Using Hydrophilic Interaction Chromatography (HILIC) for the Retention of Highly Polar Analytes
|
|
- Osborne Banks
- 5 years ago
- Views:
Transcription
1 Using Hydrophilic Interaction Chromatography (HILIC) for the Retention of Highly Polar Analytes Eric S. Grumbach,, Diane M. Diehl and Jeffrey R. Mazzeo 2003 Waters Corp.
2 Outline Introduction HILIC Definition and Benefits Important considerations for HILIC Mobile phase considerations Solvent selectivity Column performance Column reproducibility Complementary selectivity to reversed-phase Enhanced ionization in mass spectrometry Low limits of detection Direct injection of SPE eluent Summary
3 Introduction Traditional options for retaining polar analytes Ion pairing or ion exchange Only works if analytes ionize Often not compatible with mass spectrometry (MS) Manipulate ph to make compound neutral Reversed-phase columns designed for the retention of polar analytes (such as the Atlantis dc 18 ) Used under highly aqueous conditions, often resulting in poor MS sensitivity What happens when you ve tried all of these options and you STILL have inadequate retention??? Let s take a look at HILIC
4 What is HILIC? HILIC - Hydrophilic Interaction Chromatography Term coined in 1990 to distinguish from normal-phase* HILIC is a variation of normal-phase chromatography without the disadvantages of using solvents that are not miscible in water Reverse reversed-phase or aqueous normal-phase chromatography Stationary phase is a POLAR material Silica, cyano, amino, diol The mobile phase is highly organic (> 80%) with a smaller amount of aqueous mobile phase Water (or the polar solvent(s)) is the strong, eluting solvent *Alpert, A. J. J.Chromatogr. 499 (1990)
5 HILIC Retention on Silica Polar analyte partitions into and out of adsorbed water layer Charged polar analyte can undergo cation exchange with charged silanol groups Combination of these mechanisms results in enhanced polar retention
6 HILIC vs. Reversed-Phase Retention Characteristics H N O N Retention NH 3 + Cytosine % MeCN HILIC offers dramatically more retention than reversed-phase for very polar bases. HILIC RP Alden, Iraneta
7 Benefits of HILIC Retention of highly polar analytes not retained by reversed-phase Complementary selectivity to reversed-phase Enhanced sensitivity in mass spectrometry High organic mobile phases (> 80%) promotes enhanced ESI-MS response Shortens sample preparation procedure Elimination of evaporation/reconstitution step by directly injecting the organic phase
8 Outline Introduction HILIC Definition and Benefits Important considerations for HILIC Mobile phase considerations Solvent selectivity Column performance Column reproducibility Complementary selectivity to reversed-phase Enhanced ionization in mass spectrometry Low limits of detection Direct injection of SPE eluent Summary
9 Wash solvents Mobile Phase Considerations: Before You Start Seal wash: 10% acetonitrile in water Needle wash: 1:1 acetonitrile:water Purge Solvent: initial mobile phase conditions (without salt, additive or buffer) Initial conditions: 95% to 70% acetonitrile Remember, acetonitrile is the weaker solvent At least 5% should be a polar solvent (i.e., water or methanol) Where do I start? Initial scouting gradient from 95 to 50% acetonitrile If analytes are not retained, try 95% acetonitrile isocratic
10 Mobile Phase Considerations: Before You Start Buffers/additives Phosphate salt buffers are not recommended due to precipitation in the highly organic mobile phase (phosphoric acid is OK) Ammonium formate, ph 3*; ammonium acetate, ph 5*; 0.2% formic acid, ph 2.5*, 0.2% phosphoric acid, ph 1.8* For optimum performance and reproducibility it is recommended that a concentration of 10 mm buffer or 0.2% of an additive be introduced ON COLUMN To increase retention of analytes, replace some of the water with another polar solvent (i.e., methanol, isopropanol) In HILIC, these polar solvents are weaker eluters than water Peak shapes and reproducibility may be compromised by completely removing the polar solvent (e.g., water) Let s take a closer look *The actual ph of the mobile phase may be 1 ph unit higher due to the highly organic mobile phase Canals, I.; Oumada, F. Z.; Roses, M.; Bosch, E. J. Chromatogr. A. 911 (2001)
11 Solvent Strength: HILIC Solvent Water Methanol Ethanol Isopropanol Acetonitrile Acetone Tetrahydrofuran Strongest Weakest In HILIC, utilizing a less polar solvent can increase the retention of polar analytes.
