1 one-minute nmr adding integrated data processing to open-access, high-throughput nmr automation...
TRANSCRIPT
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One-Minute NMRAdding Integrated Data Processing to Open-Access, High-Throughput NMR Automation
• Easy• Fast• Efficient• Economical• Controllable• Flexible
Find the answers to…
“What is this stuff?How much is there?”
Copyright © 2006 by Protasis Corporation, All Rights Reserved
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What is it?
Everyone wants a magic wand!
Leap/CTC PALLiquid Handler
CapNMR ProbeNMR Spectrometer
ACD/Automation Server
E-Mail OrWeb Site
You Just Log In Samples
One-Minute NMRManages The Details
ACD/Web Librarian
Did I make it?
Yield?
Copyright © 2006 by Protasis Corporation, All Rights Reserved
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The One-Minute NMR System
• Optimized Liquid Handling• Application-Tuned CapNMR Probe• Exclusive Spectrometer Interfaces• Robust Software• Collaboration With Partners and End-
Users
Copyright © 2006 by Protasis Corporation, All Rights Reserved
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MRM CapNMRTM Microcoil Probes
• 5-10 microliter flow cell = 8 – 15 microliter samples• Best mass sensitivity• Short acquisition times• Concentration > 1mM works great
(10 nanomoles in 10µL)
UseCapNM
R
Copyright © 2006 by Protasis Corporation, All Rights Reserved
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One-Minute NMR Provides…
• Queue Management– Experiments, Structures, Fraction List
Import• Sample Loading
– Vials and Plates, 5 & 10 microliter• NMR Detection
– Super-sensitive CapNMR Probe• NMR Control
– Adaptive Experiments• Confirmation
– ACD/Labs 1D + 2D Structure Verification• Bonus: Quantitation
– ERETIC
Copyright © 2006 by Protasis Corporation, All Rights Reserved
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One-Minute NMR Software
• Secure Log-In• Individual sample
entry and Molecule upload
• .ZIP/.CSV Sample lists and molecule upload
• Bar-code support• Open SQL Server• Custom Fields• LDAP Login
Web-Based Open Access Sample Queue Management
Copyright © 2006 by Protasis Corporation, All Rights Reserved
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Click…
Minutes Later…In your InBox
Hot-linked E-mail reportIntegrated ACD/Web Librarian
Copyright © 2006 by Protasis Corporation, All Rights Reserved
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Adaptive ExperimentsStructure Confirmation, Elucidation Coming…
1D Proton
>= 1D Proton Acceptance Level
HSQC
ConfirmedRejected
>= AVG(HSQC + 1H1D) Acceptance Level
YESNO
Copyright © 2006 by Protasis Corporation, All Rights Reserved
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LEAP PAL
• Fast• Low Volume Pickup• Low Dead Volume• Syringe-Only
Technology• Flexible Sample
Storage • Dilute /
Reconstitute• Sample Return
Copyright © 2006 by Protasis Corporation, All Rights Reserved
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The advantages of going tubeless…
•No shimming between samples
•No washing tubes
•Minimal sample transfer
•Efficient storage
•Disposable
•Economical
Incredible Time Savings
Copyright © 2006 by Protasis Corporation, All Rights Reserved
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Safe Solvent Management
• Vacuum Degasser– Smooth
MicroFlows
• Helium Sparging / Blanketing– Protect
Expensive Deuterated SolventsH
elium
NE
ED
LE
V3
SAMPLES
SY
RIN
GE
The Deuterated Solvent Conditioning Module mounts conveniently in the door of the Discovery Tower
Vacuum
Copyright © 2006 by Protasis Corporation, All Rights Reserved
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SO
LV
. A W
AS
TEINJECT
V2
LOOP
SAMPLES
NE
ED
LE
SY
RIN
GE
High Pressure PUMP
Copyright © 2006 by Protasis Corporation, All Rights Reserved
Ca
pN
MR
PR
OB
E
High Throughput Sample Loading
Sample Return Option
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• Varian– Sun TCP/IP
Sockets
• Bruker– Windows Drive
Mapping– SGI / Linux TCP/IP
Sockets
• OthersEasy instrument
integration
Spectrometer Interfaces
Copyright © 2006 by Protasis Corporation, All Rights Reserved
Router
