University of Southern Denmark

Natural blue food color from cyanobacteria Spirulina platensis

Malwade, Chandrakant Ramkrishna; Roda Serrat, Maria Cinta; Christensen, Knud Villy;Fretté, Xavier; Christensen, Lars Porskjær

Publication date:2015

Document version:Final published version

Citation for pulished version (APA):Malwade, C. R., Roda Serrat, M. C., Christensen, K. V., Fretté, X., & Christensen, L. P. (2015). Natural blue foodcolor from cyanobacteria Spirulina platensis. Poster session presented at 6th Nordic Natural ProductsConference, Visby, Sweden. http://www.fkog.uu.se/nnpc/NNPC2015-abstractbook.pdf

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Natural Blue Food Color from Cyanobacteria Arthrospira platensis

Chandrakant R. Malwade, Maria C. R. Serrat, Knud V. Christensen, Xavier Fretté, Lars P. Christensen

Department of Chemical Engineering, Biotechnology, and Environmental Technology University of Southern Denmark, Campusvej 55, Odense M, Denmark

crm@kbm.sdu.dk

MOTIVATION

INTRODUCTION

EXPERIMENTAL

Blue colour is an important part of the food color palette used in

products such as icecream, confectionaries, chewing gum and soft

drinks. Available blue colors in the market are chemically

synthesized of which EU have the following: Patent blue V,

Indigotine, Brilliant blue FCF. Recently concerns have been raised

about the safety of synthetic blue colors [1] and together with the

growing demand among consumers for natural food colors this has

increased the need for the development of natural blue colors.

Composition of Linablue Additives removal from Linablue Cleavage of Phycocyanobilin (Methanolysis)

Patent blue V (E131)

Indigotine (E132)

Brilliant blue FCF (E133)

Aim of this project is to develop a stable natural blue food color from cyanobacteria Arthrospira platensis (syn. Spirulina platensis).

Arthrospira is a blue-green microalgae widely used as dietary supplement due to its high protein content. Arthrospira extract has already

been approved by USFDA as a food color, but its lack of stability is a problem [2]. Phycocyanobilin, responsible for blue color of the

extract, is obtained in pure form and stabilized through copigmentation and lake formation. In this project, pure phycocyanobilin (PCB) is

obtained from a commercial extract of Arthrospira (Linablue®G1).

Linablue is washed with methanol to

remove additives; methanol selectively

dissolves D-trehalose and trisodium citrate.

Procedure included stirring 50 g Linablue

in 400 mL methanol for 30 min.

Procedure is repeated 5 times; 20 g

additive free Linablue is obtained.

Methanol washing also aid in denaturation

of phycocyanins as depicted below.

10 g washed Linablue boiled in

400mL methanol for 16hrs at 65 °C

Samples are taken at regular interval

from mixture for HPLC analysis

After 16hrs, methanol solution is

analyzed with HPLC and LC-MS

Linablue powder

Methanol wash D-Trehalose & trisodium citrate

Protein in normal state

Denatured protein

Purification of methanolysis mixture

666.02

0,0 1,0 2,0 3,0 4,0 5,0 6,0 7,0 8,0 9,0 10,0

0

25

50

75

100

140

1 - 3,917

2 - 4,583

3 - 6,203

min

mAU

300 400 500 600 700

629,60 349,72

277,52

nm

300 400 500 600 700

368,71

666,02

nm

300 400 500 600 700

364,19

650,98

nm

1 2

3

Cultivation of

Arthrospira platensis

Phycobilisomes (Light harvesting complexes of

phycobiliproteins)

Phycocyanobilin (Chromophore)

Extraction

Cleavage & Purification

C-Phycocyanin (Phycobiliprotein)

PCB

Methanolysis mixture is purified by using flash

column chromatography

RP C-18 column with acetone and water is used as

mobile phase

85mg phycocyanobilin is obtained

0,0 1,0 2,0 3,0 4,0 5,0 6,0 7,0 8,0 9,0 10,0

0

125

250

375

500

625

700 1 - 4,563

min

mAU

200 400 600 800 1000 1200 1400 1600 1800 2000

m/z

0

20

60

80

587.61

1173.45

1760.33

1195.39

40

100 PCB + H+

FUTURE PERSPECTIVES

Physico-chemical properties of phycocyanobilin will be determined

Further optimization of cleavage and purification process to obtain pure

phycocyanobilin will be done.

Stabilization and intensification of phycocyanobilin color will be attempted

through its copigmentation with naturally occurring molecules.

REFERENCES

1. European Food Safety Authority (2013) EFSA Journal, 11, 2818.

2. Newsome, G. et al. (2014) J. Agric. Food Chem., 62, 6498−6511.