12 Compounds 1. 5-Fluorouracil 2. Uracil 3. 5-Fluorocytosine 4. Cytosine Increased Retention with a Less Polar Solvent Initial 90 ACN: 5 H 2 O: 5 buffer Final 50 ACN: 45 H 2 O: 5 buffer Initial 90 ACN: 5 MeOH: 5 buffer Final 50 ACN: 45 MeOH: 5 buffer Minutes Minutes Initial 90 ACN: 5 IPA: 5 buffer Final 50 ACN: 45 IPA: 5 buffer Replace H 2 O with MeOH Replace MeOH with IPA Minutes Retention increases by using a less polar solvent. Grumbach
13 Outline Introduction HILIC Definition and Benefits Important considerations for HILIC Mobile phase considerations Solvent selectivity Column performance Column reproducibility Complementary selectivity to reversed-phase Enhanced ionization in mass spectrometry Low limits of detection Direct injection of SPE eluent Summary
14 S Reproducibility of Atlantis TM HILIC Silica Injection 1 Injection 100 Injection 200 Injection 300 Analytes: S. Solvent Peak 1. 5-Fluorouracil 2. Uracil 3. 5-Fluorocytosine 4. Cytosine Injection 400 Injection 500 Injection 600 Injection 700 Injection Minutes Continuous injections of polar bases under gradient conditions yield excellent reproducibility. Grumbach
15 Retention Benefits of HILIC AU AU k = 0 V o = 0.65 min V o = 1.15 min Minutes k = 1.09 Atlantis TM dc x 50 mm, 3 µm 100% Formate Buffer, ph ml/min H 2 N Atlantis TM HILIC Silica 4.6 x 50 mm, 3 µm 95:5 ACN:Formate Buffer, ph ml/min O NH O H N N H Allantoin O Minutes HILIC offers retention when there is no retention by reversed-phase. Grumbach
16 Complementary Selectivity to Reversed-Phase HO 1 2 V o = 0.5 min Atlantis TM HILIC Silica 4.6 x 50 mm, 3 µm 90% to 50% ACN HO O H N 1. Morphine CH Minutes V o = 0.65 min 1 Atlantis TM dc x 50 mm, 3 µm 2% ACN 2. Morphine 3-ß-D-Glucuronide Minutes Grumbach
17 Reversed-Phase Conditions: Separation of Cytochrome C Tryptic Digest 100 % 204 (T2) 261 (T6,11) 361 (T18) 434 (T21) 678 (T14) 779 (T15) Peak Annotation M/Z (Fragment ID) Atlantis TM dc x 50 mm, 3 µm 634 (T4) 729 (T10) 964 (T19) 1005 (T12) 585 (T8) 0 Extremely polar peptides are not retained under reversed-phase conditions even with the aid of Atlantis dc 18, a column specifically designed for the retention of polar analytes Atlantis HILIC Silica can aid in the separation and identification of these very polar peptides Rainville
18 Complementary Selectivity to Reversed-Phase: Separation of Cytochrome C Tryptic Digest 100 % 204 (T2) 261 (T6,11) 361 (T18) 434 (T21) 678 (T14) 779 (T15) Peak Annotation M/Z (Fragment ID) Atlantis TM dc x 50 mm, 3 µm 634 (T4) 729 (T10) 964 (T19) 1005 (T12) 585 (T8) (T4) Atlantis TM HILIC Silica 4.6 x 50 mm, 3 µm % 779 (T15) 678 (T14) 434 (T21) 585 (T8) 261 (T6,11) 204 (T2) 964 (T19) 729 (T10) 1005 (T12) 361 (T18) Polar peptides not retained by reversed-phase can be retained by HILIC. Rainville
19 Enhanced Sensitivity in Mass Spectrometry LC/MS conditions were optimized individually for both HILIC and RP. 100 % 0 Reversed Phase 100 % e Time ES e5 0-50% ACN Atlantis TM dc 18 Peak Area 2.1 x 50 mm, 3 µm 10.0 µl injection volume 1. Albuterol 100 pg/µl 21 (ND) 2. Bamethan 50 pg/µl 14 (ND) 100 % 0 HILIC ES e % ACN Time Atlantis TM HILIC Silica Peak Area 2.1 x 50 mm, 3 µm 10.0 µl injection volume 2. Bamethan 50 pg/µl Albuterol 100 pg/µl HILIC requires high volatility solvents which increase sensitivity compared to high-aqueous mobile phases used in reversed-phase.* *Naidong, W.; Shou, W.; Chen, Y-L.; Jiang, X. J. Chromatogr. B. 754 (2001) Grumbach
20 Mass Spectrometer Detection Limits Approximate Limit of Detection Single Quadrupole MS (SIR) mode ES+ Triple Quadrupole MS (MRM) mode ES+ (Analyte Dependant) Reversed-phase 10 ng/ml 1 ng/ml HILIC 1 ng/ml 0.1 ng/ml [100 fg/µl] HILIC-ESI-MS will achieve at least 10x higher sensitivity with high organic mobile phases than typical RP-ESI-MS with high aqueous mobile phases.
21 Extremely Low Detection Limits on a Single Quadrupole Mass Spectrometer 100 % ACh H 3 C H 3 C N + CH 3 O CH 3 O ES e5 Atlantis TM HILIC Silica 2.1 x 50 mm, 3 µm 20.0 µl injection volume Waters ZQ single quadrupole MS SIR m/z: 104.2, 118.3, % Ch H C 3 N + H C 3 CH 3 OH ES e5 Analyte Concentration 1. Acetylcholine (Ach) 100 fg/µl 2. Choline (Ch) 100 fg/µl 3. Betaine (I.S.) 5000 fg/µl % CH 3 H C 3 H 3 C O N + O - Bet (I.S.) ES e6 86% ACN Detect 100 fg/µl on a single quadrupole MS! Time
22 Traditional SPE Methods for Reversed-Phase Chromatography Traditional SPE methods often contain an elution step that consists of high organic content To make this extracted sample compatible with your mobile phase, you must first evaporate the high organic eluent then reconstitute in some portion of aqueous Evaporation and reconstitution are often the most lengthy steps in an SPE procedure* In HILIC, the high organic eluent can be directly injected on the column, thus eliminating the need for evaporation and increasing your throughput *Jemal, M., Teitz, D., Ouyang, Z., J.Chromatogr. B, 732 (1999) 501.
23 *Oasis HLB µelution Plate Generic Oasis HLB SPE Procedure for Polar Bases Using HILIC Condition/Equilibrate* 200 µl methanol/200 µl water Load 75 µl spiked plasma sample 75 µl internal standard with 2% ammonium hydroxide Wash 200 µl 5% methanol in water Elute 75 µl 40% acetonitrile/60% isopropanol with 2% formic acid Inject eluent directly onto column Eliminate Evaporation and Reconstitution Step
24 Generic Oasis HLB SPE: Direct Injection onto HILIC Column HO HO OH Albuterol H 3 C CH 3 NH CH 3 H 3 C NH Bamethan OH OH Atlantis TM HILIC Silica 2.1 x 50 mm, 3 µm 1. Bamethan 10 pg/µl 2. Albuterol 50 pg/µl SIR of 2 Channels ES e4 % Polar species from matrix elute after analytes (normally seen in solvent front using RP) Time SPE eluent injected directly onto Atlantis TM HILIC Silica column Grumbach
25 Summary: When to Use HILIC Atlantis TM dc18 or XTerra Yes 2 Can it be retained by RP? Yes 1 Is the polar compound a base? Start Here 3 Sufficient Mass Spec sensitivity? No Yes No Atlantis TM HILIC Silica No 2 Can it be retained by RP? Yes Atlantis TM dc 18 Note: Atlantis TM dc 18 = High aqueous, Low ph XTerra = High aqueous, High ph Atlantis TM HILIC Silica = Low aqueous, Low ph No Alternate separation technique such as ion exchange, ion-pair, etc.
