting in 15 US patents.
Abstract:
Statement of the Problem: The invention of fabric phase sorptive extraction (FPSE) has begun a new era in analytical sample preparation by ingeniously combining two competing for sample preparation techniques, solid phase extraction (SPE) and solid phase microextraction (SPME) into a single sample preparation technology platform. The integrated system, FPSE utilizes a flexible, yet active fabric (cellulose, polyester and fiberglass) substrate to host a thin layer of sol-gel derived extracting sorbent. The engineered selectivity of the sol-gel sorbents and the hydrophobicity/hydrophilicity of the fabric substrate synergistically complement to the net polarity of the fabric phase sorptive extraction medium and consequently, determine its extraction efficiency. The sponge-like porous architecture of sol-gel extraction sorbent and the inherent permeability of the fabric create anextraction medium that mimics a solid phase extraction disk and allows permeating aqueous sample matrix through its body, leading to rapid sorbent-analyte interaction and subsequent successful retention of the analyte(s) onto the extraction medium. The flexibility of the FPSE medium permits direct insertion into the sample container for analyte extraction and thus minimizes the number of transfer containers used in the sample preparation process. The sol-gel coating technology allows utilization of typical functional ligands commonly used in solid phase extraction such as C8/C18 as well as polymers used in solid phase microextraction such as polydimethylsiloxane (PDMS). Unlike SPE and SPME, FPSE can be performed either in equilibrium extraction mode (as in SPME) or inexhaustive extraction mode (as in SPE). In addition, sol-gel coated sorbents demonstrate superior thermal, solvent and pH stability (1-13) compared to conventional sorbents. Due to these unmatched advantages, FPSE has gained considerable popularity in a short period and has demonstrated numerous applications in a wide variety of samples including food, biofluids, wastewater and air. In the current talk, analytical data pertaining to some fascinating applications of FPSE will be presented.
Keynote Forum
Peng Chen
Chemic Labs Inc., USA
Keynote: Applications of UPLC-MS QTOF in structural elucidation of small molecules
Time : 00:00-00:40
Biography:
Peng Chen received a Ph.D. in Analytical Chemistry from Indiana University in 1998 and a M.S. in Organic Chemistry from the University of Louisville in 1994. Hisgraduate research includes the introduction of osazones as MALDI matrices for carbohydrate analysis and the structural elucidation of fluorescent aging markers. He has been working in various chemical industry sectors in the fields of chromatography and mass spectrometry. His work in recent years at Chemic Labs Inc. involves structural elucidation of small molecules in pharmaceuticals and medical devices by high-resolution QTOF mass spectrometry.
Abstract:
The structural elucidation of small molecules by high-resolution mass spectrometry plays important roles in developmentand quality control of pharmaceuticals and medical devices. Trace amounts of small molecules can be present in forms of impurities, by-products or degradation products, etc. It is often difficult to separate and fractionate enough quantities of these analytes for conventional structural analysis by NMR and FTIR. Recent advances in instrumentation and software of UPLC-MS QTOF with MS/MS fragmentation capability can give structural insight into molecules of interest and in many cases offer structure candidates at high confidence. This presentation will use several practical examples in the analysis of synthetic compounds and identification of impurities associated with pharmaceuticals and medical devices to illustrate the convenience and power of UPLC-QTOF high-resolution mass spectrometry.
- Chromatography
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 whichtheoretical 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.
- Principles and Applications of HPLC
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.
- Polymer and Material Chemistry
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.
- Good Pharmacovigilance Practice
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
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
- Disease Detection and Formulation Development
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
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
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.
- Advances in Chromatography & HPLC Techniques
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
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.