Journal of Applied Pharmaceutical Science Vol. 7 (03), pp. 024-030, March, 2017
Available online at http://www.japsonline.com
DOI: 10.7324/JAPS.2017.70305
ISSN 2231-3354
Cytotoxic and antioxidant capacity of extracts from Vaccinium
meridionale Swartz (Ericaceae) in transformed leukemic cell lines
Margarita González1,2, Ismael Samudio3, Luis Gonzalo Sequeda-Castañeda1,2*, Crispín Celis1, José Iglesias4,
Ludis Morales4**
1
Chemistry Department, School of Sciences, Pontificia Universidad Javeriana, Bogotá, Colombia. 2 Pharmacy Department, School of Sciences, Universidad
Nacional de Colombia, Bogotá, Colombia. 3 Center of Drug Research and Development, Vancouver, Canada. 4 Biochemistry and Nutrition Department,
School of Sciences, Pontificia Universidad Javeriana, Bogotá, Colombia.
ARTICLE INFO
ABSTRACT
Article history:
Received on: 09/01/2017
Accepted on: 18/02/2017
Available online: 30/03/2017
Leukemia is one of the most common malignancies in children and represents a challenge to increase life
expectancy. Many studies have been conducted with the aim to find natural compounds to fight cancer;
however, chemotherapy is still the most widely used treatment despite side effects as harmful to patients. In this
paper the antioxidant and cytotoxic capacity of Agraz extracts (Vaccinium meridionale) in OCI AML3 and
MOLT4 cells using parallel Doxorubicin was evaluated. Extractions were made using 95% methanol, methanoldistilled water (1:1), distilled water and juice. Antioxidant capacity was performed by radical cation 2,2'azinobis-3-ethylbenzothiazoline-6-sulfonic acid and 2,2-diphenyl-1-picryl-hydrazyl-hydrate methods, and
phenols content was determined by Folin-Ciocalteu. The cytotoxic effect was evaluated on the viability of OCI
AML3 and MOLT4 cells using 96-well plates for cell seeding by hemocytometer to 96h. The antioxidant
capacity expressed as 50% inhibitory concentration was 1.7 ± 0.1 and 4.5 ± 0.4 mg/L for methanol extracts by
radical 2,2-diphenyl-1-picryl-hydrazyl-hydrate and radical cation 2,2'-azinobis-3-ethylbenzothiazoline-6sulfonic acid, respectively. The phenol content was found on 104.7 ± 4.0 and 1669.7 ± 82.6 mg AG/100g for the
methanol extract and lyophilized juice, respectively. A decrease in viability of OCI AML3 (24.4%) and MOLT4
(23.0%) cells was found when using Agraz methanolic extract, with Doxorubicin was 98.8 % and 85.0%
respectively.
Key words:
Agraz, Vaccinium
meridionale Swartz,
Antioxidant capacity,
Cytotoxicity, Leukemia,
Colombia.
INTRODUCTION
Antioxidant compounds show a great ability to
scavenge free radicals contributing to the prevention of
cardiovascular and neurological diseases among many others
chronic diseases including cancer (Sutachan et al., 2012).
Chronic diseases such as cancer, diabetes, neurodegenerative and
cardiovascular diseases are characterized by an enhanced state of
oxidative stress, which may result from the overproduction of
reactive species and/or a decrease in antioxidant defenses.
The search for new chemical entities with antioxidant
profile is still thus an emerging field on ongoing interest. The
high diversity of plant species in Colombia has proven
to be a source of many organic molecules with pharmacological
Corresponding Author
*E-mails: lsequeda@javeriana.edu.co,
**Email: ludis.morales@javeriana.edu.co
properties (Thomas et al., 2012). Studies in some fruits and
vegetables have shown to have a high content of antioxidants which
has been recommended for frequent consumption. The current trend
is the use of natural antioxidants so they can be used in food
industry since it is presumed they do not cause health problems
(Garzón et al., 2010; Abreu et al., 2014). Group of molecules
commonly found in studies phytochemicals are flavonoids, these are
characterized by their ability to scavenge free radicals which cause
oxidative stress (Gaviria-Montoya et al., 2009) attributing them the
prevention of cardiovascular, circulatory, cancer and neurological
diseases among others (Alvarez-Castro et al., 2003; SequedaCastañeda et al., 2016). In the last two decades, studies on the
antioxidant and cytotoxic capacity of plants and fruits have
considerably increased due not only to the need to improve human
health but also due to the development of new techniques for
chemical analysis.
