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Antioxidant activity of extract from elder flower (Sambucus nigra L.)



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Acta Alimentaria
DOI: 10.1556/AAlim.2014.0009
D. Mihaylovaa*, a. lanteb and a. Krastanova
aDepartment of Biotechnology, University of Food Technologies, 26 Maritza Blvd., 4002, Plovdiv. Bulgaria
bDepartment of Agronomy, Food, Natural Resources, Animals and Environment, Università di Padova,
Viale dell`Universita 16, Agripolis, 35020 Legnaro, Padova. Italy
(Submitted: 08 April 2013; accepted: 26 July 2013)
The present study was designed to investigate the antioxidant and antimicrobial activities of pressurized liquid
extracts from Haberlea rhodopensis Friv. The total phenolic content was performed using the Folin-Ciocalteu
phenol reagent. To determine the antioxidant activities of the extracts, several complementary tests were used:
ABTS and DPPH radical scavenging activities, oxygen radical absorbance capacity, and ferric-reducing antioxidant
power assay. The phenolic concentration was 15.98±0.09 and 9.42±0.06 mg GAE g–1 DW for 70 and 85% ethanol
extracts, respectively. Of all the performed methods, the highest antioxidant activity values were measured by the
ORAC assay – 224.6±6.6 and 154.0±9.9 μM TE g–1 DW for 70 and 85% ethanol extracts, respectively. Results also
showed that both extracts exhibited very weak antimicrobial activity against the examined microorganisms.
However, the 70% ethanol extract possessed higher inhibition ability, which correlated with higher total phenolic
content and antioxidant activity.
Keywords: pressurized liquid extract, Haberlea rhodopensis, antioxidant activity, antimicrobial activity
Natural antioxidants play a very important role in the prevention of different diseases, such
as cancer, arteriosclerosis, and neurodegenerative diseases (Chu et al., 2012; FernánDez-Mar
et al., 2012). Therefore, there is an increasing interest in the antioxidant effects of natural
compounds from medicinal plants and pharmaceutical products for health (raMFul et al.,
2011; ventuani et al., 2011).
Recovery of antioxidant compounds from plant materials is typically accomplished
through different extraction techniques, taking into account their chemistry and uneven
distribution in the plant matrix. Solvent extraction is the most frequently used technique for
isolation of plant antioxidant compounds. However, the extract yields and resulting
antioxidant activities of the plant materials are strongly dependent on the nature of the
extracting solvent, due to the presence of different antioxidant compounds of varied chemical
characteristics and polarities that may or may not be soluble in a particular solvent (PesChel
et al., 2006).
Alternative novel extraction procedures are now being sought after that will reduce
extraction time and solvent consumption, increase sample throughput and improve analyte
recovery. Pressurized liquid extraction (PLE) operates at high pressures and temperature
above point of the boiling point of the organic solvent. The use of higher pressure facilitates
the extraction of the analytes from samples by improving the solvent accessibility to the
analytes that are trapped in the matrix pores. The use of PLE decreases signicantly the total
* To whom correspondence should be addressed.
Phone ++ 359 32 603 645; e-mail:
Acta Alimentaria 2014
time of treatment and in addition, this method can be more effective and selective by changing
some parameters (Choi et al., 2003; ong & len, 2003).
Haberlea rhodopensis is a resurrection species and glacial relic endemic in the mountains
of the Balkan Peninsula in southeastern Europe (Djilianov et al., 2011). It has been proven
that it can survive long periods of desiccation (for up to 2 years) and quickly resume normal
growth within hours of re-watering. Despite the fact that several reliable methods were
applied to study the antioxidant (BerKov et al., 2011; Mihaylova et al., 2011) and enzyme
activity (yahuByan et al., 2005) of its different extracts, this plant is still less explored. raDev
and co-workers (2009) and Mihaylova and co-workers (2011) studied different extracts of the
plant and reported inhibition against St. aureus and Ps. uorescens.
The aim of the present study was to obtain extracts from H. rhodopensis for the rst time
using alternative PLE technique and thus to extend knowledge of this less explored plant.
The potential antioxidant properties of the extracts were studied as well and the antimicrobial
activity was measured.