RSH(a.shimFile)RPAR(a.experiment, "all")
SETCURDATAXCMD("gpro")
RGASTOREPAR("NS", a.scans)
STOREPAR("SW", a.sweepWidth)STOREPAR("o1p",
a.centerOfSpectrum)exptime = CalcExpTime();
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Challenges of Isolation and Elucidation
• Costly Sample Collection• Challenging Sample Matrix• Unknown and Complex Chemical
Structures• Quantification of Unknowns
Difficult• Purification-NMR Mismatches• Complex NMR Spectra
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Purification-NMR Mismatches
• Temporal– HPLC fast, NMR slow
• Concentration– HPLC Fractions too dilute for
good NMR• Solvent
– Protonated– Gradients– Salts
H2O
ACN
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The Smart Solution ToPurification-NMR Mismatches
Collect Fractions
Put Dry Samples OnOne-Minute NMR
•Large Capacity•Time-Optimized HPLC•No Loop Monster•No SPE Concentrator •Concentrated Sample
•No Protonated Solvent•Some salt OK
•Highest Mass S/N•Automatic Dissolution•No Solvent Supression•No Shimming
Evaporate HPLCSolvent
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Analytical Scale Purifications
Right-sized for CapNMR
Scrape off a spot into filter. Add DMSO-d6, spin.
Collect fractions at analytical scale HPLC
Automatic Dissolution with DMSO-d6
A
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Actual Use Case
Academic LaboratoryPrecious Sample AnalysisNatural Product De-replication
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Precious Sample AnalysisNatural Products Dereplication
HPLC700 µg
~ 5 µg/well
Courtesy John Blunt, U. Canterbury, and Kirk Gustafson, NCI(see March 2006 Gordon Conf. On Natural Products Presentations at
www.microNMR.com)
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Precious Sample AnalysisWell plate/Vial – centric Approach
Dionex HPLC
Foxy Jr
DADELSD
Other OptionalAnalytical Detectors
(e.g. Mass Spec.)
1
2
3
4
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Step 1: Chromatography
Dionex HPLC
Foxy Jr
DADELSD
Dionex HPLC ISCO Foxy Jr. Fraction Collector 700 ug crude extract from culture of
fungus Cladobotryum sp. C18 column with d-ACN/D20
gradient with flow of 1mL/min Eluant collected in 96-well conical
bottom polypropylene plate Receiving wells changed every 15
seconds (250uL/well)
Courtesy John Blunt, U. Canterbury, and Kirk Gustafson, NCI(see March 2006 Gordon Conf. On Natural Products Presentations at
www.microNMR.com)
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Step 2: Dry Down
Evaporation using Jouan RC10-22 centrifugal vacuum evaporator
Plate enclosed with a piercible silicone/Teflon cover
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Step 3a: Sample Reconstitution
Wells chosen to match peaks on ELSD data
Reconstitution with 16uL of methanol-d4 in each well of interest, followed by mixing step prior to sample injection
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Step 3b: NMR Data Acquisition
600MHz NMR ICG 10uL
CapNMR Probe 15uL Sample
Loop
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One-Minute NMR™Open Access, High Throughput NMR Platform
• Secure Open-Access Web Interface• Integrated LEAP/CTC Liquid Handler
– Multiple vial/plate sample formats• Safe Solvent Management
– Degassing, Sparging, Blanketing– Solvent Selection
• High-Throughput Sample Loading and Analysis
– 1-minute loading, 1-scan 1D proton• Closed-Loop Spectrometer Control
– Run -> Idle ->Run– Error handling
• Automated Structure Confirmation– Integrated 1D+2D ACD/Labs– Automated/adaptive experimental design
• E-Mail reporting– HTML Plate Maps– Error Alerts– Forgotten Passwords
Exclusive AttributesProductivity Features• Priority Samples• Night Samples• Probe Clean• Solvent Select• Sample Return• Sample List Importing• Structure Confirmation• Custom Counters• Custom Fields -> NMR Dataset
Parameters• SQL Database• Pressure Monitoring• Macros• Dilution• Reverse Rinse• Automatic instrument reboot
Copyright © 2006 by Protasis Corporation, All Rights Reserved
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Help spread the word!
www.protasis.com508-481-4163
Now NMR is easier and faster
One-Minute NMR