Polyphenols characterization and antioxidant capacities of concentrated extracts of Carapa procera D.C. and Hibiscus sabdariffa L. obtained at pilot plant scale

(1)

Polyphenols characterization and antioxidant capacities

of concentrated extracts of

of concentrated extracts of Carapa procera

Carapa procera D.C.

D.C.

and

and Hibiscus sabdariffa

Hibiscus sabdariffa L. obtained at pilot plant scale

L. obtained at pilot plant scale

Carapa procera and Hibiscus sabdariffa are 2 tropical plants traditionally used in Ivory Coast to prepare at

home level medicinal and food complement products.

A pilot plant coupled-process was developed mimicking the traditional recipes using plant leaves and flowers to prepare functional and active polyphenol extracts with colorant and antioxidant properties. These natural extracts are on high demand locally in the South or for industrial niche markets in the North, as medicinal products and as food or cosmetic ingredients to replace synthetic compounds usually used in such industrial sectors

Concentrated water-extract of plant polyphenols, were prepared at pilot plant in pilot plant. Extracted anthocyanins, phenolic acids and flavonols were characterized and their antioxidant capacities measured and compared.

The Process used to eco-friendly extract and concentrate bioactive water-soluble molecules, such as polyphenolics, from 2 tropical plants, traditionally used by village people in Ivory Coast, allowed producing natural concentrates. Mixtures of these 2 plant extracts give natural deep-red concentrates with high AOC.

 Polyphenol contents of process end-products

Contacts : yves.lozano@cirad.fr, felix.adje@yahoo.fr

1 UMR GPEB Génie des Procédés – Eau, Bioproduits, CIRAD, TA 40/16, 73 avenue J.F. Breton, 34398 Montpellier cedex 5, France

2 UMR 1083 Sciences pour l’Œnologie, Plateforme Polyphénols, INRA, 2 place Viala, 34060 Montpellier cedex 1, France

3 LAPISEN, Institut National Polytechnique Félix Houphouët-Boigny, INP-HB, BP 1093 Yamoussoukro, Côte d’Ivoire

4 _{UMR CNRS 6263, Equipe Phytochimie, Université Paul Cézanne, Avenue Escadrille Normandie Niémen, Case 461, Marseille cedex 20, France} 5 UMR A 408, Sécurité et Qualité des produits d’origine végétale, Université d’Avignon et des pays du Vaucluse, INRA, F-84000, Avignon, France

6 UFR Biosciences, Laboratoire de Biochimie et Sciences des Aliments, Université de Cocody-Abidjan, 22 BP 582 Abidjan 22, Côte d’Ivoire

Félix A. ADJE1,3,4,6_{; Yves F. LOZANO}1_{; Emmanuelle MEUDEC}2_{; Paul LOZANO}1_{; Augustin A. ADIMA}3_{; Emile M. GAYDOU}4_{; Farid CHEMAT}5

 INTRODUCTION

Giusti, M., M. and R. Wrolstad, E., (2003) Biochem. Eng. J., 14 (3): 217-225. Rodriguez-Saona et al. (1998) J. of Food Sci. 63 (3): 458-465. Einbond, L.S. et al. (2003) Food Chem. 84 (1): 23-28. Bonilla, F. et al. (1999) Food Chem. 66 (2): 209-15. Kim, D.-O. et al. (2003) Food Chem. 81 (3): 321 - 326.

 AntiOxidant Capacities (AOC) of plant extracts

 Major polyphenolic compounds identified in concentrated extracts

Material Plant Extracts Anthocyanins (µmol/L, Cya eq.)

Phenolic ac. (µmol/L, GA eq.)

Flavonols (µmol/L, Quer eq.)

C. procera

Clarified permeate 1.6 ±0.3 361 ±2 226 ±6

Concentrate end-product 28.4 ±0.3 3650 ±10 1587 ±1

Concentration Factor 18 10 7

H. sabdariffa

Clarified permeate 54 ±2 77 ±13 not detected

Concentrate end-product 503 743 ±1 not detected

Concentration Factor 9 10 not calculated

Processing plant samples of C. procera and H. sabdariffa lead to deep-red concentrated polyphenolic extracts that contained anthocyanins and phenolic-acids for both plant extracts, and flavonol glycosides, for C.

procera only. The process clarified and concentrated crude extracts 10-20

times for anthocyanin, 10 times for phenolic acid, and 7 times for flavonol compounds.

