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  • Review Article   
  • J Phytochemistry Biochem, Vol 1(1)

In vitro Antioxidant Activity and Phytochemical Screening of Flowers and Leaves of Hypericum perforatum L. Ethanolic Extracts from Tonekabon - Iran

Alimohammad Asgharian1* and Shahab Ojani2
1Department of Chemistry and Biology, Ryerson University, 350 Victoria Street, Toronto, ON M5B 2K3, Canada
2Young Researcher and Elit Club, Tonekabon Branch, Islamic Azad University, Tonekabon, Iran
*Corresponding Author: Alimohammad Asgharian, Department of Chemistry and Biology, Ryerson University, 350 Victoria Street, Toronto, On M5B 2K3, Canada, Tel: +989122958644 Exn. +981154271101-9, Email: mehranasgharian@yahoo.com

Received: 09-Nov-2017 / Accepted Date: 30-Nov-2017 / Published Date: 08-Dec-2017

Abstract

Medicinal plants are an important source of phytochemicals that offer traditional medicinal treatment of various ailments. This research set to assess phytochemicals in the ethanolic extracts of Hypericum perforatum L. leaves and flowers by quantitative and qualitative screening procedures. The Hypericum perforatum L. flowers and leaves were gathered, and the extract was provided from ethanol (5%) by microwave assisted extraction (MAE). The phytochemical assessment was done applying standard methods & the phytochemical evaluation by using standard methods. The total phenolic contents of ethanolic extracts were estimated by Folin Ciocalteau method and total flavonoids contents were determined by the Aluminium Chloride Colorimetric method. Ethanolic extracts invitro antioxidant activity was assessed by via evaluating 1,1-diphenyl-2 picrylhydrazyl (DPPH) radical scavenging activity by the standard method. Ethanolic extracts from flowers and leaves of Hypericum perforatum L. showed total phenolic contents of (15.32 ± 0.07) and (7.39 ± 0.43) mg GAE/g dry plant material respectively. Total flavonoid contents of ethanolic extracts from leaves and flowers of Hypericum perforatum L. were (1.09 ± 0.08) and (0.38 ± 0.05) mg QE/g dry plant material, respectively. The antioxidant activity of the investigated ethanolic extract of leaves and flowers of Hypericum perforatum L. were scavenging ability of DPPH radical scavenging activity and IC50 value (89.45% to 2.15 ± 0.02) and (74.77% to 1.96 ± 0.06) mg/ml respectively. The ethanolic extract of leaves and flowers of Hypericum perforatum L. contains terpenoids, flavonoids, phenols, tannins, cardiac glycosides, quinones and phlobatannins. This study may contribute to drugs development to cure different diseases.

Keywords: Hypericum perforatum L.; Microwave assisted extraction; Ethanolic extracts; Antioxidant activity; Folin-Ciocalteau; Flavonoids; Cardiac glycosides

