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Table of Contents   
ORIGINAL ARTICLE  
Year : 2018  |  Volume : 11  |  Issue : 3  |  Page : 83-86
Phytochemical screening and thin-layer chromatographic analysis of tannin from Dryopteris, Asplenium, and Davalliain, Tahura, Batu, Indonesia


Department of Biology, Faculty of Matemathic and Natural Sciences, Malang State University, Jalan Semarang 5-Malang 65145, Indonesia

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Date of Web Publication10-Dec-2019
 

   Abstract 


Context: Forest area in Tahura R. Soerjo has high moisture level with rainfall 1488 mm3/year, so it provides an optimum condition for ferns growth. Tannin is one of the secondary metabolites that have many benefits; it can be found in various amounts in plant body parts. Aim: The study aims to identify the tannin in three fern genera, namely, Dryopteris, Asplenium, and Davallia that were obtained from Tahura R. Soerjo. Materials and Methods: Both epiphytic (Davallia) and terrestrial (Asplenium and Dryopteris) fern samples were obtained from Taman Hutan Raya Raden Soerjo, Cangar, Batu city in May– August 2015. Powdered fern samples were extracted through maceration method using methanol (1:10). The methanol extract of fern were tested for the tannin presences using phytochemical screening and layer chromatography. Results: Thin-layer chromatography was done to confirm the screening results that indicate the existence of tannin. Phytochemical screening for gallotannin in methanol extracts positive results. All extracts perform a different number of spot and Rf value.Dryopteris and Davallia frond showed reddish-orange spot on the plate, while Dryopteris rhizome,Davallia rhizome,Asplenium frond, and Asplenium rhizome showed greenish brown on the plate.Conclusion: The phytochemical screening including tannin confirmation and thin-layer chromatography showed that all part (frond and rhizome) from Dryopteris,Davallia, and Asplenium contained gallotannin and tannin.

Keywords: Phytochemical screening, tannin, thin-layer chromatography

How to cite this article:
Sulasmi ES, Sari MS, Prabaningtyas S. Phytochemical screening and thin-layer chromatographic analysis of tannin from Dryopteris, Asplenium, and Davalliain, Tahura, Batu, Indonesia. Ann Trop Med Public Health 2018;11:83-6

How to cite this URL:
Sulasmi ES, Sari MS, Prabaningtyas S. Phytochemical screening and thin-layer chromatographic analysis of tannin from Dryopteris, Asplenium, and Davalliain, Tahura, Batu, Indonesia. Ann Trop Med Public Health [serial online] 2018 [cited 2020 Aug 10];11:83-6. Available from: http://www.atmph.org/text.asp?2018/11/3/83/272550



   Introduction Top


Ferns have been adapted to their environmental change, so they produce various secondary metabolites more than the other vascular plants. It has been reported that ferns contain numbers of phytochemical compounds or secondary metabolite, namely flavonoid, steroid, alkaloid, phenol, triterpenoid, various amino acid, and fat including essential oil. A study by Thite et al. showed that extract of 4 species of Filicales ferns contains saponin, phenol, tannin, phytosterol, triterpene, alkaloid, and terpenoid.[1]

Taman Hutan Raya Raden Soerjo (Tahura R. Soerjo) is one of the conservation areas that located in East Java Province. The previous study revealed that fern diversity in Tahura R. Soerjo consisted of 18 families, 30 genera, and 47 species. This area is suitable habitat for both terrestrial and epiphyte ferns due to its high humidity with rainfall 1488 mm3/year.

