Hi [[ session.user.profile.firstName ]]

Retention Mechanisms in HILIC Chromatography: Robust Method Development

Hydrophilic interaction liquid chromatography (HILIC), especially in conjunction with mass spectrometry (MS), has become a powerful tool for the analysis of a wide variety of challenging analytes. Applications of the technique have increased dramatically over the past decade, especially for the analysis of polar analytes where reversed-phase chromatography suffers. HILIC conditions employ a high percentage of acetonitrile which enables facilitated solvent evaporation in LC/MS sources and thus often an increase in analyte response when compared to more aqueous based systems. The increased retention of polar analytes afforded by HILIC provides improved selectivity and decreases the impact of endogenous species, often leading to improved qualitative and quantitative analyses.

Although HILIC has proven useful, it has also been thwarted with complications including difficulties in method development and method robustness.

In this presentation, studies investigating the underlying retention mechanisms dominant in HILIC chromatography are presented and discussed. Along with reversed-partitioning HILIC is well known to exhibit, ion-exchange and the interplay of the dominant mechanisms are unveiled and used to develop a model of overall retention and selectivity. Interactions that operate using different stationary phase chemistries and conditions are presented. The impact of analyte polarity and charge as well as the variations caused by high percentages of organic on these physiochemical parameters are highlighted. Throughout the discussion, examples of use and misuse of HILIC are employed to illustrate these important concepts to build a solid fundamental foundation for efficient and effective use of this powerful technique.
Recorded Jun 28 2012 55 mins
Your place is confirmed,
we'll send you email reminders
Presented by
David S. Bell, Ph.D.
Presentation preview: Retention Mechanisms in HILIC Chromatography:  Robust Method Development

Network with like-minded attendees

  • [[ session.user.profile.displayName ]]
    Add a photo
    • [[ session.user.profile.displayName ]]
    • [[ session.user.profile.jobTitle ]]
    • [[ session.user.profile.companyName ]]
    • [[ userProfileTemplateHelper.getLocation(session.user.profile) ]]
  • [[ card.displayName ]]
    • [[ card.displayName ]]
    • [[ card.jobTitle ]]
    • [[ card.companyName ]]
    • [[ userProfileTemplateHelper.getLocation(card) ]]
  • Channel
  • Channel profile
  • Pediatric Cancer and IHC Sep 28 2016 5:00 pm UTC 75 mins
    Jeff Gordon, Director of OEM Sales, Cell Marque Corporation
    Childhood cancer and pediatric cancer are general terms used to describe a wide range of neoplasms found in children and teenagers. Occurring in approximately 1 in 300 people under the age of 20, compared to 1 in 6 adults, pediatric cancers are more rare than adult cancers. Because less is known about pediatric cancers, diagnosis can be quite challenging for pathologists.

