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ANTIOXIDANT POTENTIAL OF THREE MEDICINAL PLANTS: SIDA RETUSA Linn., URENA LOBATA Linn. AND TRIUMFETTA RHOMBOIDEA Jacq. 6.1. Introduction Reactive oxygen species (ROS) such as superoxide (O;), hydroxyl radicals(0H ) and hydrogen peroxide (H2O2-) are considered to be important factors in the etiology of several pathological conditions such as cardiovascular diseases, diabetes. inflammation, cancer (Hemnani & Parihar, 1998; Halliwell & Gutteridge, 1985). aging and neurodegenerative diseases (Beal, 1995; Thomas & Kalyanaraman, 1997). ROS are degraded to non- reactive forms by enzymatic and non- enzymatic antioxidant defense mechanisms. Free radicals react with almost every known biological molecule in their vicinity and damage protein, cause break down of DNA strands and initiate peroxidation of various molecules. The hydroxyl radical is most reactive of all and may be considered the ultimate damaging species whenever superoxide is formed. ROS are implicated in carcinogenesis induced mutation and tumor

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Page 1: SIDA RETUSA Linn., URENA LOBATA Linn. …shodhganga.inflibnet.ac.in/bitstream/10603/357/17/17...ANTIOXIDANT POTENTIAL OF THREE MEDICINAL PLANTS: SIDA RETUSA Linn., URENA LOBATA Linn

ANTIOXIDANT POTENTIAL OF THREE MEDICINAL PLANTS:

SIDA RETUSA Linn., URENA LOBATA Linn. AND

TRIUMFETTA RHOMBOIDEA Jacq.

6.1. Introduction

Reactive oxygen species (ROS) such as superoxide (O;), hydroxyl

radicals(0H ) and hydrogen peroxide (H2O2-) are considered to be important

factors in the etiology of several pathological conditions such as cardiovascular

diseases, diabetes. inflammation, cancer (Hemnani & Parihar, 1998; Halliwell &

Gutteridge, 1985). aging and neurodegenerative diseases (Beal, 1995; Thomas &

Kalyanaraman, 1997). ROS are degraded to non- reactive forms by enzymatic and

non- enzymatic antioxidant defense mechanisms.

Free radicals react with almost every known biological molecule in their

vicinity and damage protein, cause break down of DNA strands and initiate

peroxidation of various molecules. The hydroxyl radical is most reactive of all

and may be considered the ultimate damaging species whenever superoxide is

formed. ROS are implicated in carcinogenesis induced mutation and tumor

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promotion. Antioxidants act as a masor defense against radical - mediated toxicity

by protecting the damages caused by free radicals. Inhibition of free radical

generation can serve as a facile system for identifying cancer preventive agents

(Troll et al.. 1994)

The traditional Indian medicine and the use of plant drugs against various

diseases receive considerable attention nowadays. Antioxidative components of

natural origin have attracted special interest because they can protect the human

body from free radicals (Osawa et a / , 1990) which are likely to be involved in the

pathophysiology of many diseases such as cancer (Babu et al., 2001). Many

antioxidative components of plant origin have protective activity against cancer

(Anto et a l . 1995). Free radical intermediates such as superoxide anion and

hydroxyl radicals are produced in living systems by various sources such as

ionization of water, X- rays and inflammatory phagocytes. These activated

oxygen species induce DNA strand breaks and chromosomal aberrations in

mammalian cells (C'erutii, 199 1).

It has been reported that the 'rasayanas' are rejuvenators and nutritional

supplements and possess strong antioxidant activity (Sharma et al., 1992). It has

been re-emphasized by the report of Scartezzini & Speroni (2000). A vast number

of literatures documenting the in vitro antioxidant property of polyphenol are

available. Polyphenol acts both as primary as well as secondary antioxidant, by

sequestration of metallic ion and by scavenging active oxygen species

(Morel et al., 1994). Flavones and isoflavones are hormones like phenolic

phytoestrogens of' dietary origin which influence intracellular enzymes, protein

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synthesis, growth factor action. malignant cell proliferation, angiogenesis and

possessing major role as cancer protective compounds through several

mechanisms such as antioxidant effect (Formica & Regelson, 1995; Keli et aL,

1996; Meishiang et a l . 1997).

