AZURA® Analytical HPLC/UHPLC n Stay flexible regarding flow rates and upgrade options AZURA® Analytical HPLC/UHPLC How much performance do you need? n AZURA® HPLC Plus n AZURA® Compact HPLC AZURA Analytical Available pump heads Gradient options Max back pressure system features Max. flow rate in ml/min in bar and options 5 10 Isocratic LPG HPG 400 700 1000 AZURA Compact HPLC    AZURA HPLC Plus      AZURA UHPLC     2 If you want to use HPLC columns with a smaller particle size, a higher maximum system back pressure is needed. Max. back pressure n AZURA® UHPLC n AZURA® HPLC Plus Recommended Recommended Sample Injection Detector Recommended column particle size K-Connect Manual Auto- inner diameter capillary inner injection sampler UVD MWD DAD RID diameter 4 – 4.6 mm 5 μm 0.45 mm    3 – 4 mm 3 – 5 μm       0.18 mm 3 or smaller 3 μm or smaller      0.10 mm www.knauer.net/azuraanalytical 3 AZURA® Analytical HPLC/UHPLC Efficient and adaptable to your needs The analytical HPLC and UHPLC systems of the KNAUER AZURA® liquid chromatography instruments are designed to support and facilitate your work. Whether doing routine analysis or demanding separation tasks, AZURA systems are the right tool to overcome your analytical challenges. Choose between different gradient forming technologies and maximum flow rates to find the best configuration for your task. n AZURA Analytical HPLC/ UHPLC features • Isocratic, binary high pressure gradient or quaternary low pressure gradient pump • Pump heads allowing flow rates up to 10 ml/min at 700 bar or 1000 bar up to 2 ml/min • Autosampler with 0.1 µl sample aspiration at max. 1200 bar with zero sample loss • Choice of highly sensitive UVD, MWD, DAD, or RID detectors with intelligent temperature control • Wide range of flow cells available, including remote cells • Finger-tight high pressure stainless steel capillary connections “K-Connect” Injection • Extensive safety features such as leak management Automatic or manual injection and sensors (page 10) • Frontal access of detector lamp and pump head for easy maintenance 4 Detection Various detector types (UV/VIS, DAD, RI) and flow cells (page 14) Mobile control Optional system control via tablet (page 21) Tempering Column thermostats and Eluent delivery column switching solutions A choice of pumps available (page 12) in different flow rates and pressure ratings (page 6) www.knauer.net/azuraanalytical 5 Eluent delivery For highly accurate and precise HPLC and UHPLC n AZURA Pump P 6.1L AZURA Pump Filter The analytical pump AZURA P 6.1L combines all essential components of a first class HPLC pumping system. Three Protects the column different configurations are available: A 2 x 2 binary high from contamination pressure gradient (HPG) for high accuracy blending of (2 µm filter disc) up to two eluents from two selectable solvents each; a low pressure gradient (LPG) for reliable blending of up to four eluents and a very cost-effective isocratic version for easy analyses. Excellent separations with small particle size columns can be achieved with the high-performance pump heads featuring an extended back pressure. Choose between pump heads with a maximum flow rate of 10 ml/min and 700 bar1 backpressure, or pump heads with a flow rate of 5 ml/min and 1000 bar2 backpressure. Special pump heads for Normal Phase applications will help to deliver robustly even demanding eluents like heptane or hexane. Pumps without a degasser offer a cost effective alternative. Choose your HPLC pump according to your application’s needs. Gradient formation, mixer size and pulsation compensation will AZURA Mixer have an extensive influence. Highly efficient microfluidic solvent mixer Available pump versions Pump head (type): P 6.1L P 6.1L P 6.1L material, max. pressure Isocratic HPG LPG 5 ml: stainless steel, 1000 bar2   10 ml: stainless steel, 700 bar1   10 ml normal phase: stainless steel, 700 bar1   10 ml normal phase: stainless steel, 700 bar1 without degasser   10 ml: stainless steel, 700 bar1 without degasser    1) > 5 ml/min: max. 400 bar Optional: P 6.1L with 50 ml pump head available (max. 50 