HyperShear TM HPLC and UHPLC Mixers

Transcription

1 HyperShear TM HPLC and UHPLC s HyperShear TM HPLC and UHPLC Features: Micro Flow, Low Flow, Analytical Flow, High Flow and Combo Series Reduce baseline noise, increase sensitivity and improve gradient accuracy UHPLC mixers are available in volumes ranging from 0.5 µl to 1.5 ml and pressure to 15,000 PSI Available for In-line, and formats in (SS) or PEEK Ideal for microbore HPLC, UHPLC and LC/MS Compact design is easily integrated into any HPLC and UHPLC system Increased reaction efficiency for post column derivatization Decreased mixing and delay volume without sacrificing mixing efficiency Mixing volume optimization is easy with interchangeable mixer cartridges 1

2 HyperShear TM HPLC and UHPLC s ASI manufactures a wide range of Static s to solve the most demanding high pressure mixing problems. ASI HyperShear s incorporate a highly efficient cross-flow shearing mechanism which produces vortex shear mixing over a wide flow range. This mixing technology typically delivers between 25% to 200% better mixing efficiency compared to conventional packed bed or tortuous path mixers. HyperShear s are available in a variety of housing formats including: inline, binary, and ternary inlets. s are constructed from stainless steel or Biocompatible PEEK with volumes ranging from 0.5 µl to 1.5 ml. Within a given mixer flow series, mixer cartridges are interchangeable. The ability to swap mixer cartridges within a given mixer series makes selecting the optimum mixer volume easy and economical. This is not the case with UHPLC mixers. Since we warranty the pressure rating of 15,000 PSI on the factory tested mixer assembly (housing and cartridge), UHPLC mixers are only sold as complete mixer assemblies. Optimize mixing volume to match HPLC flow rate Decrease gradient delay time Use HyperShear s to Optimize mixing volume for UHPLC Decrease mixing noise Improve gradient profile Pressure Rating Standard Specifications HPLC UHPLC Biocompatible PEEK 6,000 PSI 15,000 PSI 3,000 PSI F emale Port Geometry 10/32 Parker(1/16 CPI) 10/32 Parker(1/16 CPI) 10/32 Parker(1/16 CPI) Wetted Materials and PEEK and PEEK PEEK Micro Flow Series 0.5, 1, 2, 5, 10 and 25 µl Low Flow Series 50, 150 and 250 µl Analytical Flow Series 350 and 500 µl High Flow Series 800 µl, 1.0 and 1.5 ml Combo Series 1 to 100 µl available available available available available available available available a vailable available () a vailable available () a vailable () a vailable () available () 2

3 Static s Static Application Notes Selecting the Right Size Cartridge ASI offers static mixers with volumes that range from 0.5 microliters to 1,500 microliters. Choosing the right size mixer is a trade off between delay volume, mixing noise, gradient fidelity and chromatographic performance. Please refer to the Charts on pages 64 and 65 for more information on gradient accuracy as a function of flow rate and mixing volume. The following observations will provide some guidelines to help choose the right size mixer. For any given flow rate, the more the mixing volume the better the mixing, and the lower the baseline noise. The smaller the mixing volume, the better the definition and sharpness of linear gradients. Multi-pump high pressure gradient systems typically require far less mixing volume than low pressure single pump gradient systems when running linear gradients. An ASI 150 µl in-line static mixer can be added in addition to the standard onboard mixer to further reduce mixing noise. Multi-pump High Pressure Gradient Systems Linear Gradients If a larger mixing volume can be tolerated for a particular flow rate, the larger the volume will lower the mixing noise. The upper limits to mixing volume will be the maximum delay time that can be tolerated, and possible distortion (tailing) of the gradient at the beginning and end of the gradient. The lower limit will be defined by the amount of mixing noise that can be tolerated. Please refer to a table, page 59 for specific recommendations. or Steady State Composition Always select the largest volume that will still provide an acceptable delay volume. In general, the more mixing volume, the better the mixing will be. For most pump systems, a 150 µl cartridge will provide adequate mixing. Examples of this type of pump system include: Shimadzu LC-10AD and LC-10ADvp Beckman System Gold Gilson Model 305 Agilent Model

