The questionnaire raised a question, what are the common used ant-diabetic plants? Sample study included traditional herbs markets from different locations in north, south and middle of Jordan. The selected medicinal plant s Table 1 were collected from south and north regions of Jordan during the spring and summer The taxonomic identification was authenticated by Dr.
Al-Quran, biology department, Mutah University. The collected plants were cleaned, dried for 10 days under shade at room temperature, then grounded and extracted. Quercus calliprinos fruit, Q. Extracts preparation: The extracts were prepared according to Lapornik et al. The extract was filtrated through four layers of gauze. The final volume of the filtrate was brought to 50 mL. Total phenols: The total polyphenol content of plants extracts was estimated by the Folin-Ciocalteau phenol reagent according to Hagerman et al.
After addition of 0. The total phenolic content was expressed as Gallic Acid Equivalents GAE in milligrams per gram of dry matter of sample, using a standard curve generated with different concentrations of gallic acid. The control was prepared as above using methanol solvent instead of extract. The decrease in the absorbance of the formed blue to violet reagent was determined after 20 min at nm and the percentage inhibition activity was calculated from the formula:.
In presence of antioxidants, the pre-formed radical cation is reduced to ABTS, proportionally to the antioxidant activity. Statistical analysis: All data analyses were performed using the statistical package of statistical product and service solution SPSS version There was great variation between phenolic contents of these plants. However, the phenolic contents of the m ethanolic extract s ranged from 6. Table 2. The high antioxidant group involved Punica granatum peel, Quercus calliprinos leave, Quercus calliprinos fruit, Cinchona ledgeriana and Juniperus communis leave.
Low antioxidant plants: plants with the low antioxidant capacity involved M. Correlation between antioxidant activity and total phenolic content s: The correlation coefficients between antioxidant activity and total phenolic content of the tested plants were calculated Fig. There was a high. It has long been recognized that plants contain many natural substances. The phenolic compounds are widely distributed, sometimes present in surprisingly high concentrations, in plants and have an antioxidant activity Lapornik et al.
They have the ability to scavenge free radical s such as the Reactive Oxygen Species ROS which are determined by their reactivity as hydrogen- or electron donating agents Fernandez-Pachon et al. In present study, the total antioxidant effect of methanol and aqueous extracts of some medicinal plant s that are traditionally used in Jordan, were measured and compared to total phenolic content s. Extreme variations in antioxidant activity and total phenols were found between tested extracts. The large variation in plant antioxidant activity may result from differences in total phenolic content s Table 2.
Such observation agreed with several previous findings Velioglu et al. Moreover, Singleton and Rossi noticed that various phenolic compounds have different responses to Folin-Ciocalteau assay. The molar response of this method is roughly proportional to the number of phenolic hydroxyl groups in a given substrate, but the reducing capacity is enhanced when two phenolic hydroxyl groups are oriented in ortho or para-position. Since these structural features of phenolic compounds are responsible for antioxidant activity Katalinic et al.
Thus, polyphenols measurements in extracts may be related to their antioxidant activities. DPPH and ABTS methods have been used by many researchers to evaluate the free radical scavenging activity of antioxidant molecules and plant extracts. DPPH does generate strongly colored solutions with methanol which is eliminated in presence of antioxidants Blois, ; Matsukawa et al. On the other hand, the decolorization of the ABTS radical cation also reflects the capacity of an antioxidant species to donate electrons or hydrogen atoms to deactivate these radical species Pellegrini et al.
The data obtained from the two radical scavenging methods suggest high accuracy and constancy between them. They established that the reactions of phenols with ABTS radical cation are usually rapid, but the reactions with DPPH radical differ from compound to compound. Furthermore, the affinities of different plants toward the two above radicals were sometimes significantly altered due to variation of ABTS and DPPH solubility in aqueous medium.
This study established that the methanol extracts were significantly higher in phenolic contents and antioxidant activity than aqueous extracts. However, it was found that methanolic plant extracts are the most effective scavenger of DPPH radical Miliauskasa et al. Lapornik et al. Hence, it was suggested that methanol is more efficient solvent for cell walls and seeds degradation, that have unpolar character causing the release of polyphenols from cells.
In other literature Moure et al. Other explanation for such decrease is ascribed to the activity of polyphenol oxidase, which degrade polyphenols in water extracts, but neutralized in methanol medium Zhang et al.
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In present study, the decrease in antioxidant activity among the aqueous extracts was in accordance with the amount of plant phenolic contents. This verifies that the amount of phenolic compounds was responsible for their antioxidant activities. Therefore, the differences between antioxidant activities of methanol and aqueous extracts might reflect differences in polyphenolic contents of these extracts.
