Using a long term experiment (20 and 11 years of organic cultivation on the same soil), sunflower was cultivated on these soils under organic management and in a different part of the same farm under conventional management. Kernels, teguments and oils were analyzed for their polyphenols content. Five caffeoylquinic acids were identified. No qualitative differences were found in the three cases, while quantitative differences have been pointed out and discussed

Heimler, D.

Moschini, V.

Pinelli, P.

Romani, A.

Firenze University Press: E-Journals

85
1. Introduction
Sunflower (Helianthus annuus L.) is one of the
most important oilseed crops. Besides soy and rape-
seed oil, sunflower oil is ranking third with a world-
wide production of about 44 million tons each year
from 2012 to 2016, 28 mill ion tons in the EU
(Committee for the Common Organization of
Agricultural Markets, 2016). Sunflowers prove to be a
protein source of great interest for human nutrition;
moreover, the residues originating from oil extrac-
tion are rich in phenolic antioxidants, which account
for 1-4% of the total mass with chlorogenic acid
being the predominant component (Weisz et al.,
2009). Polyphenols in sunflower seeds were identi-
fied and quantified after HPLC analysis (Aramendia et
al., 2000; Pedrosa et al., 2000). The main phenolic
compounds present in both the kernel and hull
besides chlorogenic acid are caffeic acid and caf-
feoylquinic derivatives. Phenolic compounds in sun-
flower seeds have been shown to exert a high antiox-
idative potential, which might be beneficial from a
biofunctional point of view and may be used as effec-
tive antioxidants for sunflower oils (De Leonardis et
al., 2003, 2005; Anjum et al., 2012).
Organic agriculture is most widely used and its
benefits concern overall the environment and the
health of food, which is not contaminated with pesti-
cides and synthetic fertilizers. However organic fertil-
izer of biological origin may lead in the long term to
the “conventionalization of organic farming”
(Darnhofer et al., 2010). In order to go deeper in the
organic management, the Montepaldi Long Term
Experiment (MOLTE) trial in central Italy has been set
up to compare three agro-ecosystems with different
management: two organic (old organic since 1992
and young organic since 2001) and one conventional
(Migliorini, 2014). The aim of this research is to fol-
low the fate of polyphenols in teguments, kernels
and oil from sunflower seeds grown under the
above-mentioned three different conditions (one
conventional and two organic) in order to assess
whether such condition may affect the polyphenols
profile and which compounds may be regarded as
innovative parameters in order to deeper investigate
the relation among soil condition, agricultural man-
agement and seed characteristics.
2. Materials and Methods
The Montepaldi Long Term Experiment (MOLTE)
has been active since 1991 in the farm of the
University of Florence (location Montepaldi, San
Adv. Hort. Sci., 2017 31(2): 85-88 DOI: 10.13128/ahs-20608
Seeds and oil polyphenol content of sunflower (Helianthus
annuus L.) grown with different agricultural management
A. Romani 1, P. Pinelli 1, V. Moschini 2, D. Heimler 2 (*)
1 Dipartimento di Statistica, Informatica, Applicazioni “G. Parenti”, Università degli Studi di Firenze, Viale
Morgagni, 40, 50134 Firenze, Italy.
2 Dipartimento di Scienze delle Produzioni Agroalimentari e dell’Ambiente, Università degli Studi di Firenze,
Piazzale delle Cascine, 18, 50144 Firenze, Italy.
Key words: organic management, polyphenol content, sunflower.
Abstract: Using a long term experiment (20 and 11 years of organic cultivation on the same soil), sunflower was cultivat-
ed under organic management and in a different part of the same farm under conventional management. Kernels, tegu-
ments and oils were analyzed for their polyphenols content. Five caffeoylquinic acids were identified. No qualitative dif-
ferences were found in the three cases, while quantitative differences have been pointed out and discussed.
(*) Corresponding author: daniela.heimler@unifi.it
Received for publication 2 May 2017
Accepted for publication 15 May 2017
Copyright: © 2017 Author(s). This is an open access article distributed under the terms of the Creative Commons
Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, 
provided the original author and source are credited.
Adv. Hort. Sci., 2017 31(2): 85-88
86
Casciano, Val di Pesa, Long. 11°09’08’’ E, Lat.
43°40’16’’ N) covering a slightly sloping surface of