© 2017 Margarita González et al. This is an open access article distributed under the terms of the Creative Commons Attribution License -NonCommercialShareAlikeUnported License (http://creativecommons.org/licenses/by-nc-sa/3.0/).
González et al. / Journal of Applied Pharmaceutical Science 7 (03); 2017: 024-030
Cyanidin 3-arabinoside
ACD/IUPAC Name: 2-(3,4-Dihydroxyphenyl)-5,7dihydroxy-3-chromeniumyl β-L-arabinofuranoside
Cyanidin 3-glucoside
ACD/IUPAC Name: 2-(3,4-Dihydroxyphenyl)5,7-dihydroxy-3-chromeniumyl β-Dglucopyranoside
Cyanidin 3-galactoside (Ideain)
ACD/IUPAC Name: 2-(3,4-Dihydroxyphenyl)-5,7-dihydroxy-3-chromeniumyl
β-D-galactopyranoside
025
Petunidin 3-glucoside
ACD/IUPAC Name: 2-(3,4-Dihydroxy-5methoxyphenyl)-5,7-dihydroxy-3-chromeniumyl βD-glucopyranoside
Delphinidin 3-O-glucoside (Myrtillin)
ACD/IUPAC Name: 5,7-Dihydroxy-2-(3,4,5-trihydroxyphenyl)-3chromeniumyl β-D-glucopyranoside
Fig. 1: Compounds from Vaccinium meridionale Swartz (Ericaceae).
Table 1: Anthocyanin’s present in Vaccinium meridionale Swartz (Eriaceae).
Metabolite
Quantity
201 mg per 100 g fresh weight
Cyanidin 3-galactoside, cyanidin 3-glucoside,
329 mg per 100 g fresh weight
cyanidin 3-arabinoside, delphinidin 3-O151 mg per 100 g fresh weight
glucoside, petunidin 3-glucoside.
539 mg per 100 g dry weight
Regarding to different species of blueberries, several
researches have determined the antioxidant capacity in different
species (Canter et al., 2004; Prior et al., 1998) specifically in
Vaccinium floribundum Kunth. (Vasco et al., 2009) in wild
blackberries (Deighton et al., 2000), unripe fruits (Garzón et al.,
2010) and other different kinds of berries (Kähkönen et al., 2001).
One of the main compounds found with cytotoxic effect in them
are anthocyanin’s (Figure 1, Table 1) (Garzón et al., 2010;
Gaviria-Montoya et al., 2009; Kamei et al., 1995; Lopera et al.,
2013; Maldonado-Celis et al., 2014; H. Wang et al., 1999). Agraz
has in its phytochemical composition cyanidin 3-glucoside at
levels up to 329 ± 28 mg/100 g (Garzón et al., 2010) compound to
which it owes its high antioxidant potential of 45.5 ± 2.3 mol
TE/g; in addition to this antioxidant effect. The Vaccinium
uliginosum agraz has showed high cytotoxicity in colon cancer
strains (Zu et al., 2010). Similarly, studies realized with
cranberries have showed antioxidant-cytotoxic synergistic
effect on ovarian cancer cells (Singh et al., 2009);
using berries of French forests similar results have being achieved
Method
UV-Vis
HPLC-DAD. HPLC-ESI/MS-MS
UV-Vis
HPLC-UV
Reference
(Gaviria-Montoya et al., 2009)
(Garzón et al., 2010)
(Maldonado-Celis et al., 2014)
(Lopera et al., 2013)
(Bendaoud et al., 2010), demonstrating the association between
the antioxidant potential and cytotoxicity (Wang et al., 2007) due
to reactive oxygen species are involved in common pathways such
as AP-1 and NF-kB , showing proapoptotic, antiinflammatory and
anti-proliferative effects (Circu et al., 2010; Yang et al., 2001).
Some anthocyanins and their glicones such as cianidine,
pelargonin, delfidin exhibit antiproliferative and proapoptotic
activities on gastric HT-29 adenosarcoma and colon Caco-2 cancer
(Yi et al., 2005). Even though many studies have been conducted
to find natural methods to fight cancer, chemotherapy keeps being
the most common treatment for cancer despite the side harmful
effects. It is known that using Doxorubicin provides a diminution
of cell response between 40 and 50%, but in turn causes nausea,
mucositis and neutropenia risk of congestive cardiomyopathy
(Winchester, 2001). In this research, we use Doxorubicin to
compare and the aim was to evaluate the antioxidant capacity and
potential cytotoxic effects of Agraz extracts on OCI AML3 and
MOLT-4 cells and its possible application as an alternative
treatment to prevent and / or fight cancer.