Introduction

Free radicals are chemical species one or more unpaired electrons due which are extremely unstable and destroy other molecules by extraction of electrons to gain stability. The typical reactive oxygen species (ROS) are nitric oxide (NO), peroxyl radical (ROO) highly reactive hydroxyl radical (OH), hydrogen peroxide (H2O2), superoxide anion (O2(-)), and peroxynitrite anion (ONOO). Free radicals are in the body, as they are needed to supply detoxification, energy chemical signaling, and immune function [1-3]. Free radicals can trigger the bio molecules oxidation such as protein, amino acids, lipid, and DNA, causing cell damage and stimulating many illnesses. Oxidative stress can start by an imbalance between antioxidants and reactive oxygen species causing cellular damage; oxidative stress is the origin of numerous illnesses like age related diseases, cancer, cataracts, and Parkinson’s. Antioxidants reduce the oxidative stress in cells and may improve many illnesses like cancer, cardiovascular illnesses, and inflammatory diseases [4-6]. Oxidative stress can be caused by an imbalance between reactive oxygen species and antioxidants resulting in cellular damage, and oxidative stress is the major of many diseases as cataracts, cancer, age related diseases, and Parkinson’s disease. Antioxidants decrease the oxidative stress in cells and are beneficial for the improvement of many diseases such as cardiovascular diseases, cancer and inflammatory diseases. This activity occurs because of the capability of antioxidants to decrease oxidative stress through scavenging or neutralizing of reactive species via hydrogen donation [7-9]. The medicinal plants, as potential sources of drugs are rich sources of secondary metabolites including glycosides, steroids, alkaloids and flavonoids. Almost, one third of the pharmaceuticals are plant origin. Since all plants can synthesize a great value of organic molecules /phytochemicals, they are called secondary metabolites [10]. Plants derived compounds have crucial impact different clinically beneficial drugs. Phytochemicals are bioactive compounds which are found in plants that work with dietary fiber and nutrients to protect the body against diseases. They are non-nutritive compounds. These phytochemicals are the secondary metabolites which are found in small quantities in higher plants and contain flavonoids, terpenoids, alkaloids, steroids, tannins etc. [11]. Many phytochemicals possess antioxidant activity and decrease the risk of diseases. It is important to be acquainted with the type of phytochemical constituent, hence, knowing the biological activity type possibly exhibited by the plant [12,13]. Medicinal plants and the impact of phytomedicine on the health of a great number of the world’s population have attracted the attention of many researchers. Hypericum perforatum L. belongs to the Hypericaceae family [14-16]. Hypericum perforatum L. is a crucial medicinal plant, and has many medicinal usages, such as antidepressive medication [17-20], anti-viral [21], Hypo-cholesterolemic effects [22], antioxidant [23,24], anti-microbial [25], anti-HIV activity [26], burns, eczema, skin wounds, anti-inflammatory, diabetes mellitus, anti-cancer, migraine headaches and etc. [27-32]. This study evaluated the antioxidant activities and phytochemical screening of leaves and flowers of Hypericum perforatum L. ethanolic extracts from Iran.

Materials And Methods

All chemicals and reagents had analytical grade with utmost purity. Fresh plant flowers and leaves of Hypericum perforatum L. were collected from Se hezar (N 36° 37′, E 50° 50′; at 2650 m altitude) Tonekabon, Iran, in April 2015 and the plant was identified by Fariba serpooshan in Hebarium (Islamic Azad University - Tonekabon Branch). Then leaves and flowers dried and made powder. 10 g of powders of leaves and flowers of Hypericum perforatum L. were combined with 5% ethanol exposed to microwave irradiation at 300 W; suspension was radiated in microwave oven (model GE280S) at regular intervals (10 min irradiation and 3 min off). After extracting, the extract was cooled down to room temperature to keep room temperature. Then extract was filtered use Whatmann’s Filter No. 1 filter paper. The solvent was evaporated under vacuum in a rotary evaporator (model LABOROTA 4001) and finally, stored in refrigerator at 40°C till further use.

The ethanolic extracts of leaves and flowers of Hypericum perforatum L. was following subjected to various chemical experiments to detect various phytoconstituents by standard methods [33-37]. Qualitative phytochemical analysis of ethanolic extracts of leaves and flowers of Hypericum perforatum L. were done after the standard methods. Few drops of 1% NH3 solution was mixed with the extract in a test tube. Yellow coloration was seen for flavonoids. 2-3 ml of the extract was treated with 10% aqueous FeCl3 drops, and blue green color was detected.

In this experiment, 2-3 ml of extract was treated with drops of 10% aqueous FeCl3 and emergence of blue green color was observed. 1 ml of the extract and 2 ml of chloroform were mixed. Nearly 3 ml of conc. H2SO4 was transferred with care from the sides of test tube. Reddish brown coloration at interface showed terpenoids presence. For this purpose, the mixture including 0.5 ml of extract solution, 1 ml of distilled water prepared and 1-2 drops of ferric chloride solution were added and brownish green or a blue black coloration checked. 2 ml of extract and 3 drops of copper acetate solution were mixed. Emerald green solution shows the existence of Di-terpenes. 1 ml of extract, was added to 10% NaOH 1 ml. Yellow color formation showed the coumarins presence. 1 ml of extract and 1 ml of concentrated sulfuric acid were added. Red color indicated the presence of quinones.

