Department of Post Graduate-Studies and Research in Botany, Sir Syed College, Taliparamba, Kannur, 670142
Received: 10 Nov 2019, Revised and Accepted: 14 Jan 2020
Objective: The present study includes the qualitative and quantitative estimation of phytochemical constituents and HPTLC (High-performance thin layer chromatography) analysis of phenolic acids in the fruit extract of Artocarpus hirsutus.
Methods: Qualitative and quantitative analysis of Artocarpus hirsutus methanol fruit extract using standard methods. HPTLC analysis was performed by the CAMAG HPTLC system (Switzerland) for gallic acid, caffeic acid and ferulic acid determination.
Results: The Preliminary phytochemical analysis revealed the presence of phytoconstituents such as flavonoids, phenols, tannins, steroids, glycosides, alkaloids, carbohydrate and protein. The methanol fruit extract has high carbohydrate content (267±0.02 mg/g FW). And good amount of protein, phenol, flavanoid were also found. The results of mineral studies shows that elements such as potassium (1.601%) and nitrogen (1.4%) were present in higher quantity. The result showed the presence of caffeic acid within peak 7 with an Rf (Retension factor) value 0.49, ferulic acid within peak 8 with an Rf value 0.60, and gallic acid within peak 5 with an Rf value 0.25.
Conclusion: The results indicated that the Artocarpus hirsutus fruit contains an appreciable amount of bioactive compounds.
Keywords: Artocarpus hirsutus, Bioactive compounds, Ethnomedicine, Wild, Mineral, HPTLC, Endemic, Moraceae, Phenolic acid, Phytochemicals
© 2020 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open-access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/)
DOI: http://dx.doi.org/10.22159/ijcpr.2020v12i2.37488. Journal homepage: https://innovareacademics.in/journals/index.php/ijcpr
In India, the indigenous fruits collected from the wild play a significant role in the food and nutrient security of tribals and rural poor. Wild edible plants are the precious gift of nature of the country and most of the ethnic communities strongly depend on it for their day‐to‐day life .
However, there is no precise information about the nutritional potential of many wild edible plants. Nowadays many exotic and introduced fruits are used by modern people and indigenous species are discarded from dining tables. Food plants serve as alternatives to staple food during periods of food deficit and are the valuable supplements for a nutritional balanced diet, one of the primary alternative source of income for many resource-poor communities, and the source of species for domestication . Apart from their traditional use of food, potentially these wild plants have many advantages. They are used as medicine, fodder, and for rituals and other functions. They are immune to many diseases and often used in different formulations of traditional medicine in India. It is considered that special attention should be paid to maintain and improve this important source of food supply .
Fig. 1: Mature fruits of Artocarpus hirsutus
The Artocarpus hirsutus fruit locally called Anjili Chakka belongs to the Moraceae family. Aini (Anjili) is a tall evergreen tree, generally 20-25 m in height and up to 5 m in girth, fruits are edible, bright yellow (Fig.1-2), ovoid or globosem covered with spines, seeds ovoid and white. Artocarpus hirsutus is an endemic tree species of the southern Western Ghats of peninsular India and Maharashtra Sahyadris .
Fig. 2: Ripened fruit without pericarp
Natural products from plants called secondary metabolites are the end products of primary metabolites such as carbohydrates, amino acid, chlorophyll, lipids and so on. They are synthesis a large variety of chemical substances known as secondary metabolites which include alkaloids, steroids, flavonoids, terpenoids, glycoside, saponin, tannins, and phenolic compounds. The secondary metabolites are very good antioxidant compounds. The richness of phytochemical bioactive compounds possess antioxidant, antitumor, antimutagenic, anticarcinogenic, and antiparasitic activities .
High-performance thin-layer chromatography (HPTLC) is becoming a routine analytical technique because of the advantages that include the small amount of mobile phase required, the speed of the method and the possibility of analysing several samples simultaneously (i.e. on the same plate), unlike HPLC. It thus reduces analysis time and cost per analysis. Cloudy samples and suspensions can also be analyzed directly by HPTLC. Automatic sample application is possible and repeated scanning can be performed on the same plate, so scanning conditions can be changed .
Plants have developed efficient protective defense mechanisms against various diseases where pre-existing and induced secondary metabolites play a significant role to protect them against various adverse climatic conditions. The biosynthetic pathway of phenolic compounds is derived from the shikimate pathway and the backbone of phenolic compounds are aromatic amino acids. Role of plant phenolics in inducing resistance in plants is well documented in the literature. Results of the present experiments reveal the presence of some phenolic acids in the fruit pulp of Artocarpus hirsutus .
Preparation of fruit pulp extract
Mature fruits of Artocarpus hirsutus were collected from various localities of Kannur and Kozhikode district, Kerala. The dried material was powdered. From this powder, 10g was weighed and dissolved in methanol and kept in a shaker for 24 h. The slurry was filtered. Methanol extract was screened for the presence of secondary metabolites, Quantitative analysis of primary metabolites and minerals.