026
González et al. / Journal of Applied Pharmaceutical Science 7 (03); 2017: 024-030
MATERIALS AND METHODS
Selection of plant material
Ripe fruits of the Colombian blueberry (Vaccinium
meridionale Swartz) commonly Agraz were selected without
mechanical or microbiological changes, with good color
characteristics, fleshy and edible appearance (Obtain commercially
in Department of Boyacá into Santa Bárbara Zone in Tinjacá
region). The National Herbarium of Colombia has classified this
species under the voucher COL208724.
Preparation of the Agraz Extract
100 grams of ripe fruits were steeped in 200 mL of
MeOH (95%, acidulated with clorhidric acid 0.5 N to hydrolyze
flavonoids glycosides), 200 mL MeOH : H2O (1:1) and 200 mL of
distilled H2O for 48 hours. The extracts were filtered using a
cotton cloth, whatman filter paper # 1 and concentrated by rotary
evaporation (90 rpm and 45 °C) to syrup, which is subsequently
lyophilized and stored at -80 °C. The juice obtained by crushing
the fruit was filtered, one part was preserved and another
lyophilized (Raaman, 2006; Thangaraj, 2016).
Preliminary phytochemical analysis
The methanol extract (568 mg), methanol-water extract
(485 mg), aqueous extract (402 mg) and juice (333 mg) were
dissolved in 5 ml of ethanol (EtOH) to perform the LiebermanBurchard test for steroids and sterols, Salkowski to terpenes, Baljet
to
terpenes
and
sterols,
ferric
hydroxamate
for
sesquiterpenlactones, Shinoda and ferric chloride for flavonoids,
anthrone to flavonoid glycosides and Dragendroff for alkaloids
(Raaman, 2006; Thangaraj, 2016).
Total phenols
The determination of total phenols was performed using
the colorimetric method of Folin-Ciocalteu proposed by Singleton
and modified by Ortiz-Ardila (Singleton et al., 1999; Ortíz-Ardila
et al. 2017). Two milligrams of each separately lyophilized extract
were placed in 50 mL volumetric flasks, adding deionized water
until dissolved. Then a 0.5 ml aliquot of each of these solutions
were added 0.75 ml of Folin-Ciocalteu 1.0 N, allowed to stand at
room temperature for 5 minutes and then added 0.75 ml of sodium
carbonate 20%. The final solution was stored at room temperature
in dark for 90 minutes. After this time the absorbance was
measured at 760 nm using a spectrophotometer Thermo Genesys
10s. Results were expressed as milligrams of Gallic Acid per 100
grams of extract (mg GA/100 g).
Antioxidant activity
The antioxidant capacity was evaluated by the methods
DPPH (1,1-Diphenyl-2-picryl-hydrazyl) and ABTS (2,2'-Azinobis(3-ethylbenzothiazoline-6-sulphonic acid). 10 mg of the radical
DPPH were diluted in 10 mL of MeOH and analytical absorbance
at 516 nm was adjusted to 0.75 ± 0.05 by dilution (Brand-Williams
et al., 1995; Ortíz-Ardila et al., 2017). The lyophilized extract was
diluted in deionized water to obtain a 5000 ppm stock and
prepared 5 different concentrations: 50, 500, 1500, 3000 and 4500
ppm. From each solution, 25 μL were added to 975 μL of radical
DPPH previously prepared to reach a final volume of 1 mL. The
steady state was reached in 30 minutes, and the inhibition % was
calculated as follows: % I = [(Ao-Ae)/Ao] x 100, where Ao
corresponds to the absorbance of the solution of the radical
without extract and Ae is referred as the absorbance to the radical
plus the extract.
With the percentages of inhibition, scatterplots were
obtained (% inhibition vs. concentration of the extract), in order to
get the equation of the line (Y=mX+b) from which the 50%
inhibitory concentration (IC50) was calculated: (50-b)/m, being b
the cut point in the Y axis, and m is the slope. The results were
compared using Trolox and vitamin C (Kuskoski et al., 2005). For
the Agraz juice (not lyophilized extract) the antioxidant capacity
was determined by adding different volumes (10μL, 20μL, 30μL,
etc). To calculate the values of the density and total solids in the
juice AOAC 920.151 method and AOAC 950.28 method
respectively were used (AOAC, 2011a, 2011b; RodríguezRodríguez et al., 2012). From the value of the total solids and
density of the juice the IC50 was found.
