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Phytochemical constituents and antioxidant properties of extracts from the leaves of Chromolaena odorata

July 2007
Scientific Research and Essays 2(6):191-194

Authors:

Afolabi Clement Akinmoladun

Federal University of Technology

Emmanuel Ibukun

Federal University of Technology

Aqueous and methanolic extracts of Chromolaena odorata were screened for phytochemical consti-tuents. The evaluation of the antioxidant potential of the methanolic extract was also carried out. Tests for tannins, steroids, terpenoids, flavonoids and cardiac glycosides were positive in both methanolic and aqueous extracts. Alkaloids were detected only in the methanolic extract. The total phenolic content, reducing power and percent DPPH scavenging effect were 0.01 ± ± ± ± 0.00 mg/g GAE, 0.22 ± ± ± ± 0.01 and 28.85 ± ± ± ± 0.99%, respectively. Against the backdrop of many known medicinal properties of this plant, results from the present work suggest that relatively low values of antioxidant indices may not imply a low medicinal value.

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Scientific Research and Essay Vol. 2 (6), pp. 191-194, June 2007

Available online at http://www.academicjournals.org/SRE

Full Length Research Paper

Phytochemical constituents and antioxidant properties

of extracts from the leaves of Chromolaena odorata

Afolabi C. Akinmoladun

, E.O. Ibukun

and I.A.

Dan-Ologe

Biochemistry Department, Federal University of Technology, Akure, Nigeria.

Department of Fisheries, College of Environmental Sciences, University of Agriculture, Abeokuta, Nigeria.

Accepted 16 April, 2007

Aqueous and methanolic extracts of Chromolaena odorata were screened for phytochemical consti-

tuents. The evaluation of the antioxidant potential of the methanolic extract was also carried out. Tests

for tannins, steroids, terpenoids, flavonoids and cardiac glycosides were positive in both methanolic

and aqueous extracts. Alkaloids were detected only in the methanolic extract. The total phenolic

content, reducing power and percent DPPH scavenging effect were 0.01 ±

±±

± 0.00 mg/g GAE, 0.22 ±

±±

± 0.01

and 28.85 ±

±±

± 0.99%, respectively. Against the backdrop of many known medicinal properties of this plant,

results from the present work suggest that relatively low values of antioxidant indices may not imply a

low medicinal value.

Key words: Chromolaena odorata, phytochemicals, antioxidant activity, reductive potential, medicinal plants

INTRODUCTION

Chromolaena odorata (L) King and Robinson Asteraceae

commonly known as Siam weed, is a fast-growing pere-

nnial and invasive weed native to South and Central

America. It has been introduced into the tropical regions

of Asia, Africa and other parts of the world. It is an aggre-

ssive competitor that occupies different types of lands

where it forms dense strands that prevents the establish-

ment of other flora. It is a menace in plantations and

other ecosystems. It suppresses young plantations, agri-

cultural crops and smothers vegetation as it possesses

allelopathic potentialities and growth inhibitors (Ambika

and Jayachandra, 1980; Ambika and Jayachandra, 1982;

Muniappan and Marutani, 1988). The economic value of

C. odorata is low. Consequently, there is a relative pau-

city of research works on it. It is a perennial shrub native

of South and Central America. In recent decades, it has

become a serious pest in the humid tropics of South East

Asia, Africa and Pacific Islands. It spreads rapidly in

lands used for forestry, pasture and plantation crops such

as rubber, coffee, coconut, cocoa and cashew. The plant

can be poisonous to livestock as it has exceptionally high

level of nitrate (5 to 6 times above the toxic level) in the

*Corresponding author. E-mail: akinmoladunfc@gmail.com.

leaves and young shoots; the cattle feeding on these die

of tissue anoxia (Sajise et al., 1974).