Based on the protocol, in a test tube, 5 ml of extract was mixed with 5 ml of distilled water and heated. The stable foam formation indicated of the presence of saponins. 5 ml of the extract was treated with 2 ml of glacial acetic acid containing one drop of ferric chloride solution. It was underlayed with concentrated H2SO4 (1 ml). Brown ring formation into interface layer, manifested a deoxysugar feature of cardenolides. A violet ring may be seen below the brown ring; however, a greenish ring can be made gradually across thin layer in the acetic acid layer. To determine phlobatannins, 2 ml of extract was mixed with 2 ml of 1% HCl, and then it was boiled. A red precipitate showed the presence of phlobatannins. The phytochemicals which are present in the ethanolic extracts of flowers and leaves of Hypericum perforatum L. were determined and quantified by standard procedures.

This experiment was done using the Folin- Ciocalteau procedure with little changes [38]. Total phenolic contents were indicated using the Folin- Ciocalteau procedure with little changes. The 0.25 ml extract and Folin Ciocalteu reagent (2.5 ml, 1:10 diluted with distilled water) were mixed for 5 min 2 ml of aqueous Na2CO3 (1 M) was also added. The mixture was stood for 15 min; it was read by colorimetric method by spectrophotometry (model Unico 2100) at 765 nm. Gallic acid was applied to provide standard curve; the overall phenolic content of extract was indicated as gallic acid equivalent (mg/g of dry mass), a popular reference compound.

Based on AlCl3 method, overall flavonoid was indicated by aluminum chloride colorimetric method [39]. 0.5 ml of plant extract (1 mg/ml) and 0.1 ml of 1 M potassium acetate, 0.1 ml of 10% AlCl3, 1.5 ml of methanol, and 2.8 ml of distilled water were mixed. The reaction mixture absorbance was obtained at 415 nm on UV– Visible spectrophotometer after incubation at room temperature for 30 minutes. The overall flavonoid content was indicated according to quercetin equivalent which is typical reference criterion.

DPPH radical-scavenging activity was calculated using the Shimada et al. (1992) method with some changes [40]. One ml of 0.1 mM DPPH0 solution in methanol and 1 ml of every sample were mixed; afterwards, the mixture was vigorously shaken and put in the dark for 30 min at room temperature. The resultant solution’s absorbance was obtained at 517 nm. The activity of DPPH0 radical-scavenging was measured as following:

Scavenging activity (%) = [(A0 - (A - Ab)) / A0] × 100%

Where A0 was DPPH0 value without sample; A was the sample value and DPPH0; Ab was the sample value without DPPH0. The ascorbic acid was employed as positive control [41,42].

Inhibition Concentration (IC50) was applied to interpret the outcomes from DPPH0 method [43]. The sample discoloration was plotted against the concentration of sample to measure the value of IC50. It is des as the sample content needed to reduce the DPPH0 absorbance by 50%. All antioxidant activity assessments were done three times. The data were shown as means ± standard deviation.

Result

The initial phytochemical experiments may chemical constituents in the plant material, inducing their quantitative estimation and locating the origin of pharmacologically active chemical compound. Qualitative analysis of phytochemical compounds like saponins, tannins, flavonoids, terpenoids, phenols, coumarins, Di-terpenes, quinones, cardiac glycosides, quinones and phlobatannins were examined in the ethanolic extracts of flowers and leaves of Hypericum perforatum L. employing standard procedures. In ethanolic extracts of flowers of Hypericum perforatum L. 8 tests were positive. And 9 tests were positive in ethanolic extracts of leaves of Hypericum perforatum L. In present study, we have found that most of the biologically active phytochemicals were present in the ethanolic, extracts of flowers and leaves of Hypericum perforatum L. (Table 1) [44-48].

No. Phytochemical Constituents Result
Flowers Leaves
1 Terpenoids + +
2 Flavonoids + +
3 Phenols + +
4 Tannins + +
5 Di-Terpenes - +
6 Coumarins - +
7 Saponins + *
8 Cardiac glycosides + +
9 Quinones + +
10 Phlobatannins + +
(+): positive, (-): Negative, (*): Few and (ND): Not defined

Table 1: In vitro qualitative phytochemical analysis of ethanolic extract of flowers of Hypericum perforatum L.

