Number 4 Meditory (The Journal Of Medical Laborator.
PHYTOCHEMICAL COMPOUND LEVELS IN BAMA LEAF
EXTRACT AND ITS INHIBITORY ACTIVITY AGAINST
Salmonella Thypimurium ATCC 19430 Putu Ayu Suryaningsih1*.
IB.
Oka Suyasa1.
Komang Dewi Fridayanti2 Poltekkes Kemenkes Denpasar RSU Payangan *Jalan Sanitasi No 1 Sidakarya Denpasar Selatan *corresponding author, e-mail: suryaningsihputuayu@gmail.
Article history Posted.
Sep 14th, 2023 Reviewed.
Okt 20th, 2023 Received.
Dec, 1st, 2023 Abstract Background: Diarrhea is one of the intestinal infection symptoms that can cause poor absorption to loss of nutrition if not treated immediately.
Salmonella spp.
is 1 of the 4 fundamental global causes of diarrhea diseases and antibiotics have been widely used to treat infections caused by this group of bacteria.
WHO reported that the antibiotic resistance of Salmonella spp.
has increased in past years.
For those reasons, efforts have been made to discover new antimicrobial compounds from various sources, including the ethanol extracts of Bama leaves.
Aims:
to determine the phytochemical compounds .
lavonoids, tannins, and phenol.
in the ethanol extract of Bama leaves and their inhibitory effect against the bacteria Salmonella typhimurium which causes diarrhea.
Methods:
Design method used is RAL 5 x 4, 5 treatments were concentration of Bama leaf extract 6,25%, 12,5%, 25%, 50% and 75% and 4 replicates for each test.
The Result: The phytochemical quantitative analysis showed bama leaf extract had 520,99 mgQE flavonoid / 100 gram, 1465,19 mgTAE tannin/100 g and 16531,78 phenol/100 g extract.
Conclusion: All contrentration of bama leaf extract could not inhibit the growth of Salmonella thypimurium ATCC Keywords: Bama leaf extract, phytochemical.
Salmonella thypimurium.
Sensitivity Test Introduction 88%, equivalent to around 1,516,438 Stunting is the impaired growth and This development that children experience from poor nutrition, repeated infection and approximately 1,591,944 cases in toddlers .
Diarrhea is one of the infectious diseases In Clinically, the causes of diarrhea that can lead to disturbances in nutrient can be categorized into six major groups absorption and the loss of essential such as infection, malabsorption, allergies, nutrients if not treated immediately.
poisoning, immunodeficiency, and other According to data from the Indonesian Cases of diarrhea in Indonesia are Ministry of Health, the prevalence of typically caused by bacteria such as Vibrio diarrhea in 2018 was approximately cholerae.
Shigella spp.
Escherichia coli.
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Putu Ayu Suryaningsih, et all : Phytochemical Compound Levels In Bama Leaf Extract And Its Inhibitory Activity Against Salmonella Thypimurium ATCC 19430 Salmonella spp, and Campylobacter jejuni effect of crude extract of bama leaves in .
Among these.
Salmonella is the most common bacterial group responsible for Staphylococcus aureus and Candida had food poisoning.
Salmonella contamination been observed and discovered that crude is usually the result of poor hygiene and extract of bama leaves could inhibit and improper food handling practices .
had potential antimicrobial activity .
Antibiotics have been widely used in Research Bacillus Dhale the treatment of intestinal infections.
Markandeya, revealed that Bama leaf Excessive use of antibiotic can pose risks extract has antimicrobial activity against such as bacterial resistance, the retention Staphylococcus aureus.
Bacillus subtilis, of toxic substances, and antibiotic residues coli, and Pseudomonas aeruginosa.
The .
This has triggered a trend in society to back to nature in the field of healthcare, chloroform, and alcohol .
Therefore, this study aims to determine the effect of antibiotics .
Indonesia as a tropical country, has a Bama inhibiting Salmonella typhimurium which wealth of flora that can be utilized as cause diarrhea.
One of these is the Bama plant Method (Plumbago zeylanica L.
The leaves and The type of this study was true roots are efficacious as medicine for experimental study with a posttest-only various diseases, including the use of control design.
