Food Chemistry 115 (2009) 113–118

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Food Chemistry
journal homepage: www.elsevier.com/locate/foodchem

Cytoprotective and antioxidant activity studies of jaggery sugar

M.A. Harish Nayaka a,*, U.V. Sathisha b, M.P. Manohar a, K.B. Chandrashekar a, Shylaja M. Dharmesh b
a
Department of Studies in Sugar Technology, Sir. M. Vishvesvaraya Post Graduate Center, University of Mysore, Tubinakere, Mandya 571 402, Karnataka, India
b
Department of Biochemistry and Nutrition, Central Food Technological Research Institute, Mysore 570 020, Karnataka, India

a r t i c l e i n f o a b s t r a c t

Article history: Jaggery and other sugars namely white, refined and brown sugars were evaluated for cytoprotectivity on
Received 14 August 2008 NIH 3T3 fibroblasts and erythrocytes, DPPH radical scavenging activity, reducing power and DNA protec-
Received in revised form 4 October 2008 tion. In addition, total phenol content and phenolic acid composition were also determined. Results indi-
Accepted 19 November 2008
cated a total phenolic content of 26.5, 31.5, 372 and 3837 lg GAE/g for refined, white, brown and jaggery,
respectively. The HPLC analysis revealed the presence of different phenolic acids in brown sugar and jag-
gery. On NIH 3T3 cells oxidation, at 4 mg/ml concentration, jaggery showed 97% protection compared to
Keywords:
brown sugar, and both sugars effectively reduced erythrocyte oxidation. A dose dependent reducing
Cytoprotective
Brown sugar
power and DPPH radical scavenging activity was also observed for jaggery and brown sugar. An EC50
Jaggery of 7.81 and 59.38 lg/ml were observed for jaggery and brown sugar in the DPPH scavenging assay. In
Reducing power DNA oxidation studies, higher protection was observed in jaggery followed by brown, white and refined
Oxidation sugar treated samples.
Ó 2008 Elsevier Ltd. All rights reserved.

1. Introduction typically sold as granulated sugar, which has been dried to prevent
clumping. Raw sugar is comprised of yellow to brown sugars made
Sugars, often called culinary sugars (used in cooking) are an from clarified cane juice boiled down to a crystalline solid with
important foodstuff consumed all over the world, and are manu- minimal chemical processing, which helps in retaining more min-
factured either from sugarcane (70%) or sugar beet (30%). Its con- eral salts and phytochemicals. Manufacturers sometimes prepare
sumption remains high despite increase in synthetic sweeteners, raw sugar as loaves called jaggery in India rather than as a crystal-
and has become an essential nutrient in the world diet for its nutri- line powder. Brown sugar comes from late stages of sugar refining,
tional, sweetening and preservative properties (Chen & Chou, when sugar forms fine crystals with significant molasses content or
1993). The culinary sugars are of different types based on their from coating refined sugar with cane molasses syrup. In terms of
method of production, and also there is difference in the nature, sucrose purity, refined sugar is more pure than blanco directo fol-
size of the crystals, colour and taste of these sugars. The most com- lowed by brown sugar and jaggery sugars.
mon types of sugars that are widely consumed are white, refined, In recent years, plant and plant products have been the main fo-
brown and raw sugar. White sugar, also called blanco directo is cus in the search for nutraceuticals to combat oxidative stress in-
common in India and other south Asian countries, comes from pre- duced diseases (Saxena, Venkaiah, Anitha, Venu, & Raghunath,
cipitating many impurities out of the cane juice by using the phos- 2007). Free radicals are generated during normal cellular metabo-
phatation technique. Refined sugar is the most common form of lism and their effect is neutralised by antioxidant molecules pres-
sugar in North America as well as in Europe and is made by dissolv- ent in the body. However, this balance between the oxidants and
ing brown sugar and purifying it with a phosphoric acid method antioxidant molecules is disturbed by free radicals derived from
similar to that used for blanco directo. It is then further decolour- exogenous sources like ozone, exposure to UV radiations and ciga-
ised by filtration through a bed of activated carbon or bone char rette smoke (Gutteridge & Halliwell, 2000). The free radical produc-
depending on where the processing takes place. Refined sugar is tion in cells can be significantly increased by certain toxic redox
cycling compounds such as drugs and carbon tetrachloride (Wang,
Abbreviations: DPPH, 1,1-diphenyl-2-picrylhydrazyl; EC50, effective con- Ma, Liu, Tian, & Fu, 2007). Importantly, the main biomolecules like
centration for 50% radical scavenging activity; GAE, gallic acid equivalent; ABTS, DNA, lipids and proteins are vulnerable to free radical damage
[2,20 -azinobis(3-ethylbenzothiazoline-6-sulphonic acid); MTT, 3-[4,5-dimethylthi- resulting in cell destruction. Damaged cells lead to abnormal func-
azol-2-yl]-2,5-diphenyl tetrazolium bromide; PBS, phosphate buffer saline; SD,
tioning and results in oxidative stress induced diseases. A potent
standard deviation.
* Corresponding author. Tel.: +91 8232 291112; fax: +91 821 2419363/2419301. scavenger or quencher of these free radical species may serve as a
E-mail address: [email protected] (M.A. Harish Nayaka). possible preventive measure for free radical mediated diseases.