The plants screening for medicinal value has been done using many experiments and preliminary phytochemical analyses. Phytochemical screening is very important in identification of novel sources of industrially and therapeutically valuable compounds with medicinal significance, to efficiently use natural wealth available. These phytochemicals have anti-oxidants, anti-inflammatory, anticancer, anti-bacterial, anti-diabetic, anti-pyretic, allergenic, anticoronary, antiseptic, analgesic, anti-arthritic, sedative, anesthetic, hypo-cholesterolemic, and hepatoprotective activities [49-58]. Total phenol contents of the extracts was indicated based on Folin- Ciocalteu procedure and stated in mg of gallic acid equivalent per g (mg GAE/g) of extract, assuming the standard curve (y=26.64x - 0.042 and R2 =0.998). Total phenol contents in ethanolic extracts of flowers of Hypericum perforatum L. was 7.39 ± 0.43. But, total phenol contents in ethanolic extracts of leaves of Hypericum perforatum L. was 15.32 ± 0.07 (Table 2). Total flavonoid contents of the extracts was indicated based on aluminum chloride colorimetric procedure and stated in mg of quercetin equivalent per g (mg QE/g) of extract, assuming the standard curve curve (y=6.944x+0.070 and R2=0.996). Total flavonoid contents in ethanolic extracts of flowers of Hypericum perforatum L. was 0.38 ± 0.05. But, total flavonoid contents in ethanolic extracts of leaves of Hypericum perforatum L. was 1.09 ± 0.08 (Table 2).

Test Sample Total Phenol Contents (mg GAE/g) Total Flavonoid Contents (mg QE/g) Radical Scavenging Activity (%) IC50 values
Flowers 7.39 ± 0.43 0.38 ± 0.05 74.77 1.96 ± 0.06
Leaves 15.32 ± 0.07 1.09 ± 0.08 89.45 2.15 ± 0.02

Table 2: In vitro quantitative phytochemical analysis of ethanolic extract of flowers of Hypericum perforatum L.

The radical scavenging activity (%) and IC50 ethanolic extracts of flowers of Hypericum perforatum L. for DPPH radical scavenging activity were 74.77%, 1.96 ± 0.06 mg/ml. But, the radical scavenging activity (%) and IC50 ethanolic extracts of leaves of Hypericum perforatum L. for DPPH radical scavenging activity were 89.45% and 2.15 ± 0.02 mg/ml, while that of ascorbic acid used as the reference control was (0.0116) mg/ml (Table 2).

Discussion

Medicinal plants have become very popular because they have very few side effects as compared to synthetic drugs. Phytochemicals compounds are widely studied because they are highly abundance in nature and often used as parts of defense mechanisms in plants. The screening of plants usually involves several approach; ethno botanical approach is one of the common methods that are employed in choosing the plant for pharmacological study. The initial phytochemical experiments may chemical constituents in the plant material, inducing their quantitative estimation and locating the origin of pharmacologically active chemical compound [59-61]. Terpenoids have been implicated in antibacterial and anti-neoplastic functions hence the use of the plant to treat burns, skin diseases and insect stings [62-64]. The initial screening experiments can be useful beneficial in detecting the bioactive compounds and can bring about the drug development and discovery (phytoestrogens related to relief of menopausal signs, reducing osteoporosis, improving blood cholesterol levels, and reducing the risk of some hormone-related cancer and coronary heart illnesses. Also, several flavonoids and cardiac glycosides possess hypoglycemic activities [65]. Our results showed Hypericum perforatum L. from Tonekabon has high level of flavonoids which can be used for herb based therapy. In addition, Polyphenols can treat stomach and kidney ailments and protect and prevent against numerous degenerative illnesses and pathological processes like coronary heart disease, ageing degenerative illnesses, neurodegenerative disorders, Alzheimer’s disease, and atherosclerosis cataracts [66]. Tannins may have potential value such as cytotoxic, anti-cancer agents and hasten the healing of wound and inflamed mucous membranes [67,68]. Coumaris raise the flow of blood in the veins and reduces capillary permeability and has anti-apoptotic feature [69]. Saponins can precipitate and coagulate red blood cells; they also possess cholesterol binding features. Formation of foams in aqueous solutions, hemolytic activity [70] and traditionally saponins are widely employed as molluscicides and detergents, in addition to their industrial usages as surface active agents and foaming, and they also have useful health impacts [71], Saponins are applied in hyperglycemia, anti-oxidant, anti-inflammatory, anti-cancer, and hyper-cholesterolemia weight loss etc. [72,73]. Cardiac glycosides have severe toxicity since they may influence the heart and atrial fibrillation [74]. Among many quinone compounds such as natural compounds; the naphthoquinone derivatives in particular, possess a broad variety of bioactivities. Potency as anti- inflammatory agent, anti-asthma medicine, anti-allergic agent, medicine, anti-dragon gore medicine, bronchodilator, thrombus prevention, and hypotension are seen in their derivatives [75]. Also, plants generate numerous secondary metabolites with antioxidant capacity. Antioxidants obstruct the function of free radicals implicated in in the aging process and the pathogenesis of several illnesses. A crucial role is played by free radicals in monitoring different biological processes essential for the body. They can implicate cell-signaling mechanism in the body. Revealing the free radicals are essential, but at the same time, they are detrimental harmful for the body. Thus, it has several mechanisms to reduce free radical stimulated injury. The injury was repaired by many enzymes such as superoxide dismutase, catalase and etc. [76-78]. Additionally, antioxidants are critical for these defense mechanisms; some compounds containing vitamin A, vitamin E, vitamin C, and polyphenols basically show these tasks.

