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Agarose–Heated Transfer Lines
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| By
Wyatt Technology Corporation
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Agarose is a polysaccharide produced from seaweed that forms a gel at room temperature. The characteristics of agarose gels depend on the agarose concentration, solvent, weight-average molar mass, and molar mass distribution. Powdered agarose, or agarose gel, needs to be heated in order to dissolve properly. Reheating a sample as well as other processing steps can change the agarose gel characteristics. This note describes the characterization of agarose by size-exclusion chromatography (SEC) and multi-angle light scattering (MALS) at an elevated temperature.
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Extraction of Testosterone and Other Steroid Hormones from Human Plasma Using ISOLUTE® SLE+ 96-Well Plates
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| By
Adam Senior
,
Lee Williams
,
Rhys Jones
,
Helen Lodder
,
Gavin Jones
,
Biotage, LLC
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A new, simple, and sensitive sample preparation application for the extraction of a range of steroid hormones from human plasma at significantly low levels using ISOLUTE SLE+, an efficient alternative to traditional liquid-liquid extraction (LLE) for bioanalytical sample preparation, providing high analyte recoveries, no emulsion formation, and significantly reduced sample preparation time.
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UHPLC Analysis of Underivatized Amino Acids
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| By
Christopher Crafts
,
Bruce Bailey
,
Ian Acworth
,
Dionex, now part of Thermo Fisher Scientific
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The quantification of derivatized amino acids by (U)HPLC with UV detection is one of the most widely used methods to date. Unfortunately, separation and detection of underivatized amino acids is difficult, as many of them are similar in structure, and few possess adequate chromophores.
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Separation of Glycans by Shodex™ NH2P Amino HILIC Column
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| By
Kanna Ito
,
Shodex/Showa Denko America, Inc.
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In this application, we present the feasibility of NH2P-50 4E, polymer-based amino HILIC column, for the structure analysis of N-glycans in biological samples. Characterization of glycan is important since different biological properties of glycoprotein play unique roles on pharmacokinetics, stability, and antigenicity.
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Aggregation Analysis of Biosimilar EPO (Erythropoietin) Using a BioSep 2000 GFC Column
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| By
Michael McGinley
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Phenomenex, Inc.
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Gel filtration chromatography is the primary method used to analyze the amount of aggregate and dimer present in a therapeutic protein sample. A BioSep 2000 column is used to analyze EPO samples to determine difference in the amounts of aggregate present.
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Two Novel Multi-Mode Columns, SS-C18 and SW-C18, Provide Effective Separation of Catecholamines and siRNA
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| By
Bhavana Verma
,
Itaru Yazawa
,
Imtakt USA (formerly Silvertone Sciences)
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HPLC analysis of polar ionic compounds, such as catecholamines and siRNA, can be challenging due to the nature of their structures. Small polar compounds like catecholamines can be diffcult to retain and siRNA can be difficult to elute in reverse phase mode. The Scherzo Family of HPLC columns (SS-C18, SM-C18, and SW-C18) can overcome these challenges.
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Reduce Oligosaccharide Analysis Time with TSKgel SuperOligoPW Size Exclusion Chromatography Columns Packed with Three Micron Particles
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| By
Roy Eksteen
,
Tosoh Bioscience LLC
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At the 2011 Pittsburgh Conference, Tosoh Corporation introduced a new line of high efficiency columns for the analysis of water-soluble polymers by gel filtration chromatography.
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Going Faster, Going Greener with High Efficiency Regis' Chiral Stationary Phases in Supercritical Fluid Chiral Chromatography (SFC)
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| By
Ted Szczerba
,
Terry Berger
,
Regis Technologies, Inc.
