Novel Formulations and Process for Development of Microencapsulated Krill Oil

CSIRO ANIMAL, FOOD AND HEALTH SCIENCES

L Sanguansri*, Z Shen, S Bhail, LJ Cheng, DY Ying and MA Augustin

AAOCS Omega 3 symposium - Omega-3 Hot Topics: Science, Supply and Sources6-8 November 2013, Newcastle, NSW, Australia

Outline

Introduction

Why Krill Oil

Our strategy

Results

Summary

Luz Sanguansri2 |

Why Krill Oil

Krill oil (KO) is a premium source of marine omega-3 fatty acids

KO has a unique combination of omega-3 phospholipids and

astaxanthin

KO has inherent emulsifying power due to its phospholipids

Long chain omega-3 fatty acids in KO is susceptible to oxidation

KO has strong flavour and odour-might be unpleasant to some people

(challenge)

Market for KO is expected to be the fastest growing segment in the

global omega-3 ingredient market

Luz Sanguansri3 | *Frost & Sullivan, June 2011

Previous experience with different ω-3 oils - using standard emulsion preparation procedure- using heated protein-CHO as encapsulant

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Powder with different oil

TG PL Emulsion size

(D, 0.5) µm

Powder IP (hr)

@ 80°C

Omega-3 (DHA/ EPA)(g/100 g oil)

Total fat(g/100 g powder)

Surface fat(g/100 g powder)

P1 0.35 23.2 42 (15/27) 50 0.37

P2 0.12 11.8 55 (22/33) 50 0.31

P3 1.20 ~1.4 30 (10/20) 50 16.12

P4 0.45 60.5 30 (25/5) 50 0.62

Oil rich in PL has larger emulsion particle size and higher powder surface fat than triglyceride oils using the same formulation and process conditions

Typical major lipid components of krill oil, algal oil and fish oil

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Krill Oil Algal Oil Fish Oil

Triacylglycerol 40% 94.0% 99%

Diacylglycerol 1.2% 3.0%

Monoacylglycerol <1% <1%

Free fatty acids 4.2% <1% <1%

Total neutral lipids 46.8% 97.0% 99.0%

Phosphatidylcholin 35%

Lyso-phosphatidylcholin 3.1%

Phosphatidyletanolamin 2.1%

Total polar lipids 40.0%

Astaxanthin >200 ppm

KO has ≤40% TAG and ≤40% phospholipids compared to other marine oils (algal oils and fish oils) with >90% TAG

EPA

DHA

Phospholipid Triacylglycerol

Our strategy

Capitalise on the inherent emulsifying properties of krill oil phospholipids

Capitalise on the anti-oxidative properties of heated protein-carbohydrates as encapsulant

Apply chemistry and emulsion/material science capabilities to design the oil-water interface and microcapsule properties

Luz Sanguansri6 |

CSIRO’s Krill Oil Microencapsulation Technology- Emulsification and Spray Drying Process

Luz Sanguansri7 | CSIRO’s Patented Technology

Protein + carbohydrate solution

Krill oil-in-water Emulsion

Adjust pH as required

± Heat Treatment

Add krill oil and homogenise

Spray dry at 180/80°CInlet/outlet temperature

Spray dried Powder:Krill Oil embedded in Protein-

carbohydrate Matrix

Strategy 2Krill Oil

Spray dried Powder:Krill Oil embedded in Protein-

carbohydrate Matrix

Mix water and Krill Oil using high shear mixer

Add protein-carbohydrate solution (60°C) into emulsion

Adjust pH as required

Mix until homogeneous

Spray dry at 180/80°CInlet/outlet temperature

Krill Oil-in-water Emulsion

Neat Krill Oil-in-water Emulsion

Strategy 1

Preparation of phospholipid rich interface

Standard emulsion preparation steps

Particle size distribution and zeta-potential of KO in H2O emulsions (Strategy 1)

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Zeta-potential (mV)

KO emulsions (pH 8) pH 8.0

10% KO neat -50.40±3.3

10% KO in 1% Protein1 -25.9±1.0

10% KO in 2.5% Protein1 -19.3±0.6

10% KO in 3% Heated Protein2-CHO -49.6±3.03

For comparison:10% fish oil in 3% Heated Protein2-CHO -36.5±1.0

• Emulsion stabilised by Protein1 has weaker charge of the oil-water interface

• Emulsion stabilised by Heated protein2 -CHO (MRP) as encapsulant maintained the strong charge of the oil-water interface

10% KO emulsion: neat or in 1% Protein, 2.5% Protein or 3% Heated Protein-CHO

0

2

4

6

8

10

12

14

16

0.01 0.1 1 10 100

Volu

me (

%)

Particle size (µm)

1Isolectric Point = 7-9 (almost no charge at pH8)2Isoelectric Point = 4-5 (negative charge at pH8)

Stability of Krill Oil emulsions during accelerated storage at 40°C

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• pH has no significant effect on oxidation of neat KO in water emulsion during storage

• Encapsulation significantly improved the stability of krill oil-in-water emulsions during storage

• Heated protein-CHO (MRP) encapsulant provided the best protection from oxidation

Analysis of remaining EPA during storage Analysis of remaining DHA during storage Propanal Headspace Analysis during storage