4 Static s Single-pump Low Pressure Mixing Gradient Systems Linear Gradients These systems generally require more mixing volume to perform linear gradients than multipump high pressure systems. The following will explain why this is the case. In a low pressure system the composition can only be changed once every pump stroke. Since the pump stroke volume of most pumps is 100 µl, and it takes a mixer volume that is about 3 times the batch volume to provide adequate mixing, we need 350 µl of mixer volume, at least, to do adequate mixing. More insoluble combinations may require even more mixing volume. In general, choose the largest size mixer cartridge that will still provide an acceptable delay volume. For most applications this will be at least 350 µl. or Steady State Composition Always select the largest volume that will still provide an acceptable delay time. In general, the more mixing volume, the better the mixing will be. For most applications this will be at least 350 µl. Examples of this type of pump system include: Agilent 1090 Perkin Elmer series 200 TSP Spectra Vision Waters model 626 Varian Star 9000 Cartridge Selection Guide for High Pressure Mixing Linear Gradients - High Pressure Mixing Larger mixing volume can be tolerated for a particular flow rate, with the larger the volume the lower the mixing noise. The upper limits to mixing volume will be the maximum delay time that can be tolerated, and possible distortion (tailing) of the gradient at the beginning and end of the gradient. The lower limit will be defined by the amount of mixing noise that can be tolerated. The following cartridge volumes are a compromise between these two limits. Table 1. Flow 0-5 micro liter/min Cartridge Volum e 5 micro liter 4

5 Static s Static Comparison mvolts Less Noise - Lower Detection Limits No - Valco Tee & 10 µl Tubing mvolts Lee - 10 µl mvolts ASI HyperShear - 10 µl Figure 21 HPLC System Configuration: ASI Model 500G Gradient System ABI Model 785 on-column Varian Star Data Acquisition Minutes HPLC Conditions: MP: A = H2O, B = H2O/Acetone Flow Rate: 50 µl/min. 50% B Pressure: 1,700 PSI with ASI resistor cartridge mvolts Superior Mixing with Less Gradient Dispersion No - Valco Tee & 10 µl Tubing HPLC System Configuration: Above HPLC Conditions: Above Isocratic Flow Rate: 50 µl/min. 100% A One inlet port plugged Injection: 600 nl Acetone ASI HyperShear - 10 µl Figure 22 Lee - 10 µl Minutes 5

6 Static s Agilent Static Optimization mvolts -2.2 Micro Flow Step Gradients No ASI HyperShear - 50 µl Agilent - 80 µl -2.8 Figure HPLC System Configuration: Agilent 1100 Pump Packed bed mixer (approximate volume: 500 µl) replaced with union and/or ASI HyperShear Static Varian Star Data Acquisition Minutes HPLC Conditions: MP: A = H2O, B = ACN doped with 0.01% Acetone Flow Rate: 250 µl/min. Pressure: 2,000 PSI with ASI resistor cartridge Gradient: 0.0 = off%b 7.5 = 20%B 2.5 = 10%B 12.4 = 20%B 7.4 = 10%B 12.5 = 0.0%B and hold mvolts No or BP ASI HyperShear - 50 µl ASI HyperShear µl ASI HyperShear µl ASI HyperShear µl ASI HyperShear µl Agilent Packed Bed HPLC System Configuration: Above Agilent 1100 Pump Packed bed mixer (approximate volume: 500 µl) replaced with union and/or ASI HyperShear Static Varian Star Data Acquisition 15.0 Minutes HPLC Conditions: MP: A = H2O, B = IPA doped with 0.01% Acetone Flow Rate: 1.0 ml/min. Pressure: 2,000 PSI with ASI resistor cartridge Gradient: Above Figure

7 Static s Shimadzu Static Optimization Micro Flow Step Gradients Noise (mm) Shimadzu µl ASI HyperShear µl Composition (% B) Figure 25 mvolts Micro Flow Step Gradients 18 Shimadzu µl ASI HyperShear µl Minutes HPLC System Configuration: Shimadzu: LC-10ADvp : Connected at Shimadzu Tee Detector: Knauer HPLC Conditions: MP: A = H2O, B = H2O + 35% 2-Propanol % Acetone Flow Rate: 250 µl/min. Pressure: 2,000 PSI with ASI resistor cartridge Figure

