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Journal of Agroalimentary Processes and
Technologies 2013, 19(1), 111-115
Journal of
Agroalimentary Processes and
Technologies
_______________________________________________
Corresponding author: e-
mail:
mihaela.cirimbei@ugal.ro
Study on herbal actions of horseradish (Armoracia rusticana)
Mihaela Roxana Cirimbei*, Rodica Dinică, Liliana Gitin, Camelia Vizireanu
”Dunărea de Jos” University of Galati, Faculty of Food Science and Engineering, Domnească Street, 47,
RO-800008, Galati, Romania
Received: 07 February 2013; Accepted: 09 March 2013
______________________________________________________________________________________
Abstract
Horseradish - a strong spice which was successfully applied in phytotherapy for its valuable effects on health.
Although used for a long time in traditional medicine, its therapeutic properties had not been documented
until recently, confirming in part the utilization of this vegetal product in indigenous medicine of different
nations. The volatile oil, by its components and sinigrin transmits its revulsive, rubefacient, diuretic and
antiseptic properties.
Horseradish consumption normalizes the arterial pressure and prevents the risk of blood clots formation.
Simultaneously, sulfurous substances from horseradish enhance the elasticity of cerebral and coronary blood
vessels, thus reducing the risk of an infarct or cerebrovascular accident.
The present article shows the investigation results of antioxidant potential in fresh, lyophilized and extract
obtained by supercritical fluid extraction from horseradish. Fresh horseradish has the highest antioxidant
capacity, followed by supercritical fluid extraction and lyophilized horseradish.
Keywords: horseradish, supercritical fluid extract, antioxidant capacity
______________________________________________________________________________________
1. Introduction
Horseradish, a
strong spice which was successfully
applied in phytotherapy for its valuable effects on
health, is little demanding regarding the
environmental factors,
being resistant to both low
temperatures and long periods of drought.
Perennial vegetable plant of Brassicaceae family,
the horseradish is native to southeastern Europe
and western Asia.[1]
Therapeutic properties of the horseradish had been
recently documented, although it has been used in
traditional medicine for a long time, confirming in
part the use of this vegetable product in traditional
medicine of different nations. Thus, due to the
vitamins and especially vitamin C, it has
antiscorbutic action. The volatile oil, by its
components and sinigrin transmits its revulsive and
rubefacient properties; the diuretic and antiseptic
properties of horseradish are also due to these
products, eliminated through the respiratory and
renal systems.
Certain substances that horseradish contains have
beneficial effects on peripheral blood flow.
Horseradish consumption normalizes the blood
pressure and prevents the risk of thrombosis. Also,
sulfurous substances from horseradish improve the
elasticity of cerebral and coronary blood vessels,
therefore reducing the risk of an infarct or
cerebrovascular accident.
Horseradish root is rich in vitamin C and B1,
minerals (iron, potassium, calcium and magnesium),
phytoncide and essential oils, a glycoside (sinigrin)
which releases a volatile aglycone (allyl
isothiocyanate) identical with the essence of mustard
plant.[2] The pungent smell of the roots is due to
allyl sulfide, a substance present in garlic and onion.
Likewise, compounds showing antimutagenic and /
or anticarcinogenic activity were identified. Sinigrin,
the main component present in horseradish and
vegetables from Brasicaceae family, is degraded by
the myrosinase enzyme complex to the allyl
isothiocyanate which has antimicrobial effects
Mihaela Roxana Cirimbei et. al. / Journal of Agroalimentary Processes and Technologies 2013, 19(1)
112
against Escherichia coli, Listeria monocytogenes,
Salmonella typhimurium și Staphylococcus aureus.
[3].
Recent research has shown that hydrolyzed form
sinigrin causes apoptosis in tumor cells in vitro and
suppresses the formation of NO in macrophages.
[4].
Some researchers have shown that derivatives of
isothiocyanate type 2-thiohydantoin can form
during cooking or even in the gastrointestinal tract,
although myrosinase enzyme complex is thermally
labile and is inactivated during cooking processes
like boiling, steaming, baking. [5].
However, the use of high concentration of
horseradish is limited by the strong organoleptic
properties that can lead to serious side effects. [4]
Horseradish peroxidase is one of the most used
peroxidase due to wide application in various
fields such as analytical chemistry, environmental
chemistry or clinical trials. The enzyme is used for
many purposes and applications are found at
reasonable prices.
Generally, the enzyme shows a number of features
that make its use beneficial to the common
catalysts, namely the ability to operate under
conditions of mild reactions, as the processes are
ecological in terms of environmental development.