0

50000

100000

150000

200000

250000

300000

GC

are

a o

f p

rop

anal

(x1

0-4

)

Day 11

Day 18

Day 25

50

60

70

80

90

100

% R

em

ain

ing

of

EPA

Day 11

Day 18

Day 25

50

60

70

80

90

100

% R

em

ain

ing

of

DH

A

Day 11

Day 18

Day 25

KO emulsion 17.5 months at 4°C

CSIRO’s Krill Oil Microencapsulation Technology- Strategy 1 vs. Strategy 2

Luz Sanguansri10 | CSIRO’s Patented Technology

Protein + carbohydrate solution

Krill oil-in-water Emulsion

Adjust pH as required

± Heat Treatment

Add krill oil and homogenise

Spray dry at 180/80°CInlet/outlet temperature

Spray dried Powder:Krill Oil embedded in Protein-

carbohydrate Matrix

Strategy 2Krill Oil

Spray dried Powder:Krill Oil embedded in Protein-

carbohydrate Matrix

Mix water and Krill Oil using high shear mixer

Add protein-carbohydrate solution (60°C) into emulsion

Adjust pH as required

Mix until homogeneous

Spray dry at 180/80°CInlet/outlet temperature

Krill Oil-in-water Emulsion

Neat Krill Oil-in-water Emulsion

Strategy 1

Same formulation,Different process

Different in zeta potential (different oil-water interface)

Stability of Krill oil powder during storage at 40°C

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P1 10% KO neat + maltodextrin (Strategy 1)P2 10% KO in 1% protein + maltodextrin (Strategy 1)P3 10% KO in 3% heated protein-carbohydrates + maltodextrin (Strategy 1)P4 10% KO in 3% heated protein-carbohydrates + maltodextrin (Strategy 2)

50

60

70

80

90

100

EPA DHA EPA DHA EPA DHA

Day 0 day 17 Day 28Pe

rce

nta

ge r

em

ain

ing

of

LC-n

-3 P

UFA

s

50% KO P1 50% KO P2 50% KO P3 50% KO P4

0

2

4

6

8

10

12

14

16

50% KO P1 50% KO P2 50% KO P3 50% KO P4

Pro

pan

al G

C a

rea

(x1

0-4

)

Day 0 Day 17 Day 28

Heated protein-CHO (MRP) encapsulant provided the best protection from oxidation during storage

CSIRO’s Microencapsulation Technology- Embedding and Extrusion process

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Add into protein-carbohydrate

High temperature &

high shear Mixing

Extrude through a die

Dry

Apply secondary coating (optional)

Extruded Pellet: Krill Oil embedded in Protein-

carbohydrate Matrix

Krill Oil

Neat Krill Oil-in-water Emulsion

Spray dried Powder:Krill Oil embedded in Protein-

carbohydrate Matrix

Mix water and Krill Oil using high shear mixer

Add protein-carbohydrate solution (60°C) into emulsion

Adjust pH as required

Mix until homogeneous

Spray dry at 180/80°CInlet/outlet temperature

Krill Oil-in-water Emulsion ± additives

CSIRO’s Patented Technology

Extruded samples with Krill Oil (non-expanded)

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KO emulsion embedded in different matrices

KO Content(% dry basis)

Ratio of DHA/EPA

LF1: KO in water emulsion(protein-CHO blend matrix)

8.82 0.51

LF1: KO in water emulsion (heated protein-CHO matrix)

14.14 0.52

LF1: KO in water emulsion (protein matrix)

15.16 0.52

LF1: KO in water emulsion(carbohydrate matrix)

11.88 0.50

LF2: KO in MRP emulsion + PP(carbohydrate matrix)

9.73 0.50

LF3: KO in MRP emulsion (carbohydrate matrix)

7.10 0.50

Up to 15% krill oil was embedded in protein matrix Krill oil emulsions were stable during high temperature & high shear extrusion

Extruded snacks with Krill Oil (expanded)

Luz Sanguansri14 |

KO emulsion embedded in a snack formulation

KO Content(% dry basis)

Ratio of DHA/EPA

KO in water emulsion 7.76 0.51

KO in MRP emulsion + PP 8.53 0.51

KO in MRP emulsion 3.92 0.50

Up to 8.5% krill oil was embedded in extruded expanded snacksKrill oil emulsions were stable during extrusion of expanded snacks

Summary

Microencapsulation enhanced the oxidative stability of krill oil emulsions and spray dried KO powder during storage

The encapsulant formulation and order of processing affect the oil-water interface and stability during storage

Protein or protein-carbohydrate encapsulants enhanced the oxidative stability of KO during storage

Heated protein-carbohydrate encapsulant provided the best protection from oxidation in emulsion and powder

Stabilised KO emulsions are stable to high temperature and high shear extrusion

Luz Sanguansri15 |

CSIRO ANIMAL, FOOD AND HEALTH SCIENCES | PREVENTATIVE HEALTH FLAGSHIP

Thank you

Luz Sanguansri | Research Team Leader, Food Systems EngineeringTel: +61 3 9731 3228 | Email: luz.sanguansri@csiro.auWeb: www.csiro.au

Acknowledgement

Keith Pitts – for the production of extruded samples

Michael Mazzonetto – for production of krill oil powders