INTERNATIONAL JOURNAL OF RESEARCH CULTURE SOCIETY ISSN: 2456-6683 Volume - 4, Issue - 9, Sept – 2020 Monthly, Peer-Reviewed, Refereed, Indexed Journal Scientific Journal Impact Factor: 5.743 Received on : 21/08/2020 Accepted on : 11/09/2020 Publication Date: 30/09/2020

Phytochemical screening and estimation of total phenolic and flavonoid content in selected weeds

1 Patel Shivani S., 2 Modi Nainesh Department of Botany, Bio-informatics and Climate Change Impact Management School of Science , Gujarat University Narvangpura, Ahmedabad , Gujarat, India- 380009 E-mail:- [email protected], [email protected]

Abstract: Plant is an important source of medicine and plays a key role in world health . Medicinal herbs or plants have been known to be an important potential source of therapeutics or curative aids. Dichanthium annulatum, Boerhaavia diffusa L. and Digera muricata L. has a wide range of pharmacological properties like antimicrobial, antiviral, antioxidant, anticancer, anti-inflammatory and antiplasmodial. In present study, the effects of three different solvents such as methanol, acetone and aqueous were investigated to determine the presence of phytochemicals , total phenolic content and total flavonoid content in some weed plants like Dichanthium annulatum, Boerhaavia diffusa L. and Digera muricata L. The result concluded that, in TPC, the highest content was observed in acetone extract of stem and lowest content was observed in aqueous extract of stem while in TFC, the highest content was observed in acetone extract of stem and lowest content was observed in methanol extract of leaves. Key Words: Dichanthium annulatum, Boerhaavia diffusa, Digera muricata, Phytochemical screening, Total

phenolic content , Total flavonoid content.

1. INTRODUCTION: In the present day, the medicinal plants occupies the secure position in field of modern and advanced medicine and pharmaceutical industries showing special interest for using or synthesizing natural drugs which are extracted from the plants (Jamshidi et al., 2018). Traditional medicines are in higher demands in whole world among peoples because it is a direct source of therapeutic agents and affordable or cheaper for people (Oladeji ., 2016). Traditional medicines also provides taxonomic markers for the discovery and production of new compounds. (Samy and Gopalakrishnakone., 2007) Phytochemicals are chemicals which are produced by plants through primary or secondary metabolism (Mendoza., 2018). Phytochemical screening refers to the extraction, screening and identification of the medicinally bioactive compound which are present in plants like flavonoids, alkaloids, carotenoids, tannin, antioxidants and phenolic compounds (Dey .,2011) Dichanthium annulatum (Forsk.) Stapf, belong to Poaceae (Graminae) family (Awad et al., 2015). It is perennial and generally known as ‘marvel grass’ and classified in the Poales order. Sheda grass is a perennial grass, up to about 25 cm tall (Ram et al.,2015). It is used as forage due to its simple and cheaper development from seed. (Kumar et al., 2008). In India, these species are promising for grazing or hay-making after reseeding into degraded grassland (Fatima et al.,2018). It was also used for rough lawns (Singh et al.,2015). Dichanthium annulatum is a good quality fodder that can resist temporary flooding or drought.

Boerhaavia diffusa L. belongs to Nyctaginaceae family which have species of flowering plant in the four o'clock family and it is commonly known as 'Punarnava' in the Indian system of medicine ( Bhardwaj et al., 2014). Punarnava or Hogweed was literally means ‘bring back to life’ or ‘renewer’. Boerhaavia diffusa is an herb with anti-cancer, anti-diabetic, diuretic, anti-oxidant, antihepatotoxic, antiasthamatic, diaphoretic, anthelmintic, febrifuge, antiscabies and antiurethritis and anti-inflammatory properties (Murti et al., 2010, Majgaine and Verma ,. 2017, Rajpoot and Mishra ,. 2011).

