Beruflich Dokumente
Kultur Dokumente
CIRAD, UMR GPEB Gnie des Procds, Eau et Bioproduits, TA 40/16, 73 Avenue J.F. Breton, 34398 Montpellier Cedex 5, France
INP-HB Institut National Polytechnique Flix Houphout-Boigny, Laboratoire de Procds Industriels, de Synthse et de lEnvironnement (LAPISEN),
Unit Chimie de lEau et des Substances Naturelles, BP 1313 Yamoussoukro, Cote dIvoire
c
Universit dAbidjan-Cocody, UFR Biosciences, Laboratoire de Biochimie et Sciences des Aliments, 22 BP 582 Abidjan 22, Cote dIvoire
d
Universit dAvignon et des pays du Vaucluse, INRA, UMR A 408, Scurit et Qualit des produits dorigine vgtale, F-84000 Avignon, France
e
Institut des Sciences Molculaires de Marseille (ISM2), Equipe AD2EM (Groupe Phytochimie), UMR 6263, Universit Paul Czanne,
Avenue Escadrille Normandie Nimen, 13397 Marseille Cedex 20, France
b
a r t i c l e
i n f o
Article history:
Received 9 February 2010
Received in revised form 8 June 2010
Accepted 9 June 2010
Keywords:
Anthocyanins
Flavonols
Polyphenols
Extraction
Ultrasound
Maceration
Delonix regia
Biodiversity
a b s t r a c t
Flowers of Delonix regia trees from Ivory Coast are traditionally macerated in water to prepare beverages
with benecial health properties mainly due to their polyphenolic contents. Different maceration procedures, mimicking the traditional process, were compared on the basis of polyphenol content followed by
HPLC-DAD analysis. Under optimized maceration ratio (R = plant sample weight/maceration media volume), plant extractions were compared, including acidied-water-based media and assisted-ultrasound
or stirring extraction procedures. Diffusion kinetics of polyphenol families (anthocyanins, avonols and
phenolic acids) were followed by HPLC and lead to nd optimum ratio (R = 1/100). Flavonol contents were
28.5, 31, and 33.5 mol g1 (expressed as quercetin equivalent), using acidied-water (0.01N) media with
sulphuric or citric acids. Anthocyanin and phenolic acid contents were within the same range for these
water media (5.6 mol g1 as cyanidin equiv. and 27.5 mol g1 as gallic acid (GA) equiv., respectively).
Ultrasound-assisted procedure shortened maceration time (three times shorter) but did not show higher
level of total polyphenol than stirring-assisted procedure (39 mol g1 GA equiv.). This eco-friendly and
low-cost extraction process will provide to African people, with safer quality and longer availability,
polyphenol-rich bio-products.
2010 Elsevier B.V. All rights reserved.
1. Introduction
Polyphenol compounds, including anthocyanins, avonols, and
phenolic acids, are among the most bioactive natural molecules
found in the plant kingdom because of their antioxidant activities
(Bruneton, 2002; Teow et al., 2007; Duh and Yen, 1997; Einbond et
al., 2003; Shahidi, 1997). Plant anthocyanins are also well-known
compounds for their functional property as natural colorants (Espin
et al., 2000; Mazza et al., 2004). In tropical countries, particularly in
Ivory Coast, Delonix regia (Flame tree), also called locally Flamboyant, is often used for traditional medicine (Ali et al., 1999)
prepared from various parts of the tree. Red owers contain anthocyanins (Nabieh and Moheb, 1976), which are generally extracted
at lab scale using hydro-alcoholic solvents (Campos et al., 2008;
Chirinos et al., 2007; Neungnapa et al., 2008), and at pilot plant
scale using acidied-water media (Adj et al., 2008). During the
440
441
Table 1
Polyphenol contents of dried D. regia ower extracts obtained under M1/R3 and
M2/R3 extraction conditions.