,
Aurora SFC Systems
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Chiral method development and chiral separations have been the main contributors to the popularity of supercritical fluid chromatography (SFC). The low viscosity and high diffusivity of the mobile phase in SFC allows for low pressure drops and fast chromatography, making it an attractive alternative to LC, especially for pharmaceutical applications. SFC is often 3–5 times faster than HPLC and often produces higher efficiency. SFC is now regarded as a standard separation support tool for chemical synthesis and development.
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Direct Determination of Fluoroacetate in Water by IC-MS
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| By
Marcus Miller
,
Leo (Jinyuan) Wang
,
William C. Schnute
,
Dionex, now part of Thermo Fisher Scientific
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Fluoroacetate (Compound 1080) is a strong metabolic poison commonly used as a rodenticide and predacide (1). Due to the high level of toxicity when ingested (2), and no known antidotes, its use has been banned or restricted in many countries.
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Combining Hardware, Software, and Chromatography to Improve the GC/MS Analysis of Semi-Volatile Compounds
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| By
Jessie Butler
,
David Steiniger
,
Trisa Robarge
,
Eric Phillips
,
Thermo Fisher Scientific Inc.
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Analysis of semi-volatile organic compounds by GC–MS according to well-established methodologies requires integration of a complete system of GC–MS instrumentation and software for data interpretation, analysis, and reporting. While the overall process of analyzing these target compounds is a mature technique, there are continuous innovations that allow laboratories to meet lower detection limits and analyze new compounds to comply with changing regulations, with higher throughput and improved quality.
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Accurate Mass Screening For Pesticide Residue Testing
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| By
James Hillis
,
Petra Decker
,
Ian Sanders
,
Carsten Baessmann
,
Bruker Daltonics Inc.
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The update to the SANCO guidelines in 2009 recognized the power and potential of TOF mass spectrometry for pesticide screening applications. When compared with historical screening tools (such as the use of triple-quad mass spectrometry), high performance accurate mass TOF systems offer several advantages for screening applications. These advantages include better resolution, unlimited multiple target capabilities, and the possibility to perform comprehensive and/or retrospective analysis on samples in silico at will.
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Determination of Haloacetic Acids and Dalapon in Drinking Water by Solid-Phase Extraction and GC-ECD Detection
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| By
Xiaoyan Wang
,
Don Shelly
,
UCT, LLC
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Method 552.1 is a GC method for determination of haloacetic acid and Dalapon in drinking, ground, raw, and other waters at any stage of treatment. This application uses a silica-based quaternary amine anion exchange cartridge, with exceptional recoveries.
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A New Automated Solid Phase Extraction Approach for Troublesome Water Samples by Method 8270D
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| By
Michael Ebitson
,
David Gallagher
,
Horizon Technology, Inc.
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EPA Method 8270D is used to determine a list of up to 243 semi-volatile organic compounds that are neutral, acidic, basic, and are soluble in methylene chloride. Semi-volatile water samples can contain varying amounts of suspended particulates and/or sediment due to either the source of the water, or the collection technique. By elevating the pH of semi-volatile samples, precipitates can form and result in an emulsion that can create problems for an efficient extraction with traditional techniques. These types of samples, due to their sediment content, have historically proven very challenging when using solid phase extraction (SPE).
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IC-ICP–MS Analysis of Gadolinium-Based MRI Contrast Agents
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| By
Peter Pfundstein
,
Christian Martin
,
Wolfgang Schulz
,
Katinka Meike Ruth
,
Andrea Wille
,
Thomas Moritz
,
Alfred Steinbach
,
Dirk Flottmann
,
Hochschule Aalen
,
Leibniz-Zentrum für Marine Tropenökologie (ZMT)
,
Zweckverband Landeswasserversorgung
,
Metrohm International Headquarters
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The use of contrast agents in noninvasive diagnostics provides a specific and high-detailed picture of organs and makes it possible to differentiate between healthy and diseased tissue. Whilst radiocontrast agents (RCA) change the X-ray absorption of the radiographed organ relative to its surrounding tissue, contrast agents for magnetic resonance imaging (MRI) are based on the use of magnetic fields caused by charged elementary particles such as protons and electrons. Compared to traditional X-ray imaging, MRI uses no ionizing radiation. MRI is based on the same principles as NMR spectroscopy and thus takes advantage of the absorption and emission of energy in the radiofrequency range.