ml/min; 300 bar) 2) > 2 ml/min: max. 700 bar 6 n AZURA® Pump P 6.1L Binary HPG Online Degasser Reduces baseline noise resulting in higher Piston back flush sensitivity This standard feature of the P 6.1L will wash away contaminations for prolonged seal lifetime Solvent Selection Valve Integrated in every binary HPG pump, each channel can be used with two different solvents www.knauer.net/azurap6 7 High or low pressure gradient? The choice of gradient formation technology LPG HPG is essential for a HPLC/UHPLC system. AZURA systems can be equipped with either a high or a low pressure gradient pump. Both techniques Low investment costs Very fast gradients have advantages and disadvantages. A low pressure gradient (LPG) module dynami-Four channel blend Smallest dead volume cally composes the eluent on the inlet-side or low pressure side of the pump head, by quickly switching between the different solvent channels. 250 The AZURA pump technology adapts to the mixer volume and changes the valve switching cycle time accordingly. The eluent in a binary high pressure gradient (HPG) system is composed by combining the solvent flows of two pump heads. 25 % 24 % 23 % 22 % 150 21 % mAU 20 % Excellent gradient reproducibility of 0.3 % RSD. Overlay of 6 repetitions at 1 ml/min run with pump 100 P 6.1L low pressure gradient version 50 Time Choose a solvent mixer according to your application n AZURA® Mixer Microfluidic solvent mixer The sophisticated micro fluidics of the AZURA Mixer combine high mixing performance with a low dead volume. The user-changeable mixer is available in different volumes (50 / 100 / 200 µl) allowing best possible adaption to any application or flow rate. 8 n AZURA® Pump P 6.1L Quaternary LPG AZURA Pump Filter Piston back flush Protects the column from Washes away contaminations contamination for longer seal life (2 µm filter disc) (standard feature of P 6.1L) AZURA Mixer Multi-proportioning valve 4-Channel Online Degasser Highly efficient microfluidic Precise blending of up Reduces baseline noise enabling solvent mixer to 4 eluents higher sensitivity A small mixer volume will improve Magnified LPG step: system dead volume, but might Mixing depending on decrease mixing efficiency. mixer volume mAU –– 100 μl mixer –– 050 μl mixer –– without mixer Time www.knauer.net/azurap6 9 Sample injection n Manual Injection Valve Available valves 6-Port 8-Port Manual injection valves are the most cost-effective alternative to introduce samples. KNAUER valves for actuator 2-position 2-position feature a backpressure range of up to 1000 bar with a 0.3 mm bore size. for actuator multiposition multiposition All valves can be equipped with an automatic valve drive or can be integrated into an assistant module for manual injection 2-position 2-position housing AZURA ASM 2.1L Comparison manual vs. automatic injection 10 autosampler injections 10 manual injections 0.05 % RT RSD 0.1 % RT RSD mAU mAU Minutes Minutes 10 n Autosampler 3950 & Autosampler 3950 Basic The Autosampler 3950 is a high precision Autosampler • Up to 1000 bar (Basic: 700 bar) available for maximum backpressures of 1000 bar. This • Cooling/heating option (4 – 40 °C) autosampler can inject from up to 768 positions when equipped with microtiter plates or from up to 108 • 0.1 μl – 5000 μl sample injection volume standard 2 ml sample vials. This device is equipped with • Up to 768 samples (microtiter plates) or 108 an ILD™ valve, consisting of a rotor-stator combination standard vials with a central port for depressurizing the sample loop before receiving the sample. This way, the sample is not • Intermediate Loop Decompression – ILD™ diluted with solvent. The integrated intermediate loop decompression valve reduces sample dilution and increases measurement reproducibility Rack for 108 standard 2 ml vials www.knauer.net/azurainjection 11 Column tempering n Column Thermostat CT 2.1 The AZURA CT 2.1 is a forced air column thermostat capable of heating or cooling from 5 to 85 °C. The powerful fan and robust peltier element prevent a column from overheating while keeping it at a very stable temperature. • Temperature