Research Article |
Corresponding author: Tita Juwitaningsih ( juwitaningsih@unimed.ac.id ) Academic editor: Georgi Momekov
© 2022 Tita Juwitaningsih, Destria Roza, Saronom Silaban, Elvira Hermawati, Neneng Windayani.
This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Citation:
Juwitaningsih T, Roza D, Silaban S, Hermawati E, Windayani N (2022) Phytochemical screening, antibacterial, antioxidant, and anticancer activity of Coffee parasite acetone extract (Loranthus ferrugineus Roxb). Pharmacia 69(4): 1041-1046. https://doi.org/10.3897/pharmacia.69.e91427
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This study aimed to perform a phytochemical screening and test the antibacterial, antioxidant, and anticancer activities of acetone extracts of the Coffee parasite (Loranthus ferrugineus Roxb). A phytochemical screen was performed using specific reagents. Antimicrobial testing was performed using the paper disc diffusion method. The antioxidant activity test used the 2,2-diphenyl-1-picrylhydrazyl (DPPH) method. Anticancer activity test against HeLa and A549 cells based on the 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) assay method. Acetone extract L. ferrugineus Roxb contains alkaloids, flavonoids, triterpenoids, and tannins compounds. The acetone extract of L. ferrugineus Roxb showed activity against all the bacteria tested, with the inhibition zone diameter ranging from 6.2 mm - 11.1 mm. Acetone extract of L. Ferrugineus Roxb had a very strong antioxidant activity with a value of IC50 = 48.7122 µg/mL. The anticancer activity test showed cytotoxic activity against HeLa cells with a value of IC50 = 47.62 µg/mL and for A549 cells with a value of IC50 = 192.83 µg/mL.
antibacterial, antioxidant, anticancer, Loranthus ferrugineus Roxb, phytochemical screening
North Sumatra is the main coffee-producing province in Indonesia. Many parasitic plants grow on coffee plants. The parasite is a parasitic organism that lives on or in its host plant branches and twigs and absorbs nutrients, minerals, and water from its host. The parasite was originally considered a harmful plant because it damaged its host plant. However, several countries such as Thailand, Korea, Japan, Indonesia, Malaysia, China, Saudi Arabia, and Nigeria have used several species of parasites in ethnomedicine to treat various diseases (Kwanda et al. 2012;
One of the most widely used parasite plants is from the Loranthaceae family.
The use of parasites in traditional medicine is popular. Therefore it attracted researchers to research it. Several studies published regarding parasite plant activity of the genus Loranthus, including L. micranthus, have found antihypertensive, antimicrobial, hypolipidemic immunomodulatory activity, antidiabetic antioxidant, and antidiarrheal (
Meanwhile, L. regularis Steud ex Sprague has antiinflammatory and antioxidant activity (
The research was carried out from April - November 2020; antibacterial testing and phytochemical screening were carried out at the Universitas Negeri Medan laboratory, while antioxidant and anticancer testing was carried out at the Central Lab of Universitas Padjadjaran Bandung. A 200 g of L. ferrugineus Roxb sample was macerated using acetone as a solvent to obtain a crude extract. This process is carried out for 3 × 24 hours at room temperature. The extract was filtered with Whatman filter paper, then evaporated at low pressure using a rotary evaporator at a temperature of 50 °C (
Phytochemical screening was carried out, referring to the method that was used by
Antibacterial activity test was carried out by using the paper disc diffusion method (
The antioxidant test used the 2,2-diphenyl-1-picrylhydrazyl (DPPH) method (
The anticancer activity test was carried out as in previous studies, namely the MTT method (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) Assay (
The IC50 value obtained from the linear regression equation y = a + bx. The graph was made with the concentration of the test sample (ppm) as abscissa (x-axis) against the percent Viability as the ordinate (y-axis) - data processed with Excel version 2016.
Based on the phytochemical screening result against L. ferrugineus Roxb, the results are summarized in Table
Secondary Metabolites | Test Method | Test Results |
---|---|---|
Flavonoids | FeCl3 5% | + |
NaOH 10% | + | |
Alkaloids | Dragendorff’s reagent | + |
Wagner’s reagent | + | |
Mayer’s reagent | + | |
Saponins | Aquades | + |
Triterpenoids | Liebermann-Burchard’s reagent | + |
Steroids | Liebermann-Burchard’s reagent | - |
FeCl3 1% | + |
The results obtained were similar to L. ferrugineus Roxb from Bener Meriah Regency Aceh, which was extracted with ethanol (
Previous research has shown that the chemical compound content includes flavonoids, alkaloids, lectins, polypeptides, arginine, histamine, tannins, terpenoids, acid compound steroids, glycosides, gallic acid (
In this study, the acetone extract of L. Ferrugineus Rox was tested against three gram-positive bacteria, namely P. acnes ATCC 27853, S. mutans ATCV 35668, B. cereus ATCC 1178, and one gram-negative bacterium, S. enterica ATCC 14028, so chloramphenicol was used as control positive which is able to inhibit the growth of bacteria with a broad spectrum. The acetone extract of L. ferrugineus Roxb has antibacterial activity against all tested bacteria, with a clear zone diameter between 6.2–11.1 mm. The results of the observations are summarized in Table
Mean Value of Inhibition Zone (mm) | ||||
---|---|---|---|---|
P. acnes ATCC27853 | S. mutans ATCC35668 | S. enterica ATCC14028 | B. cereus ATCC1178 | |
Negative control (DMSO 10%) | 0 | 0 | 0 | 0 |
Chloramphenicol positive control | 22.2 | 26 | 30.7 | 25.9 |
Acetone extract of L. ferrugineus Roxb | 7.6 | 11.1 | 6.2 | 7.2 |
Based on the results of observations on the in vitro antibacterial activity test, the acetone extract solution of L. ferrugineus Roxb showed strong activity against Streptococcus mutans, and other bacteria showed moderate activity. This is based on the following criteria: an extract has very strong antibacterial activity if the inhibition zone diameter is ≥ 20 mm, strong if the inhibition zone diameter is 10–20 mm, and moderate if the inhibition zone is 5–10 mm (Davis et al. 971).
