Agilent Technologies, Inc. · NYSE:A
Agilent Technologies — Technical Presentation (October 2016)
48 slides from Agilent Technologies's technical presentation, Oct 24, 2016
Slide 1: Making LC Connections
Making LC Connections Mark Powell 24 October 2016 Confidentiality Label October 24, 2016 1
Slide 2: LC Connections
LC Connections • Problems with improper connections • Source of leaks • Mistaken for chromatography issues • Making connections can vary with technique • Different manufacturers supply different types of fittings • Capillary tubing choices • Contribute to dispersion and extra-column volume Confident
Slide 3: Dispersion Reduces HPLC Performance
Dispersion Reduces HPLC Performance What is dispersion? • Original sample concentration being diluted as it is carried through the system plumbing (extra-column volume) What increases dispersion? • Connecting tubing that is too long • Connecting tubing that is too large in diameter • Connections tha
Slide 4: Extra-Column Volume
Extra-Column Volume • Flow cell • Heat exchanger • Switching valve • Needle seat • Connecting capillaries Confidentiality Label October 24, 2016 4
Slide 5: Extra-Column Volume
Extra-Column Volume 1 1 10 µL extra-column volume 50 µL extra-column volume 3 3 2 2 4 4 0.0 0.5 1.0 1.5 2.0 0.0 0.5 1.0 1.5 2.0 Time (min) Time (min) Column: StableBond SB-C18, 4.6 x 30 mm, 3.5 mm Mobile Phase: 85% H2O with 0.1% TFA : 15% ACN Flow Rate: 1.0 mL/min Temperature: 35°C Sample: 1. Phenyl
Slide 6: Aris-Taylor Equation
Aris-Taylor Equation peak dispersion in cylindrical tubing Confidentiality Label October 24, 2016 6
Slide 7: Tubing Volume
Tubing Volume Tubing Length 10mm 50mm 100mm 150mm Tubing ID Volume Volume Volume Volume 0.17mm (green) 0.227 uL 1.1uL 2.27 uL 3.3 uL 0.12mm (red) 0.113 uL 0.55uL 1.13 uL 1.65 uL
Slide 8: As Efficiency Increases Peak Width Decreases
As Efficiency Increases Peak Width Decreases System Dispersion Becomes More of a Factor Columns: Eclipse Plus C18, as described below. Mobile Phase: A: water, B: MeOH, (15:85) Injection volume: 6uL Temperature: 25°C Flow: 1 mL/min. Detection: 310, 4 nm, 0.5 s response time, semi-micro flow cell, Sam
Slide 9: Approximate Extra Column Volume Limits for This 4.6mm i.d. Column Example
Approximate Extra Column Volume Limits for This 4.6mm i.d. Column Example 5um 3.5um 1.8um Peak (1) Width in uL 181 136 45 Injection (1/10 P.V.) 18 14 5 Extra Column (1/3 P.V.) 60 45 15 ECV minus Inj. Vol. 42 21 10 ECV Estimate for 8 4 2 2.1mm i.d. Column
Slide 10: Optimizing Connecting Tubing Volume For UHPLC Columns
Optimizing Connecting Tubing Volume For UHPLC Columns Resolution 0.961 mAU System Tubing Volume Not Optimized 400 0.17mm i.d. tubing Peak width 0.037 min 300 200 Peak width 0.038 min 100 0 0.5 1 1.5 2 2.5 min mAU Resolution 1.902 600 System Tubing Volume Optimized 550 0.12mm i.d. tubing Peak width 0
Slide 11: Stainless Steel vs. Polymeric Fittings
Stainless Steel vs. Polymeric Fittings Stainless Steel: • Agilent uses Swagelok type fittings with front and back ferrules • Also available with a long nut PEEK (<400 bar system pressure): • Connections are changed frequently • Connecting columns • Pressure is less critical • Fits on SS or PEEK tubi
Slide 12: Type of Fittings
Type of Fittings Swagelok • Two piece ferrule • Used on Agilent LCs • Short nut 0.090 in. • Also available with long nut Parker • One piece ferrule • Short nut 0.090 in. • Very similar to Swagelok Waters • Longer nut • Used on Alliance systems 0.130 in. Confidentiality Label October 24, 2016 12
Slide 13: Types of Fittings
Types of Fittings Confidentiality Label October 24, 2016 13
Slide 14: Potential Fittings Issues
Potential Fittings Issues • Leak • Peak shape problem • No dead volume Confidentiality Label 10/24/2016
Slide 15: Fitting Mismatch
Fitting Mismatch Confidentiality Label 10/24/2016
Slide 16: Proper Fit
Proper Fit Confidentiality Label 10/24/2016
Slide 17: Fitting Mismatch
Fitting Mismatch Confidentiality Label 10/24/2016
Slide 18: Proper Fit
Proper Fit Confidentiality Label 10/24/2016
Slide 19: Polymeric Fittings
Polymeric Fittings Confidentiality Label 10/24/2016
Slide 20: Peak Shape
Peak Shape Confidentiality Label 10/24/2016
Slide 21: Peak Shape
Peak Shape DAD1 A, Sig=254,4 Ref=360,100 (MERCURY20... DATA; MERCURY2010 ECLIPSE PLUS 3X100,1.8\USUYB0145300037.D) mAU 40 30 20 10 0 0 2 4 6 8 10 min DAD1 A, Sig=254,4 Ref=360,100 (MERCURY20...AND PRESS LOAD ECLIPSE PLUS C18 3X100, 1.8\USUYB0145500037.D) mAU 40 30 20 10 0 0 2 4 6 8 10 min Confidenti
Slide 22: Swaging Fittings
Swaging Fittings Confidentiality Label October 24, 2016 22
Slide 23: Swaging Fittings
Swaging Fittings Confidentiality Label October 24, 2016 23
Slide 24: Swaging Fittings
Swaging Fittings Confidentiality Label October 24, 2016 24
Slide 25: Swaging Fittings