However there are a number of constraints in using
the enzyme, being sensitive, unstable and having
to be used in water, features that are ideal for a
catalyst but undesirable in most syntheses. [6]
Great importance has to be granted to functional
supplements based on horseradish used in
cardiovascular diseases because cardiovascular
diseases are the leading cause of death and
disability worldwide, accounting for 17 million
deaths each year. Globally, Romania stands in the
first 4 places in terms of cardiovascular mortality.
In horseradish, seven isoenzymes were identified
of horseradish peroxidase (HRP), among which the
c isoenzyme of HRP (HRPc) is the most abundant
and has been successfully isolated, purified and
characterized [7]. It has a cardiotonic effect and is
recommended to the people that suffer from high
blood pressure.
Considering all these, the paper presents the results
of our research related to the antioxidant activity of
horseradish, which are part of a larger study that aims
to obtain dietary supplements with anticholesterol
and hypoglycemic action for people with
cardiovascular diseases.
2. Materials and Methods
Materials. The horseradish used in the experiments
was purchased from producers in Galaţi. It was
further submitted to lyophilization process and
supercritical fluid extraction. The equipment adapted
for the experiment was designed, installed and
operated by Natex Prozesstechnologie GembH
Ternitz, Austria.
Analysis methods performed on three samples of
horseradish (fresh, lyophilized and extract) were as
follows:
• determination of the water content according
to the AOAC - 1995 method
• determination of polyphenols - Folin
Ciocalteu method
• determination of flavonoids –
spectrophotometric method, reference
substances are rutine and quercetin
• determination of antioxidant capacity by
DPPH method.
3. Results and discussion
Using horseradish for therapeutic purposes involves
getting a horseradish extract in which the bioactive
compounds remain in native state and in a
concentration as large as possible.
Our research aimes to establish a method to fulfill
these conditions, knowing that any thermal
processing would distort the biologically active
principles.
Thus, after horseradish extract by supercritical fluids
extraction (CO
2
) three categories of horseradish were
studied: fresh horseradish (FH), lyophilized
horseradish for supercritical fluid extraction (LyH)
and horseradish extract (EH) in terms of antioxidant
compounds content and antioxidant activity.
Phenolic compounds, the most important
antioxidants, include two groups of substances which
show strong antiradical action: flavonoids and
phenolic acids, which are both present in horseradish.
The results obtained from the analysis of horseradish
samples on flavonoid content are presented in Figure
1.
Mihaela Roxana Cirimbei et. al. / Journal of Agroalimentary Processes and Technologies 2013, 19(1)
113
From Figure 1 it is noticeable that, depending on
the condition of horseradish samples, the content
of flavonoids increases except lyophilized sample,
which indicates an even lower level than the fresh
sample. According to research conducted by
Abascal et al. (2005), who tested several methods
of drying plants for preserving bioactive
compounds, the use of microwave drying and
lyophilization gives a greater stability to the
antioxidants than other conventional methods. But
in economic terms, lyophilization is not
recommended, being an energy intensive
technique.[8]
Horseradish extract obtained by supercritical fluid
extraction showed the highest values for both
flavonoids (rutine) and flavones (Quercetin). This
was explained by Huang Z., et al. (2012) as the
result of the mechanical action that the material
undergoes before extraction (mincing) and of the
higher surface extraction which is created this way
[9].
These pretreatments promote releasing the solutes
from cells, facilitating solvent flow through the
packed bed, increasing substrate extraction quantity
into units and improving the rate and yield of the
extraction process. As a consequence, superficial
horseradish cells may be considerably broken,
rendering the free analyte to be extracted on the
surface of the solid material.
The content of polyphenols in the samples followed
the same trend as flavonoids (Fig. 2).
As it can be seen, in horseradish polyphenols are
found more as tannic acid than gallic acid, the highest
content being recorded in the case of horseradish
extract obtained by supercritical fluid extraction.
The antioxidant capacity of horseradish samples,
illustrated in Figure 3, shows a similar trend in
antioxidant compounds content, higher values being
recorded on the horseradish extract.
Figure 1. Evolution of flavonoids in the obtaining of horseradish extract
FH - fresh horseradish; LyH - lyophilized horseradish, HE - horseradish extract
Mihaela Roxana Cirimbei et. al. / Journal of Agroalimentary Processes and Technologies 2013, 19(1)
114
Figure 2. Evolution of polyphenols in the obtaining of horseradish extract
FH - fresh horseradish; LyH - lyophilized horseradish, HE - horseradish extract
Figure 3. Evolution of antioxidant capacity in horseradish extract
FH - fresh horseradish; LyH - lyophilized horseradish, HE - horseradish extract
To test the perishability of the antioxidant capacity
in horseradish, the samples were stored at 2-4°C
for 1 week before being analyzed. It was found
that antiradical power of horseradish decreased,
the variation being 8.46% for fresh horseradish,
32.13% for lyophilized horseradish and 77.68% for
horseradish extract. This drastic reduction in
antioxidant capacity may be related to supercritical
fluid extraction method, the data being consistent
with those found in the specialty literature [10].