ABTS radical was prepared by dissolving 10 mg of the
compound in 10 mL of water and then 2.4 mg of potassium
persulfate were added. The prepared solution was left at room
temperature 16 hours in dark until find an absorbance of 0.72 ±
0.02 at a wavelength of 735 nm (Re et al., 1999; Ortíz-Ardila et
al., 2017). Five different concentrations of each extract between
100 ppm and 4000 ppm from a 5000 ppm stock; the absorbance
vs. time graphic was performed to determine the steady state. The
inhibition % was calculated and the IC50 was obtained from the
equation of line. For the Agraz juices the same procedure was
carried out.
Measurement of the effect on OCI-AML3 and MOLT4 cells
viability
Cell lines OCI-AML3 and MOLT4 were maintained in
RPMI-1640 medium containing 5% FCS, 1% glutamine and 100
units/mL penicillin incubated at 37 °C and 5% CO2.
Characterization of the viability effect of the Agraz extracts and
Doxorubicin was measured at the third day of culture. From the
homogenized contents a 1000 µL aliquot of the cell suspension
were plated and the cells were counted by hemocytometer (Phelan
et al., 1997). Tests were performed in 96-well plates by seeding
100 L of RPMI plus cells, adjusting its concentration to 106
cells/well (Handbook, 2011; Licor, 2010).
The lyophilized agraz methanol extract was applied to
the wells at concentrations of 10, 50 and 100 g/mL (using water
as dissolvent) for both cell lines, as a control a concentration of 0
g/mL was used. The results were correlated with viability test
using Doxorubicin at concentrations of 10, 25 and 50 ng/mL, as a
control concentration 0 ng/mL was used. After dosing, each plate
was stored in an incubator at 37 °C with 5% CO2 and the live cells
González et al. / Journal of Applied Pharmaceutical Science 7 (03); 2017: 024-030
027
RESULTS AND DISCUSSION
Mean
233.4
1669.7
104.7
StdDev
7.6
82.6
4.0
RSD. %
3.3
4.9
3.8
* mg Gallic Acid per 100 g of sample. 3 determinations
Water (1:1)
Agraz extracts
The extracts of different polarities were obtained from
100 g of fresh fruit. The extract yield for each polarity was 2.3 ±
0.5 %, 1.8 ± 0.4 % and 1.2 ± 0.3 % for methanol extract,
methanol-water extrac (1:1) and water extract, respectevely.
Preliminary phytochemical analysis
The results of qualitative chemical tests (Table 2) show
high similarity between the compounds present in the Agraz juice
and extracts using solvents such as methanol, methanol-water (1:1)
and water. The compounds detected correspond to phenols and
tannins, flavonoids, terpenes, terpene lactones, coumarins and
steroids.
Phenols and tannins
Gelatin - salt
Ferric chloride
Saponins
Foam
Alkaloids
Dragendorff
Flavonoids
Shinoda
Cardiotonics
Baljet
Triterpenes and steroids
Salkowski
Lieberman - Burchard
Terpene lactones and coumarins
Ferric hydroxymate
*
MeOH-H2O = Methanol-Water (1:1).
H2 O
MeOH-H2O*
MeOH
Test
Extract in
liophylized
no liophylized
Table 2: Preliminary phytochemical analysis.
Juice
+
+
+
+
+
+
+
+
-
-
-
-
-
-
-
-
-
-
+
+
+
+
+
+
+
+
-
-
+
+
+
+
+
+
+
+
+
+
+
+
+
+
(+): Presence. (-): Absence.
+
Antioxidant capacity and total phenols content
The results show the largest amount of total phenols for
the not lyophilized and lyophilized Agraz juice 233.4 ± 7.6 and
32.9
3.5
10.7
average.
H2 O
mg GA/100g *
MeOH:H2O a
Table 3: Total phenolic content in Agraz extracts and juice.