about 15 ha at 90 m a.s.l. The MOLTE experiment has
a system approach and includes the following three
different micro agro-ecosystem managements:
a) “Old Organic” (OldO) with an area of 5.2 ha, divid-
ed into 4 fields under organic since 1992;
b) “YoungOrganic” (YngO) with an area of 5.2 ha,
divided into 4 fields (integrated farming) from
1992 to 2000 and converted into organic manage-
ment since 2001;
c) “Conventional” (Conv) area of 2.6 ha divided into
2 conventional fields, where farming techniques
used were those normally used in the territory of
the study area for conventional management.
On October 28, 2013, after harvesting the whole
sunflower plant, seeds were put in an oven at 60°C
for 48 hours and then preserved in paper bags. Seeds
were manually dehulled in order to obtain kernel and
tegument. Phenolic compounds were extracted for
30 min from kernels (about 2 g) and from teguments
(about 1 g) with 70:30 ethanol/water (25 and 15 ml,
respectively). The extracts were evaporated under
vacuum at room temperature and finally dissolved in
10 ml ethanol/water (70:30). Cold-pressed seeds oils
(25 ml) were extracted with 3x25 ml of 70:30
ethanol/water, adjusted to pH 2.0 with formic acid;
each step involved an extraction for 30 min at room
temperature. The extracts were combined and defat-
ted with 3x50 ml hexane. The defatted extracts were
evaporated to dryness under vacuum at room tem-
perature and finally dissolved in ethanol/water
(70:30) to a final volume of 4 ml.
Solvents and reagents
All the solvents (HPLC grade) and formic acid (ACS
reagent) were purchased from Aldrich Chemical
Company Inc. (Milwaukee, Wisconsin, USA).
Chlorogenic acid was obtained from Extrasynthese
S.A. (Lyon, Nord-Genay, France). The HPLC-grade
water was obtained via double-distillation and purifi-
cation with a Labconco Water Pro PS polishing sta-
tion (Labconco Corporation, Kansas City, USA).
HPLC analysis
HPLC/DAD Analysis. The HPLC/DAD analyses were
performed with an HP 1100L liquid chromatograph
equipped with HP DAD (Agilent Technologies, Palo
Alto, CA, USA). A Kinetex C18 column 100×2.1 mm, 5
μm (Phenomenex) operating at 30°C was used. The
eluents were H2O adjusted to pH 3.2 by formic acid
and acetonitrile. A four-step linear solvent gradient
was performed starting from 100% water up to 100%
acetonitrile, with a flow rate of 0.2 ml/min for a 30-
min period (Table 1).
HPLC/ESI-MS Analysis. The HPLC-MS analyses
were performed using an HP 1100L liquid chromato-
graph equipped with a DAD and 1100 MS detectors.
The interface was an HP 1100 MSD API-electrospray
(Agilent Technologies). Mass spectrometer operating
conditions were the following: gas temperature
350°C at a flow rate of 10.0 l/min, nebulizer pressure
30 psi, quadrupole temperature 30°C and capillary
voltage 3500 V. The mass spectrometer operated in
positive and negative ionization mode at 80-120 eV,
for both ionization modes.
3. Results and Discussion
In figure 1, the chromatographic profile of organic
sunflower kernel, registered at 330 nm, maximum
wavelength of absorbance of caffeic acid and its
derivatives, is reported. Eight compounds have been
identified: 3-O-caffeoylquinic acid, chlorogenic acid,
4-O-caffeoylquinic acid, three derivatives of caffeic
acid, 3,5-O-dicaffeoylquinic acid, and 4,5-O-dicaf-
Table 1 - Linear solvent gradient system used in HPLC-DAD and
HPLC-MS analysis
Time (min) A B
0.10 100.0 0.0
5.00 80.0 20.0
7.00 80.0 20.0
13.00 70.0 30.0
18.00 70.0 30.0
22.00 30.0 70.0
26.00 30.0 70.0
30.00 0.0 100.0
Fig. 1 - HPLC chromatogram of sunflower kernel extract record-
ed at 330 nm. Peaks= 1. 3-O-caffeoylquinic acid; 2.
chlorogenic acid; 3. 4-O-caffeoylquinic acid; 4-6. caffeic
acid derivatives; 7. 3,5- O-dicaffeoylquinic acid; 8. Ac.
4,5- O-dicaffeoylquinic acid.
Solvent A=  H2O adjusted to pH 3.2 by HCOOH. Solvent B= CH3CN.
Romani et al. - Seeds and oil polyphenol content of sunflower 
87
feoylquinic acid. In the shells the same compounds
with the exception of 4-O-caffeoylquinic acid and the
two dicaffeoylquinic acids were identified. The same
caffeoyl and dicaffeoylquinic acid derivatives were
already found by Aramendia et al. (2000), Pedrosa et
al. (2000), and Weisz et al. (2009). The qualitative
pattern does not change along with the agricultural
management. As concerns quantitative data, Table 2
reports the quantitative data of kernel and tegu-
ments. Total polyphenols amount is the lowest, in
the case of kernel, when the plant grows on the old
organic field, while it is almost the same with the