Anthocyanin contents of H. sabdariffa concentrated end-products were 17 times more concentrated than those of C. procera, which showed higher concentrations in phenolic acids (5 times) and in flavonols.

 RESULTS and DISCUSSION

Calyx of H. sabdariffa and leaves of C. procera were harvested in the centre part of Ivory Coast, around the Yamoussoukro area. The harvested samples were locally dried at 40°C and sent to CIRAD-Fr. A pilot plant multi-step process was applied to non-crushed plant samples to obtained concentrated water-extracts.

Polyphenol extracts were analyzed by HPLC-DAD (Agilent 1100 model equipped with a RP 18 column: 250x4.6 mm, 5 µm) and their chemical structures were determined by LC-MS2_{-ESI. AntiOxidant Capacities (AOC) of the plant extracts obtained during the process were measured as Trolox} eq. by 2 different methods: DPPH and ABTS.



PROCESSING AND ANALYSES

 CONCLUSION

Hibiscus Carap

a

International Congress on Polyphenols – ICP-2010, Montpellier, France

 REFERENCES

--- C. procera processed extracts

---Anthocyanins generally showed lower AOC than othe r

polyphenolic families. C. procera extracts showed the strongest AOC due to flavonol and phenolic acid contents, that were found higher than in H. sabdariffa extracts.

OH OH O O COOH HO HO OH 1 6 5 4 3 2 COOH HO HO Protocatechuic acid 3-caffeoylquinic acid

Quercetin 3-O-rutinoside : R = rutinose Quercetin 3-O-galactoside : R = galactose Quercetin 3-O-glucoside : R = glucose

Anthocyanins Phenolic acids Flavonols

OH OH O O COOH HO HO OH 1 6 5 4 3 2 COOH HO HO Protocatechuic acid 3-caffeoylquinic acid

Anthocyanins Phenolic acids

O H O H C O O H H O O O H 1 6 5 4 3 2 O H O 5-caffeoylquinic acid OH OH OH COOH HO HO O 1 6 5 4 3 2 O 4-caffeoylquinic acid O O OH OH HO OH R O O+ OH R2 OR O H OH R1 O+ OH R2 OR O H OH R1

Delphinidin 3-O-glucoside : R1=OH, R2=OH, R=glucose Delphinidin 3-O-sambubioside : R1=OH, R2=OH, R=sambubiose Cyanidin 3-O-glucoside : R1=H, R2=OH, R=glucose Cyanidin 3-O-sambubioside : R1=H, R2=OH,R=sambubiose Cyanidin 3-O-rutinoside : R1=H, R2=OH, R=rutinose Cyanidin 3-O-glucoside : R1=H, R2=OH, R=glucose

Cyanidin 3-O-rutinoside : R1=H, R2=OH, R=rutinose

0 5 10 15 20 Crude extract Clarified permeate Concentrated end-product AOC (mM TEAC) ABTS Carapa DPPH Carapa ABTS Hibiscus DPPH Hibiscus RO Ultrasound- Assisted Extraction Nylon cloth Prefiltration Cross-Flow Microfiltration Reverse Osmosis Concentration Plant Material

Ultrasounds pilot plant capacity = 30 L Plant/water ratio =1/100 , w/v

Process

Flow diagram Raw material

Products

line

Crude Extract _{clarified permeate}

(slight red colour)

Concentrate

(deep red colour)

Ceramic membrane P19-60 / 0.2 µm Polymeric membrane SW 30 C. procera

leaves

H. sabdariffa

flowers

Volume Reduction Factor = 20-25 times

--- H. sabdariffa processed extracts

---OH OH OH COOH HO HO O 1 6 5 4 3 2 O OH OH OH COOH HO HO O 1 6 5 4 3 2 O