0308-8146/$ - see front matter Ó 2008 Elsevier Ltd. All rights reserved.
doi:10.1016/j.foodchem.2008.11.067
114 M.A. Harish Nayaka et al. / Food Chemistry 115 (2009) 113–118

Apart from the nutritional and sweetening aspects of sugars, samples (n = 3) were preserved in dry condition at room
very little has been studied on their nutraceutical role. The interest temperature.
in polyphenols, including flavonoids and phenolic acids, has con-
siderably increased in recent years because of their possible role 2.3. Determination of total phenol content
in the prevention of oxidative stress induced diseases such as car-
diovascular complications, diabetes, ulcers and cancer (Halliwell, The total phenol content of refined, white, brown and jaggery
2007; Repetto & Llesuy, 2002; Sachidanandam, Fagan, & Ergul sugars were determined colorimetrically using the Folin–Ciocalteu
2005; Shah, Baliga, Rajapurkar, & Fonseca, 2007). Sugarcane (Sac- method (Singleton & Rossi, 1965). A sample aliquot of 100 ll was
charum officinarum) contains phenolic compounds (Fontaniella added to 900 ll of water, 5 ml of 0.2 N Folin–Ciocalteu reagent
et al., 2003) and these compounds have also been found in sugar and 4 ml of saturated sodium carbonate solution (100 g/l) and
products such as syrup or molasses and in brown sugar (Palla, mixed in a cyclo mixer. The absorbance was measured at 765 nm
1982). However, the presence of these phytochemicals in sugar- in Shimadzu UV-160 spectrophotometer (Kyoto, Japan) after incu-
cane juice is often undesirable, as they influence the quality and bation for 2 h at room temperature. The total phenolic content was
colour of final product sugar and hence these phytochemicals are expressed as micrograms of gallic acid equivalent (GAE) per gram
removed through various purification procedures in the sugar sample.
industry. Jaggery and brown sugar are the least processed sugars
containing polyphenols. The brown sugar was also known to pos- 2.4. Extraction of phenolic acids
sess ABTS [2,20 -azinobis (3-ethylbenzothiazoline-6-sulfonic acid)
and DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging The phenolic acids of various sugars were extracted as per the
activity (Payet, Cheong Sing, & Smadja, 2005; Takara, Matsui, protocol followed by Liyana-Pathirana and Shahidi (2006) with
Wada, Ichiba, & Nakasone, 2002). White and refined sugars under- slight modification. Two grams of sugar sample was solubilised
go extensive purification procedures for the removal of phenolic in 50 ml distiled water (in triplicates, n = 3) at room temperature
compounds. The bioactivity of these sugars can be anticipated, as (25 ± 2 °C) with constant stirring. The solution was then centri-
they contain phytochemicals to different extent depending on their fuged at 4000g for 20 min (Sigma 3-16K, USA) and supernatants
manufacturing process. Jaggery is the main source of sugar in rural were collected and combined. The solution was acidified to pH 2
India and has been considered by many Ayurveda practitioners as a with 6 M hydrochloric acid and extracted six times with diethyl
wholesome sugar. Indian Ayurvedic medicine considers jaggery to ether. The ether extracts were then combined and evaporated to
be beneficial in treating throat and lung infections. Sahu and Sax- dryness at 30 °C under vacuum (Buchi 011, Switzerland). The ex-
ena (1994) have found that jaggery can prevent lung damage from tracted phenolic acids were dissolved separately in 2 ml of metha-
particulate matter such as coal and silica dust in rats. However, nol and stored at –20 °C until used within 1 week.
there are no reports available in the literature on cytoprotective