range 5 – 85 °C Heating/cooling rate 2 °C/min Temperature accuracy ± 0.2 °C Temperature stability ± 0.1 °C • Optional pre-column eluent tempering • Up to 6 columns with 350 mm max. length • Max. 16 mm inner diameter • Column switching option with ASM 2.1L • Operation via software or in stand-alone mode Using an HPLC column thermostat is highly recommended, since a stable temperature has an immense positive influence on retention time reproducibility Optional eluent pre-column tempering This device will converge solvent temperature with column temperature before entering the column and will therefore reduce temperature gradients within the column. Available with 0.1 or 0.25 mm inner diameter (5 µl / 30 µl inner volume). This device can be retrofitted to the oven chamber of the CT 2.1. 12 n Column Switching Assistant ASM 2.1L Using two valves in one ASM 2.1L to establish column switching with up to 6 columns Heating the column from outside only results in a temperature gradient in the column bed that can negatively influence separation results. Eluent pre-column Column oven tempering Heating the column and the eluent ensures a uniform temperature distribution, which is the closest-possible to ideal separation conditions. www.knauer.net/azuraanalytical 13 Detection How sensitive do you need to measure? We provide a choice of UV/VIS detectors, ranging from single variable wavelength to 8-channel diode array detector with LightGuide total reflection technology. Wavelength range 190 –1000 nm 190 –700 nm 190 –700 nm 190 –700 nm 190 – 500 nm AZURA detector UVD 2.1S UVD 2.1L MWD 2.1L DAD 2.1L DAD 6.1L Compact and Reliable UV/VIS detector Robust multi- Versatility High-end diode array versatile for a wide spectrum of channel UV/VIS through a wide detector with outstanding UV detector applications detector flow cell range performance Channels 1 1 8 8 8 3D data acquisition   Data acquisition rate 50 Hz 50 Hz 100 Hz 100 Hz 100 Hz Fiber optics available      LightGuide Flow Cells      PressureProof Flow Cells      The AZURA DAD 6.1L is equipped with revolutionary KNAUER Polka-Dot technology providing maximum light intensity and low noise over the whole UV/VIS spectrum. This technology provides more light than previous shine-through lamp configurations. A halogen lamp is needed to detect analytes with a maximum absorption in the VIS range. Intensity over wavelength comparison of Polka-Dot (blue) vs. Shine-through (grey) light mixing 14 Both AZURA diode array detectors DAD 6.1L and DAD 2.1L provide high spectral resolution enabling optimized identification and sample characterization. Spectral resolution comparison for pyrene –– AZURA DAD 6.1L Normalized Absorbance –– AZURA DAD 2.1L –– Vendor A 0210 230 250 270 290 310 330 350 Wavelength (nm) Accessible Sensitive frontal lamp and LightGuide flow cells and flow cell exchange Polka-Dot technology Flexible Robust wide flow cell range intelligent temperature control for high application for minimal drift versatility www.knauer.net/detectors 15 Detection Which flow cell suits your application best? The KNAUER PressureProof flow cells are available for all AZURA UV/VIS detectors in cartridge and classic design. These highly robust and price attractive flow cells feature an increased back pressure stability (up to 300 bar) and extended flow rates range (up to 20 ml/min). 400 Standard HPLC 200 mAU PressureProof 0 principle 2.0 4.0 6.0 8.0 Minutes HPLC separation performed with an analytical PressureProof flow cell (blue) vs. a Standard LightGuide flow cell (red) The Standard LightGuide Flow Cell Cartridge combines total reflection technology with small cell volume for high throughput and high resolution applications. 400 Fast HPLC LightGuide 200 mAU principle 0 0.5 1.0 1.5 Minutes UHPLC separation performed with an analytical PressureProof flow cell (blue) vs. standard LightGuide flow cell (red) 16 The High Sensitivity LightGuide Flow Cell Cartridge is available for the entire AZURA DAD series. With the lowest dispersion volume on the market and long path length this total refection flow cell is the ultimate choice for demanding trace analysis. 