In order to contribute further to the knowledge of Indian traditional plants,

in the present study three medicinal plants were screened to determine their

antioxidant activity in vitro. Sida retusa Lhn. belonging to the family Malvaceae

is known a Kurumthotti in Malayalam and Bala in Sanskrit. The useful parts are

root system, aerial parts and seed. Of these the root system mainly used in

Ayurvedic formulations like Kshirabala prescribed for the treatment of

rheumatism. Acording to Shastri (1968) S. retusa is one of the ingredient of

famous Ayurvedic formulations such as Aravinda Asava, Bala Arishta. Kumary

Asava, Amrita Pvasha Ghrita, Agastya Rasayana and Chavana Prasha Lehya.

The plant is used in the treatment of tuberculosis, rheumatism and in combination

with other drugs and as an antidote to snake venom. Leaves and roots are useful in

urinary complaints. fever. heart disease. burning sensation, piles and all kind of

inflammation. Root is also used in treatment of leucorrhoea and rheumatism

( Nadkami, 1976; Krthikar & Basu, 1981).

Urena lohara Linn. is considered as a substitute of S. retusa, belonging to

the Malvaceae fsmily is known as Uram in Malayalam and Vanabenda in

Sanskrit. The root system of this plant is used as an external application for

lumbago and rheumatism (Nadkami. 1976). Triumfetta rhomboidea Jacq. is

considered as an adulterant of S. retusa is coming under the family Teliaceae.

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Fruit, flowers and leaves are used in medicine. Bark and fresh leaves are used for

diarrhoea. Flowers rubbed with sugar and water are given in gonorrhoea to stop

the burning caused by urine (Nadkami. 1976).

Lipid peroxidation was measured by the method of Ohkawa et al. (1979)

by measuring the colour of thiobarbituric acid reactive substance (TBARS)

formed at the end of the reaction. Hydroxyl radical scavenging activity was

measured by studying the competition between deoxyribose and the test

compound by the method adopted by Elizabeth & Rao (1990). Superoxide radical

scavenging activity was determined by the NBT reduction method of Mc Cord &

Fridovich (1 969).

6.2.1. Review of literature

Sida species are indiscriminately used for the treatment as antituberculosis,

stomachics, nervous, urinary and cardiac diseases (Chopra et a/., 1958 &

Attygalle, 197 1 ) S. acuta Burm.f. and S. rhomboidea Roxb. are cultivated for the

production of fiber as a substihlte for jute. Extract of S. retusa showed sedative

effect as it decreased alertness, wakefulness and reactivity in mice (Thanam &

Kumari, 197 1 ) It has antirheumatic and antipyretic properties (Iyer & Kolammal,

1993). The various kinds of biological activities viz. antimicrobial and depression

of blood pressure in cats and dogs (Chopra, 1930), immunostimulant (Ghosal

et a[., 1988) antitumour. anti HIV, hepatoprotective and may serve for

hypothyroidism (Kotoky & Das, 2000), abortifacient, antipyretic, adaptogenic

hypoglycaemic (Aiyer & Koldmmal. 1962; Alam et al. 1991; Franzolti et al.,

2000) and antimalarial (Karou et a / , 2003) have generated interest in the

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chemistry of this genus. which resulted in isolation and characterization of various

classes of chemical constituents from different species.

Ihe methanolic extract of roots of S. cordijolia Linn. checked growth of

dental plaque ~ i t h IC' 50 of 10-30 pg Iml. (Namba, et a/ . , 1985). The shoots

with black pepper are useful in excessive menstrual flow (Borthakur, 1992). A

familiar indigenous formulation, Shalparnydi churna, consisting of S. cordijolia.

as an ingredient has been maluated pharmacologically showed positive response

to irritable bowl syndrome (Sharma & Mishra, 1993). Oil from the plant is

beneficial for human skin (Pal, 1994).

Alcoholic extract of S. rhombifolia Linn. was found to be effective

antibacterial activity (tioyal & Rani, 1988 a; Alam et al., 1991; Muanza et al.,

1994). The leaf exhibited antitumour and anti H N activities as it was found

cytotoxic to 60 human cell lines (Muanza et. al. , 1995). Shylaraj (1998) observed

the quantification of ephedrine in in vitro culture of Sida species. It has anti-

inflammatory activity (Venkatesh el al.. 1999 & Franzolti et a/., 2000). Sankar

et at. (2000) studied the quantification of ephedrine in in-vitro cultures of Sida

species. A recent review by Khare et a/. (2002) on chemistry and pharmacology

of genus Sida revealed the medicinal properties of this 'wonderful herb'.

Mazumder et a/ (2001) studied the antibacterial activity of U. lobata.