Swaging Fittings Confidentiality Label October 24, 2016 25
Slide 26: Inspect the Position of the Ferrule
Inspect the Position of the Ferrule Confidentiality Label October 24, 2016 26
Slide 27: Tools
Tools Confidentiality Label October 24, 2016 27
Slide 28: Tightening Fittings into a Column
Tightening Fittings into a Column Confidentiality Label October 24, 2016 28
Slide 29: Overtightened Fittings
Overtightened Fittings Confidentiality Label October 24, 2016 29
Slide 30: Overtightened Fittings
Overtightened Fittings Confidentiality Label October 24, 2016 30
Slide 31: A-Line Fitting: Unique Spring-loaded Design
A-Line Fitting: Unique Spring-loaded Design • The unique spring-loaded design applies a constant force to eliminate dead volume • Fingertight to 1300 bar Confidentiality Label October 24, 2016 31
Slide 32: A-Line Quick Connect Fitting
A-Line Quick Connect Fitting Connection problems can lead to: • Downtime • High cost of operation • Poor chromatography results • Broad or tailing peaks • Loss of resolution A-Line Quick Connect Fitting • Expensive maintenance cost • Overtightening • Column damage • Leaks, added troubleshooting A-Li
Slide 33: A-Line Fittings Last
A-Line Fittings Last Chromatogram overlap before and after 200 reconnections • No visible change of chromatogram after 200 reconnections. Confidentiality Label October 24, 2016 33
Slide 34: A-Line Fittings Last
A-Line Fittings Last • Tailing factors and theoretical plates stayed constant within the experimental allowance through the reconnection procedures. Confidentiality Label October 24, 2016 34
Slide 35: UHPLC Fingertight Fitting from Another Vendor
UHPLC Fingertight Fitting from Another Vendor Confidentiality Label October 24, 2016 35
Slide 36: Using the Quick Connect fitting
Using the Quick Connect fitting Finger tighten the fitting until you feel resistance, then close the lever. Leak tight to 1300 bar Confidentiality Label October 24, 2016 36
Slide 37: Using the Quick Turn fitting
Using the Quick Turn fitting Finger tighten for 600 bar Wrench tighten for 1300 bar Confidentiality Label October 24, 2016 37
Slide 38: Quick Connect/Turn Fittings Use Specific Capillaries
Quick Connect/Turn Fittings Use Specific Capillaries A wide variety of capillary lengths and inner diameters are available to meet HPLC and UHPLC needs Confidentiality Label October 24, 2016 38
Slide 39: Zero Dead Volume Connections
Zero Dead Volume Connections Confidentiality Label October 24, 2016 39
Slide 40: Not Just for Agilent Columns
Not Just for Agilent Columns Confidentiality Label October 24, 2016 40
Slide 41: A-Line Fittings Tips
A-Line FittingsTips A ZDV union can be used to adapt to heat exchangers but…. Confidentiality Label October 24, 2016 41
Slide 42: A-Line Fittings in Action
A-Line Fittings in Action Low Dispersion Heat Exchanger G1316-80022 Works great with the latest low dispersion heat exchanger for G1315B/C TCC Confidentiality Label October 24, 2016 42
Slide 43: A-Line Fittings in Action
A-Line Fittings in Action Confidentiality Label October 24, 2016 43
Slide 44: Replacing Capillaries and Ferrules
Replacing Capillaries and Ferrules If the capillary is damaged, or another dimension is needed, just use a plier to remove the capillary and insert a new one. The ferrules can also be replaced; just use pliers to remove the ferrule and insert a new one. Confidentiality Label October 24, 2016 44
Slide 45: Quick Connect
Quick Connect A-Line Quick Connect Assemblies 5067-5957 A-Line Quick Connect Assy ST 0.12x105mm 5067-5958 A-Line Quick Connect Assy ST 0.12x150mm 5067-5959 A-Line Quick Connect Assy ST 0.12x220mm 5067-5960 A-Line Quick Connect Assy ST 0.12x280mm 5067-5961 A-Line Quick Connect Assy ST 0.075x105mm 506
Slide 46: Quick Turn
Quick Turn Capillaries for Quick Turn fitting 5500-1188 Capillary ST 0.12mm x 105mm long socket 5500-1189 Capillary ST 0.12mm x 150mm long socket 5500-1190 Capillary ST 0.12mm x 200mm long socket 5500-1191 Capillary ST 0.12mm x 280mm long socket 5500-1192 Capillary ST 0.12mm x 500mm long socket 5500
Slide 47: A-Line Fittings
A-Line Fittings Agilent A-Line Quick Connect fittings Getting a perfect column connection, every time, A-Line Quick Turn Fitting A-Line Quick Connect Fitting by any operator. No tools - Truly finger tight up to 1300 bar. Use a Quick Connect fitting for the column inlet and a Quick Turn fitting for t
Slide 48: Who can use the Quick Connect Fitting?
Who can use the Quick Connect Fitting? • Any lab with a range of users, including newer chromatographers who may not have the experience to do traditional fittings well • Any lab that is changing columns often • Labs who are seeing variation in chromatographic results – broad or tailing peaks, and a
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