Chiral Recognition detected by

Mass Spectrometry
CHEN Ping
2013.12.06
1

Outline
I. Introduction

II. Hyphenated Mass Spectrometric Techniques for

III. Mass Spectrometric Chiral Recognition Mechan

1. Host-Guest (H-G) Associations
2. Guest Exchange Ion-Molecule Reactions
3. Chiral Recognition Based on Complex Dissociation

IV. Application of Chiral Recognition

Organocatalytic Asymmetric Conjungate Addition of Aldehydes to Ni

V. Summary

Introduction
More than half of the currently approved drugs are
chiral molecules.
Develop single enantiomer drugs
Reducing the required dose
Increasing the potency
Improving the safety profile

Asymmetric synthesis
(Catalysts screening)

7 of the 10 best-selling US
pharmaceutical products are single
enantiomer.

Mass Spectrometry
Chiral Recognition
Chiral analysis
(Quality control)
Ranking of the top 10 best-selling US pharmaceutical products in 2011 was obtained
from webpage: http://www.imshealth.com/.

Introduction

Traditionally, MS has been considered a chiral-blind tech

Enantiomers: Same mass and show identical mass
spectra
Two strategies to differentiate a pair of
enantiomers with MS
1. Coupling of chiral sensitive analytical tools
with MS
Liquid Chromatography-Mass Spectrometry (LC-MS)
Gas Chromatography-Mass Spectrometry (GC-MS)
2.
MS is used solely in chiral analysis based on
different methods of chiral recognition
Host-Guest (H-G) Associations
Guest Exchange Ion-Molecule Reactions
Chiral Recognition Based on Complex Dissociation
H. Awad, A. EI-Aneed, Mass Spectrom Rev, 2013, 32, 466483

Hyphenated MS techniques
Coupling of chiral sensitive analytical tools with MS
Liquid Chromatography-Mass Spectrometry (LC-MS)
Gas Chromatography-Mass Spectrometry (GC-MS)
Capillary Electrophoresis-Mass Spectrometry (CE-MS)
Capillary Electrochromatography-Mass Spectrometry
(CEC-MS)
Supercritical Fluid Chromatography-Mass Spectrometry

New detector:
Mass Spectrometer (MS)
(SFC-MS)

Advantages:

Limitations:

Nonpolar solvents were incompatible with

Sensitive
Accurate
ESI or APCI
Salts and other nonvolatile compounds in
Speed
High throughputthe mobile phase were incompatible with
5
ESI
H. Awad, A. EI-Aneed, MassSpectrom
Rev, 2013,
32, stationary
466483
Choosing
chiral
phase is a

Hyphenated MS techniques

Two options for chiral analysis using hyphenated MS

Indirect approach:

Direct approach:

Analysis of covalent

Analysis of noncovalent

diastereomeric

diastereomeric

complexes

complexes

Separated by
conventional methods

Derivatized
by the CS to
form
covalent
complexes

Direct approach is preferred

Indirect
approach:
Need more time
for the reaction
step

Form
transient
bond with
CS

CS: chiral derivatization reagent

CS: chiral mobile phase additives
H. Awad, A. EI-Aneed, Mass Spectrom Rev, 2013,(CMPAs)
32, 466483

Hyphenated MS techniques

The HPLC-MS chromatograms of (S ,R) ifosfamide

(IF)

R. V. Oliveira, et al, J. Pharm. Biomed. Anal. 2007, 45,

Chiral Recognition Mechanisms

Two strategies to differentiate a pair of
enantiomers with MS
1. Coupling of chiral sensitive analytical tools
with MS
Liquid Chromatography-Mass Spectrometry (LC-MS)
Gas Chromatography-Mass Spectrometry (GC-MS)

2.
MS is used solely in chiral analysis based on
different methods of chiral recognition
Host-Guest (H-G) Associations
Guest Exchange Ion-Molecule Reactions
Chiral Recognition Based on Complex Dissociation

Chiral Recognition Mechanisms

1. Host-Guest (H-G)
Associations
Ion abundance ratio:
The affinity of each
enantiomer towards the
CS

CS (host)

One of the two enantiomers

(guest) tagged with deuterium
atoms

J. Kim, et al, Bull. Korean. Chem. Soc. 2008, 29, 1069-1072.

Chiral Recognition Mechanisms

2. Guest Exchange Ion-Molecule Reactions
Unlabeled analyte
enantiomers (guest)
react with the CS (host)
forming identical
diastereomeric
complexes
Principle:
Depends on the
different exchange
behavior of
R
enantiomers
with a
foreign reagent

Cant be separated in a single stage MS

Different intensity ratio

The complex ions are mass selected and

allowed to react with a neutral gas-phase
reagent
R

J. Ramirez, et al, J. Am. Chem. Soc. 1998, 120,

73877388.

10

Chiral Recognition Mechanisms

2. Guest Exchange Ion-Molecule Reactions

Relative abundances based on two factors:

the enantiomeric ratio of the used chiral analyte
the time of the exchange reaction

Solely varying the enantiomeric ratios of the

chiral
J. Ramirez,
et al,analytes
J. Am. Chem. Soc. 1998, 120,
73877388.

