PRODUCTION OF PURE ENANTIOMERS AT HIGH YIELDS BY INTEGRATING - - PowerPoint PPT Presentation

1

PRODUCTION OF PURE ENANTIOMERS AT HIGH YIELDS BY INTEGRATING CHROMATOGRAPHY, ISOMERIZATION AND MEMBRANE FILTRATION

Sebastian Nimmig, Malte Kaspereit Institute of Separation Science and Technology, University Erlangen-Nürnberg

PIN-NL & NL-GUTS 9 April 2014

2

Motivation and objectives

3

Enantiomers

• Stereoisomers ("mirror images")
• Basically identical physico-chemical properties
• Often produced as racemate (50/50 mixture)
• Usually only one enantiomer has the desired physiological effect
• Separation?  Chromatography

Motivation a C b c d a C b c d

4

Motivation

Separation of enantiomers

• Yield limited to 50% only (conventional approach)
• Reaction required to convert E2 into E1+E2  isomerization
• Increase yield to 100% by recycling E2

E1+E2 E1 E2 E1: Enantiomer 1 (product) E2: Enantiomer 2 Single-column Chromatography Single-column Chromatography fresh feed E1 (product) solvent

E2 E1

E2 (recycle)

Recycle always diluted!

[1] Bechtold et al., J Biotechnology 124 (2006) 146-162

5

Objectives

• Inhibit dilution by solvent removal (here: nanofiltration)

Challenges

• Design specifications
• Required parameters and models
• Analysis and process behaviour
• Fully continuous implementation
• Experimental validation

Single-column Chromatography Membrane- filtration fresh feed E1 (product) solvent solvent

E2 E1

E2 (recycle)

6

Theoretical investigations I

7

Concentration c / g·L-1 c1 c2 Time t / min

t1

1

t1

2

t1

3=t2 1 t2 2

t2

3=t3 1 t3 2

t3

3=tk+1 1

Theoretical investigations I

Shortcut process design[2]

• Reproduce a given chromatogram (simulated or experimental) in each cycle
• Main design parameters:
• Feed flow rate Qfeed
• Permeate flow rate Qperm
• Chrom. flow rate Qchrom
• Injection width Δtinj
• Fractionation times t1 , t2, t3

[2] Nimmig, Kaspereit, Chem Eng Process 67 (2013) 89– 98

8

Theoretical investigations I

Shortcut process design[2]

• Simple explicit equations
• Easy performance prediction

C R M

0.2 0.4 0.6 0.8 1 20 40 60 80 100 Membrane rejection R / - Yield Y / % Shortcut R=1 Shortcut R<1 0.2 0.4 0.6 0.8 1 10 20 30 40 50 Membrane rejection R / - Solvent consumption SC / L·g-1 0.2 0.4 0.6 0.8 1 0.5 1 1.5 2 2.5 Membrane rejection R / - Productivity PR / g·h-1·L-1

9

Theoretical investigations I

Detailed process design

• Chromatography :

Equilibrium dispersive model

• Reaction:

First order kinetics, CSTR

• Nanofiltration:

Simplified solution diffusion model

• Implementation in MatLab

R C M

10

Theoretical investigations I

Detailed process design

• Fully continous connection
• Performance prediction:

0.2 0.4 0.6 0.8 1 20 40 60 80 100 Membrane rejection R / - Yield Y / % 0.2 0.4 0.6 0.8 1 20 40 60 80 Membrane rejection R / - Solvent consumption SC / L·g-1 0.2 0.4 0.6 0.8 1 0.5 1 1.5 2 Membrane rejection R / - Productivity PR / g·h-1·L-1 40mL 120mL 200mL 280mL 360mL 440mL

11

Experimental validation

12

Experimental validation

C R M

F UV

F UV

TI PI

13

Experimental validation

Experiment 1 – Design via detailed simulation

5 10 15 20 80 85 90 95 100 Fraction # Purity PUR / % 5 10 15 20 0.02 0.04 0.06 0.08 0.1 Fraction # Concentration c / g·L-1 Simulation Experiment 1 15 20 25 30 35 0.05 0.1 Time t / min Concentration cout / g·L-1