Family compounds
c
Anthocyanins
Flavonolsd
Phenolic acidse
a
b
c
d
e
Fig. 1. Effect of dried powdered D. regia ower weight versus volume of acidiedwater media on total polyphenol extraction (mol L1 as gallic acid (GA) equiv.)
using maceration without stirring at room temperature (R1 = 1/250, R2 = 1/200,
R3 = 1/100, R4 = 2/100, R5 = 4/100 g mL1 , extraction conditions: M1: watersulphuric
acid 0.005 mol L1 media and M2: watercitric acid 0.0033 mol L1 media, P1:
powdered-ower maceration 3 h maceration time).
M1/R3a
M2/R3b
5.5 0.2
28.5 0.8
26.5 0.7
5.6 0.2
31.0 0.9
28.5 0.8
Fig. 2. HPLC proles of polyphenol water-extracts of D. regia dried owers (Extraction conditionsM2/R3/P1: watercitric acid medium/R3 = 1/100 g mL1 /powdered-ower
maceration, A1: cyanidin 3-O-glucoside, A2: cyanidin 3-O-rutinoside, F1: rutin, F2: quercetin-3--d-glucoside, AC1: gallic acid and AC2: protocatechuic acid).
442
Fig. 3. Polyphenol contents of D. regia extracts obtained with P1/M1 and P1/M2 using various R ratios (mean of duplicate) (P1: powdered-ower maceration, M1:
watersulphuric acid medium, M2: watercitric acid medium, () anthocyanins as cyanidin equiv.; other polyphenols: () phenolic acids as gallic acid equiv., () avonols
as quercetin equiv.).
180 min maceration time, then decreased, slowly for anthocyanins and drastically for avonols. The maximum concentration
obtained was 5.5 mol g1 as cyaniding equiv. for anthocyanins
and 33.5 mol g1 as quercetin equiv. for avonols.
Liu et al. (2000) have reported that solvents with high polarity
and low strength, such as water, are not efcient to correctly extract
plant polyphenols. Comparing polyphenol contents of extracts (P1)
obtained with M3/R3 (alcohol-based medium) with M1/R3 and
M2/R3 (water-based media), it was found that anthocyanin contents were about the same amount (5.55.6 mol g1 (as cyaniding
equiv.)) for all media, and avonols contents slightly increased
from 28.5 up to 31 mol g1 (as quercetin equiv.), for M1/R3M2/R3, to 33.5 mol g1 (as quercetin equiv.), for M3/R3. Our
results showed that the 2 water-based maceration media, M1 and
M2, lead to extracts with the same polyphenol contents than those
of the alcohol-base maceration medium, M3, using in all cases
the same R3 ratio. It clearly appeared that polyphenol extraction,
using plant maceration in water-based media will be preferred
to methanol-based media to process large quantities of plant
polyphenol extracts. Water-based extraction will be suggested
for up-scaling extraction of natural and health-benet polyphenol
compounds from D. regia owers at pilot plant level for small-scale
local enterprises.
3.3. Procedure efciency within M1/R3 and M2/R3 maceration
conditions
Extraction polyphenol from D. regia dried owers (plant material B) were compared using 3 extraction procedures, including
extraction by simple maceration P2, stirring-assisted P3 and
Table 2
Effect of extraction procedures on total polyphenol content in dried D. regia ower
extracts.
Extraction proceduresa
M1/R3b
M2/R3b
Total polyphenolc
P4 (UAE)
P3 (mechanical stirring)
P2 (maceration)
39 2
40 1
27 1
33 2
39 2
26 1
a
P2: entire ower maceration, 3 h extraction time; P3: entire ower extraction
with mechanical stirring, 3 h extraction time; P4: entire ower extraction assisted
by ultrasounds, 1 h extraction time.
b
Extraction conditionsM1: watersulphuric acid medium; M2: watercitric
acid medium, R3 = 1/100 g mL1 .
c
mol g1 as gallic acid equiv.
443
Fig. 5. Variation of polyphenol content (Ct ) in dried D. regia ower extracts (a) and rst order law plotting kinetic (b) (Extraction conditions: M1/R3 and ultrasound-assisted
procedure P4).
equiv.) than P3, but P2 did not perform well (26 1 mol g1 GA
equiv.).