Tariq (1985) studied the anti-inflammatory effect of Grewia populnifolia.

Cytotoxic activity of G trl~aefolia was observed by Badami et al. (2003).

Alam et a1 (1 99 1) obsewed the anti-inflammatory and antipyretic activities of

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Triumfetta rotuntifolia. Pradhan et a1 (2003) studied the antiulcer activity of

roots of T. rhomboidea Jacq.

A vast number of literatures documenting the antioxidant property of

plants is listed below. (Jose & Kuttan . 1995; Anto et aL, 1995; Anto et al., 1996;

Anto eta/ . , 1998: Joy et al.. 1999; Shylesh & Padikkala, 1999; Martinez, et al.,

2000; Padma. et a/., 2000; Babu et al., 2001; Venkatesh, 2001; Ajaikumar, et al.,

2002; Aquino. et al.. 2002; Babu et al., 2002; Choudhary & Kale, 2002;

Ichikawa & Konishi, 2002; I'ieroni et al., 2002; Vayalil, 2002; Valentao, et al,,

2002; Auddy, et a/., 2003; Babu et a l , 2003; Bagul, et al., 2003; Gorinstein, et al.,

2003; Mary. et a/.. 2003; Soni. et a/., 2003; Velazquez et al., 2003;

Kamalakkannan. et al., 2003; Sabu & Kuttan, 2004.

Mushrooms also have antioxidant property. Some of the medicinally

important mushrooms which have antioxidant property are Pleurotus florida

(Jose et al., 2002) and Phellinus rimosus (Ajith & Janardhanan, 2002).

6.2.2. Chemical constituents of S. rhombifolia ssp. retusa

Root I aerial part consists of alkaloids, P-phenethylamine, N- methyl-P-

phenethylamine. ephedrine. Pseudo ephedrine, vasicinol, vasicinone, vasicine,

choline betainc. S-(+)-Nb5-methyl tryptophan methyl ester, hypaphorine methyl

ester and hypaphorine (Leslie et al., 1980, Prakash et al., 1981). Seed oil I leaves

contains fatty acids, sterculic acid , malval acid, linoleic acid, myristic acid,

palmitic acid, stearic acid, oleic acid and an unidentified fatty acid (Morghis et al.,

1976 & Bhatt et ul , 1983)

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Leaves contain Aminoacids: Lysine, histidine, arginine, asparagines,

glutamine, alanine, valine. phenylalanine. leucine, aspartic acid, glutamic acid

glycine, serine. threonine, tyrosine and proline (Bhatt et al., 1983;

Lakshminarayanan et a1 1987). Aerial part consists of n-alkane (CIS CT5) long

chain alcohols, C',, C,? (Goyal & Rani, 1989). Whole plant I aerial part consists of

sterols: 0-sitosterol. cholesterol. 24-methylene cholesterol, campesterol,

22-dehydro campesterol and stigmasterol (Kumari, 1984), 8- stigmasterol,

spinasterol. 22-dihydrospinasterol (Goyal & Rani, 1988 b, Goyal & Rani, 1989).

Sankar et al. (2003) studied four species of Sida viz. S. rhombifolia ssp. retusa,

S.acuta, S. rhombifolia ssp. rhombifolia and S. cordifolia for their in vitro

secondary metabolite productivity at callus and cell suspension stages.

6.3. Materials and methods

6.3.1. Plant materials

The root system of' the S. retusa, U. lobata and T. rhomboidea were

collected from Kolenchery during the month of June and July. The roots were

dried under shade and powdered. The powder was extracted with methanol, 20%

methanol and pure water using Soxhlet apparatus. These extracts were used for

testing antioxidant property. Methanolic fractions, 20 % methanolic fractions and

pure water fraction of S. retusu. U. lobata and T. rhomboidea were used for initial

screening. The methanolic fractions were found to be effective and were used for

further study.

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6.3.2. Animals

Male Wistar rats ( 1 50-200g) supplied by the small animal breeding station

of Kerala Agricultural University, Mannuthy, India were used. The animals were

maintained under standard environmental conditions. The liver of the rat was used

for preparing liver homogenate.

