Electrochemistry upfront MS

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ROXY™ - EC/LC upfront MS

With ReactorCell™ forGenerating Redox metabolitesCleavage of proteins/peptidesSignal enhancement in MSOxidative stressMetabolism

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Why Electrochemistry / MS?

Mimics Natures Redox Reactions

In Drug Development and Proteomics

Fast & CleanComplementary to existing methods

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Why Electrochemistry / MS?

Mimics Natures Redox Reactions

Drug developmentImmediate results, minutes vs weeks

No by products, no cleanupNo biohazardNo rodents

ProteomicsNo enzymes, clean without by products

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Electrochemistry / MS

Mimicking nature’s redox reactionsFast alternative for laborious, time consuming procedures

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Electrochemistry / MS

Mimicking nature’s redox reactionsFast alternative for laborious, time consuming procedures

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MSEC

Modes of operation

On-line EC/MS

On-line EC/LC/MS

- Pre injection

- Post column

Off-line EC/MS

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Electrochemical Reaction

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ΔE

Facilitates the EC reaction

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Role of electrode potential E

0

20

40

60

80

100

0 400 800 1200 1600

E (mV)

Sig

nal

(%

)

no reaction

formation of oxidation or reduction products

A B

Facilitates the EC reaction

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Role of electrode potential E

0

20

40

60

80

100

0 400 800 1200 1600

E (mV)

Sig

nal

(%

)

2000

formation of secondary products

A B C

B C

B: instable intermediate

How to Find Optimum ΔE ?

Traditional Electrochemistry: VoltammogramOn-line EC/ESI-MS: “MS Voltammogram”

180 200 220 240 260 280 300 320 340 360 380 4000

1000

2000

3000

4000

5000

1500

2500

2000

inte

nsity

[-]

m/z

E [mV]

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oxidative degradation of the substrate

oxidation products

oxidation products

Typical Reactions Catalyzed by Cytochrome P450 Simulated by EC

Allylic and aliphatic hydroxylation

Possible at high potentials

(only with ROXY)

Benzylic hydroxylation

Possible

Desalkylation of amines Possible

Desalkylation of ethers and thioethers

Ethers: Possible

Tioethers: not mimicked

Hydroxylation of aromatics

Possible

Epoxide formation Possible

Oxidation of heteroatoms (N, S)

Possible

Adapted from W. Lohmann et al. LC/GC Jan. 2010

R-CH2-CH3 R-CH-CH3

OH

RR R

OH

R

OH

N-CH2-R NH + R-CHO

R-O-CH2-R R-OH + RCHO

RRR

OH

R

OH

O

C=CR

C CR

C CR

HO OH

R C NH R R C+ N RO-

RR

H

Proteomics

Electrochemical cleavage (oxidation) of proteins/peptides

‘enzymatic digestion’ by electro-chemical cell

no enzymes, no non-specific cleavage, no auto-digestion, etc.

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Electrochemistry in Proteomics

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Electrochemistry in Proteomics

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Electrochemistry in Proteomics

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Conclusion EC/MS

Many application areasTime and cost savings...Fast and clean

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Conclusion EC/MS

Mimics Natures Redox Reactions

Drug developmentImmediate results, minutes vs weeks

No by products, no cleanupNo biohazardNo rodents

ProteomicsNo enzymes, clean without by products

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Thank you!

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