producing added value from amazonian fruits pallet...pure juice analysismethod: hplc compare acai...

Workshop EULAFF – ANGRA 03 – 05/12/2007

Producing Added Value

from Amazonian fruits- 2007 results -

Workshop EULAFFANGRA dos REIS

Dominique PALLET : [email protected]


In the forest of Amazonian fruits

• CIRAD participate of several international projects on Amazonian fruits valorization :

– SAFE NUT (FAO – OMS)

– FLOAGRI : Amazonian NTFP (E.U. Tropical Rainforest)

– PAVUC (FP6) Producing Added Value from Under-utilized Tropical fruit Crops

• 9 South American fruits : wild blackberry, tomatotree, red pitahaya, berrycactus, Naranjille, Açai, Camu-Camu, cashew apple, palm peach


Presentation based on 2007’s results

examples of 2007’s results:1) Improve local process: Influence of buriti pulp drying

conditions on the carotenoids content

2) To know biodiversity: anthocyanins content as a

parameter of Açaí intra specific biodiversity

3) To know biodiversity: characterization of wild

Blackberry micro constituents

4) New process for added value: purification of açaí and

blackberry anthocyanins using polymeric resins

5) Molecule functionality: pharmaceutical properties of

Tucumà (Astrocaryum vulgare)


Buriti fresh pulp carotenoids content :

- β-carotene (360 ±32 µg/g of dried weight, DW),

- α-carotene (80 ±9 µg/g DW)

- γ-carotene (36 ±4 µg/g DW)

Pulp is only 12-13% of the fruit (d.w.) and is difficult to extract

Improve local process:buriti (Mauritia flexuosa) pulp drying

- Mauritia flexuosa is a typical palm of the Brazilian Amazonian basin- Important source of calories, proteins and vitamins-Current valorization of the fruit consists in pulp-made juice called “Buriti wine”


Buriti pulp drying

• Objetif : influence of drying on carotenoids content

• Cooperation : with UNICAMP and Communitarian

Association of Santo Antônio de Abonari

• Activity : we measure the total carotenoid content

for diferent drying conditions (solar, air...)


Influence of buriti pulp drying conditions on carotenoids content

5309 ± 1,112

4314 ± 2,58326 ± 4,65

1Sun-drying

1312 ± 2,35314 ± 4,8190°C

4313 ± 7,49325 ± 5,77 *70°CAirflow drying

Losses (%)Dried pulpFresh pulp

Total carotenoids content (µg/g) on a dry basis

• Drying of Buriti pulp did not induce significant decreases of total carotenoids content.

• At a small scale, the development of pulp drying appears interesting to get an intermediary product, rich in carotenoids.


2) Anthocyanins content as a parameter of Açaí intra specific biodiversity

example of 2007’s result:


Açai

� Açai palm tree

Latin name : Euterpe oleraceae

Family : Palmae (Arecaeae)

Native from the Amazon region

Açai fruit is used to produce energetic snack beverages, ice cream, jelly, and liqueurs.


Açai pulp


Açai composition biodiversity

• Objetif : caracterize and valorize the intra specific açai biodiversity

• Cooperation with Embrapa (Amaz. Oriental & Cenargen)

• Activity : study the total anthocyanin content of some progenies (25) – Embrapa collection (Belem)

• Result: anthocyanin content: from 13,75 mg/100g (progênie 23) to 228,77 mg/100g (progênie 11) : high variability

QUANTIFICAÇÃO DO TEOR DE ANTOCIANINAS TOTAIS DA POLPA DE AÇAÍ DE DIFERENTES POPULAÇÕES DE AÇAIZEIRO (2007), Kelly de Oliveira Cohen; Maria do Socorro Padilha de Oliveira; Renan Campos Chiste; Dominique Pallet; Damares

de Castro Monte


3) Micro constituents of two wild Blackberry varieties. (PAVUC Project)

examples of 2007’s results in our lab:


Andeans Blackberries

-The plant is a vigorous shrub of luxuriant growth.