1300 1300 U mA Ultimately 1000 sensitive UHPLC U mA 0 0.75 Minutes 0.85 500 Rapid separation performed with standard LightGuide flow cell (blue) vs. high sensitivity 0 LightGuide (green) 0.5 1.0 1.5 Minutes Remote HPLC detection KNAUER fiber optic technology offers the possibility to separate the flow cell spatially from the detector via fiber optic cables in a distance up to 10 meters. This enables the measurement of UV/VIS absorption in flow-through mode remotely from the detector. Fiber optics enable the flow cell to be located very close to the column, mini-mizing peak broadening. DAD with remote nano flow cell www.knauer.net/detectors 17 Detection No chromophor? No problem. Universal detection Great performance The AZURA RID 2.1L is a highly competitive and sensitive refractive index detector, ideal for fast and reliable routine analysis of non-UV absorbing substances. The intelligent temperature control guarantees fast baseline stabilization and stable operation. Great stability Furthermore, the long-life LED, highly pressure resistant flow cell, improved safety features and enhanced diagnostics functions guarantee easy handling and minimal maintenance. Great versatility The wide linear dynamic range and 10 ml/min maximum flow rate make the AZURA RID 2.1L the perfect choice for most laboratory tasks. μRIU AZURA RID 2.1L (green) compared to previous model (blue) displays increased sensitivity and baseline stability min 18 A wide range of third-party detectors can be seamlessly integrated into AZURA analytical systems. Gradient compatible Sensitive universal and selective detection detection Using the unique Low Temperature technology, this The electrochemical detector DECADE Elite has been Evaporative Light Scattering detector 85 LT allows developed to meet the demands in ultra fast analysis, universal high sensitivity analysis. as to detection sensitivity, acquisition frequency and temperature specifications. Sensitive and selective fluorophor detection The fluorescence detector RF-20A provides world-class The KNAUER interface box IFU 2.1 allows highly sensitivity, excellent maintainability and diverse valida-precise analog acquisition of data and control of third tion / support functions. It supports a wide range of party modules over analog and relay outputs. applications from conventional to high-performance analysis. www.knauer.net/detectors 19 AZURA® Compact HPLC system Detection Injection Single wavelength detector Manual injection 190 – 500 nm valve Pumping Isocratic pump, including pressure sensor n AZURA® Compact HPLC system isocratic Complete small isocratic HPLC system The clearly defined system layout helps to readily understand the different functions of the HPLC system. • Degassing unit All important components are easy to access and maintain. The system is controlled by a laptop that also • Pump P 4.1S analyzes the measured data. • Detector UVD 2.1S Recommended software solutions: • Manual injector • Mobile Control • Eluent Tray • ClarityChrom® 20 Mobile Control AZURA® devices at your fingertips n AZURA® Mobile Control With the Mobile Control app you have your AZURA devices at your fingertips: Easily check the status or set parameters of several devices or several systems simul-taneously, even outside the laboratory. With the possibility of data acquisition, the Mobile Control is the perfect addition to the AZURA Compact Prep LC system. This combination represents a truly space-saving HPLC solution. No additional computer or software is needed. Control every AZURA system in your laboratory with one tablet. Mobile Control is safe: it ensures that device parameters See all parameters of can only be changed by authorized operators. your system on one The Mobile Control app can be run from a tablet or PC screen wherever you go. with Windows 8 or higher. Depending on the hardware control is possible via WLAN or Ethernet. • Control your devices or systems from your desktop • Easily switch between your AZURA systems and monitor all of them • Create different user accounts to securely manage your systems • Work effortless with the intuitive user-interface Two licences: • Optionally record data and control fraction • Mobile Control collection • Mobile Control Chrom (with data acquisition) www.knauer.net/mobilectr 21 HPLC of sugars, organic acids and alcohols: Polymer based phases Ideal for the organic solvent free H-form (USP L17) Analysis organic acids, sugars and alcohols analysis of: • Fruit beverages Ca-form (USP L19) • Soft drinks Pb-form (USP L34) Analysis of small carbohydrates (< DP4) • Wine • Beer Na-form (USP –) • Fermentation broths Ag-form (USP –) Analysis of sugar oligomers (up to DP8) KNAUER LC columns Analytical column dimensions: 8 4.6 4.0 3.0 Inner diameter / mm 2.0 HPLC of proteins and peptides: 5 30 50 100 Silica based phases Length / mm Your first choice for the analysis of: Wide pore C18 (USP L1) MW > 2000 Da • Proteins Wide pore C18A (USP L1) • Peptides • Oligonucleotides Wide pore C8 (USP L7) MW > 5000 Da Wide pore C4 (USP L26) MW > 20000 Da 22 HPLC/UHPLC of small molecules: Silica based phases The best choice for a wide range of application areas for small molecules (< 2000 Da) • Reversed phase mode for water- and water/organic solvent mix- soluble samples • Normal phase mode for water inso luble samples • HILIC mode for highly polar samples that are not retained by RP chromatography • Ion pair chromatography for acids and bases that are poorly retained in RP mode Silica (USP L3) NH2 (USP L8) Normal phase KNAUER LC columns Diol (USP L20) Cyano (USP L10) Analytical column dimensions: Silica (USP L3) HILIC NH2 (USP L8) Zwitterionic HILIC (USP –) Classical C18 (USP L1) Hydrophilic/aqueous C18 (USP L1) Hydrophobic/pH stable C18 (USP L1) C18 polar embedded group (USP L1) Classical C8 (USP L7) Hydrophilic/aqueous C8 (USP L7) 125 150 250 300 Length / mm Reversed phase Hydrophobic/pH stable C8 (USP L7) C8 polar embedded group (USP L7) C30 (USP L62) C1 (USP L13) C4 (USP L26) We offer replacements for CN (USP L10 nearly any HPLC column on the market. Phenyl (USP L11) www.knauer.net/columns 23 Solutions for your food analysis n Eurokat Key application High performance polymer phases Separation of alcohols, carbohydrates and organic acids Modifications • Eurokat H: separation of organic acids, sugars, alcohols and sugar alcohols • Eurokat Pb/Ca: Separation of carbohydrates 45 up to DP < 4 40 • Eurokat Na/Ag: Separation of sugar 35 oligomers and carbohydrates up to DP 8 30 DP = degree of polymerization 25 20 uRIU 15 10 5 Recommended Eurokat 0 column dimension for best -5 results: 300 x 8 mm ID ana- 0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40 Minutes lytical column, 30 x 8 mm ID precolumn. Device Key features n Sugar HPLC system Analytical isocratic configuration with UV and RI detection AZURA ASM 2.1L Isocratic analytical HPLC pump, 10 ml pump head including injection valve AZURA UVD 2.1S 190 – 500 nm, variable single wavelength UV detector AZURA RID 2.1L Temperature controlled, extended dynamic range AZURA CT 2.1 Column thermostat for constant temperatures and reproducible results 24 Solutions for your environmental or pharmaceutical analysis n Eurospher Key application & Eurospher II Determination of priority pollutant phenols High performance column material based on an ultra pure spherical silica gel 90 Highest flexibility 80 • Classical HPLC, Fast HPLC Plus, UHPLC 70 • 13 modification types 60 • 100 Å pore size 50 • 2 – 10 μm, up to 20 – 45 µm U mA 40 30 20 10 0 Find more key applications at -10 www.knauer.net/applications 0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 Minutes Device Key features n HPLC Plus system Analytical LPG configuration with DAD detection AZURA P 6.1L Quaternary analytical HPLC pump with degasser, 10 ml pump head AZURA DAD 2.1L 190 – 700 nm, diode array detector AZURA CT 2.1 Column thermostat for constant temperatures and reproducible results Autosampler 3950 Fast and very versatile analytical HPLC autosampler, 1000 bar, 0.1 – 5000 µl injection volume www.knauer.net/applications 25 Software Connect the KNAUER LC system to your chromatography data system (CDS) n ClarityChrom® n Chromeleon™ KNAUER