26 Summary Atlantis TM HILIC Silica columns offer: Retention of highly polar basic analytes Complementary selectivity to reversed-phase Excellent reproducibility Enhanced sensitivity in ESI-MS Shorter sample preparation procedures Elimination of the evaporation and reconstitution steps by directly injecting the final organic phase of the sample prep procedure
27 References 1. Alpert, A. J. J. Chromatogr. 499 (1990) Canals, I.; Oumada, F. Z.; Roses, M.; Bosch, E. J. Chromatogr. A. 911 (2001) Naidong, W.; Shou, W.; Chen, Y-L.; Jiang, X. J. Chromatogr. B. 754 (2001) Naidong, W.; Rapid Commun. Mass. Spectrom. 16 (2002),
28 Acknowledgements Bonnie Alden Pamela Iraneta Paul Rainville Uwe Neue Doug McCabe Tom Walter Michael Savaria Susan Karn
Waters Amide Column Technology For Food Analysis
Waters Amide Column Technology For Food Analysis Euan Ross Business Development Manager Chemistry perations Europe 2012 Waters Corporation 1 verview verview of HILIC Retention mechanisms and characteristics
More informationPerformance of an ultra low elution volume 96-well plate
Performance of an ultra low elution volume 96-well plate Claude R. Mallet, Ziling Lu, Jeff R. Mazzeo, Uwe D. Neue Waters Corporation PittCon 2003 March 10-14 2003 Orlando, Florida Today s Challenges Faced
More informationSystematic Evaluation of Solid Phase Extraction (SPE) Chemistries for the Determination of Acidic, Neutral, and Basic Drugs
Systematic Evaluation of Solid Phase Extraction (SPE) Chemistries for the Determination of Acidic, Neutral, and Basic Drugs Laura Marlatt, An Trinh, and David S. Bell Supelco, 595 North Harrison Road,
More informationIon Exchange and Reversed Phase interactions in selective Bio-SPME extractions of designer drugs
Ion Exchange and Reversed Phase interactions in selective Bio-SPME extractions of designer drugs Frank Michel, Craig R. Aurand, Robert E. Shirey, Yong Chen, Leonard M. Sidisky sigma-aldrich.com/analytical
More informationThe Raptor HILIC-Si Column
The Raptor HILIC-Si Column With Raptor LC columns, Restek chemists became the first to combine the speed of superficially porous particles (also known as SPP or core-shell particles) with the resolution
More informationHigh Throughput Extraction of Opiates from Urine and Analysis by GC/MS or LC/MS/MS)
High Throughput Extraction of Opiates from Urine and Analysis by GC/MS or LC/MS/MS) Michael Rummel, Matthew Trass, Michael Campognone, and Sky Countryman Phenomenex, Inc., 411 Madrid Avenue, Torrance,
More informationYMC HILIC Columns for Polar Analytes
YMC HILIC Columns for Polar Analytes Polar Analytes Advantages In general, reliable applications in HPLC require a minimum retention factor (k ) of 2. In the widely used reversed phase methods polar compounds
More informationSeQuant ZIC -HILIC For all who expect more...
SeQuant ZIC -ILIC For all who expect more... The better choice for PLC and LC/MS of all types of polar and hydrophilic compounds EMD Millipore Corp. is a subsidiary of Merck KGaA, Darmstadt, Germany Poor
More informationDr. Erin E. Chambers Waters Corporation. Presented by Dr. Diego Rodriguez Cabaleiro Waters Europe Waters Corporation 1
Development of an SPE-LC/MS/MS Assay for the Simultaneous Quantification of Amyloid Beta Peptides in Cerebrospinal Fluid in Support of Alzheimer s Research Dr. Erin E. Chambers Waters Corporation Presented
More informationApplication Note. Abstract. Authors. Pharmaceutical
Analysis of xycodone and Its Metabolites-oroxycodone, xymorphone, and oroxymorphone in Plasma by LC/MS with an Agilent ZRBAX StableBond SB-C18 LC Column Application ote Pharmaceutical Authors Linda L.