11

Chiral Recognition Mechanisms

3. Chiral Recognition Based on Complex
Dissociation

The chiral analyte and chiral reference compound (ref*) are

complexed with a transition-metal ion (M) to generate highorder metal ion-bound cluster ions

12

W. A. Tao, R. G. Cooks, Anal. Chem. 2003, 25-31.

Chiral Recognition Mechanisms

3. Chiral Recognition Based on Complex
Dissociation

Chiral selectivity Rchiral is define

Iref*(1) IR

Iref*(2) IS

R chiral =1 : no chiral discrimination

R chiral is more different from 1, the
chiral recognition ability is higher

W. A. Tao, R. G. Cooks, Anal. Chem. 2003, 25-31.

R. Berkecz, et al, J. Mass. Spectrom. 2010,45, 13121319.

13

Chiral Recognition Mechanisms

Mass Spectrometric Chiral Recognition
Mechanisms

1. Host-Guest (H-G) Associations

2. Guest Exchange Ion-Molecule Reactions

3. Chiral Recognition Based on Complex

Dissociation

14

Application of Chiral Recognition

B. Florian, et al, Angew. Chem. Int. Ed. 2013 , 52 ,16

15

Application of Chiral Recognition

Possible mechanisms of amine
catalyzed reaction of aldehyde with
mechanismelectrophiles

Enamine
Widely accepted
Not been validated
experimentally

Z. G. Hajos, D. R. Parrish, J. Org.

Chem. 1974, 39, 1615 1621.

16

Application of Chiral Recognition

Addition reaction between aldehydes

and nitroolefins catalyzed by H-d-ProPro-Glu-NH2
Excellent yields and stereoselectiviti
Catalyst loadings lower than 1 mol %

Proposed catalytic cycle

Enamine mechanism

Problem:
Enamine mechanism not been
validated experimentally

Experimental proof of enamine mechan

Detect an enamine intermediate by ES
17

Application of Chiral Recognition

Methodology: ESI-MS back-reaction screening
A pair of mass-labeled
quasienantiomeric conjugate addition
products
Concept: Host-Guest (H-G)
Associations
Host (Chiral Selector)

Guests

18

Application of Chiral Recognition

If En/En ratio (back reaction) = 2/ent-2 ratio (forward
reaction), it will provides strong evidence to enamine
mechanism.
G
Im

En

ent-2
En

Im

Back reaction

The stereoselectivity 2/ent-2(= k1/k2) is determined

by G of the transition state.
19
R=k1/k2= IEn/IEn = eG/RT

Application of Chiral Recognition

Back-reaction screening and enantioselectivity of the forward
reaction in DMSO

En/En (back reaction) = 2/ent-2 (forward

reaction):
Enamine mechanism

20

Application of Chiral Recognition

Catalyst Screening

Additional organocatalysts investigated in

this study

21

Summary
I. Chirality is significant
II. Concepts of hyphenated MS techniques
III. Mass Spectrometric Chiral Recognition Mechanisms
IV. An example that using chiral recognition to solve mechanistic
problem

Hyphenated MS techniques Host-Guest (H-G) Associations

Chiral Recognition Based

Guest Exchange Ion-Molecule Reactions
on Complex Dissociation

22

Mass Spectrometry

What can we do by using MS?

23

Studying Reaction Mechanism

Interesting reaction systems

Propose reaction mechanism

Combine MS with DFT calcu

24

Catalysts Screening by Mass Spectrometry

Simultaneous screening of a mixture of five
catalysts

25

C. Markert, A. Pfaltz, Angew. Chem. Int. Ed. 2004, 116, 2552-2554

Thanks for your attention!

26

Mass Spectrometry to study reaction mechanism

ESI-MS to capture reaction intermediates
Propose reaction mechnism

Combined with DFT calculation

27

H. Guo, et al, J. Am. Chem. Soc. 2005, 127, 13060-1306

Chiral Recognition Mechanisms

3. Chiral Recognition Based on Complex
Dissociation
Cu2+ (L-Trp)2 (+)-ephedrine
-Ref
-A

Metal: Cu2+
Ref: two L-Trp
Analytes: (+)-ephedrine
()-ephedrine

Chiral selectivity Rchiral :

Cu2+ (L-Trp)2 (-)-ephedrine

I+/I ref*(1) =
-A
3.8
I-/I ref*(2) =
0.91
R (+)-ephedrine
chiral = 4.7
The interaction between
and ref* is stro
-Ref

28

W. A. Tao, R. G. Cooks, Anal. Chem. 2003, 25-31.

Introduction

Asymmetric
Synthesis
Catalyst screening

Chiral
Drug

Chiral
Analysis

Chiral
Recognition

Quality control

Chiral
Resolution
Chromatography

PPT from Xinhao

Mass Spectrometry

29

Catalysts Screening by Mass Spectrometry

Screening Methodology

Mass Spectrometric Screening

of Their Racemic Forms

30

Conformation Analysis by Ion Mobility Spectrometry-Mass Spectrometry

Drift time versus m/z plot
measured by Mass Spectrometer

Conformers produced for cyclic peptide

from Molecular Dynamics simulations

Plot of Normalized MD energy versus

collision cross-section from the
simulated annealing

31
T. R. Brandon, J. Am. Soc. Mass. Spectrom. 2004, 15,

Structural Characterization of Oligomer-Aggregates of -Amyloid Polypeptide

ESI-mass spectra (LC-MS) of A(140)

32

Chiral Recognition Mechanisms

3. Chiral Recognition Based on Complex
Dissociation
Quantitative chiral analysis

[CuII(Pro)2(Tyr)-H]+ complex, Pro as the

[CuII(Phe)2(Ile)-H]+ complex, Phe as the
[CuII(Trp)2(Met)-H]+ complex, Trp as the

The relative rates of the two

competitive dissociations (kA and kref)
can be expressed as the relative
abundance ratio:

Different ratio of AR and AS

Calibration curves for chiral analy

33

W. A. Tao, R. G. Cooks, J. Am. Chem. Soc., 2000, 122,

Chiral Recognition Mechanisms

3. Chiral Recognition Based on Complex
Dissociation
Quantitative chiral analysis

34

W. A. Tao, R. G. Cooks, J. Am. Chem. Soc., 2000, 122,