    This presentation covers the basic science, as well as facts and statistics about pediatric cancer. We will discuss how the utility of immunohistochemical testing along with the application of novel antibodies can contribute to accurate diagnosis and survival rates of pediatric cancer patients.
  • Skin Cancer and Immunohistochemistry Recorded: Jun 22 2016 63 mins
    Jeff Gordon, Director OEM Sales - Cell Marque Corporation, Rocklin CA
    Skin cancer is by far the most prevalent cancer. Each year, approximately 3.4 million people in the US alone are diagnosed with some form of skin cancer. Skin cancer can be highly treatable if it is detected and classified early, and this detection and classification is often aided by immunohistochemistry. This presentation covers many of the basic science, facts, and statistics of skin cancer, as well as the utility of immunohistochemical testing with markers such as S-100, SOX-10, Ber-Ep4, and HHV-8 in the accurate diagnosis and survival rates of skin cancer. Continuing education credits for attending this webinar will be offered through the National Society of Histotechnology
  • Growing Cell Line Potential: From Lab To Licence S2 Recorded: Jun 9 2016 26 mins
    Dr. Lily Chan, Cambridge Enterprise
    A scientific overview of the portfolio of cell lines University of Cambridge has deposited at ECACC with a focus on the KARPAS 299 and KARPAS 422 cell lines; the HeLa Mitotrap cell lines; and CHO cell lines. We will also provide an overview of the process of partnering with ECACC and Sigma-Aldrich for the storage and distribution of cell lines for research purposes. Topics include: The scientific applications of the cell lines; The types of companies and institutions we license to; Advantages of partnering with culture experts and specialised distributors; Our experience of working with ECACC and Sigma-Aldrich.
  • Growing Cell Line Potential: From Lab To Licence S1 Recorded: Jun 9 2016 24 mins
    Dr. Lily Chan, Cambridge Enterprise
    A scientific overview of the portfolio of cell lines University of Cambridge has deposited at ECACC with a focus on the KARPAS 299 and KARPAS 422 cell lines; the HeLa Mitotrap cell lines; and CHO cell lines. We will also provide an overview of the process of partnering with ECACC and Sigma-Aldrich for the storage and distribution of cell lines for research purposes. Topics include: The scientific applications of the cell lines; The types of companies and institutions we license to; Advantages of partnering with culture experts and specialised distributors; Our experience of working with ECACC and Sigma-Aldrich.
  • Sample Preparation for Food Analysis- Part 3 Recorded: May 27 2016 42 mins
    Matthias Nold, Product Manager- Merck
    Part 3 of a 3-part presentation series about food analysis focused on sample preparation and including information about Reference Material
  • Sample Preparation for Food Analysis- Part 2 Recorded: May 25 2016 33 mins
    Uwe Wagner, Marketing Manager - Merck
    Part 2 of a 3-part presentation series about food analysis focused on sample preparation and including information about Reference Material
  • Sample Preparation for Food Analysis- Part 1 Recorded: May 13 2016 70 mins
    Jennifer Claus, M.S.
    Webinar recording for sample preparation on Food Analysis.
  • Colorectal Cancer and Immunohistochemistry Recorded: Mar 30 2016 58 mins
    Jeff Gordon, Cell Marque- Rocklin, CA
    Colorectal cancer is the third most commonly diagnosed cancer in men and women, and is the second leading over all cause of cancer deaths in the United States. Each year, the CDC reports that approximately 135,000 people are diagnosed with colorectal cancer, and over 50,000 succumb each year to the illness. The National Cancer Institute cites a decline in the mortality rate of colorectal cancer due to advanced screening and early diagnosis. Immunohistochemistry has served as the catalyst for these advancements in colorectal cancer diagnosis. This presentation covers many of the basic science, facts, and statistics of colorectal cancer, as well as the utility of immunohistochemical testing with markers such as cadherin¬17, CDX¬2, SATB2, and the mismatch repair proteins in the accurate diagnosis and survival rates of colorectal cancer.
  • Designing the Next Phase in Genome Editing: Advanced CRISPR Applications Recorded: Mar 1 2016 44 mins
    Jeremy Lehmann- CRISPR Product Specialist, Functional Genomics, MilliporeSigma
    CRISPR Cas9 nucleases have revolutionized the field of genome editing enabling unprecedented efficiency of gene targeting in a vast array of cell types and organisms. Even with such powerful technology at hand, researchers who are new to the field may find genome modification to be challenging and time-consuming. As CRISPR becomes a focus of the molecular biology research community, MilliporeSigma seeks to share the best approaches learned and methods applied in our years of genome editing experience. Today’s presentation will focus on practical applications of CRISPR for pristine genome editing to achieve knockout as well as specific sequence changes to include donor-mediated snps, reporter-tags and conditional knockouts. Special attention will be paid to design considerations for the donor constructs necessary to achieve specific sequence changes. Finally, the frontiers of CRISPR technology, including synthetic crRNA to fast-track genome editing experiments, whole genome screening and targeted gene activation will be explored.