abilities of jaggery and other sugars and their comparative 2.5. HPLC analysis of phenolic acid extracts
evaluation.
Hence, in the present investigation, the protective effect of jag- The phenolic acid extracts of jaggery and other sugars were ana-
gery in comparison with white, refined and brown sugars on free lysed on a HPLC (Model LC-10A. Shimadzu Corporation, Kyoto, Ja-
radical induced damage of NIH 3T3 fibroblasts, erythrocytes and pan) using a reversed phase Shimpak C18 column (4.6  250 mm)
DNA were assessed in addition to 1,1-diphenyl-2-picrylhydrazyl using a diode array UV-detector (operating at 280 nm). A solvent
(DPPH) radical scavenging ability and reducing power. Further, system consisting of water/acetic acid/methanol (Isocratic,
the total phenol content and various phenolic acids present in 80:5:15) was used as mobile phase at a flow rate of 1 ml/min
these sugars were also determined. (Subba Rao & Muralikrishna, 2002). Phenolic acid standards such
as caffeic, p-coumaric, ferulic, gallic, gentisic, 4-hydroxyphenylace-
tic acid, protocatechuic, cinnamic, syringic and vanillic acid were
2. Materials and methods
used for identification of phenolic acids. The identified phenolic
acids were quantified on the basis of their peak area and compar-
2.1. Chemicals
ison with a calibration curve obtained with the corresponding
standards.
1,1-Diphenyl-2-picrylhydrazyl (DPPH), Folin–Ciocalteu reagent,
ascorbic acid, tris–HCl, glutaraldehyde, agarose, ethidium bromide,
2.6. Cytoprotective effect on cultured NIH 3T3 fibroblast cells exposed
cell culture media (RPMI 1640), fetal bovine serum, L-glutamine,
to tert-butyl hydroperoxide
penicillin, streptomycin, 3-[4,5-dimethylthiazol-2-yl]-2,5-diphe-
nyl tetrazolium bromide (MTT), tert-butyl hydroperoxide, phenolic
Cytoprotective ability of different sugars was carried out using
acid standards such as caffeic, p-coumaric, ferulic, gallic, gentisic,
NIH 3T3 fibroblast cells. The cells were subjected to oxidative
4-hydroxyphenylacetic acid, protocatechuic, cinnamic, syringic
stress according to the method reported by Nardini et al (1998).
and vanillic acid were purchased from Sigma Chemical Co. (St.
NIH 3T3 fibroblast cells (1  106 cells/ml, maintained at 37 °C un-
Louis, MO). Lambda phage DNA was procured from Bangalore Geni,
der 5% CO2 and 95% air in complete medium (RPMI 1640 medium
Bangalore, India. NIH 3T3 fibroblast cells were purchased from Na-
supplemented with 10% fetal bovine serum, 2 mM L-glutamine,
tional Center for Cell Sciences, Pune, India. The other chemicals
100 U/ml penicillin and 100 lg/ml streptomycin) were used for
such as ferric chloride, trichloroacetic acid and solvents used in
the assay. NIH 3T3 fibroblast cells (2.8  104 cells/ml) were cul-
the experiment were purchased from Sisco Research Laboratories,
tured with or without sugar samples (20 lL, 0–20 lg/ml) dissolved
Mumbai, India.
in PBS in a 96 well microplate (180 ll suspension/well). After
30 min of incubation, cells were treated with 500 lM tert-butyl
2.2. Sample hydroperoxide and incubated for 3 h. Cell viability was assessed
by microculture tetrazolium assay (Hansen, Nielsen, & Berg,
Refined, white, and brown sugars were procured from three lo- 1989). Twenty-five microlitres of MTT solution (5 mg/ml) were
cal sugar factories (Mandya, Karnataka, India) and jaggery was pur- added to each well, and the cells were incubated at 37 °C for 4 h.
chased from a local market (Mysore, Karnataka, India). All the Then, 100 ll of lysis buffer were added to each well and the cells
 M.A. Harish Nayaka et al. / Food Chemistry 115 (2009) 113–118 115

