Corona Veo the Next Generation Charged Aerosol Detector Presenters Name Title Date 1 The world leader in serving science
Contents Introduction to Charged Aerosol Detection How Charged Aerosol Technology Works Key Features/Product Variations of Corona Veo Detectors Detection of Analytes in Different Matrices Inverse Gradient Solution for Uniform Response New Approach to a Generic Analytical Method Wrap-up Questions & Discussion 2
Introduction to Charged Aerosol Detection Used to quantitate any non-volatile and most semi-volatile analytes with LC Provides consistent analyte response independent of chemical structure Neither a chromophore, nor the ability to ionize, is required for detection Dynamic range of over four orders of magnitude from a single injection (sub-ng b to µg quantities on column) Mass sensitive detection provides relative quantification without the need for reference standards Compatible with gradient conditions for HPLC, UHPLC, and Micro LC Comparison of Charged Aerosol Detection to UV and MS 3
Charged Aerosol Detection How It Works The liquid eluent from the LC column enters the detector (1) where it undergoes nebulization by combining with a concentric stream of nitrogen gas or air (2). The fine droplets are carried by bulk gas flow to the evaporation sector (3) where desolvation occurs to form particles, while any larger droplets are drained to waste (4). Signal is directly proportional to the analyte quantity The dry particles exit from evaporation (5) and are combined with another gas stream that first passes over a high voltage Corona charger (6). The charged gas then mixes Any high h mobility species are removed by an ion trap (8) while with the dry particles, where the remaining charged particles pass to a collector, where the excess charge transfers to the charges from the passing particles are measured with a very particle s surface(7). sensitive electrometer (9). The resulting signal is conveyed to a chromatographic data software for quantitation (10). 4
Particle Charging for Charged Aerosol Detection Analyte particles with charged surface Charges remain on particles surface. Particles remain intact t and do not ionize. The more surface area, the more charge is carried by the particle Dry analyte particles Charged nitrogen gas Mixing Chamber 5
CoronaVeo Charged Aerosol Detector A novel, near universal, mass sensitive detector for routine LC determinations of any non-volatile and many semi-volatile analytes Uses the fastest growing g and most rapidly adopted universal LC detection technology since diode array detection (PDA or DAD) Provides unbiased detection for a wide variety of analyte classes New design incorporates unique, proprietary technologies only available from Thermo Fisher Scientific Thermo Scientific Dionex Corona Veo Charged Aerosol detector 6
Corona Veo Detector What's New? This entirely new detector incorporates many design and performance improvements: Radically new Thermo Scientific Dionex FocusJet concentric nebulization system improves sensitivity and precision All new aerosol conditioning and evaporation schemes widen the scope of applications to include capillary and micro LC, as well as UHPLC Usability and serviceability are enhanced by countless improvements, many of which came from our customers 7
FocusJet Concentric Nebulizer Technology The advanced concentric nebulizer design ensures uniform aerosol atomization pattern for superior droplet size selection Exchangeable design gives users ready access to improve serviceability and increase uptime Inlet accepts conventional male fittings, including Viper fingertight fittings for virtually dead-volume-free connections FocusJet Concentric Nebulizer Thermo Scientific Dionex Viper Fitting 8