  • The Analysis Workflow of the New EPA Method 325 for Fenceline Monitoring Recorded: Feb 17 2016 52 mins
    Jamie Brown - Supelco R&D Scientist , Lee Marotta - PerkinElmer Senior Field Application Scientist
    The new EPA METHOD 325 for sampling Volatile Organic Compounds from Fugitive and Area Sources was promulgated in September of 2015. The petroleum refineries have 2 years to comply with the new ruling. Benzene emissions are the focus of the new ruling, but other VOC’s can be monitored using the same sampling tube. The air samples are collected using thermal desorption tubes that are deployed along fenceline of the property and remain deployed for 14 days. The samples are collected passively, without the using of an air sampling pump. After sampling the diffusive endcap is replaced with a metal storage cap and sent to a laboratory where they’re analyzed by thermal desorption gas chromatography. This is the first of a two series presentation that focuses on the analysis side of the workflow. It provides users the steps required to comply with the new EPA fence line regulations and what is needed to optimize the method for high sample throughput. Refineries and testing laboratories will benefit by gaining valuable information on the optimum techniques to perform this method and what it takes to comply. An introduction to the theory and operation of thermal desorption will also be included.
  • Gene Synthesis as an Alternative to Molecular Cloning Session 2 Recorded: Feb 16 2016 31 mins
    Dr. Zhen (Calvin) Zhang, Scientist, Project Management, GENEWIZ
    Gene synthesis has become a quick and cost-effective way to build your DNA construct for a variety of applications - from protein engineering to bio-based circuit and pathway design. This webinar, presented by Sigma-Aldrich and GENEWIZ will explain the process of gene synthesis, how it compares to traditional cloning techniques, and review some key applications researchers are addressing with synthetic DNA.
  • Gene Synthesis as an Alternative to Molecular Cloning Session 1 Recorded: Feb 16 2016 30 mins
    Dr. Zhen (Calvin) Zhang, Scientist, Project Management, GENEWIZ
    Gene synthesis has become a quick and cost-effective way to build your DNA construct for a variety of applications - from protein engineering to bio-based circuit and pathway design. This webinar, presented by Sigma-Aldrich and GENEWIZ will explain the process of gene synthesis, how it compares to traditional cloning techniques, and review some key applications researchers are addressing with synthetic DNA.
  • High Protein Yields in Shaken Cultures by EnPresso Growth System Recorded: Feb 9 2016 62 mins
    Dr. Kaisa Ukkonen Head of Protein Expression Technologies, BioSilta & Dr. Antti Vasala,R & D Director, BioSilta
    EnPresso, a proprietary growth system from BioSilta Ltd, applies slow enzymatic glucose release to control the growth and protein synthesis rate in bacteria and yeasts. This high-cell-density growth system can be used in any shaken cultivation device at various scales. In this webinar, Dr. Kaisa Ukkonen and Dr. Antti Vasala will review the basic requirements for successful high-cell-density cultivations and discuss common reasons for failures in E. coli-based protein production, the focus being on the cultivation conditions. In particular, prerequisites for successful scale-up will be presented. The webinar also features examples on the use of EnPresso demonstrating the time and cost savings enabled by this powerful growth system, and presents an interesting alternative protein expression protocol: IPTG-based autoinduction.
  • Sulfonyl Fluorides in Chemical Biology and Drug Discovery Recorded: Jan 26 2016 61 mins
    Dr. Lyn Jones, Head of Rare Diseases Chemistry. Chemical Biology Pfizer
    Sulfonyl fluorides are privileged protein-reactive functional groups due to their context-specific reactivity with a broad range of amino acid residues (tyrosine, serine, lysine, threonine, cysteine and histidine). As a result, they are ideally suited to a myriad of applications in chemical biology and drug discovery. To highlight the growing utility of sulfonyl fluoride chemical probes, examples will be provided from the literature and work in which Dr. Jones has been involved.
  • Graphene and New Monoatomic Materials Recorded: Dec 10 2015 73 mins
    Prof. Vincenzo Palermo, Functional Organic Materials National Research Council, Rome, Italy
    Using 2-dimensional nanosheets in a 3-dimensional world:
    At the beginning of its exciting life, graphene was mostly a game for physicists. Then, chemists have learned how to “play” with this unique material by enhancing its processability and versatility. Today it is possible, using covalent or supramolecular chemistry, to tailor graphene into a wide variety of forms ranging from simple, soluble sheets to hierarchical architectures where 2-dimensional (2D) graphene sheets are assembled into three-dimensional (3D) composite materials or foams for applications in electronics, mechanics, energy storage or catalysis.
    After graphene, the exfoliation of a wide range of layered materials (BN, MoS2, MoSe2, WS2 etc.) has been demonstrated, featuring complementary conductive, semiconductive and insulating properties, opening the way to produce new composite, 2D meta-materials.