More informationRemoval of Triton X-100 from Plasma Samples Using Mixed-Mode Solid Phase Extraction (SPE)
Removal of Triton X- from Plasma Samples Using Mixed-Mode Solid Phase Extraction (SPE) Jonathan P. Danaceau, Erin Chambers, and Kenneth J. Fountain Waters Corporation, 34 Maple Street, Milford, MA USA
More informationComprehensive Study of SLE as a Sample. Preparation Tool for Bioanalysis
Comprehensive Study of SLE as a Sample Preparation Tool for Bioanalysis Wan Wang, Warren Chen, Jerry Wang Bonna-Agela Technologies 179 Southern Street, West TEDA, Tianjin, China Abstract A simple, fast,
More informationLC-MS/MS Method for the Determination of Tenofovir from Plasma
LC-MS/MS Method for the Determination of Tenofovir from Plasma Kimberly Phipps, Thermo Fisher Scientific, Runcorn, Cheshire, UK Application Note 687 Key Words SPE, SOLA CX, Hypersil GOLD, tenofovir Abstract
More informationO O H. Robert S. Plumb and Paul D. Rainville Waters Corporation, Milford, MA, U.S. INTRODUCTION EXPERIMENTAL. LC /MS conditions
Simplifying Qual/Quan Analysis in Discovery DMPK using UPLC and Xevo TQ MS Robert S. Plumb and Paul D. Rainville Waters Corporation, Milford, MA, U.S. INTRODUCTION The determination of the drug metabolism
More informationAnalysis of Rosuvastatin in Dried Blood Spot and Plasma Using ACQUITY UPLC with 2D Technology
Analysis of Rosuvastatin in Dried Blood Spot and Plasma Using ACQUITY UPLC with 2D Technology Claude Mallet, 1 Jennifer Simeone, 2 Paul Rainville 3 1 Workflow Integration Group, Separations Technologies,
More informationDevelopment of a Bioanalytical Method for Quantification of Amyloid Beta Peptides in Cerebrospinal Fluid
Development of a Bioanalytical Method for Quantification of Amyloid Beta Peptides in Cerebrospinal Fluid Joanne ( 乔安妮 ) Mather Senior Scientist Waters Corporation Data courtesy of Erin Chambers and Mary
More informationThe Development of LC/MS Methods for Determination of Polar Drugs of Abuse in Biological Samples
WA20259 The Development of LC/MS Methods for Determination of Polar Drugs of Abuse in Biological Samples Michael S. Young and Kevin M. Jenkins Waters Corporation, 34 Maple Street, Milford, MA 01757 Introduction
More informationSeQuant ZIC -HILIC For all who expect more...
SeQuant ZIC -ILIC For all who expect more... The better choice for PLC and LC/MS of all types of polar and hydrophilic compounds EMD Millipore is a division of Merck KGaA, Darmstadt, Germany Poor retention
More informationAnalysis of Testosterone, Androstenedione, and Dehydroepiandrosterone Sulfate in Serum for Clinical Research
Analysis of Testosterone, Androstenedione, and Dehydroepiandrosterone Sulfate in Serum for Clinical Research Dominic Foley, Michelle Wills, and Lisa Calton Waters Corporation, Wilmslow, UK APPLICATION
More informationUnique Retention and Selectivity of Pentafluorophenylpropyl Phases for High-Throughput LC/MS Analysis
Unique Retention and Selectivity of Pentafluorophenylpropyl Phases for High-Throughput LC/MS Analysis David S. Bell Supelco, Bellefonte, PA ASMS 22 T42119 (xxx) Overview 2 The LC/MS retention and sensitivity
More informationTaking the Complexity out of SPE Method Development
Taking the Complexity out of SPE Method Development Includes guidelines for the following: P SPE Format Selection P SPE Sorbent Selection P SPE Methods P SPE Volume Guidelines P Common Laboratory Conversions
More informationDevelopment of a High Sensitivity SPE-LC-MS/MS Assay for the Quantification of Glucagon in Human Plasma Using the ionkey/ms System
Development of a High Sensitivity SPE-LC-MS/MS Assay for the Quantification of Glucagon in Human Plasma Using the ionkey/ms System Mary E. Lame, Erin E. Chambers, Sukhdev S. Bangar, and Kenneth J. Fountain
More informationCorticosteroids. Optimizations. Jens Hinge Andersen. National Food Institute Danish Technical University
Corticosteroids Optimizations Jens Hinge Andersen National Food Institute Danish Technical University jhia@food.dtu.dk Optimizations - corticosteroids : Minimizing ion-suppression from bovine urin Liver:
More informationThe Development of LC/MS Methods for Determination of MDMA (Ecstasy) and Metabolites in Biological Samples
WA20714 The Development of LC/MS Methods for Determination of MDMA (Ecstasy) and Metabolites in Biological Samples Michael S. Young and Kevin M. Jenkins Waters Corporation, 34 Maple Street, Milford, MA
More informationLC/MS Analysis of Various Hydrophilic Compounds Using a Polymer-Based Amino Column - Shodex TM HILICpak TM VG-50 2D
LC/MS Analysis of Various Hydrophilic Compounds Using a Polymer-Based Amino Column - Shodex TM HILICpak TM VG-50 2D Introduction Components of pharmaceutical products and food products often include high
More informationAnalytical and Preparative SFC Columns
Analytical and Preparative SFC Columns Best Performance for Supercritical Fluid Chromatography Sepax SFC-Cyano... Sepax SFC-Amino... Sepax SFC-Pyridine... Sepax SFC-SCX... 5 Sepax SFC-Diol... 6 Sepax SFC-Silica...