Punica granatum peel m ethanolic extract exhibited the highest polyphenolic contents and antioxidant capacity among all tested extracts. The total phenolic content of the Punica granatum peel methanol extract was Previous work found that the HPLC analysis of the m ethanolic extract of Punica granatum peel have detected some polyphenols, like gallic acid These polyphenols present in the m ethanolic extract of Punica granatum peel may be responsible for its high antioxidant activity. Moreover, the antioxidant activity of ethanol, methanol and water extracts of Punica granatum peels and seeds have been reported in various in vitro models.
Among all extracts, the methanol extract possessed the highest antioxidant activity in various models Singh et al. In conclusion, present study indicated that some Jordanian medicinal plant s are promising sources of natural antioxidants. Total phenol content and total antioxidant capacity differs significantly among 21 selected plant extracts. There was significant linear correlation between phenolics concentration and antioxidant capacity of extracts.
The antioxidant capacity and radical scavenging agreed with their uses as traditional ant-diabetic remedy. The authors thanks the Deanship of Academic Research at Mutah University for financial support, grant Abuharfeil, N. Maraqa and S. Von Kleist, Augmentation of natural killer cell activity in vitro against tumor cells by wild plants from Jordan.
Abu-Irmaileh, Herbal medicine in Jordan with special emphasis on less commonly used medicinal herbs. Atta, Pharmacological screening of the anti-ulcerogenic effects of some Jordanian medicinal plants in rats.
Role of oxidative stress in development of complications in diabetes. Diabetes, Antioxidant determinations by the use of a stable free radical. Nature, Cerami, The biochemistry of the complications of diabetes. Cai, Y. Luo, M. Sun and H. Corke, Antioxidant activity and phenolic compounds of traditional Chinese medicinal plants associated with anticancer. Life Sci. Lissi, Kinetics of the reaction between 2,2-azinobis 3.
Kinetics, Chanwitheesuk, A. Teerawutgulrag and N. Rakariyatham, Screening of antioxidant activity and antioxidant compounds of some edible plants of Thailand. Food Chem. Plant products as antimicrobial agents. Yu, R. Bateman and G. Brock, Oxidative stress and antioxidant therapy: Their impact in diabetes-associated erectile dysfunction. Ke, M. Yang and C. Rice-Evans, Methods Enzymol.
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[Full text] Antioxidant compounds and activities of the stem, flower, and leaf ext | DDDT
Prosek and A. Wondra, Comparison of extracts prepared from plant by-products using different solvents and extraction time. Food Eng. Amar, Ethnopharmacological survey of traditional drugs sold in the Kingdom of Jordan. El-Oqlah, Antimicrobial activity of extracts of herbal plants used in the traditional medicine of Jordan. A survey of medicinal plants of Jajarkot district, Nepal. Dubinsky, E.
Kishimoto, K. Moringa tree is also growing in south-east of Iran. The antioxidant activity of M. Moreover, total phenolic content of the leaf extract was determined and using 1 HNMR, mass and spectroscopic methods, the structure of the isolated flavonoid glycoside, rutin, as one of the compounds responsible for reported antioxidant activities was identified. Antioxidants provide protection against degenerative diseases including cancer, coronary heart, and Alzheimer's diseases 1.
Reactive Oxygen Species ROS , contribute to cellular aging, mutagenesis, carcinogenesis, and coronary heart disease, likely through destabilization of membranes, DNA and protein damage, and oxidation of low-density lipoprotein LDL 2. Mechanism of action of antioxidants includes the suppression of ROS formation, the inhibition of enzymes or chelating of elements involved in free-radical production. Furthermore, antioxidants scavenge reactive species, and upregulate antioxidant defences 3. Plants are rich sources of natural antioxidants, the best known are tocopherols, carotenoids, vitamin C, flavonoids, and different other phenolic compounds 1.
Recently, among natural antioxidants,flavonoids have received increasing attention. As compared with vitamin C and E, dietary flavonoids are considered to be more powerful antioxidants 4. Flavonoids are known to be highly effective antioxidants by scavenging oxygen radicals, by having interesting anti-cancer, hypolipidemic, anti-ageing, and anti-inflammatory activities 5. Moreover, the protective effects of flavonoids in biological systems are attributed to their capacity to scavenge free radicals, chelate metal catalysts, activate antioxidant enzymes, reduce alpha-tocopherol radicals, and inhibit oxidases 2.
Furthermore, phenolic compounds have phenolic hydroxyl groups which can dissociate to negatively charged phenolates. Dissociated phenolics can form hydrogen and ionic bonds with various proteins, which lead to a disturbance of their 3D-structures and in consequence to a change in their bioactivity 6. Moringaceae occurs in the Middle East to India.
