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Thermal stability, antioxidant activity, and photo-oxidation of natural

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Article  in  Chemical Papers- Slovak Academy of Sciences · January 2014

DOI: 10.2478/s11696-013-0417-6

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 Chemical Papers 68 (1) 121–129 (2014)
DOI: 10.2478/s11696-013-0417-6

ORIGINAL PAPER

Thermal stability, antioxidant activity, and photo-oxidation

of natural polyphenols

Irina Volf, Ioana Ignat*, Mariana Neamtu*, Valentin I. Popa

y
Faculty of Chemical Engineering and Environmental Protection, “Gheorghe Asachi” Technical University of Iasi,
73 Prof. dr. docent Dimitrie Mangeron Bd., 700050 Iaşi, Romania

op
Received 12 January 2013; Revised 25 March 2013; Accepted 5 April 2013

The thermal stability (60 ◦C, 80 ◦C, 100 ◦C), antioxidant activity, and ultraviolet C light (UV-C)
stability of standard polyphenols solutions (catechin, gallic acid, and vanillic acid) and of vegetal
rc
extracts from spruce bark and grape seeds were investigated. Exposure of the standard solutions
and vegetal extracts to high temperatures revealed that phenolic compounds were also relatively
stable (degradations ranged from 15 % to 30 % after 4 h of exposure). The highest antioxidant
activity was obtained for ascorbic acid and gallic acid followed by catechin and caffeic acid and the
grape seeds. The results show that, after 3 h of UV-C exposure, approximately 40 % of vanillic acid,
50 % of gallic acid, and 83 % of catechin were removed. Similar degradation rates were observed
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for vegetal extracts, with the exception of the degradation of catechin (40 %) from grape seeds. In
addition, the photo-oxidation of polyphenols in the presence of food constituents such as citric acid,
ascorbic acid, sodium chloride, and sodium nitrate was assessed.

c 2013 Institute of Chemistry, Slovak Academy of Sciences
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Keywords: polyphenols, photo-oxidation, thermal stability, radical scavenging activity, food addi-
tives

Introduction toalexins, anti-feedants, contributors to plant pigmen-

A

tation, antioxidants, and protective agents against ul-

There has recently been a considerable increase in traviolet (UV) light (Ignat et al., 2011a). Polyphe-
interest in finding naturally occurring antioxidants to nols have many industrial applications in fields such
replace synthetic antioxidants, some of which are be- as medicine, cosmetics, and the food industry. These
ing restricted due to their carcinogenicity. The use compounds may be used as natural colorants and
of natural antioxidants also has great potential as a preservatives for foods, or as additives in the pro-
result of consumers demanding additive-free, fresher, duction of paints, paper, cosmetics, and pharmaceu-
and more natural-tasting food (Muanda et al., 2011; tical products (Naczk & Shahidi, 2006; Giusti &
Díaz-García et al., 2013). Wrolstad, 2003). Most notably, the antioxidant ac-
Antioxidants are compounds that can delay or in- tivities of polyphenols are presumed to exert vari-
hibit the oxidation of lipids or other molecules by ous pharmacological effects such as anti-carcinogenic,
inhibiting the initiation or propagation of oxidising anti-mutagenic, and cardio-protective effects, linked to
chain reactions. These properties can play an impor- their free radical scavenging (Parr & Bolwell, 2000;
tant role in adsorbing and neutralising free radicals, Castañeda-Ovando et al., 2009; Díaz-García et al.,
quenching oxygen, or decomposing peroxides (Karou 2013).
et al., 2005). Phenolic compounds, one of the most In recent years, special attention has been focused
widely occurring groups of phytochemicals, are of con- on the isolation of phenolics from different raw materi-
siderable physiological and morphological importance als (medicinal plants, fruits, vegetables, industrial by-
in plants. Phenolics may act, among others, as phy- products, and beverages) and on exploration of their