The first group to be leaves for treating arthritis or rheumatism, treated was called the experimental group, relieving flatulence, urinary difficulties, consisting of Bama leaf ethanol extract headaches, alleviating joint pain, skin with concentrations of 6.
25%, 12.
25%, 50%, and 75%.
The second group menstruation, and urinary tract infections.
was a control group consisting of a Bama leaves are also known to be positive control which is an antibiotic that beneficial for digestion.
is commonly used to treat Salmonellosis.
The study analyzed Bama leaf extract namely chloramphenicol and a negative and obtained compound of alkaloids, control which is used as a working control.
saponins, tannins and glycosides .
The Each treatment was repeated four times Meditory | e-ISSN: 2549-1520, p-ISSN: 2338 Ae 1159.
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Putu Ayu Suryaningsih, et all : Phytochemical Compound Levels In Bama Leaf Extract And Its Inhibitory Activity Against Salmonella Thypimurium ATCC 19430 Federer Formula About 0.
01 g of the crude extract was diluted in 5 ml of citrate phosphate Salmonella typhimurium ATCC 19430 buffer according to the treatment.
was obtained from the Microbiology sample of 0.
1 ml was pipetted and mixed Laboratory Medicine, 3 ml of 70% ethanol.
Subsequently.
Udayana University.
Meanwhile Bama 4 ml of Folinciaocalteau reagent was leaves were obtained from Nusa Penida added and the mixture was incubated for 6 Island.
Klungkung.
The Bama leaves were After the incubation process, 4.
extracted at the Post-Harvest Medicinal ml of 5% Na2CO3 was added, followed by Plant vortexing and then incubated for 90 Karangasem.
Quantitative phytochemical The absorbance was read at a analysis was conducted in the Agriculture wavelength of 760 nm.
The reading results Laboratory and the sensitivity test was were compared to a standard curve carried out in the Microbiology Laboratory constructed using gallic acid.
Faculty Processing Center
(P4TO)
at Warmadewa University.
The determination of total flavonoids Bama leaves had been picked, was conducted using a spectrophotometer cleaned, and dried were pulverized into following the AlCl3 method.
About 0.
fine powder .
Subsequently, the of the extract was diluted in 5 ml of citrate Bama leaf powder was macerated using 96% ethanol for six days.
The filtrate from Then, 1 ml of the sample was the Bama leaves was collected, and the mixed with 4 ml of distilled water and 0.
extraction process was carried out using an ml of NaNO2 10% solution was added.
evaporator until a crude extract was The mixture was incubated for 5 minutes.
Crude extract of Bama leaf were and then 0.
3 ml of AlCl3 10% solution and going to be utilized for quantitative 2 ml of NaOH 1% solution were added.
The sample was immediately tested using flavonoids, and tannin.
and for testing its a spectrophotometer at a wavelength of 510 nm.
Salmonella typhimurium ATCC 19430.
The determination of total tannin The determination of total phenols extract was analyzed using the Folin-Denis was carried out using the Folin-Ciocalteau A total of 0.
01 g of the extract Meditory | e-ISSN: 2549-1520, p-ISSN: 2338 Ae 1159.
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Putu Ayu Suryaningsih, et all : Phytochemical Compound Levels In Bama Leaf Extract And Its Inhibitory Activity Against Salmonella Thypimurium ATCC 19430 was diluted in 5 ml of citrate phosphate MHA media.
This procedure was repeated buffer according to the treatment.
The four times.
The media with the disk filters diluted sample was pipetted in 0.
25 ml, were incubated at 37AC for 24 hours in an and added 0.
25 ml of Folin-Denis reagent.
The mixture was vortexed and added 2 ml reported by measuring the diameter of the of Na2CO3 5%.
The solution was vortexed inhibition zones formed using a caliper.
again and incubated for 30 minutes.
The The obtained inhibition zone diameters were analyzed using statistical tests to spectrophotometer at a wavelength of 725 assess differences among the concentration The The media used to observe the Mueller Hinton Agar (MHA).
Result and Discussion This study utilized Bama leaves (Plumbago suspension is prepared by adding 1-3 pure culture colonies into a tube containing 5 Klungkung.
Bama plants typically grow as ml NaCl 0,9%.
It is then compared to the A crude extract was obtained standard of turbidity at 0.