Juice
Extract in
MeOH
Statistical analysis
The software IBM SPSS Statistics 19 was used. All
experiments were done in triplicate (n=3) and results were
expressed as means ± standard deviations. Analysis of variance
was analysed using the Tukey HSD test based on significant
difference of p < 0.05.
liophylized
1669.7 ± 82.6 mg GA/100g respectively, compared to the extracts
obtained with solvents such as methanol (104.7 ± 4.0 mg
GA/100g), methanol-water (32.9 ± 3.5 mg GA/100g) and water
(79.4 ± 6.4 mg GA/100g) (Table 3).
no liophylized
were counted by hemocytometer with trypan blue (Phelan et al.,
1997; Thorkild et al., 1985) at 48, 72 and 96h. The viability
percentage reduction caused by the extract and Doxorrubicin was
calculated, results were expressed in g/mL of extract and in
ng/mL of Doxorubicin (Licor, 2010; Phelan et al., 1997).
a
79.4
6.4
8.1
Methanol-
Garzón et al., in 2010 reported 758.6 ± 62.3 mg GA/100g
in methanolic extract after subjecting the fruits to liquid nitrogen;
not lyophilized and lyophilized Agraz juice showed the highest
concentration of total phenolic compounds compared to other
berries such as Vaccinium floribundum of Ecuador whose amount
is 882 mg GA/100g (Vasco et al., 2009), Vaccinium myrtillus of
North America in 525 mg GA/100g and between 190-473 mg
GA/100g for Vaccinium corymbosum, Vaccinium asheiy and
Vaccinium angustifolium (Garzón et al., 2010). In Vaccinium
meridionale 86 ± 4 mg GA/100g in the methanol extract was
reported (Sequeda-Castañeda et al., 2016), whereas in this study
for the same type extract was 104.7 mg GA/100g. Among the
extracts, the methanolic has the best IC50 and has the highest total
phenolic content. The amount of total phenols in the different
extracts was lower than that found in the juice (without
lyophylized and lyophilized). Possibly due to differences in
solubility to total phenols present in the solvents used.
Additionally, the lyophilization process substantially increase the
phenol content in the lyophilized juice because the process
removes the water present in the juice and increase the content of
solutes (solids). All samples have high antioxidant capacity. The
extract with the highest antioxidant capacity expressed as IC 50
correspond to the methanol extract with values of 1.7 ± 0.1 mg/L
(DPPH method) and 4.1 ± 0.3 mg/L (ABTS method). While
controls showed values of 3.1 ± 0.2 mg/L (ABTS method) and
20.8 ± 1.2 mg/L (DPPH method) for Trolox and 2.4 ± 0.2 and 3.5
± 0.3 mg/L for ABTS and DPPH methods, respectively for
ascorbic acid (Table 4). In another study the IC50’s were 3.8 ± 0.3
and 22.9 ± 5.4 mg/L in ABTS and DPPH methods, respectively,
for methanol extract (Sequeda-Castañeda et al., 2016); these
values are lower than those reported in this study. The high
antioxidant capacity is probably to its higher content of
anthocyanins, phenols and a different polyphenols (Lopera et al.,
2013).
028
González et al. / Journal of Applied Pharmaceutical Science 7 (03); 2017: 024-030
Table 4: Antioxidant capacity for Agraz juice and extracts*
IC50 a (mg/L)
MeOH
ABTS
4.1
0.3
7.3
Mean
StdDev
RSD. %
DPPH
1.7
0.1
5.9
Agraz extracts in:
MeOH:H2O b
ABTS
DPPH
74.5
16.9
4.6
1.1
6.2
6.5
Trolox c
H2 O
ABTS
17.6
1.3
7.4
DPPH
3.9
0.3
7.7
ABTS
3.1
0.2
6.5
DPPH
20.8
1.2
5.8
Juice
Vitamine C b
no liophylized
liophylized
ABTS
DPPH
ABTS
DPPH
ABTS
DPPH
Mean
4.5
5.5
23.7
35.6
2.4
3.5
StdDev
0.4
0.5
1.3
2.7
0.2
0.3
RSD. %
8.9
9.1
5.5
7.6
8.3
8.6
* 3 determinations average. In bold number the best antioxidant capacity. a Inhibitory concentration 50%. b Methanol-Water (1:1). c Controls used in antioxidant
capacity
Number of living cells per ml
Number of living cells per ml
IC50 a (mg/L)
(a)
(b)
Dose
Number of living cells per ml
Number of living cells per ml
Dose
(c)
Dose
(d)
Dose
Fig. 2: Cell viability of (a) OCI AML3 cells with methanolic extract of Agraz. (b) MOLT4 cells with methanolic extract of Agraz. (c) OCI AML3 cells with
Doxorubicin. (d) MOLT4 cells with Doxorubicin. Three different experiment with three replicates (wells) each one (n=3). Signi ficance values (p>0.05) was
calculated with Tukey’s test.