were again incubated at 37 °C for about 16 h to dissolve dark blue Table 1

formazan crystals. The solution in each well was completely pipet- Total phenol content and phenolic acid composition of jaggery and other sugar
varieties.a
ted out and then the absorption of formazan solution at 570 nm
was measured using a microplate reader (Molecular Devices Spec- Compound Refined White Brown Jaggery
tra Max 340, Global Medical Instrumentation Inc., Minnesota, USA) sugar sugar sugar
Total phenol (lg GAE/g) 26.5 ± 3.79 31.5 ± 1.44 372 ± 1.44 3837 ± 154
and expressed as percent cell protection. Phenolic acids (lg/g) Refined White Brown Jaggery
sugar sugar sugar

2.7. Scanning electron microscopic studies of erythrocyte oxidation Gallic acid Trace Trace 14.6 ± 0.29 122 ± 6.07
Protocatechuic acid – – 1.98 ± 0.07 60.0 ± 3.47
Gentisic acid – Trace 35.2 ± 0.13 130 ± 5.49
Erythrocytes were obtained from healthy, consenting donors.
4-Hydroxyphenylacetic – – 1.75 ± 0.07 29.5 ± 2.08
Heparinized blood was centrifuged at 1000g for 15 min. After re- acid
moval of plasma and buffy coat, the erythrocytes were washed Vanillic acid – – 5.08 ± 0.20 25.6 ± 1.82
thrice with PBS (20 mM, pH 7.4, NaCl – 0.9%) at room temperature Caffeic acid – – – –
Syringic acid – – 11.2 ± 0.26 0.75 ± 0.05
and resuspended in PBS four times its volume for subsequent anal-
p-Coumaric acid – – 6.25 ± 0.17 13.0 ± 0.51
ysis (Suwalsky, Orellana, Avello, & Villena, 2007). Erythrocytes Ferulic acid – – 1.2 ± 0.08 34 ± 1.26
were preincubated with sugar samples (20 mg/ml) for 5 min, and t-Cinnamic acid – – – –
then hydrogen peroxide (30 mM), ferric chloride (80 lM) and a
Values are expressed as mean ± SD (n = 3).
ascorbic acid (50 lM) were added and incubated at 37 °C for 1 h.
The reaction mixture was gently shaken while being incubated
(Manna, Galletti, Cucciolla, Montedoro, & Zappia, 1999). Then the
cells were fixed overnight at 4 °C with glutaraldehyde in normal Bangalore Geni, Bangalore, India) on 1% agarose for 2 h at 50 volts
saline, reaching a final fixation concentration of about 2.4%. The DC. Gels were stained with ethidium bromide (0.5 lg/ml) and doc-
cells were washed in saline solution and then dehydrated using umented (Herolab, Germany).
ascending grades of alcohol (10–100%). Few drops of each sample
were placed on A1 glass cover slips, air dried at room temperature, 2.9. Statistical analysis
gold coated and examined in a scanning electron microscope at
2000 magnification. All the experiments were carried out in triplicates (n = 3) and
the results are expressed as mean ± standard deviation (SD) using
Microsoft Excel software.
2.8. Antioxidant activity