8 Static s Shimadzu Static Optimization Analytical Flow Step Gradients Noise (mm) Noise (mm) ASI 250ul µl ASI 350ul µl Shimadzu 500ul Composition (%B) Composition (% B) Shimadzu 500 µl HPLC System Configuration: Shimadzu: LC-10ADvp : Connected at Shimadzu Tee Detector: Knauer HPLC Conditions: MP: A = H2O, B = H2O + 35% 2-Propanol % Acetone Flow Rate: 1.0 ml/min. Pressure: 2,000 PSI with ASI resistor cartridge Figure 27 mvolts mvolts 3.4 Isocratic Mixing Noise Shimadzu µl ASI HyperShear µl ASI HyperShear µl Minutes HPLC System Configuration: Shimadzu: LC-10ADvp : Connected at Shimadzu Tee Detector: Knauer HPLC Conditions: MP: A = H2O, B = H2O + 35% 2-Propanol % Acetone Flow Rate: 1.0 ml/min. Pressure: 2,000 PSI with ASI resistor cartridge Minutes Figure

9 Static s Effect of Volume on Gradient Accuracy with a Constant Flow Rate ASI 50 µl mixer, 25 µl/min. ASI 25 µl mixer, 25 µl/min. ASI 10 µl mixer, 25 µl/min. ASI 10 µl mixer, 25 µl/min. Figure 29 HPLC System configuration: Shimadzu LC-10ADVP pumps with micro-flow modification Shimadzu SPD-10AVP UV-VIS detector Shimadzu Class-VP v5.03 software HPLC conditions: Gradient: 10-90% B in 10 min. steps A = H2O B = 0.3% Acetone in H2O 9

10 Static s Effect of Flow Rate on Gradient Accuracy with a Constant Mixing Volume ASI 25 µl mixer, 10 µl/min. ASI 25 µl mixer, 25 µl/min. ASI 25 µl mixer, 50 µl/min. ASI 25 µl mixer, 100 µl/min. Figure 30 HPLC System configuration: Shimadzu LC-10ADVP pumps with micro-flow modification Shimadzu SPD-10AVP UV-VIS detector Shimadzu Class-VP v5.03 software HPLC conditions: Gradient: 10-90% B in 10 min. steps A = H2O B = 0.3% Acetone in H2O 10

11 Static s Micro Flow Series Complete Assemblies - Cartridge and Housing Assembly SS 0.5 µl Assembly SS 1 µl Assembly SS 2 µl Assembly SS 5 µl Assembly SS 10 µl Assembly SS 25 µl Assemblies Port Size: CPI ( Standard HPLC Fittings for 1/16 Tube) 0.5 µl, 1 µl, 2 µl, 5 µl, 10 µl or 25 µl Cartridge Flowlock Inlet/Outlet One Each End Figure

12 Static s Tee Complete Assemblies - Cartridge and Housing Tee Assembly SS 0.5 µl Tee Assembly SS 1 µl Tee Assembly SS 2 µl Tee Assembly SS 5 µl Tee Assembly SS 10 µl Tee Assembly SS 25 µl Tee Complete Assemblies - Cartridge and Housing ASI Part Number Tee Assembly SS 0.5 µl Tee Assembly SS 1 µl Tee Assembly SS 2 µl Tee Assembly SS 5 µl Tee Assembly SS 10 µl Tee Assembly SS 25 µl / Assemblies Port Size: CPI ( Standard HPLC Fittings for 1/16 Tube) Flowlock Inlet Ports (Two or Three) Figure µl, 1 µl, 2 µl, 5 µl, 10 µl or 25 µl Cartridge Flowlock Outlet Port Cartridges / Housings Cartridge SS 0.5 µl Cartridge SS 1 µl Cartridge SS 2 µl Cartridge SS 5 µl Cartridge SS 10 µl Cartridge SS 25 µl Micro Flow Series SS Housing Tee Micro Flow Series SS Housing Tee Micro Flow Series SS Housing