1. Materials and methods
1.1. Plant material
Haberlea rhodopensis Friv. was collected from its natural habitat (Permission
№201/07.05.2009-Bulgarian Ministry of Environment and Water) at Plovdiv region,
Bulgaria. The plant leaves were air-dried at room temperature (25-28 ºC), roughly grounded,
and stored in air-tight dark containers.
1.2. Extracts preparation
About 67 and 69 g of H. rhodopensis were randomly sampled from 200 g dry plant material.
The PLE was carried out for 1.45 h in an automatic equipment (NM LAB/M Deputex 88,
Limena, Padova, Italy) with 580 and 600 ml of ethanol/water solutions (70:30 and 85:15 v/v
acidied with 0.1 M HCl, pH 3), previously deoxygenated by ushing with nitrogen as
reported by Rossetto and co-workers (2005). The volume of each sample was measured to
calculate the extract concentration (g l–1). The extraction rate was 0.140 and 0.142, resp. The
extracts were subdivided in dark glass bottles without headspace and stored at –20 ºC.
1.3. Total phenolic content (TPC)
The TPC was analyzed using the method of Kujala and co-workers (2000) with some
modications. Each extract was mixed with Folin-Ciocalteu reagent and 7.5% Na2CO3. The
mixture was vortexed and left for 5 min at 50 ºС. After incubation, the absorbance was
measured at 765 nm by room temperature. The TPC was expressed as mg gallic acid
equivalents (GAE) per g dry weight (DW).
1.4. DPPH radical scavenging assay
The ability of the extracts to donate an electron and scavenge 2,2-diphenil-1-picrylhydrazyl
(DPPH) radical was determined by the slightly modied method of BranD-WilliaMs and co-
workers (1995). Freshly prepared 4×10–4 M methanolic solution of DPPH was mixed with the
samples in a ratio of 2:0.5 (v/v). After 30 min incubation at room temperature the light
absorption was measured at 517 nm. The DPPH radical scavenging activity was presented as
a function of the concentration of Trolox. The unit of Trolox equivalent antioxidant capacity
Acta Alimentaria 2014
(TEAC) was dened the concentration of Trolox having equivalent antioxidant activity
expressed as the μM per g DW (μM TE g–1 DW).
1.5. ABTS radical cation decolourization assay
The radical scavenging activity of the extracts against 2,2´-azino-bis(3-ethylbenzothiazoline-
6-sulfonic acid) (ABTS•+) was estimated according to re and co-workers (1999). The results
were expressed as TEAC value (μM TE g–1 DW).
1.6. Ferric-reducing antioxidant power (FRAP) assay
The FRAP assay was carried out according to the procedure of Benzie & strain (1996) with
slight modication. The FRAP reagent was prepared fresh daily and was warmed to 37 °C
prior to use. 150 µl of plant extracts were allowed to react with 2850 µl of the FRAP reagent
for 4 min at 37 °C and the absorbance was recorded at 593 nm. The results were expressed as
μM TE g–1 DW.
1.7. Oxygen radical absorbance capacity (ORAC) assay
The ORAC assay measures the antioxidant scavenging function against peroxyl radical
induced by AAPH at 37 °C. The loss of uorescence of uorescein is an indication of the
extent of damage from its reaction with the peroxyl radical (ou et al., 2001; huang et al.,
2002; goMes et al., 2005). ORAC values were expressed as µM TE g–1 DW.
1.8. Determination of antimicrobial activity (AMA)
Escherichia coli ATCC 25922, Salmonella enterica subsp. enterica ATCC BAA-2162,
Pseudomonas aeruginosa ATCC 9027 and Staphylococcus aureus ATCC 25093 were
purchased from the National Bank of Industrial Microorganisms and Cell Cultures (Soa,
Bulgaria). Strains Bacillus subtilis, Saccharomyces cerevisiae, Aspergillus niger and
Rhizopus sp. were provided by the Culture collection at the Department of Microbiology,
University of Food Technologies, Klebsiella pneumoniae and Listeria monocytogenes were
isolated from clinical samples.