    More recently, polymerization of small molecules through directional 2D crystallization or cross-linking of self-assembled monolayers was used to obtain self-standing, robust nanosheets featuring a huge variety of chemical structures.
    Using these different approaches there is practically no limit to the number of possible 2D structures inspired to graphene that shall be created.
  • Navigating the Promise and Perils of Chemical Probes Recorded: Nov 17 2015 47 mins
    Bill Zuercher- Principal Investigator (UNC Eshelman School of Pharmacy) , Heather King - Senior Scientist (MBD INC)
    Chemical probes are powerful reagents with increasing impacts on biomedical research. However, probes of poor quality or that are used incorrectly generate misleading results. To help address these shortcomings, we are building a community-driven online resource to improve quality and convey current best practice.
    In this presentation, we will highlight some selected examples of promising, high quality chemical probes and probes of lesser value. We will discuss the prevalence of using lesser quality probes and the costs. We will then detail our plans to develop the Chemical Probes Portal, a web-based resource annotated by the chemical biology community that allows users to reference the most appropriate chemical probe (or probes) for a given protein target. The Portal will be an online community of scientists, reviewers, editors.
  • Sigma Pooled CRISPR Screening for Functional Genomics Recorded: Nov 10 2015 50 mins
    Dr. Gregory D. Davis R&D Manager, Molecular Biotechnology Sigma-Aldrich Biotechnology, St. Louis, Missouri
    This 30 minute webinar will describe the design and application of pooled CRISPR libraries. A Sigma-Aldrich technology expert will discuss unique features of Sigma CRISPR pooled libraries with special attention to CRISPR design and cell culture needs.
  • Breast Cancer and Immunohistochemistry Recorded: Oct 27 2015 60 mins
    Jeff Gordon, Director of Sales and Marketing, Cell Marque Corporation A Sigma-Aldrich company
    Breast cancer is the most commonly diagnosed cancer in women, and is the second leading cause of death in women. Each year, over 220,000 women (as well as over 2000 men) are diagnosed with breast cancer, and over 40,000 succumb each year to the illness. With constant advancement of treatment options, the importance of accurate diagnosis and detection of breast cancer becomes more and more relevant to the survival of the patient. Immunohistochemistry has served as the catalyst for these advancements in breast cancer diagnosis. This presentation covers many of the basic science, facts, and statistics of breast cancer, as well as the utility of immunohistochemical testing with markers such as e-cadherin, p120 catenin, mammaglobin, and GATA3 in the accurate diagnosis and survival rates of breast cancer.
  • High-capacity drug-delivery system for cancer therapy based on poly(2-oxazoline) Recorded: Oct 22 2015 71 mins
    Prof. Rainer Jordan; Chair of Macromolecular Chemistry, School of Science; Technische Universität Dresden, Dresden, Germany
    Poly(2-oxazoline) (POx) is a very promising candidate for use in polymer therapeutics. Amphiphilic POx triblock copolymers can be used as high capacity drug delivery systems for hydrophobic drugs, like paclitaxel, and exhibit synergistic effects for the delivery of multiple chemotherapeutics. In this talk, Professor Jordan will highlight the unparalleled high drug-loading capacities of POx systems, the polymer structural variability, and outline the consequences of the drug loading on the drug delivery system morphology.
  • What’s in your Flask? Global Answers from Authentication Testing (NA) Recorded: Oct 6 2015 55 mins
    Amanda Capes-Davis, Chair Person - International Cell Line Authentication Committee (ICLAC)
    Cell culture underpins research throughout the life sciences. Continuous cell lines have been used as culture models for many cell types, and assembled into panels for drug discovery and development. However, problems like cross-contamination are quite common and can render some cell lines unsuitable as research models. Authentic cells are typically overgrown by the contaminant over several passages to give a misidentified or false cell line.

    Misidentified cell lines can be detected through authentication testing. Short tandem repeat (STR) profiling was subsequently introduced as an international reference method, and a consensus standard for human cell line authentication published by the American National Standards Institute (ANSI). International agreement has allowed all laboratories to compare their cell line stocks, resulting in a new wave of misidentified cell lines being discovered.

    Funding bodies and journals are moving to set requirements for reporting of preclinical research. A good understanding of cell line provenance is also essential.
Focusing on new / innovative technologies and industry challenges
The Sigma-Aldrich Scientific Seminar channel features scientific presentations from key specialists in analytical chemistry, biology, chemistry and life sciences on the practical and technical aspects of new developments and innovations, to help advance your research.

Embed in website or blog

Successfully added emails: 0
Remove all
  • Title: Retention Mechanisms in HILIC Chromatography: Robust Method Development
  • Live at: Jun 28 2012 6:00 pm
  • Presented by: David S. Bell, Ph.D.
  • From:
Your email has been sent.
or close