*Corresponding author, e-mail: mariana.neamtu1@yahoo.de, ioana.ignat@gmail.com

122 I. Volf et al./Chemical Papers 68 (1) 121–129 (2014)

potential benefits for human health. To date, less attention has been paid to the stabil-
Although these raw materials have been exten- ity of polyphenolic compounds and their degradation
sively investigated for the isolation of polyphenols, under different conditions. However, these aspects can
special attention continues to focus on their extrac- influence their potential applications substantially and
tion from inexpensive or residual sources. It is well might elicit substantial interest in studying the photo-
known that the by-products from industrial processes oxidation and thermal degradation of phenolics.
still contain a considerable amount of phenolic com- In the present work, the antioxidant activity, ther-
pounds. Moreover, the large amounts of by-products mal stability, and photo-oxidation of standard solu-
resulting annually from vineyards and pulp and paper tions of polyphenols (gallic acid, catechin, vanillic
mills, along with the use of grape seeds and spruce acid) and natural polyphenols (spruce bark and grape
bark extracts as food supplements, and alternative seeds extracts) were investigated. The effects of food
medical products, warrant evaluation of their prop- constituents such as citric acid, ascorbic acid, sodium
erties. chloride, and sodium nitrate on the photo-oxidation
Polyphenols are widely seen as very unstable and of polyphenols were also evaluated.

y
highly susceptible to degradation (B˛akowska et al.,
2003). The stability of polyphenols under different Experimental
conditions is a very important aspect which has to be

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taken into account to ensure that phenolic compounds General
have the desired properties and maintain their activity
and structure during the different stages of processing, Spruce wood bark was provided by a Romanian
which can involve high temperatures, light, oxygen, pulp and paper company and Merlot grape seeds were
solvents, the presence of enzymes, proteins, metal- obtained from Panciu vineyard (Vrancea, Romania).
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lic ions, or association with other food constituents Standard polyphenols (gallic acid, catechin, vanil-
(Castañeda-Ovando et al., 2009). lic acid, siringic acid, p-cumaric acid, ferulic acid, and
UV holds considerable promise in relation to food sinapic acid) and methanol (HPLC grade) were pur-
processing as an alternative to traditional thermal chased from Sigma–Aldrich (UK). The other chem-
processing; however, UV treatment has received less icals and solvents used were of analytical grade ob-
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attention than other non-thermal processing methods. tained from Merck (Germany) and Sigma–Aldrich
Applications include pasteurisation of juices, post- and used without further purification. The mobile
lethality treatment for meats, treatment of food con- phases for HPLC and aqueous solutions containing
tact surface, and ways to extend the shelf-life of fresh 100 mg L−1 standards were prepared with ultrapure
produce (Koutchma, 2008, 2009; Tikekar et al., 2011a, water (conductivity of 0.056 ΩS cm−1 ) from a Milli-
2012). Any approach to evaluating UV entails con- pore Waters Milli Q purification unit (France).
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sideration of the properties and composition of the

food product to be treated, the source of the UV ra- Methods
diation, microbial effects, as well as the modelling,
commercial, and economic aspects (Koutchma, 2009). Vegetal extracts were obtained from grape seeds
A

The use of UV is well established for air, disinfection and spruce bark as raw materials. 50 g of dried ground
of water, and wastewater treatment. The wavelength material was extracted using distilled water as extrac-
of 253.7 nm used in this study is the most efficient tion solvent, at 70 ◦C in a water bath. The extrac-
in terms of its germicidal effect, since photons are tion was repeated three times, each time for approxi-
most readily absorbed by the DNA of microorgan- mately 2.5 h, and the extracts were combined and sub-
isms at this specific wavelength (Oppenlaender, 2003; jected to UV degradation. The initial concentration of
Koutchma, 2009). A significant reduction in the num- polyphenols was determined by HPLC analysis.
ber of spoilage and human pathogenic microorganisms The alcoholic extractions of the two raw materials
has been demonstrated through the use of UV process- were carried out in a Soxhlet installation, over 8 h
ing (Koutchma, 2009; Tikekar et al., 2011a). Ultravi- at 70 ◦C, using ethanol/water (ϕr = 7 : 3) as solvent.
olet C (UV-C) light processing is relatively less costly, Prior to the HPLC determinations, all the extracts
has minimal effects on product flavour, and is adapt- were concentrated under vacuum and fractioned by
able to continuous processing methods (Tikekar et al., successive liquid–liquid extractions with ethyl acetate.
2011a). The Food and Drug Administration (2000) The organic phases were evaporated to dryness and
regulations approved the use of low pressure mercury diluted in methanol, prior to HPLC determination.
(LPM) lamps for juice-processing (Koutchma, 2009). A previously developed reverse-phase high-perfor-
Little is known about the interaction of UV with com- mance liquid chromatographic method (Ignat et al.,
plex food matrices. Moreover, the effect of some essen- 2011b) was used to identify and quantify the phenolic
tial food additives such as vitamin C, nitrate ions and compounds. The HPLC analysis was performed using
salts on the absorption effects of antioxidants needs to a DionexUltiMate 3000 chromatograph (USA) cou-
be taken into account during UV treatment. pled to a PDA detector. Separations were carried out
 I. Volf et al./Chemical Papers 68 (1) 121–129 (2014) 123