The AMA of the examined extracts was analyzed by the agar-well diffusion method. To
prepare suspensions, the tested microorganisms (TM) were cultured on Luria-Bertani (Sigma-
Aldrich) agar and incubated at 37 °C overnight. Appropriate dilutions of each microbial
suspension were used to inoculate the molten plate count agar (PCA) (Sigma-Aldrich),
equilibrated in water bath, to obtain the concentrations given in Table 2. A. niger was grown
on Wort agar (Sigma- Aldrich) at 30 °C for 48 hours and a suspension was prepared to
inoculate the molten medium. The wells (6 mm in diameter) were cut from the agar and 60 µl
of the tested extracts were delivered into them. Equal volumes of the solvents were used as
controls. PCA plates were incubated at 37 ºC for 24 hours and Wort agar plates at 30 °C for
48 hours. All plates were examined for any zones of growth inhibition, and the diameters
were measured (mm).
1.9. Statistical analysis
All measurements (accept AMA) were carried out in triplicates. The results were expressed
as mean±SD and statistically analyzed using MS-Excel software.
Acta Alimentaria 2014
2. Results and discussion
2.1. Determination of TPC
The TPC in 70 and 85% ethanol extracts from H. rhodopensis were 15.98±0.09 and 9.42±0.06
mg GAE g–1 DW, respectively. Phenols and polyphenolic compounds, such as avonoids, are
widely found in many food products derived from plant sources, and they have been shown
to possess signicant AOAs (van aCKer et al., 1996). Furthermore, phenols have been
determined to play an important role in the survival of the plants under extreme conditions
(Műller et al., 1997). Consequently the presence of those compounds in the extracts
suggested their important role in the plant. Several studies have successfully correlated the
phenolic content with AOA (tePe et al., 2006; Mihaylova et al., 2011), which provoked our
2.2. Determination of antioxidant activity (AOA) of the extracts
Various methodologies are widely used for evaluation of the antioxidant potential and, in this
work, we determined the free radical scavenging capacity of H. rhodopensis extracts using
ABTS and DPPH methods, ORAC assay and their reducing capacity by the FRAP method.
The use of several methods to measure AOA may seem a redundancy, but since different
authors used various methods, comparison of properties becomes easier if a large set of data
is available.
2.3. DPPH, ABTS, FRAP and ORAC assays
In order to investigate the AOA, experiments with two stable radicals DPPH and ABTS•+
were conducted, TEACDPPHvalues were 72.42±0.18 and 30.44±0.25 μM TE g–1 DW and the
TEACABTS•+ values were 72.98±0.77 and 37.99±2.84 μM TE g–1 DW for the 70 and 85%
ethanol extracts, respectively (Table 1). Higher TEAC value indicates that a sample has
stronger AOA. The FRAP values for the investigated extracts of H. rhodopensis were as
follows: 120.54±3.57 and 58.57±1.42 μM TE g–1 DW. Using the ORAC assay the established
results were 224.6±6.60 and 154.0±9.90 μM TE g–1 DW.
The results of the total antioxidant capacity assays (Table 1.) showed that the investigated
extracts possessed AOA, which for the 70% ethanol extract was approximately two times
higher than the capacity of the 85% one. This conrmed the results obtained from the TPC
assay. Interestingly, the highest AOA values were measured by the ORAC assay. A slight
difference among the results obtained by the DPPH, ABTS, ORAC, and FRAP assays was
observed. This might be explained by the unique mechanism and the unequal sensitivity of
each method applied. The authors therefore strongly suggested that, when analyzing the AOA
of samples, it is better to use at least two methods due to the differences between the test
systems (ou et al., 2002). From the results obtained it can be concluded that the 70% ethanol
is more efcient as solvent in order to obtain extract with higher content of biologically
active substances in terms of AOA. This is in agreement with previously conducted studies
that established the effectiveness of 70% ethanol as solvent (Mihaylova et al., 2011).
2.4. Evaluation of antimicrobial activity of the extracts
The AMA was evaluated by measuring the inhibition zone against the TMs after correction
with the diameter of wells and the zones of appropriate controls. According to the results of
Acta Alimentaria 2014
antimicrobial screening given in Table 2, neither extracts showed any signicant activity
against the TMs. However, higher antimicrobial activities were obtained for 70% ethanol
extract. The 85% ethanol extract has been shown to possess antimicrobial effect only against
A. niger among the studied TMs, which corresponds well with the results of TPC and AOA
assays. The 70% ethanol extract showed weak activity against Kl. pneumoniae, A. niger,