Effect of the Agraz extract in MeOH and Doxorubicin on cell
viability of OCI-AML3 and MOLT4 cells
The methanol extract was selected to evaluate the effect
on cell viability because it presented the best antioxidant capacity
(IC50 of 4.1 and 1.7 mg per L for ABTS and DPPH methods,
respectively). The viability in OCI-AML3 cells decreased by 11.8,
20.8 and 24.4% for doses of 10, 50 and 100 g/mL respectively.
While the MOLT4 cells showed 2.4, 12.8 and 23.0% with doses of
10, 50 and 100 g/mL. The effect of Doxorubicin in inhibiting the
viability of OCI-AML3 cells was 84.1, 91.3 and 98.8% for doses
of 10, 25 and 50 ng/ml respectively. In MOLT4 cells the decrease
was 29.0, 81.0 and 85.0% when using doses of 2.0, 4.0 and 6.0
ng/mL, respectively (Figure 2). The decrease OCI-AML3
cells viability with the methanolic Agraz extract was statistically
significant (sig 0.000 p≤0.05) among all the means of the tested
concentrations. While the decrease for MOLT4 cells was not
significant (sig 0.969 p≤0.05). Observed decrease in cell viability
percentage was much lower compare to standard drug doxorubicin
in all cases. The mechanism by which the extract inhibits cell
proliferation may be due to the induction of apoptosis (Shih et al.,
2005) by NF-kB and AP1 pathways (Wang et al., 2007) or by
inhibition of certain enzymes such as cyclooxygenase (Seeram et
al., 2003), however further investigation must be addressed.
Antioxidants used in leukemia and their relation with
chemotherapy side effects
Some plants and fruits rich in antioxidant content as
ginger root, grapes, carrot, black and blueberries etc. have been
González et al. / Journal of Applied Pharmaceutical Science 7 (03); 2017: 024-030
used for leukemia treatment (Saedi et al., 2014). Different authors
reported that antioxidant use during chemotherapy decreases
mucositis (Wadleigh et al., 1992) and that a lower consumption of
vitamins and antioxidants was related to an increased risk of a
delay in therapy or of experiencing hematologic or
nonhematologic toxicity in kids with leukemia (Kennedy et al.,
2004).
However, remains to clarify the basis of the mechanisms
by which antioxidants would be mediating the reduction of some
of the side effects of chemotherapy in patients with leukemia.
Thus, our study provides a basis for continuing research on the
potential for dietary and pharmaceutical antioxidants to enhance
the effects of chemotherapy and decrease toxicity without
interfering with oncological proceeding.
CONCLUSION
The Agraz juice and methanolic extract show a high
antioxidant value per the results of the ABTS and DPPH assays.
Even though the cytotoxic tests with the Agraz methanolic extract
did not yield high percentages of decreasing viability of OCIAML3 and MOLT4 cells, the Agraz juice -both lyophilized and
non-lyophilized- might be a potentially cytotoxic source because
of its high antioxidant capacity and total phenolic content.
Regarding to Doxorubicin, this showed a greater effect in
decreasing the proliferation of OCI-AML3 (98.8%) and
MOLT4 (85%) cells than that obtained with the Agraz
methanolic extract for OCI AML3 cells (24.4%) and MOLT4
(23.0%).
ACKNOWLEDGMENTS
Administrative Department of Science, Technology and
Innovation -COLCIENCIAS-. This work was funded by the
Academic Vice-Rectory and Vice-Rectory for Research of the
Pontificia Universidad Javeriana (Project 4033 - Evaluation of
antioxidant and antitumor capacity in four fruits of commercial
interest in Colombia). Thanks to the Experimental and
Computational Biochemistry Group and to the Phytochemistry
Research Group at the Pontificia Universidad Javeriana for their
support and collaboration in the realization of this study.
Conflict of Interests: There are no conflicts of interest.
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How to cite this article:
González M, Samudio I, Sequeda-Castañeda LG, Celis C.
Cytotoxic and antioxidant capacity of extracts from Vaccinium
meridionale Swartz (Ericaceae) in transformed leukemic cell lines.
J App Pharm Sci, 2017; 7 (03): 024-030.