on a Zorbax RX C18 (USA) (4.6 × 250 mm, particle

size 5 µm) column, operating at 30 ◦C with a flow-rate
of 1.2 mL min−1 . The injection volume was 5 µL. The
mobile phase used was 1 vol. % acetic acid in water (A)
vs. methanol (B) for a total run-time of 40 min. The
elution conditions were as follows: the vol. % of sol-
vent B was increased linearly from 10 to 40 in 40 min
and then decreased to 10 and maintained for 10 min.
For quantification, standards for external calibration
were used.
Total phenolic content (TPC) was determined us-
ing the Folin–Ciocalteau reagent, with a protocol de-
veloped previously (Hainal et al., 2011). Gallic acid
was employed as a calibration standard and the re-

y
sults were expressed as gallic acid equivalents (mg of
GAE per 100 g of dried material).
The radical scavenging activity of the natural

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extracts was evaluated using the reduction of the
di(phenyl)-(2,4,6-trinitrophenyl)iminoazanium (2,2-
diphenyl-1-picrylhydrazyl, DPPH) radical. The an-
tioxidant activity of the extracts was expressed as
EC50, an equivalent amount of an extract that neu-
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tralises 50 % of the radical. The colorimetric assay was
performed using a modification of the method devel-
oped by Almela et al. (2006).
For the DPPH assay, the alcoholic and aqueous
extracts were concentrated and freeze-dried to avoid Fig. 1. Typical chromatogram at 280 nm of standard polyphe-
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interference from the solvents. Thereafter, different nols (a), of spruce bark ethanolic extract (b), of grape
dosages of a 0.5 mg mL−1 methanolic solution of ei- seeds ethanolic extract (c). Identified compounds: 1 –
ther aqueous or ethanolic freeze-dried extracts (25 µL, gallic acid; 2 – catechin; 3 – vanillic acid; 4 – syringic
50 µL, 100 µL, 200 µL, 300 µL, 400 µL, 500 µL acid; 5 – p-coumaric acid; 6 – ferulic acid; 7 – sinapic
acid.
each) were added to screw-capped glass vials contain-
ing 2 mL of the DPPH. All the volumes were adjusted
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to 3.1 mL with MeOH. After a reaction time of 30 min,

the absorbance was measured at 517 nm. The inhi- temperatures (60 ◦C, 80 ◦C, and 100 ◦C) for 4 h. Sam-
bition percentage of the free radical DPPH (I ) was ples were taken first after 30 min and every hour sub-
calculated according to the following equation: sequently and analysed by HPLC in order to establish
A