B. subtilis, and Rh. sp., while neither of the extracts showed inhibitory activity toward
L. monocytogenes. In comparison raDev and co-workers (2009) reported effect of
H. rhodopensis against S. aureus.
Table 1. Total phenol content and antioxidant activity of extracts from H. rhodopensis
TPC, mg
GAE g–1 DW
μM TE g–1 DW
μM TE g–1 DW
μM TE g–1 DW
μM TE g–1 DW
70% ethanol 15.98±0.09 72.42±0.18 72.98±0.77 224.6±6.60 120.54±3.57
85% ethanol 9.42±0.06 30.44±0.25 37.99±2.84 154.0±9.90 58.57±1.42
Table 2. Antimicrobial activity of extracts from H. rhodopensis
Test microorganism Inhibition zone diameter, mm Concentration
of TM, CFU а ml–1 in media
70% ethanol 85% ethanol
E. coli*1010
Kl. pneumoniae 1;0cn.d.b1.9*1011
S. enterica subsp. enterica*109
Ps. aeruginosa*106
L. monocytogenes*106
St. aureus*1011
B. subtilis 1;1 n.d.b1.0*105
S. cerevisiae*105
A. niger 1;0c3;3c1.0*105
Rhizopus sp. 1;1cn.d.b1.0*105
a:CFU: colony-forming units; b: n.d.–not detected; c: the results of two parallel determinations
3. Conclusions
In summary, due to the insufcient information on H. rhodopensis, we applied the pressurized
liquid extraction method for the rst time in order to obtain new extracts and to reveal the
characteristics of this resurrection plant. The extraction technique was applied with two
different solvents – 70 and 85% ethanol and the antioxidant and the antimicrobial activities
were evaluated. Based on the conducted experiments the 70% ethanol extract was found to
possess higher inhibition, which correlated with the higher total phenolic content and
antioxidant activity.
Acta Alimentaria 2014
This is a collaborative project and has been partially supported by the „Science and Business” Program at the
Ministry of Education and Science, Bulgaria, BG051PO001/3.3-05, co-nanced by the European Social Fund.
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... This effect was similar to that induced by BHT, one of the most widely used chemical antioxidants for this purpose in the food industry. The antioxidant effect of elderflower extract and BHT has been compared before, and the capacity of S. nigra as an antioxidant has been demonstrated [36]. From our results, it can be inferred that the mixture of OEO and S. nigra extract could be adequate to retard the oxidation of lipids in salmon meat. ...
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The antioxidant capacity of oregano (OEO) and clove (CLEO) essential oils and black elderberry (Sambucus nigra) flower extract (SNE) were compared with butylhydroxytoluene (BHT) regarding its protection against lipid peroxidation and microbial counts in salmon burgers stored at 4 °C for 14 days and after cooking. The content of total phenols was 5.74% in OEO, 2.64% in CLEO and 2.67 % in the SNE. The total phenolic content and the antioxidant capacity were significantly higher (p < 0.05) for SNE and OEO. Both essential oils showed a similar IC50 and inhibition percentage of lipid peroxidation to BHT. The combination of OEO and SNE reduced 29% of thiobarbituric acid reactive substances (TBARS), while BHT reduced 31% of TBARS generated during refrigeration storage in salmon burgers in relation to the control sample without antioxidants. Additionally, the microbial counts after 14 days of refrigeration were the lowest in burgers when the combination of OEO and SNE was used. This study concludes that OEO and SNE can be used as inhibitors of lipid oxidation in salmon products and as natural candidates to replace commonly used synthetic antioxidants and antimicrobials in these food products.
... The total phenolic content for decoctions for dry blossoms is similar to that determined from Stoeva et al. (194.0 mg GAE/100 ml). [20] The antioxidant activity of decoctions and infusions is mainly related to the presence of flavonoids. Linear relationships are obtained between FRAP and TFC, as well as between DPPH and TFC, DPPH and TPC, FRAP and TPC with correlation coefficients greater than 0.91. ...