More informationExtraction of 25-hydroxy Vitamin D from Serum Using ISOLUTE. PLD+ Prior to LC-MS/MS Analysis
Application Note AN842.V. Extraction of 25-hydroxy Vitamin D from Serum Using ISOLUTE PLD+ Prior to LC-MS/MS Analysis Page Extraction of 25-hydroxy Vitamin D from Serum Using ISOLUTE PLD+ Prior to LC-MS/MS
More informationInertSustain Amide 技術資料
InertSustain Amide 技術資料 InertSustain Amide Physical Properties Silica Particle Size Surface Area Pore Size Pore Volume Bonded Phase End-capping Carbon Loading ph Range USP Code :ES (Evolved Surface) Silica
More informationAlternative Method of Hybrid SPE Sample Preparation for High Recovery LC-MS Analysis of Pharmaceutical Compounds in Plasma
Alternative Method of Hybrid SPE Sample Preparation for High Recovery LC-MS Analysis of Pharmaceutical Compounds in Plasma Authors: Xiaoning Lu, and Michael Ye Presenter: Carmen T. Santasania Supelco/Sigma/Aldrich
More informationSimultaneous Quantitative Analysis of Total Catecholamines and Metanephrines in Urine Using 500 MG CLEAN UP CCX2 and LC-MS/MS
Simultaneous Quantitative Analysis of Total Catecholamines and Metanephrines in Urine Using 500 MG CLEAN UP CCX2 and LC-MS/MS UCT Part Numbers: CUCCX256 - Clean-Up CCX2 (C8 + Carboxylic Acid) 500mg/6mL
More informationQuantitative Analysis of Opiates in Urine Using RRHT LC/MS/MS. Application. Authors. Introduction. Abstract. Forensics
Quantitative Analysis of piates in Urine Using RRHT LC/MS/MS Application Forensics Authors Sheher Mohsin Agilent Technologies, Inc. 10 N. Martingale Rd., Suite 550 Schaumburg, IL 60173 USA Yanan Yang Agilent
More informationExtraction of a Comprehensive Steroid Panel from Human Serum Using ISOLUTE. SLE+ Prior to LC/MS-MS Analysis
Application Note AN890 Extraction of a Comprehensive Steroid Panel from Human Serum Using ISOLUTE SLE+ Page Extraction of a Comprehensive Steroid Panel from Human Serum Using ISOLUTE SLE+ Prior to LC/MS-MS
More informationImpact of Reversed-Phase Chiral Chromatography on the LC-MS Analysis of Drugs in Biological Fluids
Impact of Reversed-Phase Chiral Chromatography on the LC-MS Analysis of Drugs in Biological Fluids David S. Bell, Carmen T. Santasania, Jennifer Claus, Wayne K. Way, and Craig R. Aurand Supelco, Div. of
More informationLC-MS/MS analysis of Chlorates in Milk and Whey Powder using the Agilent 6470 QQQ
LC-MS/MS analysis of Chlorates in Milk and Whey Powder using the Agilent 6470 QQQ Anthony Sullivan, LC/MS Product Specialist Melanie Mülek and Christoph Müller LC-MS Applications Specialists Hewlett-Packard-Str.
More informationGlycanPac AXH-1 Columns
Thermo Scientific GlycanPac AXH-1 Columns Product Manual P/N: 065479-01 January 2013 Part of Thermo Fisher Scientific Product Manual for GlycanPac AXH-1 Columns GlycanPac AXH-1, 1.9 µm, Analytical, 2.1
More informationDetermination of β2-agonists in Pork Using Agilent SampliQ SCX Solid-Phase Extraction Cartridges and Liquid Chromatography-Tandem Mass Spectrometry
Determination of β2-agonists in Pork Using Agilent SampliQ SCX Solid-Phase Extraction Cartridges and Liquid Chromatography-Tandem Mass Spectrometry Application Note Food Safety Authors Chenhao Zhai Agilent
More informationLC-MS/MS Method for the Determination of 21 Opiates and Opiate Derivatives in Urine
LC-MS/MS Method for the Determination of 21 Opiates and Opiate Derivatives in Urine J. Jones, S. Westwood, T. Liddicoat, L. Pereira, T. Edge Thermo Fisher Scientific, Manor Park, Runcorn, UK Overview Purpose:
More informationSimultaneous Quantitative Analysis of Total Catecholamines and Metanephrines in Urine Using CLEAN UP CCX2 and LC-MS/MS
Simultaneous Quantitative Analysis of Total Catecholamines and Metanephrines in Urine Using CLEAN UP CCX2 and LC-MS/MS UCT Part Numbers CUCCX256 Clean Up CCX2 (C8 + Carboxylic Acid) 500mg / 6mL SPE Cartridge
More informationAnalysis of HMF by HPLC
COST Action 927 Training School Building Skills on the Analysis of Thermal Process Contaminants in Foods 22-26 October 2007, Ankara Analysis of HMF by HPLC Vural Gökmen O O OH Background O COOH O R 2 Carbonyl
More informationEnrichment of Phospholipids from Biological Matrices with Zirconium Oxide-Modified Silica Sorbents
Enrichment of Phospholipids from Biological Matrices with Zirconium Oxide-Modified Silica Sorbents Xiaoning Lu, Jennifer E. Claus, and David S. Bell Supelco, Div. of Sigma-Aldrich Bellefonte, PA 16823
More informationA Novel Solution for Vitamin K₁ and K₂ Analysis in Human Plasma by LC-MS/MS
A Novel Solution for Vitamin K₁ and K₂ Analysis in Human Plasma by LC-MS/MS By Shun-Hsin Liang and Frances Carroll Abstract Vitamin K₁ and K₂ analysis is typically complex and time-consuming because these
More informationLC-Based Lipidomics Analysis on QTRAP Instruments
LC-Based Lipidomics Analysis on QTRAP Instruments Junhua Wang and Paul RS Baker SCIEX LC-Based Lipidomics Analysis Topics Covered Lipid extraction techniques Hydrophilic Interaction Chromatography (HILIC)
More informationThe HPLC Preparative Scale-Up of Soybean Phospholipids Application
The HPLC Preparative Scale-Up of Soybean Phospholipids Application Food and Flavors Authors Cliff Woodward and Ron Majors Agilent Technologies, Inc. 285 Centerville Road Wilmington, DE 1988-161 USA Abstract
More informationThe LC Column Playbook
The LC Column Playbook Mark Powell Applications Engineer Technical Support 1 Overview How do we achieve our goal: Resolution? Efficiency Particle size Column length Selectivity Bonded phase Mobile phase
More informationAbout bioassay of Oximes:? New isolation alternatives from biomatrices? Chromatographic separation issues? Reserve on using MS or MS/MS detection
State of the art: xime-type AChE Reactivators: Wide use as antidotes in organo phosphorous compounds poisoning; Synthesis of new congeners; Intense studies on their transfer through BBB; Intense studies
More informationThermo Scientific. GlycanPac AXR-1. Column Product Manual. P/N: April, Part of Thermo Fisher Scientific
Thermo Scientific GlycanPac AXR-1 Column Product Manual P/N: April, 2014 Part of Thermo Fisher Scientific Product Manual for GlycanPac AXR-1 Columns GlycanPac AXR-1, 1.9 µm, Analytical, 2.1 x 150 mm (P/N
More informationProtein Precipitation for Biological Fluid Samples Using Agilent Captiva EMR Lipid 96-Well Plates
Application Note Clinical Research Protein Precipitation for Biological Fluid Samples Using Agilent Captiva EMR Lipid 96-Well Plates Authors Limian Zhao and Megan Juck Agilent Technologies, Inc. Abstract
More informationFortis Method Development Options
Method Dev Fortis Method Development Options Choice of Stationary phase functionality Based on Ultra pure silica Reversed Phase (RP) and Normal Phase (NP) options Fortis Ho - Polar endcapped C8 - Increased
More informationSimultaneous Analysis of Intact Human Insulin and Five Analogs in Human Plasma Using μelution SPE and a CORTECS UPLC Column
Simultaneous Analysis of Intact Human Insulin and Five Analogs in Human Plasma Using μelution SPE and a CORTECS UPLC Column Erin E. Chambers and Kenneth J. Fountain Waters Corporation, Milford, MA, USA
More informationSepax Technologies, Inc.