Despite the negative sides to the plant, it still has patro-

nage from practitioners of traditional medicine. It has

been reported to have antispasmodic, aniprotozoal, anti-

trypanosomal, antibacterial and antihypertensive active-

ties. It has also been reported to possess anti-inflamma-

tory, astringent, diuretic and hepatotropic activities (Watt

and Brandwijk, 1962; Feng et al, 1964; Weniger and

Robinean, 1988; Iwu, 1993). In the southern part of Nige-

ria, the leaves are used for wound dressing, skin infection

and to stop bleeding. Some specific phenolic compounds

have been isolated from the plant (Metwally and Ekejuba,

1981).

The medicinal values of plants lie in their component

phytochemicals such as alkaloids, tannins, flavonoids

and other phenolic compounds, which produce a definite

physiological action on the human body (Hill, 1952). A

systematic search for useful bioactivities from medicinal

plants is now considered to be a rational approach in

nutraceutical and drug research. Therefore, the present

work has been designed to evaluate the antioxidant pot-

ential of C. odorata with a view to contributing to the

search for beneficial uses of this invasive plant which is a

menace to farmers.

192 Sci. Res. Essays

Table 1. Phytochemicals detected in extracts of Chromolaena odorata.

Phytochemicals Methanolic extracts Aqueous extracts

Alkaloids + -

Saponins - +

Tannins + +

Phlobatannins + +

Anthraquinones - +

Steroids + +

Terpenoids + +

Flavonoids + +

Cardiac glycosides

With steroidal ring + +

With deoxy – sugar + +

+ = Present

- = Absent

MATERIALS AND METHODS

Chemicals

DPPH (2, 2-diphenyl-1-picrylhydrazyl) radical, gallic acid, ascorbic

acid and Folin-Ciocalteau reagent were obtained from Sigma-Ald-

rich, USA. All other chemicals and reagents used were of analytical

grade.

Plant materials

Leaves of C. odoarata were collected from a farmland in Akure,

South-Western Nigeria and identified at the Department of Crop,

Soil and Pest Management, Federal University of Technology,

Akure. They were air dried, packed in paper bags and stored. The

dried leaves were pulverized and 196 g of the pulverized sample

was extracted with 500 ml of 80% methanol by maceration for 72 h.

The methanolic extract was concentrated in a rotary evaporator,

lyophilized and thereafter preserved for further use. An aqueous

extract was also prepared from the pulverized sample for the

purpose of comparison of the phytochemical constituents with that

of the methanolic extract.

Phytochemical screening

Chemical tests were carried out on the aqueous and methanolic

extracts for the qualitative determination of phytochemical

constituents as described by Harborne (1973), Trease and Evans

(1989) and Sofowora (1993). Total phenolic content was

determined using Folin-Ciocalteau reagent as previously described

(McDonald et al., 2001). Total phenol value was expressed as mg/g

gallic acid equivalent.

DPPH radical scavenging activity

The ability of the extract to scavenge DPPH radical was determined

according to the method described by Mensor et al. (2001). One ml

of a 0.3 mM DPPH methanol solution was added to a solution of the

extract or standard (250 µg/ml, 2.5 ml) and allowed to react at room

temperature for 30 min. The absorbance of the resulting mixture

was measured at 518 nm and converted to percentage antioxidant

activity (AA %). Methanol (1.0 ml) plus extract solution (2.5 ml) was

used as a blank. 1 ml of 0.3 mM DPPH plus methanol (2.5 ml) was

used as a negative control. Solution of gallic acid served as positive

control.

Reductive potential

This was determined according to the method of Oyaizu (1986).

The extract or standard (100 µg/ml) was mixed with phosphate

buffer and potassium ferricyanide. The mixture was incubated at

50˚C for 20 min. Tricloroacetic acid (10%, 2.5 ml) was added to the

mixture. A portion of the resulting mixture was mixed with FeCl

(0.1%, 0.5 ml) and the absorbance was measured at 700 nm in a

spectrophotometer. Higher absorbance of the reaction mixture

indicates higher reductive potential.

Statistical analysis

Data were expressed as mean ± SEM. A one-way analysis of

variance was used to analyze data. P<0.5 represented significant

difference between means (Duncan’s multiple range test).