3. Results
2.8.1. DPPH radical scavenging assay
The effect of different sugar varieties on DPPH radical was esti-
3.1. Total phenol content
mated according to the method of Lai, Chou, and Chao (2001). Su-
gar samples (0–62.5 lg/ml) in 200 ll aliquot was mixed with
Total phenolic content as quantified by Folin–Ciocalteu method
100 mM Tris–HCl buffer (800 ll, pH 7.4) and then added to 1 ml
indicated (Table 1) higher total phenolics in jaggery followed by
of 500 lM DPPH in ethanol (final concentration of 250 lM). The
brown, white and refined sugars. Approximately 10-fold higher
mixture was shaken vigorously and left to stand for 20 min at room
phenolic content was observed in jaggery (3837 lg GAE/g) com-
temperature in the dark. The absorbance of the resulting solution
pared to brown sugar (372 lg GAE/g). The total phenolic content
was measured spectrophotometrically at 517 nm. The capability
of white and refined sugars was found to be 31.5 and 26.5 lg
to scavenge DPPH radical was calculated using the following equa-
GAE/g, respectively.
tion. An effective concentration for 50% DPPH radical scavenging
activity was also calculated (EC50).
3.2. Analysis of phenolic acids in jaggery and other sugars
Scavenging effect ð%Þ ¼ ½ðAcontrol  Asample Þ=Acontrol   100
The phenolic acids present in refined, white, brown and jaggery
was determined using HPLC. Results indicated (Table 1) the pres-
2.8.2. Measurement of reducing power
The reducing power of sugar samples were determined accord-
ing to the method of Yen and Chen (1995). The sugar sample (0–
20 mg/ml) was mixed with an equal volume of 0.2 M phosphate 100
Brown
buffer, pH 6.6 and 1% potassium ferricyanide. The mixture was
Percent cell viability

White
incubated at 50 °C for 20 min. Then an equal volume of 10% trichlo-
80
roacetic acid was added to the mixture and then centrifuged at Refined

5000g for 10 min. The upper layer of solution was mixed with dist- Jaggery

iled water and 0.1% ferric chloride at a ratio of 1:1:2 and the absor- 60

bance were measured at 700 nm. Increased absorbance of the

reaction mixture indicated increased reducing power. 40

2.8.3. DNA protection assay

20
DNA protection ability of sugar samples were performed using
lambda phage DNA (Suresh Kumar, Harish Nayaka, Shylaja, & Sal-
imath, 2006). Briefly, k phage DNA (0.6 lg) was subjected to oxida- 0
0 4 8 12 16 20
tion using Fenton’s reagent (0.3 mM hydrogen peroxide, 0.5 lM
Concentration of sugar samples (mg/ml)
ascorbic acid and 0.8 lM ferric chloride) in presence and absence
of sugar samples (0.8 mg/16 ll) for 2 h at 37 °C. The samples were Fig. 1. Cytoprotective effect of jaggery and other sugar samples on t-butyl
subjected for electrophoresis (Submarine electrophoresis system, hydroperoxide induced cell damage in NIH 3T3 cells. Values are mean ± SD (n = 3).
116 M.A. Harish Nayaka et al. / Food Chemistry 115 (2009) 113–118

Fig. 2. Scanning electron microscopic studies of erythrocyte oxidation and their protection by jaggery and other sugar samples (at 20 mg/ml concentration).

ence of gallic, protocatechuic, gentisic, 4-hydroxyphenylacetic, To substantiate the results of cytoprotectivity on NIH 3T3 fibro-
vanillic, syringic, p-coumaric and ferulic acids in both brown sugar blasts, the effect of various sugars (20 mg/ml) on erythrocyte oxi-
and jaggery. Comparatively, jaggery had higher phenolic acid con- dation was also studied. The scanning electron micrographs
tent than brown sugar. White sugar showed the presence of gallic (Fig. 2) show the protective ability of various sugars on erythrocyte
acid and gentisic acid in trace amounts and the remaining phenolic membrane oxidation. As compared to normal erythrocytes (A),
acids tested for were totally absent. Refined sugar had only trace erythrocytes treated with hydrogen peroxide showed the appear-
amounts of gallic acid, while the rest of phenolic acids were absent. ance of echinocytes indicating damage to the cell membrane. In
jaggery and brown sugar treated samples the presence of normal
3.3. Cytoprotective effect of jaggery and other sugars on NIH 3T3 cells can be seen in addition to oxidised cells indicating the protec-
fibroblasts tive role of these sugars.