Conclusion

The findings revealed that the ethanolic extracts of leaves and flowers of Hypericum perforatum L. have medicinally crucial bioactive agents, justifying its usage in the traditional medication for treating various illnesses.

Acknowledgement

We gratefully acknowledge the financial support from the research council of Tonekabon Branch Islamic Azad University.

References

  1. Halliwell B, Gutteridge JMC (1985) Free Radicals in Biology and Medicine. Oxford University Press, Oxford, UK.
  2. Ho YS, Magnenat JL, Gargano M, Cao J (1998) The nature of antioxidant defense mechanisms: A lesson from transgenic studies. Environ Health Perspect 106: 1219-1228.
  3. Tepe B, Sokmen A (2007) Screening of the antioxidative properties and total phenolic contents of three endemic Tanacetum subspecies from Turkish flora. Bioresour Technol 98: 3076- 3079.
  4. Uddin SN, Akond MA, Mubassara S, Yesmin MN (2008) Antioxidant and Antibacterial activities of Trema cannabina. Middle-East J Sci Res 3: 105-108.
  5. Jayasri MA, Mathew L, Radha A (2009) A report on the antioxidant activities of leaves and rhizomes of Costus pictus D. Don. Inter J Integ. Biol 5: 20-26.
  6. Chen FW, Shieh P, Kuo D, Hsieh C (2006) Evaluation of the antioxidant activity of Ruellia tuberosa. Food Chem 94: 14-18.
  7. Erkan N, Ayranci G, Ayranci E (2008) Antioxidant activity of rosemary (Rosmarinus officinalis) extract, Black seed (Nigella sativa) essential oil, carnosic acid, rosmarinic acid and sesamol. Food Chem 110: 76-82.
  8. Mantle D, Eddeb F, Pickering AT (2000) Comparison of relative antioxidant activities of British medicinal plant species in vitro. Ethnopharmacol 72: 47- 51.
  9. Koleva II, Van Beek TA, Linssen JPH, de Groot A, Eustatieva LN (2002) Screening of plant extracts for antioxidant activity: A comparative study on three testing methods. Phytochem Anal 13: 8-17.
  10. Madhuri S, Pandey G (2009) Some anticancer plants of foreign origin. Curr Sci 96: 779-783.
  11. Peteros NP, Mylene MU (2010) Antioxidant & cytotoxic activities & phytochemical screening of four Phillppine medicinal plants. J Med Plant Res 4: 407-414.
  12. Pourshamsian K, Ojani S (2016) Phytochemical Screening of the aqueous extract of seeds of Polylophium involucratum (Pall.) Boiss. From Ramsar-Iran. Planta Medica 82.
  13. Agbafor KN, Nwachukwu N (2011) Phytochemical Analysis and Antioxidant property of leaf extract of vitex doniana and mucuna pruriens. Bio Chem Res Int: 1-4
  14. Cakir A, Mavi A, Yildirim A, Duru ME, Harmandar M, et al. (2003) Isolation and characterization of antioxidant phenolic compounds from the aerial parts of Hypericum hyssopifolium L. by activity-guided fractionation, J. Ethnopharmacol 87: 73-83.
  15. Stark O (1969) Clinical Laboratory Analysis, Medical Book, Belgrade, Serbia.
  16. Maisenbacher P, Kovar KA (1992) Adhyperforin: A homologue of hyperforin from Hypericum perforatum. Planta Med 58: 291-293.
  17. Butterweck V, Jurgenliemk G, Nahrstedt A, Winterhoff H (2000) Flavonoids from Hypericum perforatum show antidepressant activity in the forced swimming test. Planta Med 66: 3-6.
  18. Bombardelli E, Morazzoni P (1995) Hypericum perforatum. Fitoterapia. 66: 43.
  19. Volz HP (1997) Controlled clinical trials of hypericum extracts in depressed patients--An overview Pharmacopsychiatry 30: 72.