Sepax Technologies, Inc. Sepax Technologies, Inc. develops and manufactures products in the area of chemical and biological separations, biosurfaces and proteomics. Sepax product portfolio includes ) liquid
More informationSPE-LC-MS/MS Method for the Determination of Nicotine, Cotinine, and Trans-3-hydroxycotinine in Urine
SPE-LC-MS/MS Method for the Determination of Nicotine, Cotinine, and Trans-3-hydroxycotinine in Urine J. Jones, Thermo Fisher Scientific, Runcorn, Cheshire, UK Application Note 709 Key Words SPE, SOLA
More informationRaptor HILIC-Si: Simplify the Switch to HILIC
Stationary Phase: Silica Selectivity Accelerated Raptor HILIC-Si: Simplify the Switch to HILIC Retain polar compounds without ion-pairing reagents. 2.7 µm Raptor core-shell particles provide the speed
More informationShuguang Li, Jason Anspach, Sky Countryman, and Erica Pike Phenomenex, Inc., 411 Madrid Ave., Torrance, CA USA PO _W
Simple, Fast and Accurate Quantitation of Human Plasma Vitamins and Their Metabolites by Protein Precipitation Combined with Columns Using HPLC-UV, HPLC-FLD or LC/MS/MS Shuguang Li, Jason Anspach, Sky
More informationLC/MS Method for Comprehensive Analysis of Plasma Lipids
Application Note omics LC/MS Method for Comprehensive Analysis of Plasma s Authors Tomas Cajka and Oliver Fiehn West Coast Metabolomics Center, University of California Davis, 451 Health Sciences Drive,
More informationDienes Derivatization MaxSpec Kit
Dienes Derivatization MaxSpec Kit Item No. 601510 www.caymanchem.com Customer Service 800.364.9897 Technical Support 888.526.5351 1180 E. Ellsworth Rd Ann Arbor, MI USA TABLE OF CONTENTS GENERAL INFORMATION
More informationAnalyze Barbiturates in Urine with Agilent 6430 LC/MS/MS and Poroshell 120 EC-C18
Analyze Barbiturates in Urine with Agilent 6 LC/MS/MS and Poroshell EC-C8 Application ote Forensic Toxicology Authors Elijah Steinbauer and Pat Friel Toxicology Laboratory at the Veterans Administration
More informationA Robustness Study for the Agilent 6470 LC-MS/MS Mass Spectrometer
A Robustness Study for the Agilent 7 LC-MS/MS Mass Spectrometer Application Note Clinical Research Authors Linda Côté, Siji Joseph, Sreelakshmy Menon, and Kevin McCann Agilent Technologies, Inc. Abstract
More informationSeparation of Vitamin D and Vitamin D Metabolites on FLARE C18 MM (Mixed Mode) HPLC Column
Diamond Analytics Technical Note: T131-1 Separation of Vitamin D and Vitamin D Metabolites on FLARE C1 MM (Mixed Mode) HPLC Column Introduction In this technical note, the versatility of the diamond-based
More informationFig. 1: Chemical structure of arachidonic acid COOH CH 3
Elimination of Matrix Effects Using Mixed-mode SPE Plate for High Throughput Analysis of Free Arachidonic Acid in Plasma by LC-MS/MS Wan Wang, Suzi Qin, Linsen Li, Warren Chen, Jerry Wang 179, Southern
More informationExtended Temperature Range Impact on HPLC Column Stability and Performance
Extended Temperature Range Impact on HPLC Column Stability and Performance Stephanie J. Marin Dale Felix, Jody Clark Selerity Technologies, Inc. Salt Lake City, UT www.selerity.com The Selerity Series
More informationUPLC/MS Monitoring of Water-Soluble Vitamin Bs in Cell Culture Media in Minutes
UPLC/MS Monitoring of Water-Soluble Vitamin Bs in Cell Culture Media in Minutes Catalin E. Doneanu, Weibin Chen, and Jeffrey R. Mazzeo Waters Corporation, Milford, MA, U.S. A P P L I C AT ION B E N E F
More informationRaptor HILIC-Si: Simplify the Switch to HILIC
Stationary Phase: Silica Selectivity Accelerated Raptor HILIC-Si: Simplify the Switch to HILIC Retain polar compounds without ion-pairing reagents. 2.7 µm Raptor core-shell particles provide the speed
More informationLC Columns with Liquid Separation Cell Technology
Obelisc LC Columns with Liquid Separation Cell Technology Obelisc HPLC Columns - Liquid Separation Cell Technology Introduction Obelisc HPLC columns are the latest, innovative columns from SIELC Technologies,
More informationGlycanPac AXR-1 Columns
CHRMATGRAPHY GlycanPac AXR- Columns For High Resolution Glycan Analysis Product Specifications The Thermo Scientific GlycanPac AXR- columns are highperformance, silica-based HPLC columns for simultaneous
More informationSeparation of Macrocyclic Lactones (Avermectins) on FLARE C18 MM & FLARE C18+ Columns
Separation of Macrocyclic Lactones (Avermectins) on FLARE C8 MM & FLARE C8+ Columns Introduction Diamond Analytics Technical Note: T05- Avermectins are a series of 6-membered macrocyclic lactone derivatives
More informationQuantification of lovastatin in human plasma by LC/ESI/MS/MS using the Agilent 6410 Triple Quadrupole LC/MS system
Quantification of lovastatin in human plasma by LC/ESI/MS/MS using the Agilent 641 Triple Quadrupole LC/MS system Application Note Clinical Research Author Siji Joseph Agilent Technologies Bangalore, India
More informationUNISON UK - Amino For Aqueous Elutions and Exceptional Separation Balance