The cytoprotective effect of jaggery, white, refined and brown 3.4. Antioxidant activities of jaggery and other sugars
sugars on NIH 3T3 fibroblasts indicated (Fig. 1) 97% protection by
jaggery at 4 mg/ml concentration against tert-butyl hydroperoxide Further, antioxidant activity of jaggery and other sugars was
induced cell death. At higher jaggery concentration, there was no evaluated by DPPH radical scavenging, reducing power and DNA
toxic effect of the sugar on the cells and the cytoprotection re- protection assays. The free radical scavenging ability of sugars as
mained constant up to 20 mg/ml. Brown sugar showed dose evaluated by DPPH scavenging model system indicated free radical
dependent cytoprotection with maximum activity at 20 mg/ml scavenging ability of jaggery, brown, white and refined sugars
concentration. However, white and refined sugars showed very (Fig. 3A). Both, jaggery and brown sugars showed free radical scav-
less activity and were not statistically significant. enging ability with an EC50 of 7.81 and 59.4 lg/ml, respectively.
 M.A. Harish Nayaka et al. / Food Chemistry 115 (2009) 113–118 117

increase in absorbance for jaggery and brown sugar. The increased

A 100 Brown White absorbance at 700 nm indicated the presence of reducing power. At
Refined Jaggery 20 mg/ml concentration, an absorbance unit of 2.66 and 0.248 was
Percent Radical scavenging activity

80
observed for jaggery and brown sugars, respectively compared to
white (0.008) and refined sugars (0.018).
Also, DNA protective ability of jaggery and other sugars was
evaluated on lambda phage DNA oxidation (Fig. 4). The hydroxyl
60
radical generated by Fenton’s reagent caused DNA fragmentation
with increase in its electrophoretic mobility (lane 2). This DNA
fragmentation was recovered with the treatment of jaggery and
40
other sugars (0.8 mg/16 ll) to varying extent. As evidenced by
gel documentation analysis, higher protection (70%) was observed
in jaggery treated samples, while 31%, 15% and 18% protection was
20 observed for brown, white and refined sugar treated samples,
respectively.

0 4. Discussion
0 12.5 25 37.5 50 62.5
Concentration of sugar samples (µg/ml) In the present investigation, the bioactivity studies of jaggery in
3
comparison with refined, white and brown sugars were investi-
B gated. The results include, total phenolic content, cytoprotection
Brown White
of NIH 3T3 fibroblasts and human erythrocyte, protection to DNA
2.5 Refined Jaggery oxidation, DPPH radical scavenging activity and reducing power.
Previously, the presence of phenolic compounds in sugarcane juice
Absorbance at 700 nm

as well as in brown sugar has been reported (Payet et al., 2005;