 the concentration of polyphenols previously exposed

A0 − A
I= × 100 % (1) to temperature treatment.
A0 Analysis of variance or R2 values was used to ver-
Methanolic solutions of ascorbic acid, gallic acid, ify the statistical significance between the treatments
caffeic acid, and catechin were tested as reference an- using Microsoft Excel (version 2007). Each data point
tioxidants. The different quantities of the extracts represents the average of three measurements ± stan-
tested, expressed in micrograms, were plotted on a dard deviation (5 %).
dose-inhibition curve.
The photo-degradation experiments were carried Results and discussion
out in a stirred-batch photo-reactor (volume of irradi-
ated solution = 500 mL, optical path length = 4 cm) at Identification and quantification of polyphe-
298 K. The lamp was located on the central axis of the nols by HPLC
reactor, in a quartz sleeve. The aqueous solutions, pre-
pared as above, were irradiated with a UV-immersed The chromatographic profiles of the standards and
low-pressure Heraeus mercury lamp TN 15/32 (Her- vegetal extracts are shown in Fig. 1.
aeus Nobelight, Germany) with a nominal output of The analytical polyphenolic composition of the
15 W, attaining its spectrum at the 254 nm line. The samples is given in Table 1; it should be noted that
incident photonic flux (P0 = 1.013 × 10−5 E s−1 ) was the major compounds identified in the aqueous and
measured by hydrogen peroxide actinometry. ethanolic fractions for both vegetal materials were gal-
The stability of standard solutions of polyphenols lic acid and catechin. Furthermore, in the spruce bark
and vegetal extracts was evaluated at three different extract, vanillic acid was also identified in relatively
124 I. Volf et al./Chemical Papers 68 (1) 121–129 (2014)

Table 1. Concentration of phenolic compounds (mg per 100 g of dried plant) in samplesa investigated

Raw material Type of extract Gallic acid Catechin Vanillic acid TPC/(mg of GAE per 100 g−1 )

Grape seeds Aqueous extract 6.12 ± 0.20 44.36 ± 0.10 – 506 ± 5

Ethanolic extract 12.54 ± 0.80 63.60 ± 1.70 – 1368 ± 14

Spruce bark Aqueous extract – 31.00 ± 1.90 39.40 ± 0.20 517 ± 5

Ethanolic extract 10.20 ± 0.30 71.90 ± 2.70 71.90 ± 0.80 1355 ± 12

a) Results represent average values of triplicate determination (n = 3) ± standard deviation. Caffeic acid, siringic acid, p-cumaric
acid, ferulic acid, and sinapic acid were not detected in the studied samples.

high concentrations. As expected, the concentrations Table 2. EC50 values obtained for standard compounds and
of the compounds were considerably higher in the al- vegetal extractsa
coholic extracts. Table 1 also shows the total phenolic
Samples EC50/µg R2

y
content; the high concentration values (up to 1368 mg
GAE 100 g−1 ) indicate that the identified compounds Ascorbic acid 4.60 ± 0.87 0.998
represent only a small percentage (from 9.6 % to 11 %) ±

op
Gallic acid 1.70 0.32 0.907
of the total phenolic compounds detected. Similar con- Caffeic acid 42.10 ± 1.02 0.993
centrations were reported by Neo et al. (2008) and Catechin 12.70 ± 0.98 0.999
Kirca and Arslan (2008). Grape seeds alcoholic extract 45.75 ± 1.09 0.994
Spruce bark alcoholic extract 159.15 ± 3.21 0.983
Grape seeds aqueous extract 76.25 ± 1.12 0.995
Radical scavenging activity Spruce bark aqueous extract 246.00 ± 5.56 0.997
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The radical scavenging activity of natural extracts a) Values are expressed as means ± SD of three replicate anal-
and pure compounds was evaluated by DPPH assay. yses.
The antioxidants react with the stable free radical, i.e.,
1,1-diphenyl-2-picrylhydrazyl (deep violet colour) and
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convert it to 1,1-diphenyl-2-picrylhydrazine accompa- recorded. The antioxidant activity of the plant ex-
nied with discoloration. The degree of discoloration tracts is correlated with their phenolic content (Mon-
indicates the free radical scavenging potentials of the toro et al., 2006). The DPPH assay indicated that the
sample/antioxidant (Sarikurkcu et al., 2008). grape seeds’ alcoholic extracts had high radical scav-
Determination of an absolute value for the antioxi- enging activities, which could be attributed to high
dant activity of an extract is problematic because it is levels of polyphenols (considerable amounts of gallic
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dependent on the actual concentration of the radical, acid and catechin). The results are strongly correlated
its degradation during the analysis or the matrix inter- with the total phenolic content and the HPLC deter-
ference. Accordingly, the EC50 parameter was used, minations. The alcoholic extracts show a higher rad-
which represents the equivalent amount of an extract ical scavenging activity than the aqueous ones, but a
A