Chemical compounds produced by the plant are the product of secondary metabolism, which are called as secondary metabolites. These compounds have no important role in fern life, but they protect the plant from environmental changes and herbivores. In addition, they also have been reported for benefits such as anticancer, antimicrobial, and antioxidant. Several studies have reported that secondary metabolites act as antifungi.[2],[3]

One of the secondary metabolites with many benefits is tannin. It can be found in almost all of plant body parts in different of quantity and quality. Tannin has been described as polyphenol compound with high molecule mass. It consisted of hydroxyl and carboxyl chains, so it can form macromolecule complex or protein in a certain condition. This compound is soluble in water, glycerol, alcohol, and hydroalcoholic substances. It insoluble in petroleum ether will be decomposed on temperature 210°C and will be burnt in 526°C.[4],[5]

In general, tannins are classified into hydrolysable tannin and condensed tannin. Phytochemical screening for tannin in several Pteridophyta in the Western Ghats has been conducted by Mithraja et al. using acetone, benzene, chloroform, water, ethanol, and petroleum ether as solvents.[6] There were also several ferns that have been tested such as Desmodium heterophyllum, Dicranopteris linearis, Blechnum orientale, Ceratopteris thalictroides, Hexastylis arifolia, Lindsaea ensifolia, Nephrolepis multiflora, Pityrogramma calomelanos, Pteris confuse, and Drynaria quercifolia. The results showed that all extracts contain tannin 47/66. Condensed tannin can be found in Pteridophyta and Gymnospermae and distributed in all Angiospermae plant, especially wooden plant. Based on those data, this study aims to identify the tannin in three fern genera, namely Dryopteris, Asplenium, and Davallia that were obtained from Tahura R. Soerjo.


   Materials and Methods Top


Sample preparation

Both epiphytic (Davallia) and terrestrial (Asplenium and Dryopteris) fern samples were obtained from Taman Hutan Raya Raden Soerjo, Cangar, Batu city in May–August 2015. Samples were washed and separated from their rhizome and frond, and after that, they all were left dried at room temperature. Dried samples were grounded and stored for extraction.

Extraction

Powdered fern samples were extracted through maceration method using methanol (1:10). The process was repeated three times, and then the filtrate was evaporated for about 3–4 h in order to obtain its pure extract.

Phytochemical screening for tannin

The methanol extract of the three samples was put into four reaction tubes with 0.5 ml of volume. Potassium acetate and FeCl3 1% were added for gallotannin test that would result in several colors, such as blue, purple, or black. Catechol was tested by addition of formaldehyde 3% and absolute chloric acid (2:1) into 0.5 ml of extract. Catechol was indicated by a color change of the solution into the red.

Tannin confirmation and thin-layer chromatography

Silica plate G60 F254 (20 cm × 20 cm) was used in thin-layer chromatography. The methanol extract of samples was plotted in a plate with 1 cm of distance from the bottom line and 1 cm from the sideline. Extracts were eluted in n-butanol: acetic acid: water (4:1:5). Elution was stopped when the eluent has passed the line. The plate was sprayed with FeCl3, so spots can appear. The Rf value of each spot was measured and was observed using ultraviolet (UV) light 254 nm and 366 nm.


   Results Top


Phytochemical screening results of methanol extract from fern genera, namely Dryopteris, Davallia, and Asplenium, are presented in [Table 1]. Thin-layer Rf value and spot color as chromatography results are presented in [Table 2].
Table 1: Phytochemical screening for tannin in methanol extract derived from fern genera Dryopteris, Davallia, and Asplenium

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Table 2: Spot colors that were observed using ultraviolet light

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Based on the results, it can be seen that all samples contain tannin. Further tannin test was done to confirm tannin types in each fern. All sample colors changed into blackish blue that indicated they contain gallotannin. Catechol was detected positive by red color in frond samples of all genera and in the rhizome of Dryopteris and Asplenium. Dryopteris frond and Davallia frond showed reddish-orange spot after UV366 irradiation, which indicates tannin content. Spot colors on plates are presented in [Figure 1].
Figure 1: Spot color of samples under ultraviolet 366 (from left to right) Dryopteris frond, Dryopteris rhizome, Asplenium frond, Asplenium rhizome, Davallia frond, and Davallia rhizome

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Six methanol extracts of fern have a different number of spots and Rf value [Table 3]. The number of spots represents the number of compounds that appear in different colors.
Table 3: Number of spots and Rf value of each sample

