ASSESSMENT OF TOTAL PHENOL AND FLAVONOID CONTENT OF COSTUS SPECIOSUS AND CATHARANTHUS ROSEA IN DIFFERENT GEOGRAPHICAL REGIONS OF BALAGHAT DISTRICT, PRADESH

anti-diuretic, antiulcer and anti-diarrheal activities. In the present study the total phenol and flavonoid content of root methanol extract of both plants were determined from five blocks of Balaghat district Madhya Pradesh. Folin-Ciocalteau assay was used to determine total phenol content and aluminium chloride method for flavonoid content using gallic acid and quercetin as standard respectively. The result showed highest concentration of total phenol content in Birsa block for both C. speciosus (95.85±1.56 mg GAE/g) and C. rosea (82.32±0.81 mg GAE/g). The total flavonoid content was found highest in Paraswada block for C. speciosus (97.67±0.53mg/QE) and Baihar block for C. rosea (93.20±0.54mg/QE). The ratio of flavonoid to the phenolic in each extract was also determined to as certain extracts that are rich in flavonoids. The present work explained the total phenol and flavonoid content of root methanol extracts of both plants in different location of Balaghat district Madhya Pradesh. The concentration of phytocompounds varies greatly due to environmental effects. A significant concentration of phenol and flavonoid content was found in both plants which showed its correlation with antioxidant potential. This study should be beneficial for characterization and structure elucidation of pharmaceutically lead compound.


ISSN: 2320-5407
Int. J. Adv. Res. 8 (11), 1024-1029 1025 furostanol, saponins-costusosides, dioscin, gracillin, dihydrophytylplastoquinone, diosgenin, β-sitosterol, β-D-glucoside, prosapogenins, and α-tocopherolquinone (Lijuan et al., 2011). It has been reported that the plant contain β-amyrin, camphene, costunolide, diosgenin, α-Humulene, lupeol, and zerumbone having anticancer activity (Saleem, 2009;Zhang et al., 2012). The rhizome also contains secondary metabolites like phenol, flavonoids, tannin, saponnin, steroids, and glycoside which are studied for a number of biological consequences including antimicrobial and antioxidant activities (Borkataky et al., 2014). The rhizomes of Costus speciosus are bitter in taste and exhibit various activities such as antihelmintic, astringent, expectorant and used mainly for treating diabetes. It has been reported that rhizome extract was used for treating burning sensation, constipation, leprosy, asthma, bronchitis, anaemia and other skin ailments (Bown Deni, 2008). Rhizomes of plant were used as herbal remedy for fever and its paste were used for treating boils. Leaves of the plant were used for scabies and stomach ailments. Besides rhizomes, stems are also used for treating blisters and burns and roots are used against snake bite (Gruenwald et al., 2000). Catharanthus roseus belongs to family Apocynaceae and is a dicotyledonous angiosperm plant which synthesizes two terpene indole alkaloids namely vincristine and vinblastine that act against cancer (Ajaib, 2010). More than 100 monoterpenoidindole alkaloids (TIA) are produced by these plants in different organs. Catharanthus roseus have 400 alkaloids, and alkaloids are the most active components of the plant used in pharmacological activities, flavor and fragrance, ingredients, food additives, pesticides and agrochemicals (Bernett at el., 1978). The ariel parts of the plant contain alkaloids like vindesine, vindeline tabersonine, vinblastine, vincristine, whereas raubasin, reserpine, catharanthine, vinceine, vineamine, and ajmalicine are present in basal or root parts of the plant. About 150 alkaloids of pharmacological importance are present on plant leaves. In laboratory animals significant antihyperglycemic and hypotensive activity was reported by the leaf extracts of Catharanthus roseus. The plant has good antioxidant potential and contain significant amount of volatile and phenolic compounds such as caffeoylquinic acids and flavonal glycosides which act as antioxidants against reactive oxygen species. These compounds also exhibit anti-inflammatory, antimicrobial, antioxidant, antithrombotic, anti-allergic, vasodilatory effects and cardio-protective properties (Kumar et al., 2015). In 2007 Mustafa and Verpoorte reported a number of phenolic compound such as 2, 3-dihydoxybenzoic acid, and phenylpropanoids such as flavonoids, anthocyanins and cinnamic acid derivatives in Catharanthus roseus. Phenol and flavonoid compounds are a class of antioxidant agents which act as free radical terminators and their bioactivities may be related to their abilities to chelate metals, inhibit lipooxygenase and scavenge free radicals. Currently, plant materials rich in antioxidants are used in the food industry because they decrease the oxidative degradation of lipids and maintain the quality and nutritional value of food (Kahkonen et al. 1999).The aim of the present study was to determine the total phenol and flavonoid content in methanol root extracts of Costus speciosus and Catharanthus rosea collected from five blocks of Balaghat district, Madhya Pradesh.