RESULTS AND DISCUSSION

Table 1 shows the phytochemicals detected in C. odorata

leaf extract. Tests for tannins, steroids, terpenoids, flavor-

noids and cardiac glycosides were positive in both met-

hanolic and aqueous extracts. Alkaloids were detected

only in the methanolic extract. Phenolics, alkaloids, terpe-

noids and cardiac glycosides detected in the extracts are

compounds that have been documented to possess

medicinal properties and health-promoting effects (Salah

et al., 1995; Del-Rio et al., 1997; Okwu, 2004; Liu, 2004).

The total phenolic content in the methanolic extract was

0.01 ± 0.00 mg/gGAE; a rather low value. Phenolics are

the largest group of phytochemicals and have been said

to account for most of the antioxidant activity of plant

extracts (Thabrew et al., 1998).

The result of the DPPH scavenging assay is shown in

Figure 1. The percentage antioxidant activity of C. odora-

Akinmoladun et al 193

% AA

# #

Figure 1. Antioxidant activity of CO compared with some standards. CO,

Chromolaena odorata; GA, gallic acid; and AA, ascorbic acid.

Not significantly

different, P>0.05 and *Significantly different, P<0.001.

0 0.2 0.4 0.6 0.8 1

1.2

Reductive Potential

Figure 2. Reductive potential of Chromolaena odorata. AA, Ascorbic acid; and

CO, Chromolaena odorata. *

Significantly different (P<0.001).

ta is about a third of the value for gallic acid and ascorbic

acid while the reductive potential is about a fifth of the

value for ascorbic acid. These values are appreciable

enough for a plant that has been tagged as an obnoxious

weed.

The relatively low values for the percentage antioxidant

activity (Figure 1) and the reductive potential (Figure 2)

are in harmony with the finding that a strong positive

association exists between total phenolic content and

DPPH scavenging effect and also between total phenolic

content and reductive potential (Miliauskas et al., 2004) ).

Low values in some antioxidant assays do not imply low

value in all other assays since many antioxidant assays

show no correlation (Schaich, 2006). Result of a parti-

cular antioxidant assay depends on the chemistry of the

assay and the nature and combination of bioactive princi-

194 Sci. Res. Essays

ples in the material under investigation.

Emerging trends in antioxidant research point to the

fact that low levels of phenolics (and other phytochemi-

cals) and low values of antioxidant indices in plants do

not translate to poor medicinal properties. Mineral ele-

ments, other secondary plant metabolites not detected or

evaluated and vitamins contribute to the synergy of phy-

tochemicals that confer medicinal properties on plants.

The present investigation indicates that though C. odora-

ta has been described as a plant of low economic value,

it is not worthless. Its use in traditional medicine attests to

this. There are prospects for its commercial utilization

especially in view of its abundant and widespread nature.

The toxic compounds in the plant could be removed

through appropriate extraction and processing methods

making extracts and products from the plant safe for the

utilization of animal and man. Control of this invasive and

notorious weed has been unsuccessful. Finding ways of

profitably utilizing it may be the best option left. Further

work is in progress in our laboratory along these lines.