13 Static s UHPLC Complete Assemblies - Cartridge and Housing Assembly SS 0.5 µl, HP Assembly SS 1 µl, HP Assembly SS 2 µl, HP Assembly SS 5 µl, HP Assembly SS 10 µl, HP Assembly SS 25 µl, HP HP HP HP HP HP HP UHPLC Tee Complete Assemblies - Cartridge and Housing Tee Assembly SS 0.5 µl, HP Tee Assembly SS 1 µl, HP Tee Assembly SS 2 µl, HP Tee Assembly SS 5 µl, HP Tee Assembly SS 10 µl, HP Tee Assembly SS 25 µl, HP HP HP HP HP HP HP UHPLC Tee Complete Assemblies - Cartridge and Housing Tee Assembly SS 0.5 µl, HP Tee Assembly SS 1 µl, HP Tee Assembly SS 2 µl, HP Tee Assembly SS 5 µl, HP Tee Assembly SS 10 µl, HP Tee Assembly SS 25 µl, HP ASI Part Number HP HP HP HP HP HP 13

14 Static s Micro Flow Series Biocompatible PEEK Complete Assemblies - Cartridge and Housing Assembly PEEK 0.5 µl Assembly PEEK 1 µl Assembly PEEK 2 µl Assembly PEEK 5 µl Assembly PEEK 10 µl Assembly PEEK 25 µl B B B B B B Assemblies Port Size: CPI ( Standard HPLC Fittings for 1/16 Tube) ASI Flowlock Inlet/Outlet One Each End 0.5 µl, 1 µl, 2 µl, 5 µl, 10 µl or 25 µl Cartridge Figure

15 Static s Tee Complete Assemblies - Cartridge and Housing Tee Assembly PEEK 0.5 µl Tee Assembly PEEK 1 µl Tee Assembly PEEK 2 µl Tee Assembly PEEK 5 µl Tee Assembly PEEK 10 µl Tee Assembly PEEK 25 µl B B B B B B Tee Complete Assemblies - Cartridge and Housing Tee Assembly PEEK 0.5 µl Tee Assembly PEEK 1 µl Tee Assembly PEEK 2 µl Tee Assembly PEEK 5 µl Tee Assembly PEEK 10 µl Tee Assembly PEEK 25 µl ASI Part Number B B B B B B / Assemblies Port Size: CPI ( Standard HPLC Fittings for 1/16 Tube) Flowlock Inlet Ports (Two or Three) Flowlock Outlet Port ASI Figure µl, 1 µl, 2 µl, 5 µl, 10µL or 25µL Cartridge Cartridges / Housings Cartridge PEEK 0.5 µl Cartridge PEEK 1 µl Cartridge PEEK 2 µl Cartridge PEEK 5 µl Cartridge PEEK 10 µl Cartridge PEEK 25 µl Micro Flow Series PEEK Housing Tee Micro Flow Series PEEK Housing Tee Micro Flow Series PEEK Housing B B B B B B B B B 15

16 Static s Low Flow Series Complete Assemblies - Cartridge and Housing Assembly SS 50 µl Assembly SS 150 µl Assembly SS 250 µl Assemblies Port Size: CPI ( Standard HPLC Fittings for 1/16 Tube) 50 µl, 150 µl or 250 µl Cartridge Flowlock Inlet/Outlet One Each End Figure

17 Static s Tee Complete Assemblies - Cartridge and Housing Tee Assembly SS 50 µl Tee Assembly SS 150 µl Tee Assembly SS 250 µl Tee Complete Assemblies - Cartridge and Housing Tee Assembly SS 50 µl Tee Assembly SS 150 µl Tee Assembly SS 250 µl / Assemblies Port Size: CPI ( Standard HPLC Fittings for 1/16 Tube) 50 µl, 150 µl or 250 µl Cartridge Flowlock Outlet Port Figure 36 Cartridges / Housings Flowlock Inlet Ports (Two or Three) Cartridge SS 50 µl Cartridge SS 150 µl Cartridge SS 250 µl Low Flow Series SS Housing Tee Low Flow Series SS Housing Tee Low Flow Series SS Housing

18 UHPLC Complete Assemblies - Cartridge and Housing Static s Assembly SS 50 µl, HP Assembly SS 150 µl, HP Assembly SS 250 µl, HP HP HP HP UHPLC Tee Complete Assemblies - Cartridge and Housing Tee Assembly SS 50 µl, HP Tee Assembly SS 150 µl, HP Tee Assembly SS 250 µl, HP HP HP HP UHPLC Tee Complete Assemblies - Cartridge and Housing Tee Assembly SS 50 µl, HP Tee Assembly SS 150 µl, HP Tee Assembly SS 250 µl, HP HP HP HP 18

