ResourceSeparations

Reducing Sample Volume and Increasing Sensitivity for the Quantification of Human Insulin and 5 Analogs in Human Plasma Using ionKey/MS

4 May 2016

This application note demonstrates the need for robust and sensitive analysis of peptide species challenges, both through chromatographic separation and mass spectrometry. Recombinant human insulin and its analogs are perhaps the best known and most widely sold biotherapeutics. Historically, such biologics have been quantified using ligand binding assays. However, specifically in the case of insulin and analogs, these affinity-based assays lack standardization, are subject to matrix effects, and in some cases lack adequate specificity. The use of ionKey/MS facilitated a reduction in sample volume and concomitant increase in sensitivity for the quantification of human insulin and 5 important analogs.

Oasis® µElution Plate

Waters

The Oasis® uElution plate format combined with the Oasis chemistries allow elution volumes as low as 25uL, eliminating the time consuming evaporation step, producing very clean extracts with a 5 to 10 fold increase in concentration.The Oasis® family of sample extraction products is designed to simplify and improve your sample preparation by combining the right sorbent chemistry, device format and methodology. Achieve robust, selective, and sensitive solid-phase extraction [SPE] methods without worrying about low recoveries caused by breakthrough, sorbent drying, pH limitations, and undesirable silanol activity.Oasis® SPE sorbents—covered by eight U.S. patents—are unique in their purity, reproducibility, stability, and retention characteristics. Today, they are the most widely used polymeric SPE products in bioanalytical laboratories.

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UHPLC and HPLCHigh performance liquid chromatography (HPLC) and ultra high performance liquid chromatography (UHPLC), also known as UPLC, are analytical techniques used to separate, identify and quantitate components of complex mixtures including biological samples such as proteins and lipids as well as chemical mixtures of pesticides, drugs and oils. Both techniques are liquid chromatographic methods but differ by operating pressures (HPLC < 6000 psi < UHPLC ). Components of HPLC and UHPLC systems include columns, detectors, pumps, autosamplers and column heaters. Explore a range of UHPLC and HPLC columns for your specific sample needs including reverse phase, normal phase, ion exchange, HILIC, ion exclusion and size exclusion columns. For more specialized HPLC, explore FPLC, countercurrent LC and simulated moving bed systems. Find the best UHPLC and HPLC equipment in our peer reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Sample PreparationSample preparation can improve the quality and speed of separation techniques. Products to assist sample preparation include filtration equipment, evaporators, membranes and sieves.Mass SpectrometryMass spectrometry (MS) is a powerful analytical technique used to identify and quantify molecules based on the mass-to-charge ratio of gas-phase ions. It provides detailed information about the structure, composition, and properties of compounds and is widely used across fields such as environmental monitoring, materials science, drug discovery and development, food and beverage testing, and wider chemical research. Key MS techniques include tandem mass spectrometry (MS/MS), liquid chromatography–mass spectrometry (LS-MS) and inductively coupled plasma (ICP-MS). Choosing from these wide range of techniques and technologies can be a daunting task, so keep up to date with scientific applications, performance expectations, and customer reviews here all in one place. Visit our product directory to receive quotes direct from the manufacturer. ProteomicsProteomics is the systemic bioinformatics study of proteins and amino acids, including their structure, size, function and identification. Tools used in proteomics include chromatography, blotting and gels, protein arrays, mass spectrometry and ELISA and associated analysis software. Analyzers and proteomic systems should be sensitive, high resolution, fast and may be automated for high-throughput.PeptidesPeptides are short chains of amino acids that play key roles in biological processes, including signaling, immune responses, and enzyme regulation. Peptide research is critical in drug discovery, diagnostics, and vaccine development. Browse our peer-reviewed product directory to find the best peptides and peptide-related products, compare options, check reviews, and get pricing directly from manufacturers.BiotherapeuticsBiotherapeutics are proteins and other compounds (such as nucleic acids) produced by living organisms that have uses as therapeutics or in <i>in vivo</i> diagnostics. The most well-known example of a biotherapeutic product, and the first to be approved for therapeutic use, was recombinant human insulin.2D ChromatographyInsulinHuman PlasmaHuman plasma is the liquid component of blood, which carries blood components - nutrients, hormones, proteins, and waste - throughout the body. Plasma-based studies provide insights into biomarkers, immune responses, and disease mechanisms. Compare plasma handling tools and systems in our peer-reviewed product directory.Sample PreparationProtein Quantification