1

100 200 300 0.05 0.1 0.15 0.2 Time t / min Concentration cout / g·L-1 Simulation Experiment 1

1

14

Experimental validation

Experiment 2 – Design via shortcut method

5 10 15 20 50 60 70 80 90 100 Fraction # Purity PUR / % 5 10 15 20 0.1 0.2 0.3 Fraction # Concentration c / g·L-1 Design Experiment 2 20 30 40 0.05 0.1 0.15 0.2 0.25 Time t / min Concentration cout / g·L-1 100 200 300 0.1 0.2 0.3 0.4 Time t / min Concentration cout / g·L-1 Experiment 2

1

15

Theoretical investigations II

16

Theoretical investigations II

Unit Arrangements Different setups - different performance?

C M R C M R C M R C R M C R M C R M

R: Reactor M: Membrane C: Column : Fresh Feed 1 2 3 4 5 6

17

Theoretical investigations II

Perfomance prediction - Yield

C R M

4 Best choice:

0.4 0.6 0.8 1 20 30 40 50 60 70 80 90 100 Membrane rejection R / - Yield Y / % Da = 0.3 0.4 0.6 0.8 1 20 30 40 50 60 70 80 90 100 Membrane rejection R / - Yield Y / % Da = 10 Var 1 Var 2 Var 3 Var 4 Var 5 Var 6 Batch

18

Theoretical investigations II

Perfomance prediction - Productivity

C R M

4 Best choice:

0.4 0.6 0.8 1 1.5 2 2.5 3 Membrane rejection R / - Productivity PR / g·h-1·L-1 Da = 0.3 0.4 0.6 0.8 1 1.5 2 2.5 3 Membrane rejection R / - Productivity PR / g·h-1·L-1 Da = 10 Var 1 Var 2 Var 3 Var 4 Var 5 Var 6 Batch

19

Further extensions of concept?

20

Further extensions of concept

heating jacket reaction chamber stirrer membrane

21

Further extensions of concept

On-column protein refolding

Single-column Chromatography Membrane reactor Denatured protein (D) SolvR

X D

Native protein (N) Undesired conformations (X) Solvent SolvD

D N X

22

Summary

Summary

• Proposed concept capable of significantly improving yield and performance
• Shortcut methods developed for basic design, full model for detailed design
• Performance limited mainly by membrane rejection
• Process setup influences performance
• First successful realization of such process in directly coupled operation

Outlook

• Potential application to industrial relevant compunds?

23

Thank you for your attention!

24

Racemization scheme

Chlorthaidone racemization

• Nearly insolubile in Water
• Increasing solubility in

MeOH/H2O

• MW =338 g/mol
• Kinetics known as function of

pH-value and temperature[3] Racemization under acid conditions

[3] J. G. Palacios, B. Kramer, A. Kienle, M. Kaspereit, J Chromatogr A 1218 (2011) 2232- 2239

25

Detailed process design

Reactor concentration behavior for different Volumes

VR=1mL VR=1000mL

26

Membran behavior

Reaktor Membranmodul

I

• Membran Reactor design
• Membrane unit acts as CSTR

(residence time function) N E P F B I

27

Experimental characterization

Parameter determination - Nanofiltration

2 4 6 8 50 100 150 200 250 300 350 400 450 500 Pressure ∆p [bar] Permeate flux for pure solvent [mL/min/m

2]

0.1 0.2 0.3 0.4 100 150 200 250 300 350 400 450 500 550 600 Concentration cCTD [g/L] Permeate flux [mL/min/m

2]

8.30 bar 6.38 bar 4.80 bar

Pure water experiments (k1) Batch concentration experiments (k2)

28

Experimental characterization

• Parameter determination - Racemization

Time [min] Concentration [g/L] Time [min] ln(c*)