4. Conclusion
Horseradish (Armoracia rusticana), a plant so
common and yet so important to our health, using the
most varied fresh or preserved as is customary
romanian cuisine, herbal medicine is recommended
for the treatment of many diseases and can replace
successful drug treatment for bronchitis and other
respiratory conditions.
Mihaela Roxana Cirimbei et. al. / Journal of Agroalimentary Processes and Technologies 2013, 19(1)
115
This paper intended to highlight a class of
bioactive polyphenols and flavonoids in
horseradish (Armoracia rusticana) by
qualitative and quantitative determinations
and
their antioxidant capacity.
Following the results we can conclude the
following:
• increase flavonoid content, except
lyophilized sample shows a lower even
than the fresh sample;
• horseradish extract obtained by
supercritical fluid extraction showed the
highest values for both flavonoids (rutin)
and the flavones (quercetin);
• polyphenols are found more than tannic
acid, gallic acid form, the highest content
was recorded in the case of horseradish
extract obtained by supercritical fluid
extraction;
• Horseradish antioxidant capacity of the
samples shows a similar trend in
antioxidant compounds content, higher
values being recorded horseradish extract.
In addition, the storage stability of horseradish
samples was very low. The minimum value
observed with horseradish extract obtained by
supercritical fluid extraction, within a week of its
antioxidant capacity decreasing significantly. The
results, however, allow us to seek other methods of
obtaining a stable horseradish extract for use in
therapeutic purposes.
Compliance with Ethics Requirements
Authors declare that they respect the journal’s ethics
requirements. Authors declare that they have no conflict
of interest and all procedures involving human and/or
animal subjects (if exists) respect the specific
regulations and standards.
References
1. Parvu C., Universul plantelor, Plante din flora
Romaniei, vol I, Editia a II a, Ed. Tehnica.
Enciclopedica, Bucuresti, 2002;
2. Istudor V., Farmacognozie. Fitochimie.
Fitoterapie, vol I, Oze, ozide,si lipide, Ed.
Medicala, Bucuresti, 1998;
3. Wang S.Y, Chen C.T., Yin J.J., Effect of allyl
isothiocyanate on antioxidants and fruit decay of
blueberries, Food Chemistry, 2010, 120 (1), 199–
204, doi.org/10.1016/j.foodchem.2009.10.007.
4. Patel, DK, Patel, K, Gadewar, M., Tahilyani, V., A
concise report on pharmacological and bioanalytical
aspect of sinigrin, Asian Pacific Journal of
Tropical Biomedicine, 2012, S446-S448, doi:
10.1016/S2221-1691 (12)60204-4.
5. Takahashi A., Matsuoka H., Yamada K., Uda Y.,
Characterization of antimutagenic mechanism of 3-
allyl-5-substituted 2-thiohydantoins against 2-
amino-3-methylimidazo[4,5-f]quinoline, Food and
Chemical Toxicology, 2005, 43, 521–528
6. Perazzini R., Saladino R., Guazzaroni M., Crestini
C., A novel and efficient oxidative functionalization
of lignin by layer-by-layer immobilised Horseradish
peroxidase, Bioorganic & Medicinal Chemistry,
2011, 19, 440–447, doi:
10.1016/j.bmc.2010.11.009.
7. Szigeti, K., Smeller, L., Osváth, S., Majer, Z., and
Fidy, J., The structure of horseradish peroxidase C
characterized as a molten globule state after Ca
2+
depletion, BBA-Proteins and Proteomics, 2008,
1784, 1965-1974.
8. Abascal K, Ganora L, Yarnell E., The effect of
freeze-drying and its implications for botanical
medicine: a review, Phytother. Res., 2005 Aug;
19(8), 655-660.
9. Huang Z., Shi X.H., Jiang W.J., Theoretical models
for supercritical fluid extraction, Journal of
Chromatography A, 2012, 1250, 2–26, doi:
10.1016/j.chroma.2012.04.032.
10. Fricks, A.T., Oestreichera,G.E., Filho,L.C.,
Feihrmann,A.C., Cordeiro,Y., Dariva,C., Antunes,
O.A.C., Effects of compressed fluids on the activity
and structure of horseradish peroxidase, J. of
Supercritical Fluids, 2009, 50, 162–168,
doi.org/10.1016/j.supflu.2009.04.014.