2
Takara et al., 2002). The presence of high total phenolic content
(Table 1) in jaggery and brown sugar compared to white and re-
1.5 fined sugar as per our results may be due to minimal chemical pro-
cessing in the manufacture of jaggery and brown sugar which
retains more polyphenols. The phenolic compounds impart colour
1 as well as taste to the sugar and its removal is an important prob-
lem associated with sugar manufacture (Godshall, Vercellotti, &
Triche, 2002). The different techniques used in cane processing to
0.5
remove colour and impurities affect the amount of polyphenols
in different sugars and this may explain the low phenolic content
0 of white and refined sugars.
0 4 8 12 16 20
Further, both jaggery and brown sugar indicated cytoprotective
Concentration of sugar samples (mg/ml)
abilities against tert-butyl hydroperoxide and hydrogen peroxide
Fig. 3. (A) DPPH Radical scavenging activity of jaggery and other sugar samples. (B) induced oxidative damage of NIH 3T3 fibroblasts and human
Reducing power of jaggery and other sugar samples. Values are mean ± SD (n = 3). erythrocytes, respectively. Since sucrose is a non reducing sugar,
it is less obvious to have free radical quenching abilities and pro-
tect cells from oxidative damage. Hence, the cytoprotective ability
may be attributed to the presence of polyphenolic components in
These results indicate the potential electron donating ability of jag- jaggery and brown sugars. There are no reports to indicate cytopro-
gery and brown sugars. tective abilities of polyphenols in culinary sugars. However, reports
In addition, reducing power of jaggery and other sugars was are available in the literature from other plant sources to indicate
also evaluated by their ability to reduce ferric chloride and potas- cytoprotective role of polyphenols in cell lines (Lima et al., 2007).
sium ferricyanide complex. Fig. 3B indicated a dose dependent Erythrocytes have been extensively used to study oxidative stress,
which represent a simple cell model. Oxidants produce alterations
in the erythrocyte membrane as manifested by a decreased cyto-
skeletal protein content and production of high molecular weight
proteins which leads to abnormal erythrocyte shape (Battistelli
et al., 2005). Hydrogen peroxide and ascorbate/Fe2+ induce an
echinocytic type of shape alteration, characterised by protuber-
ances over the cell membrane (Fig. 2B), indicative of oxidative
damage (Srour, Bilto, Juma, & Irhimeh, 2000). From our results
(Fig. 2 E and F) it is evident that jaggery and brown sugars were
effective in bringing down the oxidative stress induced erythrocyte
damage.
The antioxidant activity as evaluated by DPPH radical scaveng-
ing ability, reducing power and protection to DNA damage induced
by hydroxyl radicals also showed the dominant antioxidant poten-
tial of the jaggery and brown sugar. The correlation co-efficient be-
Fig. 4. Electrophoretic analysis of DNA protection by jaggery and other sugar
tween total phenolic content and cytoprotective activity
samples (at 0.8 mg/16 ll concentration). Lane 1- native DNA; lane 2-DNA + oxidant;
lane 3-DNA + jaggery + oxidant; lane 4-DNA + brown sugar + oxidant; lane 5- DNA + (r = 0.9809), DPPH radical scavenging activity (r = 0.9187), reduc-
white sugar + oxidant and lane 6- DNA + refined sugar + oxidant. ing power (r = 0.9978) and protection to DNA damage
118 M.A. Harish Nayaka et al. / Food Chemistry 115 (2009) 113–118

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From our investigation, the presence of cytoprotective and antiox- Saxena, R., Venkaiah, K., Anitha, P., Venu, L., & Raghunath, M. (2007). Antioxidant
activity of commonly consumed plant foods of India: Contribution of their
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for sweetening as well as for nutraceutical benefits. 250–260.
Shah, S. V., Baliga, R., Rajapurkar, M., & Fonseca, V. A. (2007). Oxidants in chronic
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Acknowledgement
Siddaraju, M. N., & Dharmesh, S. M. (2007). Inhibition of gastric H(+), K(+)-ATPase
and Helicobacter pylori growth by phenolic antioxidants of Curcuma amada.
The authors thank Dr. R. L. Jagadish, Lecturer, Department of Journal of Agricultural Food Chemistry, 55, 7377–7386.
Polymer Science, University of Mysore, Mysore for his keen interest Singleton, V. L., & Rossi, J. A. (1965). Colorimetry of total phenolics with
phosphomolybdic–phosphotungstic acid reagents. American Journal of Enology
in the work and encouragement. Mr. M.A. Harish Nayaka thanks and Viticulture, 16, 144–158.
SC/ST cell, University of Mysore, Mysore, for providing the Teacher Srour, T., Bilto, Y. Y., Juma, M., & Irhimeh, M. R. (2000). Exposure of human
Research Fellowship. erythrocytes to oxygen radicals causes loss of deformability, increased osmotic
fragility, lipid peroxidation and protein degradation. Clinical Hemorheology and
Microcirculation, 23, 13–21.
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