1 one-minute nmr adding integrated data processing to open-access, high-throughput nmr automation...

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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?


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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


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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


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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


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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


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Click…

Minutes Later…In your InBox

Hot-linked E-mail reportIntegrated ACD/Web Librarian


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Adaptive ExperimentsStructure Confirmation, Elucidation Coming…

1D Proton

>= 1D Proton Acceptance Level

HSQC

ConfirmedRejected

>= AVG(HSQC + 1H1D) Acceptance Level

YESNO


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LEAP PAL

• Fast• Low Volume Pickup• Low Dead Volume• Syringe-Only

Technology• Flexible Sample

Storage • Dilute /

Reconstitute• Sample Return


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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


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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


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SO

LV

. A W

AS

TEINJECT

V2

LOOP

SAMPLES

NE

ED

LE

SY

RIN

GE

High Pressure PUMP


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


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.)

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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


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Help spread the word!

www.protasis.com508-481-4163

Now NMR is easier and faster

One-Minute NMR

http://www.protasis.com/

1 one-minute nmr adding integrated data processing to open-access, high-throughput nmr automation...

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sample lists

quantitation eretic

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