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Introduction: In folk medicine, dried white flowers of Sambucus nigra L. are used to make infusions, decoctions, and juices. Aim: The present article aims to study and compare the antioxidant activity of aqueous solutions of leaves and flowers of Sambucus nigra L obtained at different exposure times and assess the antibacterial activity of these solutions against Escherichia coli ATCC 8739, Salmonella NCTC 6017, Listeria monocytogenes NCTC 11994, and Staphylococcus aureus ATCC 25093. Materials and methods: We studied the physicochemical properties of aqueous extracts of leaves (fresh) and flowers (fresh and dry) of Sambucus nigra L collected from the Rhodope region of Bulgaria. The samples from Sambucus nigra L were analyzed to determine their total phenolic content (TPC), total flavonoid content (TFC), and antioxidant activity using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP). The diameters (in millimeters) of the growth inhibition zones of four pathogens were measured, and a comparative assessment of their antibacterial activity was made. Results: The infusions of fresh blossoms and fresh leaves of Sambucus nigra L had the highest antioxidant activity at the total contact time of 30 minutes (82.7 mmol TE/100 ml) and 35 minutes (36.5 mmol TE/100 ml), respectively. The phenol-richest infusions were those made from dried flowers of Sambucus nigra L after a 30-minute contact time (86.7 mg GAE/ml). Of the four pathogens we studied, we found that the extracts affected partially only the pathogenic bacteria of Salmonella . Conclusions: The highest content of bioactive components was obtained from dried blossoms of Sambucus nigra L. for infusions with a total contact time of 30 minutes and for decoctions at a contact time of 45 minutes.
... Among other substances, there are flavonoids (mainly kaempferol, quercetin, isoquercetin, rutin, astragalin, and hyperoside), multiple phenolic acids (including caffeic, p-coumaric, ferulic, and chlorogenic acids) [7], organic acids, tannins, vitamins (B, C), and mineral salts. Moreover, the flowers also contain lipophilic active substances, e.g., triterpenes (mainly β-amyrin, α-amyrin, lupeol, cycloartenol, ursolic acid, etc.), as well as phytosterols (e.g., β-sitosterol, stigmasterol), and choline [8][9][10]. Additionally, there is 0.03-0.14% of essential oil in the elderflower [11]. ...
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Elderflower extracts are known to be a source of valuable substances that show a wide spectrum of biological activity, including antibacterial and antiviral properties, which demonstrate a degree of effectiveness against SARS CoV-2. In this work, the influence of fresh inflorescence stabilisation methods (freezing, air drying, and lyophilisation) and extraction parameters on the composition and antioxidant properties of the extracts were studied. Wild elderflower plants growing in the Małopolska Region of Poland were studied. Antioxidant activities were evaluated by 2,2-diphenyl-1-picrylhydrazyl free radical-scavenging ability and ferric-reducing antioxidant power assays. The total phenolic content was determined using the Folin-Ciocalteu method and the phytochemical profile of the extracts was analysed using HPLC. The obtained results showed that the best method for the stabilisation of elderflower was lyophilisation, and the determined optimal maceration parameters were 60% methanol as a solvent and a process time of 1–2 days.
... Elderberry fruit and flowers are a very rich source of antioxidants [18,19]. The elderberry fruit analysed in this study had an equal or higher antioxidant capacity as compared with flowers of the same cultivars or wild plants from the same locations (Figure 1.). ...
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This study compared the mineral content and bioactive properties of flowers and fruit coming from wild elderberry plants with those of flowers and fruit harvested from elderberry cultivars grown in an orchard. Elderberry fruit and flowers were analysed for the content of selected minerals, phenolic compounds and anthocyanins and for antioxidant activity. Mineral content was determined by atomic absorption spectrometry method, while antioxidant activity and the content of polyphenols and anthocyanins were determined by spectrophotometric methods. Flowers were found to contain more total ash and to have much higher content of most of minerals, except magnesium, which was present in high concentrations in fruit. Fruit showed significantly higher antioxidant activity than flowers, whereas the total phenolic content varied depending on the growing location / cultivar. The material obtained from selected cultivars growing in an orchard had higher antioxidant activity and polyphenol and anthocyanin content than the material obtained from wild plants. Fruit of the ‘Haschberg’ cultivar and flowers of the ‘Sampo’ cultivar had the best bioactive properties of the studied samples.
... Tea from fruits is intended for the treatment of colds and its' symptoms such as high temperature 11 . Abundant in tannins and polyphenols extracted from the S. nigra shows antioxidant properties, resistance to U.V. radiation, and high biological activity, which can be a base of users in cosmetology 15,16 . ...