Fruit extract was screened for the presence of secondary metabolites such as alkaloids, saponins, tannins, steroids, flavonoids, terpenoids, phlobotannins, coumarins, cycloglycosides, phenols and quinines. The method of analysis employed were those described by Kokate  for the presence of various active components.
Estimation of total carbohydrate was done by using the method of Sdasivam and manickam . And Estimation of total protein by Lowry et. al. . Makkar method was followed for phenolics . The acid digestion method of Toth et al.  has been followed for the analysis of inorganic constituents.
HPTLC analysis was performed by CAMAG HPTLC system (Switzerland). Chromatographic Conditions used are, stationary phase aluminum backed pre-coated silica gel plates Merck 60 F254 (0.2 mm thickness). Samples were applied to the plate as bands at 10 mm from the bottom of the plate by using CAMAG ATS 4. The plate was developed up to 80 mm in ascending mode with solvent system Toluene: ethyl acetate: formic acid (5:5:0.5) at room temperature (28±2 °C) in a Twin Trough Chamber (Camag, Switzerland) which previously saturated with the mobile phase. After development, the air-dried plate scanned at 254 nm 366 nm, and 550 nm after derivatizing with anisaldehyde sulphuric acid reagent in CAMAG TLC SCANNER 3 with winCATS software.
Data were analyzed using MS-Excel. Each set of data is an average of triplicates. The data represents mean±SE.
The Preliminary phytochemical analysis revealed the presence of phytoconstituents such as flavonoids, phenols, tannins, steroids, glycosides, alkaloids, carbohydrate and protein. Saponin was absent in the extract (table 1).
Table 1: Preliminary qualitative analysis of fruit pulp of Artocarpus hirsutus
|S. No.||Phytochemical compound||Artocarpus hirsutus|
The methanol fruit extract has high carbohydrate content (267±0.02 mg/g FW). And a good amount of protein, phenol, flavanoid was also found (table 2).
Table 2: Quantitative analysis of total phenol, carbohydrate and protein contents in methanol extract of fruit pulp
|Total carbohydrate||267±0.02 mg/g FW|
|Total protein||5.71±0.01 mg/g FW|
|Total phenolic content||1.91±0.02 mg GAE/g DW|
|Total flavanoid content||1.22±0.01 mg/g FW|
The results of mineral studies shows that elements such as potassium (1.601%) and nitrogen (1.4%) were present in higher quantity (table 3).
Table 3: Mineral analysis of Artocarpus hirsutus fruit
Phenolic compounds are considered as secondary metabolites that are synthesized by plants during normal development. HPTLC phenolic acid profile of methanolic extract of Artocarpus hirsutus fruit was recorded in fig. 3 and table 4.
Table 4: Peak table with Rf values, height, and area of phenolic acids
Fig. 3: Densitogram display for gallic acid, caffeic acid and ferulic acid of methanolic fruit extract of Artocarpus hirsutus
The extract was run along with the standard phenols compounds such as caffeic acid, ferulic acid and gallic acid. The result showed the presence of caffeic acid within peak 7 with an Rf value 0.49, ferulic acid with in peak 8 with an Rf value 0.60, and gallic acid within peak 5 with an Rf value 0.25.
These results were supported by Johnson et al.  who reported that saponins, alkaloids, tannins, phytates, oxalate, phenols, HCN and flavonoids known phytochemicals were present in the fruit, seed and rind of C. lanatus. The results obtained with the hexane, ethyl acetate and methanol extracts showed that hexane was more efficient solvent with a yield of 10% followed by methanol, ethyl acetate with 8% and 7% respectively. This result may be due to the high polarity of methanolic solvent which can draw a higher variety of plant constituents than the other solvents .
The presence of phenolic compounds (total phenols and flavonoids) provides pharmacological activities like anticancer , antioxidant , antimicrobial , wound healing  and anti-inflammatory activities .
Potassium is one of the important nutrients for controlling human blood pressure, therefore fig and mulberry fruits were recommended for hypertension in previous studies. Toth et. al.  were studied vitamins and mineral substances in fruit and nuts of wild siberian plants. Potassium content is found to be higher in cranberry (2760). Bilberry is rich in Sodium (794 mg/l), Magnesium (736 mg/l), and manganese (393 mg/l) and low in copper (not detected). Ayensu (1985) reports that the amounts of calcium, iron, sodium, and potassium in A. officinalis were 22 mg/100 g, 1 mg/100 g, 2 mg/100 g, 278 mg/100 g, respectively. They also reported the amounts of ash, calcium, iron, potassium, and sodium in P. oleracia to be 20 mg/100 g, 1,500 mg/100 g, 29 mg/100 g, 1,800 mg/100 g and 55 mg/100 g respectively. Potassium is essential in the maintenance of cellular water balance, pH regulation in the body and also associated with protein and carbohydrate metabolism .