  20. Stevinson C, Ernst E (1999) Hypericum for depression. An update of the clinical evidence. Eur Neuropsychopharmac 9: 501-505.
  21. Zou YP, Lu YH, Wei DZ (2005) Hypocholesterolemic effects of a flavonoid-rich extract of Hypericum perforatum L. in rats fed a cholesterol-rich diet. J Agric Food Chem 53: 2462-2466.
  22. Benedi J, Arroyo R, Romero C, Martin-Aragon S, Villar AM ( 2004) Antioxidant properties and protective effects of a standardized extract of Hypericum perforatum on hydrogen peroxide-induced oxidative damage in PC12 cell. Life Sci 75: 1263-1276.
  23. Spiteller M, Ozen T, Smelcerovic A, Zuehlke S, Mimica-Dukic N(2008): Phenolic constituents and the in vitro antioxidant activity of the flowers of Hypericum venustum. Fitoterapia 79:191-193.
  24. Radulovic N, Stankov-Jovanovic V, Stojanovic G, Smelcerovic A, Spiteller M (2007) Screening of in vitro antimicrobial and antioxidant activity of nine Hypericum species from the Balkans. Food Chem 103: 15-21.
  25. Maury W, Price JP, Brindley MA, Oh C, Neighbors JD, et al. (2009) Identification of light-independent inhibition of human immunodeficiency virus-1 infection through bioguided fractionation of Hypericum perforatum. Virol J 6: 1-12.
  26. Wang ZQ, Gorski JC, Hamman MA, Huang S, Lesko LJ, et al. (2001)The effects of St John’s wort (Hypericum perforatum) on human cytochrome P450 activity. Clin Pharm Ther 70: 317-326.
  27. Schempp CM, Windeck T, Hezel S, Simon JC (2003) Topical treatment of atopic dermatitis with St. John's wort cream a randomized, placebo controlled, double blind half-side comparison. Phytomedicine 4: 31-37.
  28. Luo L, Sun Q, Mao YY, Lu YH, Tan RX (2004) Inhibitory effects of flavonoids from Hypericum perforatum on nitric oxide synthase. J Ethnopharmacol 93: 221-225.
  29. Butterweck V (2003) Mechanism of action of St. John’s Wort in depression: What is known? CNS Drugs 17: 539-562.
  30. Jakovljevic V, Popovic M, Mimica-Dukic N, Sabo A, Gvozdenovic LJ (2000) Pharmacodynamic study of Hypericum perforatum L. Phytomedicine 7: 449-453.
  31. Barnes J, Anderson LA, Phillipson JD (2001) St. John’s wort (Hypericum perforatum L.): A review of its chemistry, pharmacology and clinical properties. J Pharm Pharmacol 53: 583-600.
  32. Harborn JB (1973) Phytochemical Methods: A guide to Modern Techniques of plants Analysis, Chapman & Hall. London, Ltd.: 49-188.
  33. Trease GE, Evans IC (1983) Textbook of Pharmacognosy. 12th Edn. Bailliere Tindall. London: 21-22.
  34. Sofowora LA (1993) Medicinal plants and traditional medicine in Africa. Spectrum Books Ltd, Ibaban: 55-71.
  35. Pan Y, Wang K, Huang S, Wang H, Mu X, et al. (2008) Antioxidant activity of microwave-assisted extract of longan (Dimocarpus longan Lour.) peel. J Food Chem 106:1264-1270.
  36. Savithramma N, Lingarao M, Suhrulatha D (2011) Screening of medicinal plant for secondary metabolites. Middle-East J Sci Res 8: 579-584.
  37. Singleton VL, Orthofer R, Lamuela-Raventos RM (1999) Analysis of total phenols and other oxidation substrates and antioxidants by means of Folin-Ciocalteu reagent. Methods Enzymol 299: 152-178.
  38. Chang C, Yang M, Wen H, Chern J (2002) Estimation of total flavonoids content in propolis by two complementary colorimetric methods. J Food Drug Analysis 10: 178-182.
  39. Shimada K, Fujikawa K, Yahara K, Nakamura T (1992) Antioxidative properties of xanthan on the autoxidation of soybean oil in cyclodextrin emulsion. J Agric Food Chem 40: 945-948.