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Nowadays, the application of alternative methods instead of clinical treatment creates a new possibility to prevent the development of diseases. Medicinal plants such as Sambucus nigra have been well known due to their extraordinary properties. The similarity to synthetic substances makes it potentially dependable; however, a high concentration of cyanogenic glycosides may exert detrimental consequences. It has been documented that Sambucus nigra extracts are used against both human and animal viruses, like influenza A and B viruses, human immunodeficiency virus (HIV), dengue virus (DENV-2), human herpesvirus type 1 (HSV-1) and human coronavirus NL63 (HCoV-NL63). Such reports are notably valuable especially considering the widespread usage of commercial drugs, which could be ineffective. This review provides insight on recent research on the health properties of plant Sambucus nigra as an antiviral medication that may help propose new therapy.
... The antioxidant effects of elder extracts are responsible for their efficacy in reducing the pain, and fever. The polyphenols (Stoilova et al. 2007;Viapiana and Wesolowski 2017), and anthocyanins in elders (da Silva et al. 2019) are strongly responsible for the antioxidant activities. Elder berries are used as expectorant and mild anti-inflammatory agents in treatment of upper respiratory ailments (Blumenthal et al. 2000). ...
Common cold and flu are caused by common respiratory viral pathogens, which results in hospitalization and death in the world. Among the viral infections, influenza viruses have worldwide spread with major effects on health of societies. Change in antigenic structures of influenza viruses is associated with the lack of effective treatments. Therefore, the use of herbal medicine as alternative choice can be used for management of flu and cold. The flowers of Sambucus nigra or black elders have been approved by commission E for cold, and flu. Although, elders are used in different herbal formulates, but there is no comprehensive study. The subject of this review article was to summarize the efficacy of black elder in treatment of cold and flu. For preparing this manuscript, the electronic resources, books, and thesis were searched by key words of Sambucus, elder, cold, flu, and viral infections. The results of investigations exhibited that there are four clinical trials for elder berries, which it reduced the cold duration and severity (fever, pain, congestion, cough), while there is no clinical trial for elder flower on common cold and flu in spite of its approval by commission E. So, evaluating the efficacy of elder flowers in comparison with its berries and standard treatment on patients with viral respiratory infections should be the subject of large clinical studies.
... Also, the effect is harnessed to obtain compounds (through biotransformation) that act in the natural control of antibiotic use [4]. The use of this substrate in the development of pharmacological products comes from the enhancement of knowledge in folk medicine [5]. ...
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Infections with Escherichia (E.) coli is an important problem in clinical practice. Finding natural, non-toxic ways to prevent infections or control the proliferation of these strains is a current topic. The purpose of this study was the antioxidant analysis and the inhibition of E. coli multiplication of the extract of four species of common edible flowers (tulip, daffodil, freesia and chrysanthemum), yellow and white chrysanthemum, by in vitro study. The petals were extracted with a mixture of three solvents (ethanol/water/acetic acid, 50/49.50/0.50). After concentration, the antibacterial effect against E. coli was determined, by the well diffusion method by measure the diameter of the inhibition zone. At a general evaluation of the results it was observed that the content in bioactive compounds was correlated with the biological effect in vitro. Also, the primary results were influenced by the degree of coloration of the petals. DPPH radical inhibition was approximately 80% for Tagetes erecta. These extracts can be successfully used to combat recurrent infections with coliform strains associated with the presence of an inflammatory process, favored by certain physiological imbalances (oxidative stress).
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GC–MS metabolic profiling of the apolar and polar fractions from methanolic extracts of Haberlea rhodopensis revealed more than one hundred compounds (amino acids, fatty acids, phenolic acids, sterols, glycerides, saccharides, etc.). Bioactivity assays showed that the polar fractions possessed strong free radical scavenging activity (IC50 = 19.95±14.11 μg ml-1 for fresh leaves and 50.04±23.16 μg ml-1 for desiccated leaves), while both the polar and apolar fractions failed to provoke any significant cytotoxic effects against the tested cell lines. Five compounds possessing antiradical activity were identified – syringic, vanillic, caffeic, dihydrocaffeic and p-coumaric acids.
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SUMMARY The present work is a preliminary study of the antibacterial activity of the total extract of the medicinal plant Haberlea rhodopensis. A total extract from Haberlea rhodopensis leaves macerated for 48 hours in 70% water-ethanolic solution with subsequent distillation of the ethanol in vacuum vaporizer to a drug/liquid phase proportion of 5:1, was used. The antibacterial activity of the extract was done on some standard and wild pathogenic bacterial strains. The testing was done by the disc- diffusion method using filter paper discs impregnated with the different concentration of the initial extract: undiluted or diluted 1:1 or 1:2. The results show that the inhibition of the bacterial growth was more pronounced on Staphylococcus aureus than on Gram -negative strains - Pseudomonas aeruginosa and Escherichia coli. A possibility of its antibacterial activity is discussed.