FocusJet Nebulizer and Spray Chamber The inlet liquid and nebulization gas streams combine coaxially into an aerosol spray of uniform droplets at the nebulizer tip The fine droplets are transported into the heated evaporation sector Any large droplets fall and are expelled by a precision micro-pump To Evaporation Sector Impactor Coaxial N 2 gas flow Aerosol Concentric Nebulizer Fused silica capillary Spray chamber FocusJet Concentric Nebulizer Tip Inlet Gas Cross-flow In (N 2 ) Nebulizer Active drain pump 9
Standard Features Corona Veo Detector Exchangeable nebulizer the FocusJet nebulizer/spray chamber system improves performance Inlet flow capability accepts flows from 0.20 to 2.0 ml/min Front access to all fluidic connections improves serviceability and maximizes uptime Touch screen user interface provides clear operating commands and displays practical diagnostic messaging Revised fluid waste management a non-pressurized waste collection scheme simplifies operation Evaporation control two selectable evaporation temperatures permit stable operation with the typical range of eluents Active detector thermostatting to maintain stable performance and improve reproducibility Expanded response linearity through integration of a selectable Power function 10
Advanced Features Corona Veo RS Detector Includes all the features of the standard Corona Veo detector with the addition of: Programmable evaporation temperature a flexible tool for further optimizing analyte response over an expanded range of eluents (ambient +5 to +100 o C) Extended inlet flow capability accepts inlet flow rates for HPLC, UHPLC, capillary, and micro LC (as low as 10 µl/min.) Electronic gas pressure regulator (EGR) ensures optimized performance over the entire flow rate range Integrated stream switching (SSV) for automated flow diversion to alternate devices or for peak elimination and collection Advanced digital signal processing for fast data acquisition speeds necessary with UHPLC (up to 200 Hz with Thermo Scientific Dionex Chromeleon 7.2 Chromatography Data System) 11
Expanding the Range of Non-Linear Response Non-linear response with charged aerosol detection Linear response utilizing the power function Use a Power Function Value (PFV) to expand the linear calibration range for a non-linear aerosol detector Enhanced calibration using the Power Function 12
Applying Power Function to a Corona Veo Detector Improves visualization of the linear calibration range Acts to improve accuracy of linear interpolation at each end Has no effect on the limit of detection (LoD) Extend the linear calibration range using the Power Function 13
Product Variations of Corona Veo Detectors Model/Feature Corona Veo Detector Corona Veo RS Detector Flow Rate Range 0.20 2.0 ml/min 0.01 2.0 ml/min Evaporation Temp 35 C or 50 C Ambient +5 C to +100 C Integrated Stream Switching Valve (SSV) Not offered 6-port, 2-position micro valve (TTL controlled) Internal Gas Regulation User Adjustable Electronic control Data Acquisition Rate Up to 100 Hz Up to 200 Hz (with CM 7.2) Inlet Gas Pressure (reqd) Standalone Interface Liquid Waste Management Enhanced Linearity Analog Signal Output (0 1 VDC) 4.8 5.5 bar (70 80 psig; 482 551 kpa) Integrated color LCD touch screen (rationalized GUI) Non-pressurized (uses general LC waste stream) Via Power Function (User applied) Installable Option 14
Electronic Gas Regulation Corona Veo RS Detector Internal Electronic Gas Regulator Provides ultra-precise gas pressure to ensure efficient nebulization and stable particle flow through the entire flow path Ensures optimum detector performance over the entire inlet flow range of the Corona Veo RS detector (10 µl 2.0 ml per min.) Both analytical and micro operation modes are selectable via remote control from a chromatography data system 15
