Epa Purification With Contichrom & DAISOGEL

© ChromaCon AG // www.chromacon.ch // EPA purification by Contichrom® 1

Enabling processing capabilities by significantly

reducing CAPEX and OPEX

EPA purification from fishoil using

Contichrom® (MCSGP)


Overview

Fatty acids are useful in for many applications, including in

pharmaceuticals, nutraceuticals, food supplements and chemical

intermediates

Often large volumes (multi-tons) at high purity (>80%) are required

for the applications

Classical large-scale purification techniques such as distillation

and precipiation are not capable of providing high purity fatty acids

Comparing available purification technologies, Contichrom® with

MCSGP seems to be the only available production technology

capable of producing high volume, high purity fatty acid products at

reasonable COG


Introduction: Fat, Oil

Oil (liquid at room temperature), fat (solid at room temperature):

Major -3 fatty acids:

Eicosapentaenoic acid (EPA): C20:5

Docosahexaenoic acid (DHA): C22:6

Example of an unsaturated fat triglyceride. Left part:

glycerol, right part from top to bottom: palmitic acid,

oleic acid, alpha-linolenic acid,

Source: Wikipedia

The first double bond is located at the third carbon from the methyl end of the fatty acid chain, known as

the n end. Thus, EPA acid is a polyunsaturated n−3 (omega-3) fatty acid

http://upload.wikimedia.org/wikipedia/commons/e/ed/DHA_numbers.svg


Case Study: Purification of ω-3 fatty acids

ω-3 fatty acids for pharmaceutical applications are

made from fish oil

Products: Omacor®/Lovaza™ (1.3 bio US$ sales)

Used as prescription medicine as a supplement to

diet for the treatment of elevated levels of

triglycerides

API of Omacor: 45% EPA-EE, 35% DHA-EE

Several companies develop biosimilar APIs

Several companies develop high purity EPA-EE

and derivatives as 2nd generation product


Case Study: MCSGP process design: 1st step

Perform analytical RP-HPLC batch chromatography

Feed purity 74%, target purity >97%

(generic fish oil feed purchased from TCI Europe N.V.)

Main impurity Docosahexaeonic acid ethyl ester (DHA-

EE)

EPA-EE DHA-EE


Case Study: MCSGP process design: 2nd step

Batch chromatography for the purification of EPA-EE, recording the

elution fraction product purity by analytical HPLC

Results: UV-signal and fraction analyses:

70%

75%

80%

85%

90%

95%

100%

0.0

2.0

4.0

6.0

8.0

10.0

12.0

14.0

0 5 10 15 20 25

Pu

rity HP

LC [%

]

UV

no

rm[m

AU

/mA

U]*

12,

Co

nc

[mg

/ml]

Time [min]

conc EPA-EE

UV

Purity EPA-EE

EPA-EE spec

B01on Daisogel 120-20 ODS-RPS


70%

75%

80%

85%

90%

95%

100%

0.0

2.0

4.0

6.0

8.0

10.0

12.0

14.0

12 13 14 15 16 17 18 19 20 21 22

Pu

rity HP

LC [%

]U

Vn

orm

[mA

U/m

AU

]*1

2, C

on

c [m

g/m

l]

Time [min]

conc EPA-EE

UV

Purity EPA-EE

EPA-EE spec

B01

Case Study: MCSGP process design: 3rd step Define sections of pure product extraction (red) and product-containing impurity

recycling (blue + green) based on prior analytical purity measurements using the

Contichrom® Software Wizard

15.8min 17.3min 19.3 min

product recycle recycle waste waste


Use Contichrom® MCSGP wizard to convert batch method into

MCSGP method by simple click on the „Write Method“ button (yellow

arrow)

Case Study: MCSGP process design: 4th step


Case Study: anatomy of one MCSGP cycle run Zoom of 1 cycle (only UV1 and tasks of column 2 shown)

W/P rec product

elution

P/S rec Cleaning and re-

equilibration

Removal of W W/P

uptake

feed P/S

uptake

Pressure

at pump 1


Results from MCSGP:

Internal UV profiles of both UV detectors shown for 5 cycles shown

Run is very consistent

Case Study: anatomy of multiple MCSGP cycles


Case Study: multiple MCSGP cycles overlay

Online evaluation of MCSGP experiment: Cycle overlay can be used to determine

if run has reached cyclic steady state and shows a consistent pattern

Example: Overlay of 5 cycles: Exact matching of product profiles in (blue) window.

Cyclic steady state: Product of constant quality and concentration is withdrawn

Product consistency shown

product

elution


Experimental verification

Performance summary:

MCSGP

(20 m

resin)

Batch

(15 m

resin)

Improvement by

MCSGP

Purity [%] >97% >97%

Yield [%] 90% 36% + 250%

Productivity (Throughput) [(g product)/(L resin)/(hr operation time)]

65 11 + 590%

Solvent Consumption [L solvent/g product]

0.8 3.2 - 75%


Purification of EPA: Scale-Up modelling Contichrom® (2 col MCSGP) Batch

Lab/prep scale

(for feasibility)

i.d. 4.5cmx15cm columns,

200kg/year

Pilot-scale

(2 t/year)

i.d. 15cmx15cm columns,

solvent: 133m3/month

i.d. 45cmx25cm column,

solvent: 530m3/month

Production-

scale

(100 t/year)

i.d. 100cmx15cm columns,

solvent: 6‘700m3/month

i.d. 317cmx25cm column,

solvent: 26‘700m3/month

Column size not

feasible in batch


Conclusions from scale modelling of EPA purification

Batch chromatography not feasible on production

scale

Contichrom using MCSGP process at production

scale: 100 t/y and beyond is feasible at competitive

COG

Other multi-column production technologies (SMB

and derivatives) are inferior as they require much

larger CAPEX and OPEX

Technology

Epa Purification With Contichrom & DAISOGEL