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Digera muricata L. is an annual herb growing upto 20-70 cm tall. It is an important medicinal herb belongs to Amaranthaceae family and found as a weed throughout India (Ghaffar et al., 2019). is used in both folk and traditional systems of medicines (Mety & Mathad., 2010). The whole plant is also commonly grazed as forage, predominantly by sheep and goats (Qureshi and Bhatti., 2009). In India, seeds and flowers are used to treat urinary disorders (Usmani et al .,2013). Leaves and young shoots of D. muricata are locally used as a vegetable and given to relieve constipation (Rahmatullah and Bhatti., 2009). It is also used internally against digestive system disorders (Reddy et al., 2010). In India, flowers and seeds are used to treat urinary discharges (Parrotta., 2001). Leaf paste is applied locally to prevent pus formation (Katewa and Galav., 2005). Boiled root infusion given to mother after child birth for lactation purpose (Grosskinsky and Gullick., 2000). 2. MATERIAL AND METHODOLOGY:  Collection of plant material:- The leaves and stem of Dichanthium annulatum, Boerhavia diffusa, Digera muricata were collected locally from the district Detroj, Gujarat, India.  Preparation of Sample:- The whole plant materials were air dried until all the water molecules evaporated. After drying, the plant material were grinded well using mechanical blender into fine powder and transferred in to air tight containers with proper labeling for future use.  Preparation of Extraction:-  Hot Extraction of extract The powder sample of leaves was weighted with the help of weighing balance (gm). 10 gm of leaves powder extracted with 100 ml solvent (90%) using a soxhlet extractor for five to six hours at 40 to 50 oC temperature. Then extracted solvent filtered through muslin cloth and transfer in to petri-plate. The filtrate was open at room temperature and allowed it for solvent evaporation. After 48 hours the dried extract was ready. The yield of both cold and hot extract was calculated in the form of percentage by given formula. Yield (%) =W1x 100/W2 Where W1 =Weight of extract after solvent evaporation; W2= Weight of the grinded fine leaf powder.  METHODS USED IN EXPERIMENTS:-  Qualitative analysis: (Harbone 1998, Kokate, Ramman 2000) In the present study, solvents like acetone, aqueous and methanol are used to extract the phytochemicals from Dichanthium annalutum, Boehaavia diffusa and Digera muricata by using standard protocols of Harbone 1998, Kokate, Ramman 2000.

 Estimation of Total Flavonoid Content in Different Parts of Plant 500 µl of the extract of different parts of plants (12.5-300 µg/ml of methanolic and ethanolic extracts) was mixed with 1500 µl of 95% methanol and then 100 µl of Aluminium chloride (10%) and Potassium acetate (1M) was added respectively and makes volume up to 10ml with distilled water and agitated. Incubation was done for 20-30 minutes at room temperature. The absorbance was assessed at 415 nm against a blank having all the reagent without the sample using spectrophotometer. Measurement was done in triplecates and the total flavanoid quantified by the standard curve of quantified by the standard curve of quercetin solution (12.5, 25, 50, 100 and 150 µg/ml, R2 =0.9963).  Estimation of Total Phenol Content in Different Parts of Plant One ml of the extracts of different parts of plants (12.5-300 μg/ml of methanolic and ethanolic extracts) was thoroughly mixed with 10 ml of distilled water, added 1.5 ml of Folin-Ciocalteu reagent. After 5 minutes, 4 ml of 20% sodium carbonate (Na2CO3) was added and adjusted with distilled water up to 25 ml and agitated. Than incubated for 30 minutes at room temperature. The absorbance was measured at 765 nm against a blank having all the reagents excluding the sample using spectrophotometer. This procedure was repeated 3 times for each extract. The total phenols were quantified by the standard curve of gallic acid solution (12.5, 25, 50, 75, 100, 150, 200, 250 and 300 μg/ml) which was prepared using the similar procedure .( R2 = 0.9993).

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3. RESULT AND DISCUSSION: The dried powder of different parts of plant were extracted with methanol and ethanol solvent to examine the phytochemical screening , total phenol content and total lavonoid content.  Phytochemicals Screening of Different plant extract The qualitative phytochemicals screening assay of Dichanthium annulatum, Boerhaavia diffusa L. and Digera muricata L. in three different solvents i.e., methanol, acetone and aqueous. Table 1:- Qualitative Analysis of Different Parts of Dichanthium annalutum, Boerhaavia diffusa L. and Digera muricata L.