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Aug 2024

The small bowel, also known as the small intestine, is a key component of the digestive system responsible for the majority of nutrient absorption from food. Positioned between the stomach and the large intestine, it receives digestive enzymes from bile and pancreatic juice via the pancreatic duct. The small intestine measures approximately 18 feet (6.5 meters) in length and is intricately folded to fit within the abdominal cavity. The Garcinia Kola plant was recognized by the Department of Pharmacology at Niger Delta University, located on Wilberforce Island, Amasomma, Bayelsa State. The seeds of this plant were collected from Amasomma and subjected to a drying process under natural sunlight for 14 days. Subsequently, they were ground into a fine powder and distributed into various beakers. After allowing the mixture to settle for three days, it was filtered using Watchman's No. 1 filter paper. For this research, thirty albino rats with weights ranging from 150g ± 3.5 to 205g ± 2.6 were utilized. These rats were sourced from the animal facility of the Pharmacology Department at Niger Delta University. They were kept in a controlled environment with temperatures maintained between 20°C and 27°C and a 12-hour cycle of light and darkness. The rats were grouped and provided with standard feed pellets from Guinea Feed Nigeria Plc and unlimited access to clean water for the duration of the study, which included a two-week acclimatization period. The handling of the animals adhered to the institution's guidelines for animal experimentation. The study spanned two months, with an initial four-week acclimatization phase. Post-acclimatization, the 30 rats were randomly assigned to seven groups (A to G). Group A (control) received a diet of pelleted growers feed (mash) and water. Group B (chronic exposure group) was given 1ml of 4% acetic acid orally, along with the standard diet, for 30 days. Group C (sub-chronic exposure group) received the same acetic acid treatment for 14 days. Groups D and E (sub-chronic exposure groups) were treated with acetic acid and varying doses of Garcinia Kola extract for 14 days. Groups F and G (chronic exposure groups) underwent a similar regimen with Garcinia Kola extract for 30 days. At the end of the treatment period, the rats were euthanized using chloroform anesthesia, and vital organs were harvested and preserved in 10% formalin. The tissues underwent standard histological processing and were stained with Haematoxylin and Eosin at the Histopathology laboratory of the Niger Delta Teaching Hospital (NDUTH) in Okolobiri. Histological examination revealed that the control group's villi and submucosal walls remained intact. However, plates 4.2 to 4.4 displayed erosion of the submucosal layer, interruption of the villi, and aggregation of white blood cells. Plate 4.5 indicated mild erosion of the mucosal layer and the presence of inflammatory cells. The findings suggest that oral ingestion of acetic acid for 30 days can cause inflammation and necrosis in the stomach tissues of lab animals. Additionally, it was noted that administering Garcinia Kola seed extract alone had minimal impact on the animals' histology.

THERAPEUTIC AND ANTAGONIST VILLI INSURGENCY OF GARCINIA KOLA ON ACETIC ACID INDUCED EROSIVE SMALL INTESTINAL ULCER OF WISTAR ALBINO RATS MODEL

Article

Full-text available

Jan 2024

The small bowel, also known as the small intestine, is a key component of the digestive system responsible for the majority of nutrient absorption from food. Positioned between the stomach and the large intestine, it receives digestive enzymes from bile and pancreatic juice via the pancreatic duct. The small intestine measures approximately 18 feet (6.5 meters) in length and is intricately folded to fit within the abdominal cavity. The Garcinia Kola plant was recognized by the Department of Pharmacology at Niger Delta University, located on Wilberforce Island, Amasomma, Bayelsa State. The seeds of this plant were collected from Amasomma and subjected to a drying process under natural sunlight for 14 days. Subsequently, they were ground into a fine powder and distributed into various beakers. After allowing the mixture to settle for three days, it was filtered using Watchman's No. 1 filter paper. For this research, thirty albino rats with weights ranging from 150g ± 3.5 to 205g ± 2.6 were utilized. These rats were sourced from the animal facility of the Pharmacology Department at Niger Delta University. They were kept in a controlled environment with temperatures maintained between 20°C and 27°C and a 12-hour cycle of light and darkness. The rats were grouped and provided with standard feed pellets from Guinea Feed Nigeria Plc and unlimited access to clean water for the duration of the study, which included a two-week acclimatization period. The handling of the animals adhered to the institution's guidelines for animal experimentation. The study spanned two months, with an initial four-week acclimatization phase. Post-acclimatization, the 30 rats were randomly assigned to seven groups (A to G). Group A (control) received a diet of pelleted growers feed (mash) and water. Group B (chronic exposure group) was given 1ml of 4% acetic acid orally, along with the standard diet, for 30 days. Group C (subchronic exposure group) received the same acetic acid treatment for 14 days. Groups D and E (sub-chronic exposure groups) were treated with acetic acid and varying doses of Garcinia Kola extract for 14 days. Groups F and G (chronic exposure groups) underwent a similar regimen with Garcinia Kola extract for 30 days. At the end of the treatment period, the rats were euthanized using chloroform anesthesia, and vital organs were harvested and preserved in 10% formalin. The tissues underwent standard histological processing and were stained with Haematoxylin and Eosin at the Histopathology laboratory of the Niger Delta Teaching Hospital (NDUTH) in Okolobiri. Histological examination revealed that the control group's villi and submucosal walls remained intact. However, plates 4.2 to 4.4 displayed erosion of the submucosal layer, interruption of the villi, and aggregation of white blood cells. Plate 4.5 indicated mild erosion of the mucosal layer and the presence of inflammatory cells. The findings suggest that oral ingestion of acetic acid for 30 days can cause inflammation and necrosis in the stomach tissues of lab animals. Additionally, it was noted that administering Garcinia Kola seed extract alone had minimal impact on the animals' histology.