Materials and Methods:-Plant collection:
The plant samples of Costus speciosus and Catharanthus rosea were collected from natural environment from five block of Balaghat district, Madhya Pradesh (Table1) and authenticated by Botanical Survey of India, Allahabad.

Preparation of extracts:
The root part of the plants was washed, dried under shade and grinded to make powder. The methanol extract of both plants were prepared using Soxhlet apparatus (MSW, India) according to method developed by Wagner et al. (1996). After extraction each extract was stored in air tight bottles and kept in refrigerators for further studies.

Determination of total phenolic content (TPC):
The total phenol content in plant extracts were determined by Folin-Ciocalteu assay method using gallic acid as standard with slight modification (Singleton et al., 1999). The reaction mixture consists of 1 ml of extract and 9 ml of distilled water was taken in a volumetric flask (25 ml) and then 1ml of Folin-Ciocalteu phenol reagent was mixed with the mixture and shaken well. After 5 minutes, 10 ml of 7 % sodium carbonate (Na 2 CO 3 ) solution was treated to the mixture. The volume was made up to 25 ml. A set of standard solutions of gallic acid (20, 40, 60, 80 and 100 µg/ml) were prepared. Incubated for 90 min at room temperature and the absorbance for test and standard solutions were determined against the reagent blank at 650 nm with an ultraviolet (uv) /visible spectrophotometer. The total phenol content was determined from calibration curve (Fig1) and expressed as mg of GAE/gm of extract.

Determination of Total flavonoid content (TFC):
Aluminium-chloride spectrophotometric method was used to determine total flavonoid content. The plant extract of 0.25mL was dissolved in 1.25mL distilled water then NaNO 3 was added. This mixture was placed in dark for 6 minutes then 0.150 ml of 10% AlCl 3 was added in the mixture and again incubated in dark for 5 minutes. Now 0.5 ml NaOH and 0.275 ml distilled water was added in the mixture. The set of standard solutions of quercetin (20, 40, 60, 80 and 100 mg/ml) were prepared in the same manner as described for the extracts. The absorbance of standard and the extracts solutions were measured against the reagent blank at 510 nm with a UV/Visible spectrophotometer. The total flavonoid content was determined from the calibration curve (Fig2) and expressed as milligram of quercetin equivalent (QE) per gram of extracts.

Statistical Analysis:
The experiments were carried out in three replicates and data were presented as mean ± S.E. in Microsoft Excel 2010 for statistical and graphical evaluations.

Conclusion:-
Costus speciosus and Catharanthus rosea are multipurpose medicinal plant used in traditional and modern medicines for treatment of many health problems. The present work explained the total phenol and flavonoid content of root methanol extracts of both plants in different location of Balaghat district Madhya Pradesh. The concentration of phytocompounds varies greatly due to environmental effects. A significant concentration of phenol and flavonoid content was found in both plants which showed its correlation with antioxidant potential. This study should be beneficial for characterization and structure elucidation of pharmaceutically lead compound.