6.3.3. Preparation of liver homogenate

25% (ulv) liver homogenate in 30 mM KC1 was prepared. The tissue was

well ground in a cold mortar and pestle or homogenizer. Homogenate was kept for -4

15 minutes under cold condition and collected the clear homogenate from the top

for studies

6.3.4. Superoxide radical scavenging activity

Superoxide radical scavenging activity was determined by the Nitroblue

tetrazolium (NBT) reduction method of Mc Cord and Fridovich (1969). The

reaction mixture contained EDTA (0.1 M) containing 0.0015 % NaCN, riboflavin

(0.12 mM). NBT (1.5mM) and various concentrations of the extract (200-1000

pg), and phosphate buffer (pH 7.8) in a final volume of 3 ml. The tubes were

uniformly illuminated under an incandescent lamp for 15 minutes and the optical

density was measured at 530 nm before and after illumination. The percentage of

inhibition of' superoxide generation was evaluated by comparing the absorbance

values of the control and experimental tubes. A known antioxidant, curcumin

(1 - 100 p g) was used as reference.

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6.3.5. Inhibition of lipid peroxide formation.

6.3.5.1. Induction by ~ e " / ascorbate system

The peroxide formation was measured by the method of Ohkawa et al.

(1 979) by measuring the colour of thiobarbituric acid reactive substance (TBARS)

formed at the end of the reaction. Malondialdehyde (MDA), which is formed, as

the end product in lipid peroxidation will react with thiobarbituric acid (TBA) to

give TBARS, which is pink ill colour, measured at 530 nm. The reaction mixture

contained rat liver homogenate (0.1 ml, 25% (wlv) in Tris-HC1 buffer

(20 mM, pH 7.0), KC1 150mM, ferrous ammonium sulphate (0.8 mM), ascorbic

acid (0.3 mM) and various concentrations of the drug (400-2000 pg) in a final

volume of 0.5 ml, was incubated for I hour at 3 7 ' ~ (Bishayee &

Balasubramanian. 1971 ).

The incubated reaction mixture (0.4 ml) was treated with 0.2 ml of 8 %

sodium dodecyl sulphate (SDS), thiobarbituric acid (1.5 ml, 8 %) and acetic acid

(1.5 ml, 20%, pH 3.5). The total volume was then made up to 4 ml by adding

distilled water and kept in a water bath at 1 0 0 " ~ for 1 hour. After cooling, 1 ml of

distilled water and 5 ml of a mixture of n-butanol-pyridine (15:l vlv) were added

and shaken vigorously and centrifuged at 4000 rpm for 10 minutes. The

absorbance of the organic layer was measured at 560 nm after centrifugation. The

percentage inhibition of lipid peroxide formation was determined by comparing

the results of the drug- treated and untreated samples. Curcumin (1 - 100 vg) was

used as reference

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6.3.6. Hydroxyl radical scavenging activity

Hydroxyl radical scavenging activity was measured by studying the

competition between deoxyribose and the extract for hydroxyl radicals generated

from the ~ e ~ ' i a s c o r b a t e l ~ , ~ l ' A M ~ O ~ system. The hydroxyl radicals attack

deoxyribose, which eventually results in TBARS formation (Elizabeth & Rao,

1990). The reaction mixture contained deoxyribose (2.8 mM), FeCI3 (0.1 mM),

EDTA (0.1 mM). H,O>(lmM), ascorbate (0.1 mM), KH2P04-KOH buffer

(20mM, pH 7.4) and various concentrations of the drug (400-2000 pg) in a final

volume of I ml The reaction mixture was incubated for 1 hour at 3 7 ' ~ .

Deoxyribose degradation was measured as TBARS by the method of Ohkawa

et a/. (1979) and percentage inhibition was calculated. Curcumin (1-100 pg) was

used as reference.

6.3.7. Statistical Analysis

Method of Finney was adopted ( 1 971) to find 50% inhibition concentration

(IC 50) of plant extracts.

6.4. Observations

The methanolic fractions were found to be effective in S. retusa, U. lobata

and T. rhomboidea. So the methanolic fractions were selected for further study.

6.4.1. Superoxide radical scavenging activity

The methanolic extracts of the root of S. retusa, U, lobata and

7: rhomboidea were found to scavenge the superoxide radical generated by

photoreduction of riboflavin. The conccntration needed for 50% inhibition of

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scavenging of superoxide was found to be 71.29 pg/ml, 470.60 pg/ml, 336.65

pglml and 6.3 pglml for S. retusa. 11. lobatn, T. rhomboidea and Curcumin

respectively (Table 27)

The methanolic extract of S. retusa root showed 40 % inhibition when

43.24 pdml of the extract was used and 60 % inhibition when 112.61 pg/ml of

the extract was used. The results are shown in Table 28.