  40. Blois MS (1958) Antioxidant determination by the use of a stable free radical. Nature 29: 1199-1200.
  41. Brand-Williams W, Cuvelier ME, Berset C (1995) Use of a free radical method to evaluate antioxidant activity, Lebensmittel- Wissenschaftund-Technologie. Food Sci Technol 28: 25-30.
  42. Kumaran A, Karunakaran R (2006) Antioxidant and free radical scavenging activity of an aqueous extracts of Coleus aromaticus. Food chem 97: 109-114.
  43. Mungole A, Chaturvedi A (2011) Hibiscus sabdariffa L a rich source of secondary metabolites. J Pharmaceutical Sci Rewiev Res 6: 83-87.
  44. Ram RL (2001) Preliminary phytochemical analysis of medicinal plants of South Chotanagpur used against dysentery. Advances Plant Sci 14: 525-530.
  45. Ni Q, Xu G, Lu G, Gao Q, Zhou C, et al. (2012) Investigation of the Stability and Antioxidant Properties of Anthocyanins-based Purple Potato Colorant After Processing. African J Biotechnol11: 3379-3387.
  46. Ongoka PR, Matini L, Moutou JM, Ngabe DY (2006) Annals Marien Nougabi Univ. 7: 138-146.
  47. Lopes-Lutz D, Alviano SD, Alviano SC, Kolodziejczyk PP (2008) Screening of chemical composition, antimicrobial and antioxidant activities of Artemesia essential oils. Phytochem 69: 1732-1738.
  48. Sakthivel K, Palani S, Santhosh K, Devi K, Senthil KB (2010) Phytoconstituents analysis by GC-MS, cardioprotective and antioxidant activity of Buchanania axillaris against Doxorubicininduced cardio toxicity in albino rats. Int J Pharma Studies Res 1: 34-48.
  49. Orhan I, Kupeli E, Sener B, Yesilada E (2007) Appraisal of anti-inflammatory potential of the clubmoss, Lycopodium cuvatum L. J Ethnopharmacol 109: 146-150.
  50. Price KR, Johnson TI, FenwickGR (1987) The chemistry and biological significance of saponins in food and feeding stuffs. Crit Rev food Sci Nutr 26: 27-135.
  51. Brian FH, Thomas-Bigger J, Goodman G (1985) The Pharmacological Basis of Therapeutics. Macmillan, New York: NY, USA.
  52. Caceres A, Giron LM, Alvarado SR, Torres MF (1987) Screening of antimicrobial activity of plants popularly used in Guatemala for treatment of dermatomucosal diseases. J Ethnopharmacol 20: 223-237.
  53. Eleazu CO, Eleazu KC, Awa E, Chukwuma SC (2012) Comparative study of the phytochemical composition of the leaves of five Nigerian medicinal plants. J Biotechnol Pharm Res 3: 42-46.
  54. Hussain I, Ullah R, Ullah R, Khurram M, Ullah N, et al. (2011) Phytochemical analysis of selected medicinal plants. Afr J Biotechnol 10: 7487-7492.
  55. Aiyegoro OA, Okoh AIR (2010) Preliminary phytochemical screening and in vitro antioxidant activities of the aqueous extract of Helichrysum longifolium DC. BMC Complement Altern Med 10: 21.
  56. Olaleye MT (2007) Cytotoxicity and antibacterial activity of methanolic extract of Hibiscus sabdariffa. J Medicinal Plants Res 1: 9-13.
  57. Patel DK, Kumar R, Prasad SK, Sairam K, Hemalatha S (2011) Antidiabetic and in vitro antioxidant potential of Hybanthus enneaspermus linn f. muell in streptozotocin-induced-diabetic rats. Asian Pac J Trop Biomed 1: 316-322.
  58. Parekh J, Jadeja D, Chanda S (2005) Efficacy of aqueous and methanol extracts of some medicinal plants for potential antibacterial activity. Turk J Biol 29: 203-210.