that neutralises 50 % of the radical. slightly lower one than the standard compounds.
The radical scavenging activity of alcoholic and
aqueous extracts was compared with that of some Photo-oxidation
standard polyphenols (gallic acid, catechin, caffeic
acid), as well as with ascorbic acid, one of the synthetic The experiments were carried out with solutions of
antioxidants commonly used in the food industry. polyphenols (gallic acid, catechin, and vanillic acid) in
The reduction in absorbance was measured at ultrapure water and also with spruce bark and grape
517 nm for different quantities of standards and seeds extracts. The selected standards of polyphenols
extracts, and the results were plotted on a dose- used in this study correspond to the compounds iden-
inhibition curve. The resulting linear calibration tified and quantified in the plant extracts.
curves were used to derive the EC50 value. The equiv- The results of the degradation of polyphenols in
alent amount of an extract that neutralises 50 % of the the presence of citric acid, ascorbic acid, sodium ni-
radical is reported in Table 2. trate, and sodium chloride are presented in Table 3
Table 2 shows that the lowest values of EC50, and Fig. 2. The UV-C exposure of standard polyphe-
which indicates the highest antioxidant activity, were nols (100 mg L−1 concentration) for 480 min led to
obtained for ascorbic acid and gallic acid, followed by the complete degradation of catechin, whereas a re-
catechin and caffeic acid. The linear regression shows moval of 85 % of gallic acid and 50 % of vanillic
a good accord with the experimental data; high R2 acid was achieved after the same irradiation time. The
values result for almost all the samples. degradation of catechin was seen to be complete after
In the natural extracts, significant differences in 8 h of irradiation, whereas a removal of 85 % of gal-
scavenging activity against the DPPH radical were lic acid and only 50 % of vanillic acid was achieved
 I. Volf et al./Chemical Papers 68 (1) 121–129 (2014) 125
Table 3. Kinetic parameters for photo-degradation of polyphenols in standard solution and vegetal extracts using a low-pressure
mercury lamp

Gallic acid Catechin Vanillic acid

Experience 10−3 k 10−5 Pabs 10−3 k 10−6 Pabs 10−3 k 10−5 Pabs

min−1 E s−1 min−1 E s−1 min−1 E s−1

Standard solutions

Without additives 5.70 1.007 1.17 7.960 3.70 1.010

In presence of NaCl 4.10 1.007 8.40 8.830 2.60 1.001
In presence of NaNO3 5.80 1.007 9.60 9.496 2.00 1.007
In presence of ascorbic acid 3.40 1.010 1.01 8.400 3.60 1.010
In presence of citric acid 1.50 1.010 4.80 9.510 1.00 1.010

Vegetal Extracts

y
Grape seeds aqueous extract 5.40 – 2.10 – – –
Spruce bark aqueous extract – – – – 3.70 –

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d[M ]
k – Pseudo-first order rate constant, − = k1 [M ]; Pabs – photonic flux absorbed by components, Pabs = P0 (1 − 10−Aλ ).
dt
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after the same irradiation time. The gallic and vanil-
lic acids presented a higher stability under UV ex-
posure, which is in agreement with their highest an-
tioxidant activity. The first order rate constant of
5.9 × 10−3 min−1 of gallic acid photolysis was reported
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by Benitez et al. (2005), comparable with our result