-High productivity of fruit

� Blackberry shrub


Two varieties of Andeans Blackberries

Family : Rosaceae

� Rubus adenotrichus : Mora Común

� Rubus glaucus : Mora de Castilla

From Mexico to Ecuador

Native from the northern Andes to the southern highlands of Mexico

� Blackberry fruit

In Latin countries they are important fresh fruits as well as ingredients in jellies, jams, juices, thick syrups and even wines.


7th International Food Data Conference. Sao Paulo, Oct.2007

Extraction for HPLC analyses

- Extraction acetone/water/ formic acid

- Qualitative (LC/MS) and quantitative analysis of major compounds

⇒⇒⇒⇒ ellagitannins and anthocyanins (blackberry)

� liq-liq extraction with ethyl acetate

- Analysis of minor compounds in blackberries

⇒⇒⇒⇒ flavonols (quercetin and kaempferol glycosides)

⇒⇒⇒⇒ ellagic acid glycosides



Blackberry Phenolic compounds characterization

ellagitannins anthocyanins

Rubus glaucus

(Ecuador)

Rubus adenotrichus

(Costa-Rica)

0

500

1000

1500

2000

2500

3000

Co

nte

nt

(mg

/10

0g

MS

)

2 major classes

0

100

200

300

400

500

600

700

Co

nte

nt

(mg

/10

0g

MS

)

cyanidin

glucoside

cyanidin

rutinosidecyanidin malonyl

glucoside

anthocyanins



Rubus glaucus

Rubus adenotrichus

0

10

20

30

40

50

60

70

80

90C

on

ten

t (m

g/1

00g

MS

)

ellagic acid

glycosides

quercetin

glycosides

kaempferol

glycosides

⇒⇒⇒⇒ Content in agreement with literature data

⇒⇒⇒⇒ Phenolic composition: discriminatory factor

Phenolic compounds (Ethyl acetate extraction)


Anthocyanins content

156,165,6Açai

121,6Blackberry

(R. adenotricus)

Cyanidin-3-rutinoside

mg / 100 mL

Cyanidin-3-glucoside

mg / 100 mL

Pure juice

Analysis method : HPLC

Compare Acai and blackberry



4) Purification of açai and blackberry anthocyanins using polymeric resins for natural red colorant purpose.


- Colour is one of the most important attributes affecting consumer acceptance of food

- Colour mainly defines aesthetic value of food

Natural colorants : problematic research

� Importance of colour in food

� Interest of natural colorants

-Real safety of synthetic colorant ?

-Worldwide tendency toward the use of natural pigments

� Drawback of natural colorant

-Low stability

-Supply of raw material


Aim of this study

� 2 raw materials riches in anthocyanins (açai & Bl Be)

� Adsorber technology (polymeric adsorbent resins)

Obtention of natural anthocyanins-enriched and purified extracts as natural food colorant.

� Optimisation of the ACN’s purification


Anthocyanin molecules

O+

OH

O

OH

OH

OH

Glucose

O+

OH

O

OH

OH

OH

Rutinoside

� Structure

-Belong to the flavonoid group of polyphenol

-Water soluble pigments

-Responsible for the red, purple, and blue colors of many fruits, vegetables, and flowers

Cyanidin-3-glucoside

Cyanidin-3-rutinoside

� Characteristics


Anthocyanins stability

ACNs are influenced by many factors as:

-pH (varying pH levels induces structural transformations)

-Storage temperature

-Presence of enzymes

-Structure and ACN content-Light

-Other compounds such as other flavonoids and phenolics.