UNISON UK - Amino For Aqueous Elutions and Exceptional Separation Balance Revolutionary aqueous durability Aqueous to non-aqueous High speed normal phase separation 3μm particle size Superior resolution
More informationMethod Development for the Analysis of Endogenous Steroids Using Convergence Chromatography with Mass Spectrometric Detection
Method Development for the Analysis of Endogenous Steroids Using Convergence Chromatography with Mass Spectrometric Detection Christopher J. Hudalla, Stuart Chadwick, Fiona Liddicoat, Andrew Peck, and
More information[ APPLICATION NOTE ] Oasis PRiME HLB Cartridge for Cleanup of Infant Formula Extracts Prior to UPLC-MS/MS Multiresidue Veterinary Drugs Analysis
Prior to UPLC-MS/MS Multiresidue Veterinary Drugs Analysis Michael S Young and Kim Van Tran Waters Corporation, Milford, MA, USA APPLICATION BENEFITS Efficient, timesaving multiclass/ multiresidue methodology
More informationfor the Identification of Phosphorylated Peptides
Application of a Data Dependent Neutral-Loss Experiment on the Finnigan LTQ for the Identification of Phosphorylated Peptides Gargi Choudhary Diane Cho Thermo Electron, San Jose, CA Abstracted from posters
More informationAnalysis of Polar Pesticides Using a Mixed-Phase Mode HPLC Column
Analysis of Polar Pesticides Using a Mixed-Phase Mode HPLC Column Narong Chamkasem, Southeast Food and Feed Laboratory (SFFL), U.S. Food and Drug Administration, Atlanta, GA Disclaimer The findings and
More informationIdentification and Quantification of Psychedelic Phenethylamine 2C Series using SPE and LC-MS/MS
Identification and Quantification of Psychedelic Phenethylamine 2C Series using SPE and LC-MS/MS UCT Part Numbers: CSXCE106-Clean Screen XCEL I 130mg/6mL SLDA100ID21-3UM - Selectra DA HPLC column, 100
More informationAuthor. Introduction. Abstract
Simultaneous Measurement of Sirolimus and Tacrolimus Concentrations in Blood by Semi-Automated Extraction and Liquid Chromatography-Electrospray Ionization Mass Spectrometry Application Author Gary L.
More informationA Definitive Lipidomics Workflow for Human Plasma Utilizing Off-line Enrichment and Class Specific Separation of Phospholipids
A Definitive Lipidomics Workflow for Human Plasma Utilizing Off-line Enrichment and Class Specific Separation of Phospholipids Jeremy Netto, 1 Stephen Wong, 1 Federico Torta, 2 Pradeep Narayanaswamy, 2
More informationSUPPORTING INFORMATION:
1S SUPPORTING INFORMATION: The discovery of the once weekly glucagon like peptide 1 (GLP-1) analog semaglutide Jesper Lau*, Paw Bloch, Lauge Schäffer, Ingrid Pettersson, Jane Spetzler, Jacob Kofoed, Kjeld
More informationDetermination of Amantadine Residues in Chicken by LCMS-8040
Liquid Chromatography Mass Spectrometry Determination of Amantadine Residues in Chicken by LCMS-8040 A method for the determination of amantadine in chicken was established using Shimadzu Triple Quadrupole
More informationBiomed Pap Med Fac Univ Palacky Olomouc Czech Repub
SUPPLEMENTAL MATERIAL Sistik P, Turjap M, Iordache AM, Saldanha H, Lemr K, Bednar P. Quantification of selected antidepressants and antipsychotics in clinical samples using chromatographic methods combined
More informationApplication Note. Author. Abstract. Introduction. Food Safety
Determination of β2-agonists in Pork with SPE eanup and LC-MS/MS Detection Using Agilent BondElut PCX Solid-Phase Extraction Cartridges, Agilent Poroshell 120 column and Liquid Chromatography-Tandem Mass
More informationA New HILIC/RP Mixed-Mode Column and Its Applications in Surfactant Analysis
A New HILIC/RP Mixed-Mode Column and Its Applications in Surfactant Analysis X. Liu, C. Pohl, Dionex Corporation, Sunnyvale, CA, USA ABSTRACT Although reversed-phase (RP) silica columns (e.g., C18 and
More informationThe Comparison of High Resolution MS with Triple Quadrupole MS for the Analysis of Oligonucleotides
The Comparison of High Resolution MS with Triple Quadrupole MS for the Analysis of Oligonucleotides Mohammed Abrar Unilabs York Bioanalytical Solutions Outline Introduction Why LC-MS/MS? Limitations of
More informationRobust extraction, separation, and quantitation of structural isomer steroids from human plasma by SPE-UHPLC-MS/MS
TECHNICAL NOTE 21882 Robust extraction, separation, and quantitation of structural isomer steroids human plasma by SPE-UHPLC-MS/MS Authors Jon Bardsley 1, Kean Woodmansey 1, and Stacy Tremintin 2 1 Thermo
More informationSimple Method (IS-MRM) to Monitor Lysophospholipids and Phospholipids During LC-MS Method Development via In-Source CID
Simple Method (IS-MRM) to Monitor Lysophospholipids and Phospholipids During LC-MS Method Development via In-Source CID James Little, Eastman Chemical Company, Kingsport, TN Overview Phospholipids and
More informationExtraction of Aflatoxin M1 From Infant Formula Using ISOLUTE Myco SPE Columns prior to LC-MS/MS Analysis