19 Static s Low Flow Series Biocompatible PEEK Complete Assemblies - Cartridge and Housing Assembly PEEK 50 µl Assembly PEEK 150 µl Assembly PEEK 250 µl B B B Assemblies Port Size: CPI ( Standard HPLC Fittings for 1/16 Tube) 50 µl, 150 µl or 250 µl Cartridge Flowlock Inlet/Outlet One Each End Figure

20 Static s Tee Complete Assemblies - Cartridge and Housing Tee Assembly PEEK 50 µl Tee Assembly PEEK 150 µl Tee Assembly PEEK 250 µl B B B Tee Complete Assemblies - Cartridge and Housing Tee Assembly PEEK 50 µl Tee Assembly PEEK 150 µl Tee Assembly PEEK 250 µl B B B / Assemblies Port Size: CPI ( Standard HPLC Fittings for 1/16 Tube) 50 µl, 150 µl or 250 µl Cartridge ASI Flowlock Outlet Port Figure 38 Flowlock Inlet Ports (Two or Three) Cartridges / Housings Cartridge PEEK 50 µl Cartridge PEEK 150 µl Cartridge PEEK 250 µl Low Flow Series PEEK Housing Tee Low Flow Series PEEK Housing Tee Low Flow Series PEEK Housing B B B B B B 20

21 Static s Analytical Flow Series, Biocompatible PEEK Complete Assemblies - Cartridge and Housing Assembly SS 350 µl Assembly SS 500 µl Biocompatible PEEK Assembly PEEK 350 µl Assembly PEEK 500 µl B B Assemblies Port Size: CPI ( Standard HPLC Fittings for 1/16 Tube) 350 µl or 500 µl Cartridge Flowlock Inlet/Outlet One Each End Figure

22 Complete Assemblies, - Cartridge and Housing Tee Assembly SS 350 µl Tee Assembly SS 500 µl Complete Assemblies, - Cartridge and Housing Tee Assembly SS 350 µl Tee Assembly SS 500 µl Static s / Assemblies Port Size: CPI ( Standard HPLC Fittings for 1/16 Tube) 350 µl or 500 µl Cartridge Flowlock Outlet Port Figure 40 Flowlock Inlet Ports (Two or Three) Cartridges / Housings Cartridge SS 350 µl Cartridge SS 500 µl Analytical Flow Series SS Housing Tee Analytical Flow Series SS Housing Tee Analytical Flow Series SS Housing Biocompatible PEEK Cartridge PEEK 350 µl Cartridge PEEK 500 µl Analytical Flow Series PEEK Housing B B B 22

23 Static s UHPLC Complete Assemblies - Cartridge and Housing Assembly SS 350 µl, HP Assembly SS 500 µl, HP HP HP UHPLC Complete Assemblies - Cartridge and Housing Tee Assembly SS 350 µl, HP Tee Assembly SS 500 µl, HP HP HP UHPLC Complete Assemblies - Cartridge and Housing Tee Assembly SS 350 µl, HP Tee Assembly SS 500 µl, HP HP HP 23

24 High Flow Series, Biocompatible PEEK Static s Complete Assemblies - Cartridge and Housing Assembly SS 800 µl Assembly SS 1.0 ml Assembly SS 1.5 ml Biocompatible PEEK Assembly PEEK 800 µl Assembly PEEK 1.0 ml Assembly PEEK 1.5 ml B B B Assemblies Port Size: CPI ( Standard 800uL, HPLC 1.0mL Fittings OR 1.5mL for 1/16 Tube) CARTRIDGE 800 µl, 1.0 ml or 1.5 ml Cartridge ASI 2.68 Flowlock Inlet/Outlet One Each End 1.43 Figure