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In Iran it is a desert tree growing in Sistan and Baluchestan province and is locally called Gas-e-rowghan or Gaz Rokh 7 , 8. Several reports on antioxidant activity of leaves and seeds of another Moringa species exist 9 , Leaves of M. To the best of our knowledge no effort has been made to evaluate in vitro antioxidant activity of leaves of M. Therefore, in the present study, the antioxidant activity of the leaf extract of M. In addition, total phenolics determination as well as isolation and identification of the flavonoid rutin as the main active antioxidant were carried out.
All solvents used for the preparation of crude samples and separation process, diphenylboric acid 2-amino ethyl ester natural product , rutine, and nitroblue tetrazolium NBT were purchased from Merck Darmstadt, Germany. Voucher specimens of the seeds, leaves and stems No. About g of air-dried leaves of M. After filtration the solvent was evaporated under reduced pressure and the dry extract stored under refrigerator until used for further analysis 9.
The DPPH radical scavenging assay is a standard procedure to determine the antioxidant activity of plant extracts and natural compounds. The free radical scavenging activity of methanolic leaf extract was measured using the method described by Blois 12 with some modifications. After 30 min, the absorbance was measured at nm The radical scavenging activity of DPPH was calculated using the following equation:. Superoxide anion scavenging ability of the leaf extract was determined by the modified method of Robak and Gryglewski Five min later, the absorption was measured at nm.
Antioxidants have the ability of inhibiting the blue NBT formation which results in the decrease of the absorbance at nm. The inhibition of superoxide anion generation was calculated as percentage using the following formula:. Antioxidant compounds generally contain phenolic groups and hence, the amounts of phenolic compounds in the methanolic extract of M. To 1 ml of the extract in methanol, 5. After shaking, it was kept for 2 h in darkness and the absorbance was measured at nm. Using gallic acid monohydrate, a standard curve was prepared.
Methanolic extract of leaves was primarily developed on TLC plates with solvent system ethylacetate: formic acid: acetic acid: H2O After developing, the plates were observed under UV at nm with and without exposure to ammonia and then sprayed with DPPH reagent 0. Further isolation and purification methods for identification of the bold spot found on TLC were designed.
More confirmation was obtained using the analytical HPLC system. Then cooled for TLC analysis which performed on cellulose with n-Buthanol: acetic acid: water and ethylacetate: pyridine: water solvent systems 17 , 19 beside standard sugars. The developed chromatograms were sprayed by aniline hydrogen phthalate reagent to detect hydrolyzed sugars.
Antioxidant activity of some plant extracts of the family labiatae
Statistical comparisons were made with two-tailed unpaired student's t test assuming equal variance. The leaf extract showed IC 50 value of 8. Anti radical activity of M. Antioxidant activity of M. The IC 50 value of leaf extract was Scavenging activity of M.
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Scavenging activity of EGCG as positive control on superoxide anion radicals. The TPC was determined following a modified Follin-Ciocalteu method and result was expressed as gallic acid equivalents. Isolated compound was obtained as pale yellow solid with positive reaction to natural product reagent and to the yellow spot in purple background after spraying with DPPH reagent.
Antioxidant activity is evaluated by different methods but the most widely used methods are those that generate free radical species which are then neutralized by antioxidant compounds. EGCG had stronger dose-dependent radical scavenging activity against DPPH and superoxide anion radicals which agrees with an earlier study However, leaf extract as well as EGCG revealed dose-dependent anti radical activity.
The phenolic compounds may contribute directly to antioxidative effect. The presence of 3-OH group as well as hydroxyl groups in ring B is related to the superoxide scavenging activity of flavonoids. Epigallocatechin with a hydroxyl group at C-3 and three hydroxyl groups in ring B shows high superoxide scavenging activity It is also demonstrated that the principal site of antioxidant reactions in EGCG and epigallocatechin EGC was the trihydroxy-phenyl B-ring, regardless of the presence of a 3-galloyl moiety Therefore, the observed in vitro antioxidant activity of the leaf extract of M.
Spraying the TLC with DPPH is a suitable method for detection of antioxidants in crude plant extracts or pure compounds isolated from plant material. This method was also used for screening the antioxidant activity of flavonoids from the leaves of Licania licaniaeflora 5. Our study demonstrates that methanolic extract of M.
The antioxidant potential of M. Thus, the free radical scavenging ability of M. We are grateful to Mr. Parvaresh Department of Natural Resources of Chabahar for providing the seeds and leaves of the plant. National Center for Biotechnology Information , U. Journal List Res Pharm Sci v. Res Pharm Sci. Wink , 2 S. Afsharypuor , 1 G. Asghari , 3 and A. Mohagheghzadeh 4. Find articles by S. Find articles by M. Find articles by G.
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