Potential synergy of phytochemicals in cancer prevention: Mechanism of action

Conference Paper

Dec 2004

R.H. Liu

Epidemiological studies have consistently shown that regular consumption of fruits and vegetables is strongly associated with reduced risk of developing chronic diseases, such as cancer and cardiovascular disease. It is now widely believed that the actions of the antioxidant nutrients alone do not explain the observed health benefits of diets rich in fruits and vegetables, because taken alone, the individual antioxidants studied in clinical trials do not appear to have consistent preventive effects. Work performed by our group and others has shown that fruits and vegetable phytochemical extracts exhibit strong antioxidant and anti proliferative activities and that the major part of total antioxidant activity is from the combination of phytochemicals. We proposed that the additive and synergistic effects of phytochemicals in fruits and vegetables are responsible for these potent antioxidant and anticancer activities and that the benefit of a diet rich in fruits and vegetables is attributed to the complex mixture of phytochemicals present in whole foods. This explains why no single antioxidant can replace the combination of natural phytochemicals in fruits and vegetables to achieve the health benefits. The evidence suggests that antioxidants or bioactive compounds are best acquired through whole-food consumption, not from expensive dietary supplements. We believe that a recommendation that consumers eat 5 to 10 servings of a wide variety of fruits and vegetables daily is an appropriate strategy for significantly reducing the risk of chronic diseases and to meet their nutrient requirements for optimum health.

Studies on product of browning reaction prepared from glucose amine

Article

Jan 1986

M. Oyaizu

Phytochemicals and Vitamin content of indigenous spices of Southeastern Nigeria

Article

Jan 2004

D.E. Okwu

The biology of Cromolaena odorata (L. King and Robison) 1. Flowering behaviour, pattern of growth and nitrate metabolism

Article

Jan 1974

Developing a rational basis for selection of antioxidant screening and testing methods

Article

Jan 2006

Karen Schaich

As pharmaceutical and nutraceutical interest in natural antioxidants has skyrocketed in the past few years, a plethora of methods have come into common use for screening antioxidant content of natural materials and for comparing antioxidant activity of various classes of compounds. These include ORAC (Oxygen Radical Absorbance Capacity), TEAC (Trolox Equivalent Antioxidant Capacity), FRAP (Ferric Reducing Antioxidant Power), TOSC (Total Oxidant Scavenging Capacity), TRAP (Total Radical-Trapping Antioxidant Parameter), DPPH reactivity, croton bleaching, LDL oxidation, liposome oxidation, and total phenolics analyses. For the most part, assay selection is based on ease of use and availability of instrumentation, and very often there is lack of correlation between is activity assays and phenolic content, between activities determined on the same material by different assays, and between activities determined by the same assay in different laboratories. The need for standardization of procedures is being addressed in a series of International Congresses on Antioxidant Methods. Perhaps more importantly, however, the assays listed above do not all measure the same chemical action. Some assays measure hydrogen atom transfer capability (classical radical quenching), some measure electron transfer propensity, and others measure a variety of other actions. The basic chemistry dominant in the assays and also active in the system of application must be considered as a basis for appropriate application of assays, accurate interpretation of results, and evaluation of antioxidant effectiveness for different end uses. This paper reviews mechanisms and effects of environment on the dominant antioxidant assays and recommends a new integrative approach using a panel of assays and conditions to provide more complete and accurate information during antioxidant testing.