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Validation of Wisconsin Gasoline Range Organics
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| By
Tyler Trent
,
Teledyne Tekmar
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The Wisconsin Department of Natural Resources (DNR) Modified Gasoline Range Organics (Wisconsin GRO) Method is used to determine gasoline and gasoline components in water and soil. The optimal hydrocarbon range is from C6-C10 with a boiling point range from 60 ?C–220 ?C (1). The analysis of Wisconsin GRO is vital in determining the "composition and the origin of such products including weathered fuels leaking from underground storage tanks, spills in pipe lines and run-off from surface transportation" (2).
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Identification of Trace-Level TO-15/TO-17 Air Toxics in Complex Air Samples
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| By
Nick Bukowski
,
Gareth Roberts
,
Damien Rosser
,
Nicola Watson
,
ALMSCO International
,
Markes International
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Air pollution remains a global concern due to its impact on human health and the environment. Compounds known as "air toxics" are of particular importance in this respect, and US EPA methods TO-15 and TO-17 describe how to test air samples for the presence of these compounds.
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Fast Chlorinated Pesticide Analysis Using Zebron™ MultiResidue™ GC Columns
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| By
Kory Kelly
,
Kristen Parnell
,
Phenomenex, Inc.
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The US EPA regulates the testing of 20 specific chlorinated pesticides under the official Method 8081. Zebron MultiResidue columns achieve baseline separation of all of these analytes with short run times, providing greater confidence in identification and increased lab productivity.
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Call for Application Notes
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LCGC is planning to publish the next issue of The Application Notebook special supplement in December. The publication will include vendor application notes that describe techniques and applications of all forms of chromatography and capillary electrophoresis that are of immediate interest to users in industry, academia, and government. If your company is interested in participating in these special supplements, contact:
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Liposome Characterization by FFF-MALS-QELS
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| By
Wyatt Technology Corporation
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Liposomes are made of lipid bilayers and are often used in drug delivery by encapsulating the core with therapeutic drugs. During liposome research, formulation, manufacturing, and quality control, it is of great importance to monitor liposome size and encapsulation.
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Liposome Characterizatio by FFF–MALS–QELS
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| By
Wyatt Technology Corporation
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Liposomes are made of lipid bilayers and are often used in drug delivery by encapsulating the core with therapeutic drugs. During liposome research, formulation, manufacturing, and quality control, it is of great importance to monitor liposome size and encapsulation.
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Significant Costs Savings Achieved Through the Development of a New Analytical Method for Residual API Analysis, with a Kinetex® 1.7 μm Core-Shell UHPLC Column
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| By
Jeremy Bierman
,
Chris Message
,
Phenomenex, Inc.
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This study demonstrates how the ultrahigh-performance of 1.7 ?m core-shell columns can be utilized in the laboratory in order to realize significant cost savings.
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Exploring UPLC Selectivity with New High Strength Silica (HSS) Cyano and PFP Stationary Phases
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| By
Christopher J. Hudalla
,
Jane Xu
,
Kenneth J. Fountain
,
Waters Corporation
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For successful chromatographic method development, it is advantageous to maximize the difference in column selectivity, or "selectivity space" investigated in order to minimize the number of critical analyte co-elutions.
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Identifying and Collecting Weak UV Absorbance and Low Level Peaks of Interest From a Complex Mixture Using Conditional Logic Purification on Mass and UV Signals
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| By
Luke Roenneburg
,
Tim Hegeman
,
Toni Hofhine
,
Gilson, Inc.