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A few genera of angiosperms are known as ‘resurrection plants’ since their leaves withstand complete desiccation. In many organisms, including some resurrection plants, desiccation tolerance is associated with the accumulation of special carbohydrates. We examined whether this is also true for the two European angiosperm genera of resurrection plants, Ramonda and Haberlea in the Gesneriaceae. Using gas chromatography, non-structural carbohydrates were determined as a percentage of the dry weight in leaves of Ramonda nathaliae subjected to various desiccation regimes. Sucrose was the predominant soluble carbohydrate in all samples, and its level steadily increased from 2 to 10% during desiccation. Starch amounted to ca 2% in control leaves and disappeared completely within 8 days of desiccation. Considerable amounts (1–2.5%) of raffinose and smaller amounts of its precursor galactinol (1-a-galactosyl-myo-inositol) were present in control leaves; these carbohydrates showed only minor changes upon desiccation. Similar results were obtained when excised leaves of Ramonda nathaliae, Ramonda myconi and Haberlea rhodopensis were subjected to desiccation. These data indicate that sucrose accumulation is connected to desiccation tolerance in Gesneriaceae; the presence of raffinose may be a pre-adaptation since this sugar prevents crystallization of sucrose during drying.
The present study investigated Haberlea rhodopensis Friv. – a less-explored Balkan endemic plant. The purpose was to determine the total phenolic content and antioxidant activity of alcohol extracts derived from the leaves of H. rhodopensis Friv. by applying reliable methods. The phenolic concentration in the examined extracts, calculated as mg gallic acid equivalent (GAE)/g leaf dry weight (DW), ranged from 99.03 to 151.24 mg GAE/g DW. The results from the total phenolics assay and the antioxidant activity tests were significantly correlated. The methanol and 70% ethanol extracts, which had the highest total phenolic content values (151.24 and 150.67 mg GAE/g DW, respectively), ranked the highest radical scavenging activity according to the 2,2′-azino-bis-3- ethylbenzothiazoline-6-sulfonic acid assay (1.451 and 1.343 mMTE/g DW, respectively). The major phenolic acids and some flavonoid aglycones and glycosides were analyzed by high-performance liquid chromatography. Luteolin (2,706.20 µg/g DW), hesperidin (2,641.46 µg/g DW), sinapic acid (1,296.78 µg/g DW) and ferulic acid (619.80 µg/g DW) were predominant in the H. rhodopensis 70% alcohol extract. The research yielded novel information about Haberlea rhodopensis, a less-explored endemic plant. The aim of the present study was to determine the antioxidant capacity of this resurrection plant with a focus on phenolic antioxidants rather than enzymes. High-performance liquid chromatography analysis was performed to quantify the major phenolic acids and flavonoids. The antioxidant potential of the extracts obtained was estimated using different approved methods. The results suggested the possibility of practical application of H. rhodopensis leaf extracts because of the established free radical-scavenging activity.
Regular moderate wine consumption is often associated with reduced morbidity and mortality from a variety of chronic diseases in which inflammation is the root cause. This review is focused on three of the numerous bioactive compounds present in wine: resveratrol, hydroxytyrosol and melatonin. Resveratrol and hydroxytyrosol are polyphenols. Melatonin, recently described in wine, is an indoleamine. Their structures, concentrations in wine, bioavailability, pharmacokinetic and health promoting properties are reviewed. Resveratrol seems to be one of the most promising compounds due to its bioactivity, with wine being the main source of resveratrol in diet. Hydroxytyrosol, which its main source in diet is olive oil has been also found in both red and white wine in considerable amounts. Melatonin has been found in wine in low amounts. However, both high bioactivity and bioavailability have been attributed to it. They show antioxidant, cardioprotective, anticancer, antidiabetic, neuroprotective and antiaging activities. However, human studies are still in the initial stages and therefore further studies are needed.