  59. Orhan N, Hosbas S, Deliorman OD, Aslan M, Ergun F (2014) Enzyme inhibitory and radical scavenging effects of some antidiabetic plants of Turkey. Iran J Basic Med Sci 17: 426-432.
  60. Doss A, Mohammed Mubarak H, Dhanabalan R (2009) Antibacterial activity of tannins from the leaves of Solanum trilobatum Linn. Indian J Sci Technol 2: 41-43.
  61. Bown Deni (1995) The Royal Horticultural Society. Encyclopaedia of Herbs and their uses. Dorling Kindersley Book.
  62. Chiej R (1988) The Macdonald Encyclopoedia of Medicinal Plants. 1984, reprinted 1988. Macdonald Orbis.
  63. Foster S, Duke JA (2000) A Field Guide to Medicinal Plants and Herbs. 2nd Ed. Boston: Houghton Mifflin Company.
  64. Malairajan P, Gopalakrishnan G, Narasimhan S, Veni KJK (2006) Analgesic activity of some Indian medicinal plants. J Ethanopharmacol 106: 425-428.
  65. Han X, Shen T, Lou H (2007) Dietary Polyphenols and Their Biological Significance. Int J Mol Sci 8: 950-988.
  66. Lu L, Liu SW, Jiang SB, Wus G (2004) Tannin inhibits HIV-1 entry by targeting gp 41. Acta Pharmacol Sin 25: 213-218.
  67. Kolodziej H, Kiderlen AF (2005) Antileshmanial activity and immune modulatory effects of tannin and related compounds on leshmania parasitized RAW 264.7 cells. Phytochemistry 66: 2056-2071.
  68. Gonzalez SL, Garciaa JP, Cazaresa HA, Guarnerosc DM, Fuentesb JAM, et al. (2004) Apoptosis and cell cycle disturbances induced by coumarin and 7- hydroxycoumarin on human lung carcinoma cell lines. Lung Cancer: 275-283.
  69. Sodipo OA, Akiniyi JA, Ogunbamosu JU (2000) Studies on certain characteristics of extracts of bark of macruceras (K schemp) pierre Exbeille. Glob J Pure Appl Sci 6: 83-87.
  70. Shi J, Kakuda Y, Yeung D (2004) Antioxidative properties of lycopene and other carotenoids from tomatoes: Synergistic effects. BioFactors 21: 203-210.
  71. Mandal P, Sinha Babu SP, Mandal NC (2005) Antimicrobial activity of Saponins from Acacia auriculiformis. Fitoterapia 76: 462-565.
  72. Manjunatha BK (2006) Antibacterial activity of Pterocarpus santalinus. Ind J Pharm Sci 68: 115-116.
  73. S Kanji, RD MacLean (2012) Cardiac glycoside toxicity: More than 200 years and counting. Critical Care Clinics 28: 527–535.
  74. Krvavych AS, Konechna RT (2014) Phytochemical research of plant extracts and use in vitro culture in order to preserve rare wild species Gladiolus imbricatus. RJPBCS 5: 240.
  75. Aruoma OI (2003) Methodological considerations for characterizing potential antioxidant actions of bioactive components in plant foods. Mutat Res 523: 9-20.
  76. Coruh N, Celep AGS, Ozgokce F (2007) Antioxidant properties of Prangos ferulacea (L) Lindl, Chaerophyllum macropodum Boiss. And Heracleum persicum Desf. From Apiaceae family used as food in Eastern Anatolia and their inhibitory effects on glutathione-S-transferase. Food Chem 100: 1237-1242.
  77. Bahar E, Ara J, Alam M, Nath B, Bhowmik U, et al. (2013) In vitro antioxidant and thrombolytic activity of methanol extract of Sida acuta. J Pharmacogn Phytochem 2: 125-133.

Citation: Asgharian AM, Ojani S (2017) In vitro Antioxidant Activity and Phytochemical Screening of Flowers and Leaves of Hypericum perforatum L. Ethanolic Extracts from Tonekabon-Iran. J Phytochemistry Biochem 1:102.

Copyright: © 2017 Asgharian AM, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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