25 Static s Complete Assemblies, - Cartridge and Housing Tee Assembly SS 800 µl Tee Assembly SS 1.0 ml Tee Assembly SS 1.5 ml Complete Assemblies, - Cartridge and Housing Tee Assembly SS 800 µl Tee Assembly SS 1.0 ml Tee Assembly SS 1.5 ml / Assemblies Port Size: CPI ( Standard 800uL, 1.0mL HPLC OR Fittings 1.5mLfor 1/16 Tube) CARTRIDGE 800 µl, 1.0 ml or 1.5 ml Cartridge ASI 2.60 Figure 42 Flowlock Inlet Ports (Two or Three) FLOWLOCK INPUT PORTS 1.43 Flowlock Outlet Port Cartridges / Housings Cartridge SS 800 µl Cartridge SS 1.0 ml Cartridge SS 1.5 ml High Flow Series SS Housing Tee High Flow Series SS Housing Tee High Flow Series SS Housing Biocompatible PEEK Cartridge PEEK 800 µl Cartridge PEEK 1.0 ml Cartridge PEEK 1.5 ml High Flow Series PEEK Housing B B B B 25

26 UHPLC Complete Assemblies - Cartridge and Housing Assembly SS 800 µl, HP Assembly SS 1.0 ml, HP Assembly SS 1.5 ml, HP HP HP HP Static s UHPLC Complete Assemblies - Cartridge and Housing Tee Assembly SS 800 µl, HP Tee Assembly SS 1.0 ml, HP Tee Assembly SS 1.5 ml, HP HP HP HP UHPLC Complete Assemblies - Cartridge and Housing Tee Assembly SS 800 µl, HP Tee Assembly SS 1.0 ml, HP Tee Assembly SS 1.5 ml, HP HP HP HP 26

27 Static s Combo Micro Series, Biocompatible PEEK Order Instruction You can choose any 2, 3 or 4 cartridge combination from 0.5 µl, 1 µl, 2 µl, 5 µl, 10 µl and 25 µl mixer cartridges to create the desired volume to fit your application. The minimum volume you can create will be 2 x 0.5 µl cartridge combination and the maximum volume will be 4 x 25 µl cartridge combination. You will be able to create almost the exact volume you need within that range. Example) For a 4 mixer assembly if you choose 1 x 5 µl, 1 x 10 µl and 2 x 25 µl combination, you will get 65 µl volume mixer. You can specify as below when you order: Complete Assembly SS: Complete Assembly PEEK: B Assemblies Port Size: CPI ( Standard HPLC Fittings for 1/16 Tube) Flowlock Inlet/Outlet One Each End Figure 43 4 s: X = s: X = s: X =

28 Static s Complete Assemblies - 2, 3 or 4 Cartridge Combination and Housing Combo Micro 2 Assembly SS Combo Micro 3 Assembly SS Combo Micro 4 Assembly SS 471-x-x 481-x-x-x 491-x-x-x-x UHPLC Biocompatible PEEK Combo Micro 2 Assembly SS, HP Combo Micro 3 Assembly SS, HP Combo Micro 4 Assembly SS, HP Combo Micro 2 Assembly PEEK Combo Micro 3 Assembly PEEK Combo Micro 4 Assembly PEEK 471-x-xHP 481-x-x-xHP 491-x-x-x-xHP 471-x-xB 481-x-x-xB 491-x-x-x-xB The application and Use of ASI Combo Micro Static s Chromatographers seeking to optimize mixing volumes at flow rates below 100 µl/min. now have the ability to customize a mixing volume which is perfectly tuned to match the separation system. Choosing the right mixing volume is always a compromise between absolute noise reduction, delay volume, and gradient distortion. Ideally the optimum mixing volume should be determined experimentally by evaluating several mixing volumes to determine which provides optimum chromatographic performance. For the separation below at 50 µl/min., a 50 µl mixer contributes too much delay volume and gradient distortion. The ASI Combo 35 µl (25 µl + 10 µl) mixer provides just the right balance between noise reduction and gradient performance. mvolts 5 Combo Micro Static Performance at 50µL/min. 4 Combo Micro 35 µl 3 2 ASI Standard In-line 50 µl HPLC System Configuration: ASI Model 500G Micro Head Detection: UV214 Varian Star Data Acquisition HPLC Conditions: MP: A = H2O, B = H2O doped with 0.05% red dye Flow Rate: 50 µl/min. Gradient 0 to 100% B in 10% Steps -2 Figure Minutes Minutes 28