5 McFarland.
through the maceration and extraction Salmonella Nusa Penida.
The antibacterial activity test was The maceration method dissolves conducted using the disk diffusion method.
active compounds due to the difference in Empty disk-shaped filters were saturated compounds inside and outside the cell concentrations : 6.
25%, 12.
5%, 25%, 50%,
causing the active compounds to exit until and 75%.
The negative control utilized the concentration equilibrium is reached.
sterile distilled water and the positive The maceration process uses 96% ethanol control used Chloramphenicol antibiotic because ethanol is more selective and mold The inoculated onto the surface of MHA concentrations of 20% and above and The negative and were then placed non-toxic.
Ethanol is semi-polar so it can on the MHA media.
The negative and dissolve active ingredients in plants that positive control were placed on separate are polar, semi-polar, and non-polar.
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Putu Ayu Suryaningsih, et all : Phytochemical Compound Levels In Bama Leaf Extract And Its Inhibitory Activity Against Salmonella Thypimurium ATCC 19430 The analysis aims to determine the total content .
lavonoids, phenol.
in the ethanol extract of Bama simplicia or crude extract.
The total leaves iare presented in the table below.
Table 1.
Total Content of Phytochemical Compounds in the Ethanol Extract of Bama Leaves Replication Average Flavonoid Test gQE/100.
493,42 Tannin Test gTAE/100.
1475,41 Phenol Test gGAE/100.
16606,17 474,33 595,24 520,99 1443,21 1476,97 1465,19 16492,20 16496,99 16531,78
Based on table 1, the total flavonoid immunoregulators, antibacterial, anticancer, concentration in the ethanol extract of antidiabetic agents, antioxidants, antiviral.
Bama leaves is 520.
99 mgQE/100g.
The estrogenic agents, and playing a role in total flavonoid concentration obtained can neurodegenerative diseases.
They also have be compared with other plants, such as function as inhibitors of AChE and BChE soursop leaves (Annona muricata L.
and have hepatoprotective .
Flavonoids which have a total flavonoid concentration act as antibacterial agents through several 82% .
mg/100.
However, the mechanisms, including inhibiting nucleic total flavonoid concentration in mulberry leaves (Muntingia calabura L.
) is 13.
mgQE/100g .
Therefore, it can be bacterial energy metabolism.
Based on the study results of Manik.
Hertiani, and concentration in Bama leaves is higher than Anshory, the antibacterial activity of that in soursop and mulberry leaves.
ethanol extract from mulberry leaves and Flavonoids pharmacological activities, namely as as steroid-genesis modulators, neuroprotective anti-inflammatory each fractions was 93% influenced by the total flavonoid concentration .
Based on table 1, the total tannin concentration in the ethanol extract of Meditory | e-ISSN: 2549-1520, p-ISSN: 2338 Ae 1159.
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Putu Ayu Suryaningsih, et all : Phytochemical Compound Levels In Bama Leaf Extract And Its Inhibitory Activity Against Salmonella Thypimurium ATCC 19430 Bama leaves is 1465.
19 mgTAE/100g.
The higher than in soursop leaves but lower tannin concentration in soursop leaf extract than in mulberry leaves.
53 mg/100g .
However the The antibacterial mechanism of tannin concentration in mulberry leaf phenolic compounds involves interfering extract is 137.
15 mg GAE/100g extract with the peptidoglycan components of the .
Therefore, it can be concluded that the cell wall of Gram-positive bacteria.
This tannin concentration in Bama leaves is interference prevents the binding of N- higher than soursop and mulberry leaves.
acetylmuramic acid to the muropeptide Tannin is one of the active secondary metabolites with several properties such as providing rigidity to the cell wall.
As a result, bacterial cell wall synthesis is anti-diarrheal.
The disrupted and does not form correctly.
This leads to the loss of the rigid cell wall in antibacterials is by inhibiting reverse vulnerable to damage and leakage.
Positive enzymes, preventing the formation of correlation analysis with a correlation bacterial cells.
This is achieved by its coefficient of 0.
577 indicates that as the ability to inactivate microbial cell adhesins, antibacterial activity increases, the phenol enzymes, and disrupt transport proteins in content in the S.
muticum simplicia also the inner cell layer .