other two managements; in the case of teguments,
on the contrary, the lowest amount is observed in
the case of conventional management. It should
however be noted that in the case of teguments,
polyphenols content is very low changing from about
10% to 2% with respect to kernel content.
Chlorogenic acid is the main compound and it
accounts for about 90% in tegument. Chlorogenic
acid in our samples, both kernel and tegument, is
about ten times more abundant with respect to the
quantitative data reported by Aramedia et al. (2000)
and Pedrosa et al. (2000). The comparison of quanti-
tative data is however difficult since different tech-
niques and sample treatments are involved (Weisz et
al., 2009). Polyphenols content along with agriculture
management has recently reviewed and in most veg-
etables grown under organic conditions, a higher
content of polyphenols has been found; on the other
hand a higher soil nitrogen availability decreases
polyphenols content (Heimler et al., 2017). Our data
support previous findings that indicate how old man-
aged organic soil was the most efficient in term of C
and N storage (Migliorini et al., 2014). Furthermore,
when the individual compounds are taken into
account the relative percentages of compounds from
new organic and conventional managed soils are sim-
ilar, while in the case of the old managed organic soil,
notwithstanding the lowest total polyphenol content,
a higher dicaffeoylquinic acids content was found
(17% with respect to 12%). Chlorogenic acid and
dicaffeoylquinic acids derive from the phenyl-
propanoid pathway. Generally, this pathway is
induced by biotic and abiotic stress such as wound-
ing, UV irradiation, or pathogen attack (Moglia et al.,
2008). No information is available on the regulation
of dicaffeoylquinic acids in any plant species, even if
the same regulation of chlorogenic acid synthesis
could be foreseen (Moglia et al., 2008).
In Table 3, the data of sunflower oil are reported.
Oil has been obtained by means of a mechanical
OldO= old organic; YoungO= young organic; CO= conventional (see experimental section). ND= not determined. Standard deviation
within brackets.
Table 2 - Polyphenols (mg/g, fresh weight) in sunflower kernel and tegument
Compound
Agricultural management
Kernel Tegument
OldO YoungO CO OldO YoungO CO
3-O-caffeoylquinic acid 2.51(0.37) 4.57(0.82) 4.74(0.76) 0.08(0.01) 0.08(0.01) 0.05(0.01)
chlorogenic acid 6.25(1.12) 11.39(1.94) 11.82(1.42) 0.95(0.18) 0.96(0.18) 0.41(0.07)
4-O-caffeoylquinic acid 0.44(0.08) 0.8(0.11) 0.83(0.13) 0.03(0.005) traces traces
caffeic acid derivatives 0.21(0.04) 0.38(0.04) 0.39(0.06) traces n.d. traces
3,5-O-dicaffeoylquinic acid 1.3(0.24) n.d. n.d. n.d. n.d. n.d.
4,5-O-dicaffeoylquinic acid 0.77(0.11) 2.36(0.34) 2.45(0.27) n.d. n.d. n.d.
Total polyphenols 11.48 19.5 20.23 1.06 1.04 0.46
OldO= old organic; YoungO= young organic; CO= conventional (see experimental section). ND= not determined. Standard deviation
within brackets.
Table 3 - Polyphenols (mg/l) in sunflower oil
Compound
Agricultural management
OldO YoungO CO
3-O-caffeoylquinic acid 0.232(0.192) 0.35(0.09) 0.176(0.057)
chlorogenic acid 1.96(0.393) 2.43(0.11) 1.936(0.12)
p-coumaroylquinic acid 2.36(0.318) 0.112(0.022) 0.592(0.079)
4-O-caffeoylquinic acid 0.048(0.005) 0.104(0.011) 0.08(0.02)
caffeic acid derivatives 2.28(0.644) 0.328(0.079) 3.216(0.415)
not identified polar compounds 0.504(0.011) 1.2(0.045) 1.456(0.429)
Total polyphenols 7.384 4.368 7.456
Adv. Hort. Sci., 2017 31(2): 85-88
88
press, which is the only technology that allows the
maintenance of polyphenols high content (Bendini et
al., 2011). Oil yields are different (22.2% for old
organic soil, 29.8% for young organic soil and 24.7%
for conventional management) and, the young
organic soil, with the highest yield, exhibits the low-
est total polyphenol content. Sunflower oil-amount
of 10 mg kg-1 has been reported (De Leonardis et al.,
2005). Polyphenols content of the three seeds oils
(Table 3) are almost in accordance with such amount.
Young managed organic soil gave rise to an oil with
the smallest polyphenols content notwithstanding
the high polyphenols content of almond and tegu-
ments.
Acknowledgements
The authors acknowledge the Regione Toscana
‘‘PRAF 2012-2015 MISURA 1.2 e)’’ program (Project
VOLATOSCA).
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Seeds and oil polyphenol content of sunflower (Helianthus annuus L.) grown with different agricultural management

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