Integrated Stream Switching Corona Veo RS Detector Stream Switching Valve (SSV) Simplify system operation by enabling access to alternate devices without the need to unplumb/replumb modules (UV, MS, etc.) Diverts flow to waste in event of gas or pump flow errors, to protect against damage Improve chromatography results for complex samples by allowing elimination of harmful salts or collection of unretained peaks 16
Detection of a Broad Range of Analytes in Different Matrices 17
Glycan Analysis for Bovine Fetuin Separation of Oligosaccharide Alditols Column: Thermo Scientific GlycanPac AXH-1 1.9 μm, 2.1 150 mm Mobile phase A: 80% Acetonitrile Mobile phase B: 80 mm Ammonium formate, ph 4.4 Gradient: 2.5 % B to 25% B from 1 to 40 min Flow rate: 0.4 ml/min Inj. volume: 5 μl Col. temp: 30 o C Evap. temp: 50 o C Native Glycans 18
Determination of Adjuvants Analysis of Plant Saponins Column: Thermo Scientific Hypersil GOLD PFP 1.9 um, 2.1 100 mm Mobile phase A: 0.1% Formic acid in water Mobile phase B: 0.1% Formic acid in 10:90 acetonitrile: reagent alcohol Gradient: 35% B to 83%B in 6 min to 90% B in 10 min Flow rate: 0.46 ml/min Inj. volume: 2 μl Col. temp: 45 o C Evap. temp: 50 o C UV @ 210 nm 19
Determination of Carbohydrates in Juice Analysis of Simple Sugars Column: Amino, 3 μm, 3 250 mm Mobile phase: Acetonitrile:water (92:8) Flow rate: 0.8 ml/min Inj. volume: 2 μl Col. temp; 60 o C Post-column temp: 25 o C Evap. temp: 75 o C Sample preparation: Add 20 ml of 85% ACN to 1 gram juice Simplified sample preparation Dilute-and-shoot method 20
Extracts from Natural Products Analysis of Hoodia Plant Extracts Column: Mobile phase A: Mobile phase B: Gradient: Flow rate: Inj. volume: Col. temp: Evap. temp: Thermo Scientific Accucore C18 2.6 μm, 2.1 150 mm Water Acetonitrile 35%B to 90%B in 10 min 0.5 ml/min 2 μl 40 o C 35 o C Hoodigosides are oxypregnane steroidal glycosides abundant in Hoodia gordonii 21
Separation of Herbal Extracts Analysis of Milk Thistle Extracts Column: Thermo Scientific Acclaim 120 C18 2.2 µm, 2.1 100 mm Mobile phase A: 80:20 (v/v) 3mM Ammonium formate, 0.3% formic Acid:methanol Mobile phase B: 20:80 (v/v) 3mM Ammonium formate, 0.3% formic Acid:methanol Gradient: 15% B to 45% B in 4.4 min Flow rate: 0.5 ml/min Inj. volume: 2 μl Col. temp: 40 C Evap. temp: 35 C 22
Characterization of Algae-Based Biofuels Analysis of Algal Oils Column: Accucore C18, 2.6 μm, 3.0 150 mm Mobile phase A: Methanol/water/acetic acid (600:400:4) Mobile phase B: Tetrahydrofuran/acetonitrile (50:950) Mobile phase C: Acetone/acetonitrile (900:100) Gradient: Time FlowRate %A %B %C (min) (ml/min) -10.0 1. 00 90 10 0-0.1 1. 00 90 10 0 0. 0 0. 25 90 10 0 20.00 0. 50 15 85 0 35.0 0. 50 2 78 20 60.0 0. 50 2 3 95 65.0 0. 50 90 10 0 Flow rate: 1.0 ml/min Inj. volume: 2 μl Col. temp: 40 o C Evap. temp: 40 o C 23
Analysis of Industrial Descaling Agents Measurement of Polyacrylic Acid In Boiler Water Column: Acclaim SEC-300, 5 μm, 4.6 300 mm Mobile phase A: Acetonitrile Mobile phase B: Water Isocratic: 10:90 Flow rate: 0.35 ml/min Inj. volume: 100 μl Col. Temp: 30 o C Evap. temp: 55 o C 24
Active Ingredient Composition Analysis of Gentamicin standard (200 μg/ml) Column: Acclaim RSLC PolarAdvantage II, 2.2 μm, 2.1 100 mm Mobile phase A: 0.025:95:5 HFBA:water:acetonitrile Mobile phase B: 0.3:95:5 TFA:water:acetonitrile Gradient: - 0to15min1to 1.5min,1 10%B - 1.5 to 7min,10 to 100%B - 7 to 10min,100%B - 4 min. pre-injection equilibration Flow rate: 0.45 ml/min Inj. volume: 1 μl Col. temp: 15 o C Evap. temp: 80 o C 25