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Purification of active pharmaceutical ingredients (APIs) often yields high numbers of fractions, leading to a bottleneck of fractions to dry-down and subsequently analyze. UV-based fraction collection based on slope or threshold values offers some specificity and reduction in the number of fractions collected compared to basic time- or volume-based collection.
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Identification of Metabolites of Doxylamine and Dextromethorphan Using UHPLC with High-Performance Time-of-flight Mass Spectrometry
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| By
Kevin Siek
,
Joe Binkley
,
Jeffrey S. Patrick
,
LECO Corporation
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Ultrahigh-performance TOF–MS is utilized for the identification of a common pharmaceutical compound's metabolites. Mass spectral analysis is combined with UHPLC and post-source fragment ion generation to facilitate analysis.
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Organic Acid Counter Ion Analysis using a Bonded Zwitterionic Stationary Phase
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| By
Gora Sharangi
,
Sanjay Poman
,
Patrik Appelblad
,
EMD Millipore
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Using a bonded zwitterionic stationary phase, ZIC?-HILIC, polar organic acid counter ions such as methylsulfonic acid can easily be retained, to aid simultaneous analysis of basic pharmaceutical APIs together with their counter ions.
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Improved Clean Up and Recovery of Pharmaceutical Compounds From Plasma using Strata™-X Solid Phase Extraction (SPE) vs. Traditional Liquid-Liquid Extraction
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| By
Deborah Jarrett
,
Phenomenex, Inc.
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Diclofenac is a slightly acidic (pKa = 4.0) non-steroidal anti-inflammatory drug (NSAID) that has been used as a post operative pain reliever in adult and pediatric patients.
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Ion Chromatography Method for Determination of Risedronate in Pharmaceutical Preparations
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| By
Avanthi Kanmatareddy
,
Brian De Borba
,
Jeffrey Rohrer
,
Dionex, now part of Thermo Fisher Scientific
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Risedronate, a pyridinyl bisphosphonate, is one of the most popular first-line drugs in the treatment of osteoporosis, Paget's disease, and other bone fragility conditions. In October 2010, the United States FDA approved a second-generation risedronate in a delayed-release formulation.
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Rapid Test for Content Uniformity of Coenzyme Q10 in Soft Gel Capsules by HPTLC
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| By
Anita Ankli
,
Daniel Handloser
,
Valiera Widmer
,
Eike Reich
,
Eliezer Ceniviva
,
CAMAG Laboratory
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Coenzyme Q10, found naturally in the body, is involved in the production of body energy. Thus, heart, lung, and liver have the highest concentration of Q10.
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Optimized Sunscreen Method Used for Impending 2012 Enforcement of FDA Regulation in Characterizing Their Active Ingredients
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| By
Deborah Jarrett
,
Sky Countryman
,
Jeff Layne
,
Phenomenex, Inc.
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On June 14, 2011, the FDA issued stricter regulations on how sunscreen manufacturers can label over-the-counter sunscreen products. In the summer of 2012, manufacturers with an annual sale of $25,000 or more will have to characterize the active ingredients found in their sunscreens (1).
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Separation of Vitamin K Isomers with Enhanced Selectivity
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| By
Mark Tracy
,
Xiaodong Liu
,
Dionex, now part of Thermo Fisher Scientific
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Vitamin K is a group of structurally similar, fat-soluble vitamins that are needed for the post-translational modification of certain proteins, mostly required for blood coagulation but also involved in metabolic pathways in bone and other tissue.
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Highly Efficient Separation of Bath Salts Using Hydrophilic Interaction Chromatography (HILIC)
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| By
David S. Bell
,
Craig R. Aurand
,
Wayne K. Way
,
Supelco/Sigma-Aldrich
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According to law enforcement and health officials, "bath salts" are powerful synthetic stimulants, designed to be comparable in effect to cocaine or methamphetamine, carrying similar risks. Unlike cocaine or methamphetamine, these stimulants are currently legal in many parts of the United States and available at a low cost in many convenience stores and head shops.