With both extraction conditions, P3 was found to have the highest polyphenol extraction capability among all procedures tested.
However, extraction time to reach the maximum polyphenol content was shorter for P4 (1 h) than for the 2 others (3 h), whatever
the maceration media used. Maximum level of polyphenol contents
reached by the 3 procedures was about 40 mol g1 GA equiv. On a
process duration basis, P4 appeared to be the most suitable extraction procedure for both maceration conditions (M1/R3 and M2/R3)
to obtain optimal polyphenol content in the shortest time. P4 did
not enhance polyphenol extraction yields, but only shortened drastically extraction duration time.
Moreover, Table 2 shows that maceration conditions M1/R3 are
better than M2/R3 on a basis of total polyphenol content of plant
extracts. So, sulphuric acid will be preferred to citric acid for acidication of water maceration medium.
Polyphenol content was monitored during extraction process
with P4/M1/R3 conditions. The kinetic curve of total polyphenol
content with time is given in Fig. 5a.
It was shown that ultrasound-assisted extraction of plant
polyphenol was a linear representation of the polyphenol content
versus time during the course of extraction, by plotting Ln(A) versus
time for the rst 30 min, where A = (C Ct )/(C C0 ) and C0 , Ct ,
and C are respectively extract polyphenol contents at start time t0 ,
at t, and at t , which was in reality set at 1 h extraction
time. According to Fick rst law, A = (C Ct )/(C C0 ) =
a exp (kt),
i=1
this relation can be transformed into the linear equation ln(A) = kt.
Plotting experimental data according to this last equation lead to
determine k values for every extraction conditions tested, which
was represented by the slope of the line obtained with a good
regression coefcient (R2 = 0.99) (Fig. 5b). This coefcient is also the
diffusivity Deff = 48 103 min1 of polyphenols in this maceration
conditions (P4/M1/R3).
For every other extraction conditions studied, a similar linear
regression was found, as shown in Table 3. Polyphenol content in
the extracts still followed a rst order kinetic law for the rst 30 min
maceration with P4/M1/R3 conditions and for the rst 60 min maceration using P2 and P3 procedures. One could notice that Deff
was found at least more than twice higher with P4 procedure
(Deff = 3545 103 min1 ) than with P3 and P2 procedures tested
(Deff = 821 103 min1 ).
These results raised the fact that ultrasound technology added
in P4 procedure increased the specic polyphenol Deff . The total
Mediumb
Equation
Regression coefcient
P4
M1
M2
Y = 0.048X + 0.060
Y = 0.035X + 0.021
r2 = 0.991
r2 = 0.988
P3
M1
M2
Y = 0.021X 0.039
Y = 0.020X + 0.01
r2 = 0.993
r2 = 1
P2
M1
M2
Y = 0.011X 0.118
Y = 0.008X 0.08
r2 = 0.91
r2 = 0.975
a
P2: entire ower maceration; P3: entire ower extraction with mechanical stirring; P4: entire ower extraction assisted by ultrasounds.
b
M1: watersulphuric acid 0.005 mol L1 media; M2: watercitric acid
0.0033 mol L1 medium.
Table 4
Comparative extraction efciency of polyphenols using maceration, stirring and
ultrasound procedures in either sulphuric or citric acids (HPLC polyphenol
quantications).
M1/R3a
Anthocyaninsc
Flavonolsd
Phenolic acidse
M2/R3a
P2b
P3b
P4b
P2b
P3b
P4b
1.3
10.1
40.2
1.4
28.9
39.4
2.0
12.5
42.6
1.3
1.1
25.5
3.7
11.0
27.0
3.2
14.0
26.5
a
Extraction conditionsM1: watersulphuric acid medium; M2: watercitric
acid medium, R3 = 1/100 gm L1 .
b
ProceduresP2: entire ower maceration; P3: entire ower extraction with
mechanical stirring; P4: entire ower extraction assisted by ultrasounds.
c
mol g1 as cyanidin equiv., max std error 7%.
d
mol g1 as quercetin equiv., max std error 5%.
e
mol g1 as gallic acid equiv., max std error 5%.
444
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