(5.7 × 10−3 min−1 ).
In order to evaluate the impact of preservatives
usually used in food products on the antioxidant com-
pounds under UV exposure for 90 min, citric acid,
ascorbic acid, sodium nitrate, and sodium chloride, in
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ratios similar to those used in food, were employed as

additives to the polyphenol standard solutions.
After 90 min of irradiation, the removals of gallic
acid amounted to 27 % and 28 % in the presence of
A

sodium chloride and ascorbic acid and, respectively,

to 40 % in the presence of sodium nitrate and in the
absence of additives. The use of citric acid as additive
led to a lower degradation of the gallic acid (16 %).
A similar tendency was also observed in the case of
catechin and vanillic acid. The corresponding catechin
removals after 90 min of irradiation were 58 % in the
presence of sodium nitrate and ascorbic acid, 53 % in
the presence of sodium chloride and less than 40 %
when citric acid was used as an additive. In compari-
son, the removal rate constituted approximately 65 %
without additives. Vanillic acid presented a higher sta-
bility against UV exposure, even in the presence of
other compounds.
The effect of food additives on the UV stability
of polyphenols was rather low. Nitrate had a low ab-
sorption rate in the UV-C range (200–280 nm), while
Fig. 2. Influence of different additives on catechin (a), gallic
the nitrate molar absorption coefficient at 254 nm
acid (b), and vanillic acid (c) upon exposure to UV. Ini-
tial conditions: 100 mg L−1 of compound; × – 0.5 g L−1 was only 4 L−1 M−1 cm−1 . The photolysis of nitrates
NaCl;
– 0.5 g L−1 NaNO− 3 ; – 0.5 g L
−1 citric acid; leads, in an overall reaction, to the formation of ni-
• – 0.5 g L −1 ascorbic acid;  – without additives. trite and oxygen. The photolysis of nitrite in the 200–
126 I. Volf et al./Chemical Papers 68 (1) 121–129 (2014)

400 nm region results in the generation of NO. and O. .

At pH < 12 O.− protonates, to form the OH radical
(Mack & Bolton, 1999). The photolysis proceeds via
complex reaction sequences, involving the formation
of intermediary hydroxyl radicals (Mack & Bolton,
1999; Neamtu & Frimmel, 2006; Schindelin & Frim-
mel, 2000; Warneck & Wurzinger, 1988). The yield of
hydroxyl radicals generated upon exposure to UV ra-
diation of nitrate at 253.7 nm is low.
In theory, dissolved chloride ions react with OH
radicals and lead to the generation of ClOH.− . This
species can decompose to yield chlorine atoms. The
couple Cl. /Cl− has a reduction potential E from 2.2 V
to 2.6 V. Chlorine atoms can add to the C—C dou-

y
ble bonds of the compounds present, thus generating
chlorinated hydrocarbons (Openlaender, 2003). Sajiki
and Yonekubo (2004), reported chemical degradation

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of bisphenol A (BPA) in the presence of chloride ions
by reactive oxygen species (ROS) such as hydroxyl
radicals. They also suggested that NaCl could enhance
the degradation in the presence of ROS.
Ascorbic acid, also known as vitamin C, is natu-
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rally present in some fruit juices or is added as an
antioxidant to minimise losses in colour, flavour, and
nutrients during processing and storage (Koutchma,
2009; Tikekar et al., 2011b, 2012). It is a unique radical
scavenger and is known to enhance the oxygen uptake
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(Φ−O2 ). When exposed to UV radiation, molecular ex-

citation is induced and the photochemical degrada-
tion reactions occurring include multiple free radical
reactions through the formation of ascorbyl radicals.
The ascorbyl radicals generated upon exposure to UV
radiation have a long half-life (≈ 50 s) and persist
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even during storage in the dark for a certain period,

continuing to degrade the compounds (Tikekar et al.,
2011b, 2012). The present results are consistent with
this. The presence of ascorbic acid has been shown to
A

slightly accelerate the oxidation of catechin.