Sarma and Singh , Maurya et. al.  reported that ferulic acid has potent antimicrobial properties. Strong antioxidant activities of jackfruit pulp extracts were reported earlier and the same is correlated with the total phenolic and flavonoids content in the samples.
Soong and Barlow  also reported that the seeds of jackfruit are rich in total phenolic acid content and have strong antioxidant properties. Presence of phenolic compounds in food enhances food quality as they provide various benefits to human health.
The results indicated that the Artocarpus hirsutus fruit contain an appreciable amount of bioactive compounds. Fruit is a good source of carbohydrate, protein, phenol, and alkaloid. The presence of phenolic acids was confirmed by HPTLC analysis. Medically the presence of these phytochemicals especially the phenols and flavonoids explains the use of this plant in ethnomedicine for the management of various ailments.
The authors are extremely thankful to the council of scientific and industrial research for providing financial assistance.
All the authors have contributed equally.
The authors declare that there is no conflict of interest regarding the above article.
Deb D, Sarkar A, Deb BB, Datta BK, Deb D, Sarkar A, et al. Wild edible plants and their utilization in traditional recipes of Tripura. Northeast India. Adv Biol Res 2013;7:203-11.
Shrestha PM, Dhillion SS. Diversity and traditional knowledge concerning wild food species in a locally managed forest in Nepal. Agroforestry Syst Kluwer Academic Publishers; 2006.
Ajesh TP, Naseef SAA, Kumuthakalavalli R. Ethnobotanical documentation of wild edible fruits used by muthuvan tribes of idukki, Kerala-India. Int J Pharm Bio Sci 2012;3:479–87.
Dibinlal D, Sheethadevi B. Anti-ulcer activity of the bark of Artocarpus hirsutus. J Pharm Res 2013;8:3-5.
Beecher. Phytonutrients' role in metabolism: effects on resistance to degenerative processes. Nutr Rev 1999;57:S3-S6.
Ananda S, Tragoolpua K, Chantawannakul P, Tragoolpua Y. Antioxidant and anti-cancer cell proliferation activity of propolis extracts from two extraction methods. Asian Pac J Cancer Prev 2013;14:6991-5.
Kokate CK. Practical pharmacognosy. Vallabh Prakashan, Delhi; 2000.
Sadasivam S, Manickam A. Biochemical methods. 3rd edn. New Age International Publishers, New Delhi, India; 2008.
Lowry OH, Rosenbrough NJ, Farr AL, Randall RJ. Protein measurement with the folin’s phenol reagent. J Biol Chem 1951;193:265-75.
Makkar HPS. Quantification of tannins in tree and shrub foliage: a laboratory Mannual. Dondrecht. The Netherlands: Kluwer Academic; 2003.
Toth SJ, Prince AL, Wallace A, Mikkelsen DS. Rapid quantitative determination of eight mineral elements in plant tissue systematic procedure involving use of a flame photometer. Soil Sci 1948;66:459-66.
Johnson JT, Iwang EU, Hemen JT, Odey MO, Efiong EE, Eteng OE. Evaluation of anti-nutrient contents of watermelon Citrullus lanatus. Ann Biol Res 2012;3:5145-50.
Paulsamy S, Jeeshna MV. Preliminary phytochemistry and antimicrobial studies of an endangered medicinal herb Exacum bicolour Roxb. Res J Pharm Biol Chem Sci 2011;2:447-57.
Lin Y, Shi R, Wang X, Shen HM. Luteolin, a flavonoid with potential for cancer prevention and therapy. Curr Cancer Drug Targets 2008;8:634-46.
Hendra R, Ahmad S, Sukari A, Shukor MY. Flavanoid analyses and antimicrobial activity of various parts of Phaleria macrocarpa (Scheff.) Boerl fruit. Int J Mol Sci 2011;12:3422-31.
Nayak BS, Pereira LM. Catharanthus roseus flower extract has wound-healing activity in s prague dawley rats. BMC Complement Altern Med 2006;6:1-6.
Rathee P, Chaudhary H, Rathee S, Rathee D. Mechanism of action of flavonoids as anti-inflammatory agents: a review. Inflamm Allergy Drug Targets 2009;8:229-35.
Onibon VO, Abulude FO, Lawal LO. Nutritional and antinutritional composition of some Nigerian fruits. J Food Tech 2007;5:120-2.
Sarma BK, Singh UP. Ferulic acid may prevent infection by Sclerotium rolfsii in Cicer arietinum. World J Micro Biotech 2003;19:123-7.
Maurya S, Rashmi S, Singh DP, Srivastava J, Singh SP. Phenolic compounds of sorghum vulgare in response to sclerotium rolfsii infection. J Pl Inter 2007;2:25-29.
Soong YY, Barlow P. Antioxidant activity and phenolic content of selected fruit seeds. Food Chem 2004;88:411-7.