Methanolic extract of U. lobata showed 40% inhibition when 405.15

pg/ml of the root extract was used. However, 539.69 pg/ml of the root extract of

U. lobata showed 60% inhibition, 234.76 pg/ml extract of T rhomboidea

inhibited 40 % reaction while 468.10 pglml extract of 7: rhomboidea extract

inhibited 60 %reaction (Table 29,30).

6.4.2. Hydroxyl radical scavenging activity

Hydroxyl radical scavenging activity was calculated by hydroxyl radicals

generated by the ~ e " l a s c o r b a t e l ~ ~ T ~ 1 ~ ~ system was also found inhibited by

S. retusa, ti. lobata and 1: rhomboidea methanolic root extract. The

concentration of root extract needed for 50% inhibition was 1763.22 pg/ml,

1627.35 pgiml and 1346.03 pdml in S retusa, U, lobata and T. rhomboidea

respectively and that of Curcumin was 2.7 pg/ml (Table 27).

40% inhibition was obtained by 1178.09 pg Iml, 1074.71 pglml and

920.04 pg/ml root extract of S. retusa, CI. lobata and T rhomboidea respectively

(Table 3 1,32, 33)

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6.4.3. Inhibition of lipid peroxidation

The generation of lipid peroxides by ~e~~ lasco rba t e in rat liver homogenate

was found inhibited by the addition of the root extract of S. retusa, 0. lobata and

T. rhomborderr fhe concentration of extract needed for 50% inhibition was

1130.24 pglml. 1 109.24 pglml and 1004.22 pglml in S. retusa, U. lobata and

T. rhomboidea respectively and that of Curcumin was 8.9 pg (Table 27).

40% inhibition was obtained by 713.55 pg/ml, 502.34 pg/ml and 565.19

pg/ml root extract of S. retusa, U. lobata and T rhomboidea respectively. 60%

inhibition was obtained by 172 1.16 &ml, 1792.59 pg/ml and 1698.61 pg/ml root

extract of S. retusa. 11. lobata and T rhomboidea respectively (Table 34,35,36).

6.5. Discussion

Oxygen derived free radicals such as the superoxide anion and hydroxyl

radicals are cytotoxic and promote tissue injury (Salim, 1987). Free radicals are

produced endogenously as a by-product of aerobic metabolism. They have

considerable chemical reactivity with biological molecules. Oxygen in high

concentrations can damage the brain, lungs and other organs. Inadequate dietary

intakes of antioxidant micronutrients, such as flavinoids and phenolic compounds,

which prevent free radical-mediated damage in vivo, contribute to disorders such

as atherosclerosis, cancer. ulcer and inflammatory diseases in living organisms

(Halliwell & Gutteridge. 1989). 95% of the oxygen taken in by the aerobic

organisms is fully reduced to I-1202 during the process of mitochondria1

respiration, a small percentage ( 4 % ) of the oxygen consumed is converted to

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semi reduced species i.e. superoxide anion radical (OZ-), hydrogen peroxide

(H202-) and hydroxyl radical (OH-) (Kehrer & Smith, 1994).

Some disorders in which free radicals are implicated are Alzheimer's,

arthritis, Hemorrhoids. Parkinson's, Rheumatism, Heart attack, AIDS, Cataract,

Stroke, Cancer. Stress, Varicose veins, Heart diseases, Immune system disorders

and a long list of degenerative diseases, including aging (Hemnani & Parihar,

1998; Tiwari, 200 1; Pillai & Pillai, 2002). Over production of superoxide radical

takes place in various chronic inflammatory cases, induced by drug, toxin, stress,

tissue injury and heavy exercise. Hydroxyl radical is involved in inflammatory

processes. The inflammation is mainly caused by the generation of free radicals.

Hence the administration of .antioxidants may have a protective role in these

conditions. The tindings of present study give emphasis to this factor 71.29

pg/ml extract of S retusa root provide 50% inhibition of superoxide radical

(Table 28). U. lohota the substitute of S. retusa provide 50% inhibition of

superoxide radical scavenging at a concentration of 470.60pg/ml (Table 29).

While its adulterant, T rhomhoidea has IC 50 (50% inhibition concentration) of

336.65pgml (Table 30) in the case of superoxide scavenging activity (Text

fig. 34). Even though T. rlzomboidea is an adulterant of S. retusa, it has its own

medicinal property

The antioxidants are also known as free radical scavengers. For 50%

inhibition of reaction (hydroxyl scavenging activity) 1763.22 pglml. root extract

of Sida retusa is needed ('Table 31). However, 1627.35 pg/ml. root extract of

Urena labata Linn. is required for IC 50 (Table 32). 1346.03 pg/ml. root extract

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of Triumfetta rhomboidea Jacq, is required for 50% scavenging of hydroxyl

radical(Tab1e 33).