Previous studies of the genus Loranthus showed antibacterial activity, such as L. micranthus, which was extracted with methanol, ethanol, chloroform, and petroleum ether solvent, which showed antibacterial activity against the test bacteria of Bacillus subtilis, Escherichia coli, and Klebsiella pneumonia (
Based on these observations, the antibacterial activity is related to the content of secondary metabolites of L. ferrugineus Roxb, namely alkaloids, terpenoids, flavonoids, and tannins. These secondary metabolites have an antibacterial effect with different mechanisms of action. The alkaloid action mechanism is by disrupting the peptidoglycan component of bacterial (
The test parameter for antioxidant activity is Inhibition Concentration (IC50), which is the concentration of an antioxidant substance that causes 50% DPPH to lose its radical character or its concentration antioxidant substance, which gives an inhibitory percentage of 50% (Nursid et al. 2013). The amount of antioxidant activity is inversely proportional to the IC50 value, meaning that the greater the antioxidant activity, the smaller the IC50 value is obtained. The acetone extract of L. ferrugineus Roxb has antioxidant activity with an IC50 value of 48.7122 (µg/ml), which was the mean value of two repetitions. The results of IC50 calculations can be seen in Table
Sample | IC50 (µg/ml) | ||
---|---|---|---|
1st repetition | 2nd repetition | Mean | |
Acetone extract of L. ferrugineus Roxb | 49.0650 | 48.3593 | 48.7122 |
If the IC50 value is < 50 µg/ml, the antioxidant activity is categorized as very strong. If the value is 50–100 µg/ml is classified as strong, and if the IC50 value is 101–250 µg/ml, it is categorized as moderate (
In Indonesia, through the Global Burden of Cancer Study (Globocan) report from the World Health Organization (WHO), the number of cancer patients reached 396,914 cases of cancer in 2020, and 54% of cases occurred in women. Cervical cancer ranks second with 36,633 cases, and lung cancer with 34,783 cases out of total cases. The anticancer activity test was carried out in vitro using the MTT assay method. This test aims to determine the toxicity of a compound. The IC50 value of L. ferrugineus Roxb acetone extract against Hela cells was 47.62 (µg/mL), and the A549 cell was 192.83 (µg/mL). These values were obtained based on the linear regression equation (Figs
NCI (National Cancer Institute) has established criteria for anticancer activity, namely an extract is declared active to have anticancer activity if it has a value of IC50 < 30 μg/mL, moderate active if it has a value of IC 50 ≥ 30 μg/mL and IC 50 < 100 μg/mL, it is said to be inactive if the value of IC 50 > 100 μg/mL (Suffness et al. 1990). Based on these provisions, the acetone extract of L. ferrugineus Roxb has strong anticancer activity against cervical cancer cells with an IC50 value of 47.62 µg/mL. It is the result of the observations of HeLa cells and A549 cell morphology. L. ferrugineus Roxb. Acetone extract at the concentration of 62 µg/mL could kill all HeLa cells and against A549 cells at a concentration of 500 g/mL (Figs
In the cell morphology (Figs
In Fig.
In many studies carried out, phenolic, terpenoids, and alkaloids group compounds are known to have activities that can inhibit growth and kill cancer cells. Flavonoids such as Epicatechin, quercetin, and catechin are also isolated from L. micranthus (
The acetone extract of parasitic L. ferrugineus Roxb in coffee trees contains flavonoids, alkaloids, triterpenoids, and tannins, and exhibits antibacterial, antioxidant, and anticancer activity. Acetone extract of L. ferrugineus Roxb is the potential as a source of antibiotic, antioxidant, and cervical anticancer compounds.
Acknowledgments are conveyed to the Chancellor of the State University of Medan for funding this research through the Public Service Agency Fund (BLU) of the Universitas Negeri Medan with the Decree of the Head of LPPM Unimed No. 104/UN33.8/KEP/PPKM/PT/2022.