Thousands of tons of crude caffeine are produced annually in the decaffeination of coffee. Crude caffeine is further purified to obtain pure caffeine, and the non-caffeine residue is typically discarded as waste. In the present study, we discovered that crude caffeine possessed unexpected bioactive properties. Crude caffeine had potent hydrophilic antioxidant activity (145 μmol Trolox equivalent (TE)/g) and lipophilic antioxidant activity (66 μmol TE/g). It also inhibited cyclooxygenase-2 with a higher potency (IC50, 20 μg/ml) than 2-acetoxybenzoic acid (aspirin, IC50, 190 μg/ml). Crude caffeine increased glucose uptake 1.45-fold in cultured human skeletal muscle cells and 2.20-fold in adipocytes. In contrast, pure caffeine, which accounts for approximately 90% of the crude caffeine mass, was found to possess negligible antioxidant activity and did not inhibit cyclooxygenase-2, nor stimulate glucose uptake. We believe crude caffeine has potential health benefits and may serve as a novel functional ingredient in the food industry.
The distribution of total phenolics and main betacyanins in red beetroot (Beta vulgaris) root was determined. Also, the subsequent effects of cold storage on the content of total phenolics, main betacyanins (betanin and isobetanin), and the main known ferulic acid ester (β-d-fructofuranosyl-α-d-(6-O-(E)-feruloylglucopyranoside) were determined in the peel, which is the root part containing the largest amount of total phenolics. The content of total phenolics in the red beetroot water extracts was determined according to a modification of the Folin−Ciocalteu method and expressed as gallic acid equivalents (GAE). The compounds of interest were identified by HPLC−ESI−MS and NMR techniques, and the contents of compounds were determined by HLPC analyses. The total phenolic contents in various root parts were found to decrease in the order peel, crown, flesh. Significant differences in the contents of total phenolics and individual compounds were found when the effect of cold storage (5 °C, 0−196 days) on the constituents of the peel from intact roots was examined. In addition to the betacyanins of red beetroot peel found in our earlier study, tentative identifications of betanidin and feruloylamaranthin were made. Keywords: Beta vulgaris; red beetroot; phenolics; betacyanins; distribution of phenolics; distribution of betacyanins; effect of cold storage
Pressurized hot water extraction (PHWE) using a laboratory made system was applied for the extraction of thermally labile and reasonably polar components such as berberine in coptidis rhizoma, glycyrrhizin in radix glycyrrhizae/liquorice and baicalein in scutellariae radix. PHWE was carried out dynamically at a flow of 1ml/min, temperature between 95 and 140°C, an applied pressure of 10–20bar and extraction time of 40min. Extraction by PHWE was found to give efficiencies comparable to Soxhlet extraction for baicalein in scutellariae radix and sonication for berberine in coptidis rhizoma, and glycyrrhizin in radix glycyrrhizae. Effects of ethanol added into the water used in PHWE were explored. Pressurized liquid extraction (PLE) with methanol as solvent was used for extraction of baicalein in scutellariae radix. The marker compounds present in the various medicinal plant extracts were determined by gradient elution HPLC.
The polyphenolic profile of a leaf extract of the Mauritian endemic plant, Eugenia pollicina, was assessed as a source of natural antioxidants. The amounts of flavan-3-ol derivatives determined by HPLC, were in the order of (−)-epicatechin (EC) > (−)-epigallocatechin gallate (EGCG) > (+)-catechin (C) > (−)-epicatechin gallate (ECG) with the levels of Procyanidin B2 and B1 dimers ranging from 1 to 3 mg g−1 FW. The trolox equivalent antioxidant capacity and ferric reducing antioxidant power values were 796 μmol g−1 FW and 302 μmol g−1 FW respectively. E. pollicina extracts also strongly inhibited the FeCl3 and ascorbate-dependent microsome lipid peroxidation, a function that is linked to their flavonoid contents. The extent of DNA damage induced by the extract under study in the copper-phenanthroline assay was lower than the effect of a reference of 240 μM ascorbate. E. pollicina extracts also inhibited lipid autoxidation in the 30% (v/v) olive oil and soybean oil oil-in-water (O/W) emulsions and was effective in slowing down the formation of hydroperoxides in the emulsions during 13 days storage at 40 °C as determined by the peroxide, conjugated diene and para-anisidine values. The high levels of total phenolics, flavonoids and procyanidins measured indicate that E. pollicina is a potential source of antioxidants relevant to the maintenance of oxidative stability of the food matrix, cosmetics and/or pharmaceutical preparations.