Sr. Test Dichanthium annulatum Boerhaavia diffusa L. Digera muricata L. No Methanol Acetone Aqueous Methanol Acetone Aqueous Methanol Acetone Aqueous L S L S L S L S L S L S L S L S L S Alkaloids 1 Mayer’s test ------2 Wager’s test ------3 Hager’s test ------4 Dregendroff ’s ------test Carbohydrates 1 Molish test + + + + + + + + + + + + + + + + + + 2 Fehling test + + + + + + + + + + + + + + + + + + 3 Barford’s test + + + + + + + + + + + + + + + + + + 4 Bendict’s test + + + + + + + + + + + + + + + + + + Glycosides 1 Borntrager’s ------test 2 Chloroorm test ------3 Keller-Killiani ------test Protein 1 Millon’s test + + + + + + ------2 Biuret test + + + + + + ------Phenolics 1 Ferric chloride + + + + + + + + + + + + + + + + + + test 2 Lead acetate + + + + + + + + + + + + + + + + + + test 3 Folin- + + + + + + + + + + + + + + + + + + Cioculten test Flavanoids 1 Alkaline test + + + + + + + + + + + + + + + + + + 2 Lead acetate + + + + + + + + + + + + + + + + + + test Saponins 1 Froth test + + + + + + + + + + + + + + + + + + Terpenoids 1 Salkowski test + + + + + + + + + + + + + + + + + + 2 Copper acetate + + + + + + + + + + + + + + + + + + test Fixed oils & fats 1 Filtered paper + + + + + + + + + + + + + + + + + + test Cardiac Glycosides 1 Legal test ------Steroids 1 Liebermann + + + + + + + + + + + + + + + + + + Burrchard’s test 2 Salkowaski’s + + + + + + + + + + + + + + + + + + test

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 Total Phenol Content of different plant extract in different solvent In Dichanthium annalutum , the total phenol content was found highest in the the leaves of acetone extract and lowest was found in the leaves of aqueous extract. In Boerhaavia diffusa , the total phenol content was found highest in the the leaves of acetone extract and lowest was found in the stem of aqueous extract. In Digera muricata , the total phenol content was found highest in the the stem of acetone extract and lowest was found in the leaves of methanol extract. Table 2 :- Total Phenol Content of different plants in different solvents Sr. Plant Name Total Phenol Content No Methanol Acetone Aqueous Leaves Stem Leaves Stem Leaves Stem 1 Dichanthium annalutum 246 ± 0.003 260 ± 0.005 313 ± 0.006 327 ± 0.006 90 ± 0.002 100 ± 0.006 2 Boerhaavia diffusa 223 ± 0.003 136 ± 0.003 448 ± 0.006 412 ± 0.004 80 ± 0.003 61 ± 0.008 3 Digera muricata 103 ± 0.005 119 ± 0.009 352 ± 0.007 371 ± 0.007 133 ± 0.009 172 ± 0.009

 Total Flavanoid Content of different plant extract in different solvent In Dichanthium annalutum , the total lavonoid content was found highest in the the stem of acetone extract and lowest was found in the leaves of methanol extract. In Boerhaavia diffusa , the total lavonoid content was found highest in the the stem of acetone extract and lowest was found in the stem of methanol extract. In Digera muricata , the total lavonoid content was found highest in the the stem of acetone extract and lowest was found in the leaves of acetone extract. Table 3 :- Total Flavanoid Content of different plants in different solvents Sr.No Plant Name Total Flavanoid Content Methanol Acetone Leaves Stem Leaves Stem 1 Dichanthium annalutum 191 ± 0.008 238 ± 0.009 248 ± 0.022 262 ± 0.012 2 Boerhaavia diffusa 252 ± 0.007 241 ± 0.004 236 ± 0.016 275 ± 0.002 3 Digera muricata 233 ± 0.018 202 ± 0.013 186 ± 0.010 246 ± 0.006

4. CONCLUSION: The current study revealed that the whole plant extracts in three different solvent of Dichanthium annulatum, Boerhaavia diffusa L. and Digera muricata L. The phytochemistry is regarded as the heart of herbal therapy and the phytochemical research plays an important role in the development of green medicines, which are safer to use than the crude drugs . The major class of bioactive compounds like alkaloids, flavonoids, lignans, phenols, tannins, terpenes and volatile oils has been isolated from the plants.

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