Phytochemical Methods

Chapter

Jan 1984

J. B. Harborne

The macromolecules of plants are distinguished from all other constituents by their high molecular weight. This may vary from 10000 to over 1000000, whereas in other plant metabolites the molecular weight is rarely above 1000. Chemically, macromolecules consist of long chains of small structural units or ‘building blocks’, linked covalently in a number of different ways. Chemical characterization in the first instance therefore depends on identifying these smaller units. Proteins, for example, are long chains of amino acids (up to twenty different ones) joined together through peptide (—CO—NH—) links. Polysaccharides are similarly derived from the union of simple sugar units, such as glucose, joined through ether (—O—) links. The nucleic acids, by contrast, are more complex and have three types of structural unit: purine and pyrimidine bases, pentose sugars and phosphate groups. The three main classes of macromolecules found in plants are thus proteins, polysaccharides and nucleic acids. However, mixed polymers are also known. such as the glycoproteins, which contain both sugars and amino acids in covalent linkage.

Antioxidant activity ofOsbeckia aspera

Article

Jun 1998
PHYTOTHER RES

Plants of the Osbeckia family have been shown to possess hepatoprotective properties, which could be due to the presence of antioxidant compounds. The plant extract was shown to inhibit significantly in a dose-dependent manner, the activities of the DPPH (1,1-diphenyl-2-picrylhydrazyl) free radical (EC50 of 27.5 μg/mL), xanthine oxidase (EC50 of 1.16 mg/mL) and demonstrate a scavenging effect on hydroxyl radical mediated damage to deoxyribose (EC50 of 140 μg/mL). The plant extract possessed some prooxidant activity from the effect on bleomycin-induced DNA damage, but this was less than that shown by comparable concentrations of (+)-catechin or silymarin. © 1998 John Wiley & Sons, Ltd.

The Handbook of African Medicinal Plants

Article

Jan 1993

Mmaduakolam Iwu

Uses and Properties of Citrus Flavonoids

Article

Dec 1997

Flavonoids are a widely distributed group of polyphenolic compounds with health-related properties, which are based in their antioxidant activity. These properties have been found to include anticancer, antiviral, antiinflammatory activities, effects on capillary fragility, and an ability to inhibit human platelet aggregation. The antioxidant capacity of any flavonoid will be determined by a combination of the O-dihydroxy structure in the B-ring, the 2,3-double bond in conjugation with a 4-oxo function and the presence of both hydroxyl groups in positions 3 and 5. Flavanones, flavones, and flavonols are the flavonoids present in Citrus, and although flavones and flavonols have been found in low concentrations in Citrus tissues, in relationship to flavanones, these types of compounds have been show to be powerful antioxidants and free radical scavengers. Some Citrus flavonoids can be used directly as repellents or toxins or be used in plant improvement programs to obtain more resistant crops. In addition, some Citrus flavonoids and their derivates, in the field of food technology, are principally known for their ability to provide a bitter or sweet taste and as bitterness inhibitor. Keywords: Free radicals, antioxidant; anticarcinogenic; antiinflammatory; platelet aggregation; antiallergic; analgesic; antimicrobial; food additives

Phenolic content and anti-oxidant activity of olive extract

Article

Apr 2001
FOOD CHEM

The phenolic component of freeze-dried olive fruit was fractionated by high-performance liquid chromatography using ultraviolet, atmospheric pressure chemical ionisation (APCI) and electrospray ionisation (ESI) detection. The fractions together with several standards were tested for antioxidant activity in an aqueous and a lipid system. The negative ion mode of APCI and ESI showed less fragmentation than positive ion mode. The latter was generally more useful in obtaining fragmentation data and hence structural information. Some olive phenolics notably tyrosol exhibited a low ionisation efficiency in both APCI and ESI. There was no simple relationship between antioxidant activity and chemical structure. The ranking of antioxidant activity was strongly dependent on both the test system and on the substrate demonstrating the need to examine activity in both aqueous and lipid systems. Significant antioxidant activity was seen in most olive fractions and this was related to phenolic content. The kinetics of the oxidation process are complex and suggest that multiple pathways may be involved at different antioxidant concentrations.