Impurity Testing at Low Flow Rate Analysis of Amikacin Sulfate USP (1.0 mg/ml) Column: C18,.2μm, 1.0 35 mm Mobile phase A: 0.2% Pentafluoropropionic acid in water Mobile phase B: Methanol Gradient: Time %A %B (min) -3.0 90 10 0.0 90 10 0.3 90 10 45 4.5 40 60 Flow rate: 50 µl/min Inj. volume: 0.5 μl Col. Temp: 55 o C Evap. temp: 35 o C 26
Determination of Drug Discovery Mass Balance Charged Aerosol UV Column: Acclaim 300 C18, 3 μm, 4.6 150 mm Mobile phase A: 20 mm Ammonium acetate, ph 4.5 Mobile phase B: Acetonitrile Gradient: 2% B to 98% B in 30 min, Inverse Gradient Flow rate: 0.8 ml/min Inj. volume: 2 μl Col. temp: 30 o C Evap. temp: 35 o C 27
Formulation Testing Analysis of Diclofenac-Sodium Salt (1 mg/ml) Column: Mobile phase: Flow rate: Inj. volume: Col. temp: Evap. temp: Acclaim Trinity P1, 3μm, 3.0 50 mm 75% Acetonitrile:25% 200mM Ammonium acetate ph 4 0.8 ml/min 5 μl 30 o C 60 o C Measurement of Chloride Impurity 28
Inverse Gradient Solution for Uniform Response with Charged Aerosol Detection 29
Inverse Gradient Compensation for Uniform Response Conventional Gradient Elution Inverse Gradient Compensation 30
The Effects of Gradient and Mass on Calibration 7 Standard Gradient (Single Pump) Inverse Gradient (Dual Pump) 7 6 6 5 5 Peak Area (CAD) 4 3 2 Sulfanilamide Famotidine Perphanzine R² = 0.9998 R² = 0.9995 R² = 0.9999 4 3 2 R² = 0.9997 R² = 0.9999 R² = 1 Sulfanilamide Famotidine Perphanzine 1 1 0 0 500 1000 1500 2000 2500 0 0 500 1000 1500 2000 2500 Mass on Column (ng) Mass on Column (ng) Inverse gradient extends the consistency of response 31
Solution for Uniform Response with Gradients A dual-gradient pump is the heart of this exclusive solution Inverse gradient kits with Viper connections provide easy installation Kit contains a media CD with eworkflows for Chromeleon CDS software Thermo Scientific Dionex UltiMate 3000 X2 Dual Gradient Pump Analytical Flow path Make-up Flow path Inverse Gradient Configuration A unique solution provided only by Thermo Scientific 32
A Radically New Approach to a Generic Method HILIC Excellent for polar molecules Reversed Phased Excellent for non- polar molecules Universal Charged Aerosol Detection 33
Combination of Composition and Flow Gradients Programmed gradients on both pumps mau RP+HILIC/UV 6 3 4 7 UV method misses peaks 5 8 Resulting gradient on HILIC column 0 5 10 15 20 23 min RP+HILIC/Charged Aerosol 4 5 6 7 Additional peaks found, more Constant flow at 1.5 ml/min pa 1 2 3 8 consistent response for others 0 5 10 15 20 23 min 34
Next Steps Toward a True Generic Method The high organic mobile phase used in HILIC separations offer increased run speeds under UHPLC conditions. If volatile substances with chromophores are present, a UV-vis or DAD could be coupled before the charged aerosol detector to give complementary data. Combining this technique with LC-MS as an orthogonal detector would enable simultaneous confirmation of the identity of degradation products. RP-HILIC-UV-Charged Aerosol has great potential for generic screening 35
Wrap-Up A Corona Veo charged aerosol detector is the only universal LC detector that you need in your lab! Can provide consistent inter-analyte responses across a wide range analytes An ideal replacement for older, less capable HPLC detectors (RI, ELSD, etc.) An outstanding complementary detector to UV, DAD, and MS Inverse gradient techniques offer a unique solution for even better mass balance determinations Opens up exciting new possibilities for development of truly generic analytical methods 36
Thank You for Your Attention For a Cleaner, Healthier, Safer World 37