ClarityChrom® is a powerful, yet easy-to-use Chromeleon is one of the most wide-spread chromato-chromatography software for instrument control, data graphy data systems. Its intuitive handling benefits acquisition and data processing. ClarityChrom is de-laboratory workflow and the highly developed algo-signed for smaller laboratories. It is an economical rithms simplify data processing. It offers a broad solution com pared to other more complex chromato-range of third-party drivers and can be easily used graphy software while still offering FDA 21 CFR Part 11 with existing HPLC systems. KNAUER offers drivers compliance. for a lot of its devices. It is as an analytical chromatography software solution: n OpenLAB® CDS • Complete LC and autosampler control package • Support for a wide range of instruments EZChrom Edition • Scalable from 1 to 4 LC systems (instruments), OpenLAB CDS EZChrom Edition is latest technology each with up to 12 detectors (channels) and successor of former ChromGate CDS. It provides • Optional extensions, e.g. for PDA data handling, chromatography data acquisition, processing and GPC analysis, SST for automated system tests and control of LC systems. It is scalable from single user/ GC control single instrument to multi-user/multi-instrument laboratories. Devices from KNAUER and many other ClarityChrom supports all KNAUER devices that can be manufacturers are supported. controlled by software. Please refer to the supported instruments list at www.knauer.net/cds-devices. More-over, devices and systems from more that 45 manufacturers can be controlled. Additionally, data acquisition Every software solution offers can also be performed with any detector providing a different functionality. Choose the voltage output by simply connecting a KNAUER IFU 2.1 right one that fits your needs. interface box or any other supported A/D converter. www.knauer.net/software 26 Your liquid chromatography specialists With profound application knowledge of analytical and preparative HPLC and FPLC, our team is at your service around the world. Our experts are pleased to receive your inquiries and requests, and will offer attractive customized solutions. Purification Academy Application development LC LAB Rent a lab Rent an expert Analysis on demand Research Method transfer and optimization Method transfer Rent a lab/ & optimization Rent an expert Transfer your methods to Rent lab space with a one of our HPLC systems suitable HPLC or an or make your analyses expert for assistance in faster, more efficient and your lab cost effective Academy Analysis on HPLC trainings and demand specia lized courses Fast and reliable custom- for beginners and tailored analytical HPLC experienced users service Application Purification development Complete solutions for your HPLC, FPLC, SMB Proof of concept and or continuous processes support in selecting a HPLC / FPLC method for your separation task www.knauer.net/lc-lab 27 AZURA UHPLC systems AZURA HPLC Plus systems AZURA UHPLC: AZURA HPLC Plus: advanced systems for the complete solution for highest demands and your daily HPLC routine most complex tasks up with a “Plus”-feature of to 1000 bar. 700 bar max. pressure. www.knauer.net/uhplc www.knauer.net/hplcplus AZURA Compact HPLC AZURA Educational system systems The AZURA Educational system is a complete HPLC AZURA Compact HPLC: system, specially designed the basic HPLC system with for the education of future ultimate flexibility on HPLC professionals. minimal space! www.knauer.net/azuracompact www.knauer.net/azuraedu Distributed by: AZURA Mobile Control With the AZURA Mobile Control app by KNAUER, you can control selected AZURA devices from your tablet. Technical data are subject to change without notice. www.knauer.net/mobilectrl All trademarks and registered trademarks are the property of their respective owners. www.knauer.net HPLC · SMB · Osmometry KNAUER Phone: +49 30 809727-0 Wissenschaftliche Geräte GmbH Telefax: +49 30 8015010 Hegauer Weg 38 E-Mail: info@knauer.net 14163 Berlin, Germany Internet: www.knauer.net © KNAUER 2016 V7852-2US/2.0/05.16/www.studio-w.net