Free radicals react with the cell membrane lipid and cause peroxidation of

polyunsaturated fatty acids and cause generation of further free radicals. In lipid

peroxidation, the 1C 50 is achieved by 1130.24 pglml root extract of S. retusa

(Table 34), 1109.24 pgrnl. root extract of U. lobata (Table 35) and 1004.22

pg/ml root extract of T. rhomboidea (Table 36).

The antioxidant property of the three taxa studied revealed that the

original, substitute and adulterant have medicinal properties and it varies from

taxa to taxa. 'The substitute U. lobata has its own medicinal properties.

Phytosterols, alkaloids and fatty acids are present in S. retusa Linn. (Khare et al.,

2002). This may be responsible for its antioxidant property. Such chemical

constituents present in adulterant may also have medicinal properties. The

chemical constituent responsible for antioxidant property is present in all the three

plants in varying concentrations. This study validates the traditional use of

S. retusa for the treatment of rheumatism.

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Table 27. Effect of the methanolic extract of plant materials on oxygen derived free radical generation

Test material I C 50 (pg/ml) Superoxide hydroxyl Lipid peroxidation radical radical

Sida r e t ~ s u 71.29113.61 1763.221211.43 1130.24i112.08

Urenn loburc~ 470.60-1: 17.17 1627.35+182.46 1109.241141.39

Triumfettu 330.65143.1 8 1346.03*116.76 1004.22*125.48 rhomboideu

Curcumin* 6.310.06 2.7.tO.07 8.910.05 - Values are mean * SE (n=4)

* Reference - Mary et a/. , 2003

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Table 28. Data of superoxide radical scavenging of Sida retusa

~

~ ~ Table of Percentiles 7-.--.-- ~

Standard 95.0% Fiducial C1

Error Lower Upper 0.06937 0.095 16 0.001 184 0.5 168

4~ ~

0.006691 1.2148

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Table 29. Data of superoxide radical scavenging of Urena lobata

1 Table of Percentiles 1 1 Standard 95.00% Fiducial C1 I

Percent Error Lower Upper

12.726 34.8123 83.9817 15.0955 53.3178 1 1 1.9056

3 -,,;>,,-. 16.4406 68.4921 132.5136

- 17.3234 81.875 149.5147

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Table 30. Data of superoxide radical scavenging of Triumfetta rhomboidea

r-- - -

- -- .- Table of Percentiles

Standard 95.00% Fiducial C1 I

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Table 32. Data of hydroxyl radical scavenging of Urena lobata

1 Table of Percentiles I 1 Standard 95.00% Fiducial C1 I

Error Lower Upper

5.4894 / 0.1781 1 23.4404

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Table 33. Data of hydroxyl radical scavenging of Triumfetta rhomboidea

~ --

-- --- Table of Percentiles

Standard 95.00% Fiducial CI -- -~~

Percent I Percentile Lower 7--

Upper 1 I 0.502 25.4133 -.f - - .

2 1 - - -.L!!L402 1.8541 48.866 27.3647 3.9963 7 1.8002

4 1 23.3869 6.9094 94.4975 __i -

52.4039 28.5706 10.587 117.0902

33.4888 15.0294 139.6569

___C _ 38.1541 20.241 1 162.2508

8 1 90.0337 42.577 26.2295 184.9115 ___+ - -

9 i 111.9719 46.7661 33.004 207.67

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Table 34. Data of lipid peroxidation inhibition of Sida retusa

- - - --- - - Table of Percentiles

Standard 95.00% Fiducial C1

Percent Percentile I - - ' 1. --- L-JRO~ Lower upper

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Table 35. Data of lipid peroxidation inhibition of Urena lobata

-

Table of Percentiles

Standard 95.00% Fiduci

-.

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Table 36. Data of lipid peroxidation inhibition of Triumfetta rhon~boide~

~~. ~~ - - ~- ~ -- -

--

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1 2 3 Name of tam

1. Supemxi& radical 2. Hydroxyl radical 3. Lipid peroxidation

Tex Figure 34. Bar chart showing IC 50 (wml) of S.retwa U lobato a d I: rhomboidea