Application Note AN807 Extraction of Aflatoxin M From Infant Formula Using ISLUTE Myco Page Extraction of Aflatoxin M From Infant Formula Using ISLUTE Myco SPE Columns prior to LC-MS/MS Analysis This application
More informationChallenges in Developing an Ultra-Sensitive Bioanalytical Method for Ethinylestradiol in Human Plasma
Challenges in Developing an Ultra-Sensitive Bioanalytical Method for Ethinylestradiol in uman Plasma Jessalynn P. Wheaton, Erin E. Chambers, and Kenneth J. Fountain Waters Corporation, 34 Maple Street,
More informationPHOTOCATALYTIC DECONTAMINATION OF CHLORANTRANILIPROLE RESIDUES IN WATER USING ZnO NANOPARTICLES. DR. A. RAMESH, Ph.D, D.Sc.,
PHOTOCATALYTIC DECONTAMINATION OF CHLORANTRANILIPROLE RESIDUES IN WATER USING ZnO NANOPARTICLES DR. A. RAMESH, Ph.D, D.Sc., raamesh_a@yahoo.co.in 1 OBJECTIVES Determination of persistence and photolysis
More informationUnequalled durability against water elution. % tr
Unequalled durability against water elution 2 Revolutionary aqueous durability for aminopropyl phase 1 3 4 1. fructose (2.5mg/mL) 3. sucrose (2.5mg/mL) 500 hr 2. glucose 400 hr Aqueous to non-aqueous Normal
More informationDetermination of N-Nitrososarcosine (NSAR) in tobacco
JTI-Ökolab Vienna, Austria Determination of N-Nitrososarcosine (NSAR) in tobacco Madeleine Werneth, Jutta Pani, Bernhard Mayer-Helm 2014 CORESTA CONGRESS - ST46 Québec City, Canada 12-16 October 2014 Background
More informationIntegration of steroids analysis in serum using LC-MS/MS with full-automated sample preparation
PO-CON69E Integration of steroids analysis in serum using LC-MS/MS with full-automated sample preparation MSACL 6 EU Stéphane Moreau, Daisuke Kawakami, Toshikazu Minohata Shimadzu Europe GmbH, Duisburg,
More informationNoora Perkola Finnish Environment Institute. Nordic MS Symposium November 9, 2011, Båstad, Sweden
Noora Perkola Finnish Environment Institute Nordic MS Symposium November 9, 2011, Båstad, Sweden Artificial Sweeteners Used as additives in various food products to replace sugar 4 non-caloric sweeteners
More informationHigh Throughput simultaneous analysis of 39 amino acids in various foods without derivatization using LC-MS/MS
PO-CON1532E High Throughput simultaneous analysis of 39 amino acids in various foods without derivatization using LC-MS/MS ASMS 2015 ThP 409 Manabu Yukiyama 1, Keiko Matsumoto 1, Jun Watanabe 1, Itaru
More informationDirect Analysis of Urinary Opioids and Metabolites by Mixed-Mode µelution SPE Combined with UPLC/MS/MS for Forensic Toxicology
Direct Analysis of Urinary Opioids and Metabolites by Mixed-Mode µelution SPE Combined with UPLC/MS/MS for Forensic Toxicology Jonathan P. Danaceau, Erin E. Chambers, and Kenneth J. Fountain Waters Corporation,
More informationEfficient Quantitative Analysis of THC and Metabolites in Human Plasma Using Agilent Captiva EMR Lipid and LC-MS/MS
Application Note Forensic Analysis Efficient Quantitative Analysis of THC and Metabolites in Human Plasma Using Agilent Captiva EMR Lipid and LC-MS/MS Authors Joan Stevens and Limian Zhao Agilent Technologies,
More informationExtraction of Multiple Mycotoxins From Nuts Using ISOLUTE Myco prior to LC-MS/MS Analysis
Application Note AN784 Extraction of Multiple Mycotoxins from Nuts Using ISOLUTE Myco Page 1 Extraction of Multiple Mycotoxins From Nuts Using ISOLUTE Myco prior to LC-MS/MS Analysis This application note
More informationIdentification and Quantification at ppb Levels of Common Cations and Amines by IC-MS
Application Note 269 Identification and Quantification at ppb Levels of Common Cations and Amines by IC-MS INTRODUCTION Ion chromatography (IC) has been extensively used as the preferred separation technique
More informationSOLID PHASE EXTRACTION: Fundamentals and Recent Developments
SOLID PHASE EXTRACTION: Fundamentals and Recent Developments Olga I. Shimelis, Ph.D Principal R&D Scientist sigma-aldrich.com/analytical Time Spent on the Analytical Process R.E. Majors, LC/GC Magazine,
More informationIC-MS Environmental Applications - Water Testing. Application Notebook
IC-MS Environmental Applications - Water Testing Application Notebook Table of Contents Environmental Sample Using IC-MS 3 Environmental Applications: Water Testing Acrylamide in Water 4 Trace Urea in
More informationRapid and Robust Detection of THC and Its Metabolites in Blood
Rapid and Robust Detection of THC and Its Metabolites in Blood Application Note Forensics/Doping Control Author Stephan Baumann Agilent Technologies, Inc. Santa Clara CA 95051 USA Abstract A robust method
More informationAnalytical Method for 2, 4, 5-T (Targeted to Agricultural, Animal and Fishery Products)
Analytical Method for 2, 4, 5-T (Targeted to Agricultural, Animal and Fishery Products) The target compound to be determined is 2, 4, 5-T. 1. Instrument Liquid Chromatograph-tandem mass spectrometer (LC-MS/MS)
More information