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Determination of Sulferin Natural Gas by ASTM Method D 6228-11
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| By
Laura Chambers
,
Gary Engelhart
,
OI Analytical
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Natural gas and other gaseous fuels contain varying amounts and types of sulfur compounds which can be corrosive to equipment and can inhibit or destroy gas processing catalysts. Small amounts of sulfur odorants are added to natural gas and liquefied petroleum gases (LPGs) for safety purposes. Accurate measurement of sulfur species ensures proper process operation and odorant levels for public safety.
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Rapid Determination of Anthocyanins in Bilberries
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| By
Pranathi R. Perati
,
Brian De Borba
,
Jeffrey Rohrer
,
Dionex, now part of Thermo Fisher Scientific
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Bilberry (Vaccinium myrtillus L.) extracts are high in anthocyanin content and are widely used in nutritional supplements for improving visual acuity and treating circulatory disorders.
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"Green" Analysis of Diet Soft Drinks Containing Caffeine and Aspartame
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| By
Dwight R. Stoll
,
Dr. Clayton V. McNeff
,
ZirChrom Separations, Inc.
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The popularity of diet soft drinks containing aspartame requires the analysis of thousands of samples for quality control purposes each year. These analyses are typically carried out with some fraction of organic modifier in the eluent, leading to the generation of thousands of gallons of hazardous organic waste. This note shows the "green" analysis of several commercial diet soft drinks in combination with a 100% water mobile phase.
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Using the iDQuant™ Standards Kit for Pesticide Analysis to Analyze Residues in Fruits and Vegetable Samples
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| By
André Schreiber
,
Brent Lefebvre
,
AB SCIEX
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The iDQuant? Standards Kit for pesticide analysis of certified reference material (ISP Guide 34, IOS/IEC 17025, and ISO 9001:2008) includes 204 pesticides in 10 mixes at a concentration of 100 ?g/mL. This kit has been designed to eliminate the need to source pesticides individually, and measure each one manually. This allows users to setup, verify, and validate the performance of pesticide screening methods faster.
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Analysis of Sugars in Feeds by HPLC with Post-column Derivatization and Fluorescence Detection
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| By
Maria Ofitserova
,
Sareeta Nerkar
,
Pickering Laboratories, Inc.
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The types and amounts of sugar in animal feeds are as important as the amount of protein, minerals, and fats in the determination of nutritive value. We developed a simple and sensitive HPLC method for analyzing six sugars in animal feeds – sucrose, fructose, glucose, galactose, maltose, and lactose. Post-column derivatization reagents convert reducing and nonreducing sugars into fluorescent derivatives, which greatly improves the sensitivity and selectivity of the detection.
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HPLC Stevia Sweetener Analysis with CAD and UV Detection
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| By
Deanna C. Hurum
,
Jeffrey S. Rohrer
,
Dionex, now part of Thermo Fisher Scientific
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In December 2008, the United States FDA recognized rebaudioside A (reb. A) purified from Stevia rebaudiana (Bertoni) as Generally Recognized as Safe (GRAS) as a sugar substitute in foods. As a result, stevia products have become popular tabletop and beverage sweeteners. Two steviol glycosides, stevioside and reb. A, are largely responsible for the desired sweet flavor, with reb. A preferred for sweeteners (1).
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Rapid Screening and Quantitation of 534 Pesticides From Food By LC–MS-MS
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| By
Sky Countryman
,
Andre Schreiber
,
David Lavarato
,
Phenomenex, Inc.
,
AB SCIEX
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With more than 800 pesticides currently in use there is growing concern by government agencies about the effect of pesticide exposure on human health. While analytical techniques like GC or GC–MS have traditionally been used for pesticide analysis, LC–MS-MS has become the technique of choice due to speed, increased sensitivity, and reduced sample preparation. The following describes an LC–MS-MS method for screening of 534 pesticides in food products.
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