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   Discussion Top


Phytochemical screening of six samples from fern genera, namely Dryopteris, Davallia, and Asplenium, showed that it contained tannin. Harborne stated that tannin content can be detected using short-wave UV light by the appearance of reddish orange color.[7] The other samples (Dryopteris rhizome, Davallia rhizome, Asplenium frond, and Asplenium rhizome) showed brownish green color after UV366 irradiation. This indicated that the samples contain tannin. It is correlated to Averrhoa bilimbi thin-layer chromatography extract which showed brownish green color in eluent.[8]

Rf value also represents the detected compound; it is calculated from the comparison between spot high and eluent distance. Active compounds that obtained from thin-layer chromatography are not always appeared as specific spot; it depends on the label.[9] Compounds that are suspected as tannin in thin-layer chromatography of Belimbing Wuluh using the same eluent have the same Rf value of 0.61. This is similar to Rf value of Davallia frond extract. In addition, mimosa extract that has been eluted in the same eluent has Rf value of 0.62, which is similar to Dryopteris frond. The other extracts that are also suspected contain tannin due to their Rf value that close to 0.62.

Gallotannin is showed to be a potential antifungal. It has been reported in another research about the antifungal effect of hydrolysable tannin to dermatophytes, slime molds, and yeast. A study with microdilution has revealed that ellagitannin corilagin (minimum inhibitory concentration [MIC], 0.8 nM) has the similarity of activity with amphotericin B (MIC, 0.5 nM) and sertaconazole (Zalain; MIC 0.9 nM) against Torulopsis glabrata strains.[10]


   Conclusion Top


In summary, the phytochemical screening including tannin confirmation and thin-layer chromatography in three fern genera, namely, Dryopteris, Asplenium, and Davallia that were obtained from Tahura R. Soerjo, Batu, Indonesia showed that all part (frond and rhizome) from Dryopteris, Davallia, and Asplenium contained gallotannin and tannin.

Acknowledgment

The authors would like to thank to Malang State University for supporting this research.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
   References Top

1.
Thite SV, Datil DT, Gurav KD, Kadam AS, Thoke KB, Kore BD. Qualitative analysis of secondary metabolites fern some filicales members. Int J Res Pharm Chem 2013;3:300-2.  Back to cited text no. 1
    
2.
Parihar P, Bohra A. Antifungal efficacy of various pteridophyticplant parts extracts: A study in vitro. Adv Plant Sci 2002;15:35-8.  Back to cited text no. 2
    
3.
Candra S, Srivastava M, editors. Pteridology in the New Mellenium. London: Kluwer Academic Publication; 2003.  Back to cited text no. 3
    
4.
Jayalaksmi A, Mathew AG. Chemical composition and processing the arecanut palm (Areca catechu L). India: CPCRI Kasaragod; 1982.  Back to cited text no. 4
    
5.
Sax I, Lewis RJ. Condensed Chemical Dictionary. 11th ed. New York: Van NostradReinhold Companya; 1989.  Back to cited text no. 5
    
6.
Mithraja MJ, Marimuthu J, Mahesh M, Paul ZM, Jeeva S. Chemical diversity analysis on some selected medicinally important pteridophytes of Western Ghats, India. Asia Pac J Trop Biomed 2012;2:S34-9.  Back to cited text no. 6
    
7.
Harborne JB. Phytochemical Methods: Guides for Modern Way of Plant Analyzing. Bandung: ITB Press, Translated by K. Padmawinata and Isoediro; 2006.  Back to cited text no. 7
    
8.
Hayati EK, Fasyah AG, Sa'adah L. Fractionation and identification of tannin compounds on Belimbing Wuluh leaves (Averrhoa bilimbi L.). J Kimia 2010;4:193-200.  Back to cited text no. 8
    
9.
Sutrisno B. Reverse Approach. Jakarta: Pancasila University; 1986.  Back to cited text no. 9
    
10.
Latté KP, Kolodziej H. Antifungal effects of hydrolysable tannins and related compounds on dermatophytes, mould fungi and yeasts. Z Naturforsch C 2000;55:467-72.  Back to cited text no. 10
    

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Correspondence Address:
Dr. Murni Sapta Sari
Deparment of Biology, Faculty of Matemathic and Natural Sciences, Malang State University, Jalan Semarang 5, Malang 65145
Indonesia
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/ATMPH.ATMPH_36_17

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