Use of polymeric adsorbents (resins) for the purification of ACNs

-Nonthermal and no chemical anthocyanins purification

� Interest of resins

-Approved by for food contact use, by the European code of regulation

-Most of the impurities are removed

� Physical characteristics

� 2 steps process

specific surface area, porosity, and pore radius

It’s a styrene-divinylbenzene copolymerisate polymer with a

1. Adsorption of the ACNs on resins

2. Desorption


Process for the purification with resins

Step1. Adsorption of ACN on resins

Hydrophobic interaction, hydrogen bonding


Step 2. Desorption

With an organic solvent which can weaken the attractive forces between the solute and the polymer for removing remove ACNs

At the beginning At the end

Process for the purification with resins


Result of ACN’s purification with resin

8%

19,46%

Purity of final extract

(ACN/DM)

5154,4VI / BI

34,124,6VI / BI

31,32

6,05

Yield DM % (extract/juice)

87

54,331,2ACN contents(mg/100mL)

4817,8E1%1cm

364E1%1cm

96

130,8 40,8ACN contents(mg/100mL)

Yield ACN % (extract/juice)

Final Extract

Juice

BBerry

Açai

Strength colour : correspond to the Abs of (x g) pigment in 100mL buffer pH3E1%1cm=

ACN = Anthocyanins

DM = Dry matter

VI = Violet Index ; BI = Brown Index


Results 2007

The use of resin prove to be an interesting process: Simplicity,

Great % of ACN’s recovery, Increase of the colour strength, No

colour degradation

�Natural food colorant stability test for industrial purposes:

2/ Compare different combined natural pigments stability: Açai, blackberry, black carrot, blackberry+grappe,

bberry+açai…

1/ Measure the stability of red color in a soft drink solution, at 4°C, 20°C, 30°C, 50°C during 2 months

Methodology to know if the natural pigment (Andean and Amazonian fruit) could be use as natural food colorant



5) Pharmaceutical properties of Astrocaryum

vulgare


Tucumà : Astrocaryum vulgare

• Family: Arecaceae, Palmaceae• Common name: Awara, Tucumà…• Latin name: Astrocaryum vulgare C.

Martius• Origin: Amazon rainforest; it grows

principally in French Guiana, Surinam, Brazil and neighbouring areas (savannas, lowlands).

• The production of oil fruit is used in majority for the diet of the population.


Characteristics of the fruit from Astrocaryum vulgare

• Ovoid orange red fruit, more or

less globose

• Fibrous and fleshy fruit

• Weight : 20,1 +/- 2,3 g ; pulp

mass : 8,1 +/- 0,4 g ; kernel mass:

4,0 +/- 0,8 g; core mass: 12,0 +/- 2,1

g

• Rate moisture: 38 +/- 10%.

• Composition: proteins (3,5%);

carbohydrates (19,1%); fats

(16,6%); The fruit contains edible

oil with a vitamin A content of

50,000 i.u. per 100 g of pulp;

Epicarpe

Mesocarpe

Kernel

endocarpe


Traditional uses of Astrocaryum vulgare

• Root: in a decoction to remedy furonculosis and syphilis• Seed: oil is used for rheumatism, pain, laxative and earache,

and its preparation for furoncles, also swabbed onto aching feet and rubbed on feverish person to assist perspiration.

• Fruit: pericalp pulp is used to calm colicky babies. Mesocarp is used to treat coughs and as a breath freshener. The fruit is used against the eye disease xerophthalmia. The immature endosperm gives a juice used for a drink or in culinary preparations. The oil of pulp is used for rheumatism, cramps and aches. The oil of kernel is used for dermatits, furonculosis, skin ache….


Work in progress on Astrocaryum vulgare with/by

Montpellier Pharmacology University

• Anti-inflammatory characteristics from fractions of

Astrocaryum vulgare

• Bactericide effect of Astrocaryum vulgare fractions

• Oxidative stress analysis for Astrocaryum vulgare entire oil

• Anti-elastase studies for fractions of Astrocaryum vulgare

• Studies concerning the benign prostatic hypertrophy (with

dogs: protocol)


Thank you for your attention

Maison de la Technologie CIRAD – Montpellier - France

producing added value from amazonian fruits pallet...pure juice analysismethod: hplc compare acai...

Documents