Fig. 3 shows the results obtained for the UV-C ex-
posure of vegetal extracts versus the standard com-
pounds aqueous solutions. Over 180 min of irradia-
tion, approximately 60 % of the gallic acid and more
Fig. 3. Stability against UV irradiation of catechin (a), gallic
than 50 % of the vanillic acid was degraded.
acid (b), and vanillic acid (c) from vegetal aqueous ex-
Some differences were observed for catechin. Af- tracts: Initial conditions: 161.69 mg L−1 of gallic acid;
ter 3 h of irradiation, the removal of catechin from 275.55 mg L−1 of catechin; and 149.11 mg L−1 of vanil-
the grape seeds extract was 40 % whereas, in the lic acid;  – spruce bark extract; – standard.
case of the standard compound, it reached 90 %.
The results may be explained by the high ini-
tial concentrations of catechin in the analysed ex- other authors. B˛akowska et al. (2003) previously in-
tract. The initial concentration of catechin in the vestigated the influence of UV irradiation on the sta-
analysed grape seeds extract was 275 mg L−1 , bility of cyanidin and anthocyanin (AC) – natural
while the concentration in the standard solution was polyphenol co-pigments responsible for some colours
100 mg L−1 . of fruit, vegetables, and other plant tissues. They
The results show that the polyphenols’ standard found that the presence of co-pigments inhibited the
solutions degraded faster under UV than the vege- degradation effect of UV. The presence of co-pigments
tal extracts. This can be explained by the presence in natural extracts prevented the UV degradation
of complex matrices in the plant extracts, such as and stabilised the polyphenols in the vegetal ex-
co-pigments. Similar results have been reported by tracts.
 I. Volf et al./Chemical Papers 68 (1) 121–129 (2014) 127

y
op
rc
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ut
A

Fig. 5. Thermal degradation of natural polyphenols; gallic

Fig. 4. Thermal degradation of standard polyphenols; gallic •
acid ( ), catechin ( ), and vanillic acid (
) at 60 ◦C
(a), 80 ◦C (b), and 100 ◦C (c). Initial conditions:
•
acid ( ), catechin ( ), and vanillic acid (
) at 60 ◦C (a),
161.69 mg L−1 of gallic acid and 275.55 mg L−1 of
80 ◦C (b), and 100 ◦C (c). Initial conditions: 100 mg L−1
catechin from grape seeds extract; 149.11 mg L−1 of
of gallic acid; catechin; and vanillic acid.
vanillic acid from spruce bark extract.

Thermal degradation stable compound, its degradation rate being approx-

imately 20 % at 60 ◦C, increasing to 32 % at 100 ◦C.
The thermal stability of polyphenols is crucial and Gallic acid and vanillic acid exhibited a higher sta-
may be correlated with both the extraction and char- bility, their degradation rates being almost similar at
acterisation methods, and the recommendations on 60 ◦C and 80 ◦C, with degradations of 15 % and 25 %,
their fields of use. Figs. 4 and 5 show the stability respectively. On the other hand, at 100 ◦C over 4 h of
of gallic acid, catechin, and vanillic acid at different exposure, the removal of gallic acid (30 %), catechin
temperatures. (32 %), and vanillic acid (37 %) was more visible.
The data obtained showed a good correlation with In the case of vegetal extracts, the degradation of
the UV stability. Thus, catechin was the most un- the phenolic compounds surveyed was lower for all the
128 I. Volf et al./Chemical Papers 68 (1) 121–129 (2014)

temperatures used. Similar results were reported by Díaz-García, M. C., Obón, J. M., Castellar, M. R., Collado,
Fischer et al. (2013) in investigation of the thermal J., & Alacid, M. (2013). Quantification by UHPLC of total
stability of ACs in three pomegranate juices. The re- individual polyphenols in fruit juices. Food Chemistry, 138,
938–949. DOI: 10.1016/j.foodchem.2012.11.061.
sult may be explained by the higher concentrations of
Fischer, U. A., Carle, R., & Kammerer, D. R. (2013).
phenolics in the natural samples. However, their com- Thermal stability of anthocyanins and colourless pheno-
plex chemical composition should not be disregarded. lics in pomegranate (Punica granatum L.) juices and
model solutions. Food Chemistry, 138, 1800–1809. DOI:
Conclusions 10.1016/j.foodchem.2012.10.072.
Food and Drug Administration (2000). Irradiation in the pro-
duction, processing and handling of food. Federal Register,
The antioxidant activity, thermal and UV-C stabil-
65, 71056–71058.
ities of standard polyphenols solutions and of vegetal Giusti, M. M., & Wrolstad, R. E. (2003). Acylated antho-
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