DNA
Based on table 1, the total phenol The concentration in the ethanol extract of activity was conducted by measuring the Bama leaves is 16,531.
78 mgGAE/100g.
diameter of the inhibition zone around the The total phenol concentration in soursop disks containing the extract.
Based on leaf extract is 27.
14 g GAE/g, which is figure 1, ethanol extracts of Bama leaves at equivalent to 2.
714 mg GAE/100g13.
The concentrations of 6.
25%, 12.
5%, 25%,
total phenol concentration in mulberry leaf 50%, and 75% have not able to inhibit the mgGAE/100g10.
growth of Salmonella typhimurium ATCC
22,389
Therefore, it can be concluded that the total 19430 bacteria.
phenol concentration in Bama leaves is Meditory | e-ISSN: 2549-1520, p-ISSN: 2338 Ae 1159.
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Putu Ayu Suryaningsih, et all : Phytochemical Compound Levels In Bama Leaf Extract And Its Inhibitory Activity Against Salmonella Thypimurium ATCC 19430 Figure 1.
Inhibition zones of chloramphenicol, negative control, and ethanol extracts of Bama leaves at concentrations of 6.
25%, 12.
5%, 25%, 50%, and 70%.
Several Salmonella typhi .
This is because of antibacterial activity include the type of the differences in the cell wall structure bacteria being inhibited, the content of between gram-positive and gram-negative antibacterial compounds, the concentration of the extract, and the diffusion capacity of The phytochemical compounds in an extract.
Furthermore, variations in cell the ethanol extract of Bama leaves, wall structure also determine the activity, including flavonoids, tannins, and phenols penetration, and binding of antibacterial have antibacterial properties but have not Salmonella typhimurium is a been able to inhibit the growth of Gram-negative bacterium with an outer Salmonella typhimurium.
This may be due membrane composed of three layers, including lipopolysaccharides, lipoproteins, typhimurium bacterial cell membrane which and phospholipids, with porins formed from consists of three layers.
Therefore, active This outer membrane acts as a ingredients have difficulty penetrating the The study results of Lio.
Useng, and enzymes, and dry conditions.
This aligns Ramang indicate that a 60% concentration with the study findings of Jamili.
Hidayat, and Hifizah which showed that Gram- resistance to Salmonella typhimurium .
positive bacteria, such as Staphylococcus This suggests that the cell wall of aureus produced larger inhibition zones Salmonella typhimurium is indeed more Salmonella compared to Gram-negative bacteria like Meditory | e-ISSN: 2549-1520, p-ISSN: 2338 Ae 1159.
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Putu Ayu Suryaningsih, et all : Phytochemical Compound Levels In Bama Leaf Extract And Its Inhibitory Activity Against Salmonella Thypimurium ATCC 19430 Jurnal Surya Medika.
: 23-31
ingredients in the extract.
Suhirman.
Fatimah, dan M.
Sukmasari.
Analisis kimia daun, batang, dan akar daun encok (Plumbago zeylanica ) secara kualitatif.
hlm 350Ae357.
Dalam Supriadi.
Januwati.
Balfas.
Bermawie, dan M.
Rahardjo (Ed.
Prosiding Seminar Nasional XXVi dan Pameran Tumbuhan Obat Indonesia Bogor, 15Oe16 September 2005.
Hiradeve S.
Danao K.
Kharabe V.
Mendhe Evaluation of anticancer activity of Plumbago zeylanica Linn leaf extract.
International Journal of Biomedical Research.
: 01-09
Dhale.
Markandeya.
Antimicrobial screening of Plumbago zeylanica Linn (Plumbaginacea.
Journal Experimental Sciences.
: 04-06
Mukhriani, et al.
Analisis kadar flavonoid total pada ekstrak daun sirsak (Annona spektrofotometri uv-vis.
Jurnal Farmasi UIN Alaudin Makassar.
: 37Ae Conclusion Based on this study, it can be concluded that the ethanol extract of Bama leaves (Plumbago zeylanica L.
) has a total flavonoid content of 520.
99 mgQE/100g, a total tannin content of 1465.
19 mgTAE/100g, and a total phenol content of 16531.
78 mgGAE/100g.
However, the ethanol extract of Bama leaves has not been able to inhibit Salmonella typhimurium ATCC 19430.
Reference