The document summarizes a study that investigated the antibacterial activities of methanol extract and chloroform fraction of soursop (Annona muricata L.) leaves against Escherichia coli and Staphylococcus aureus. The methanol extract showed inhibition zones of 14.1 mm against S. aureus and 13.1 mm against E. coli at a concentration of 150 mg/ml. At 250 mg/ml, the methanol extract inhibited E. coli with a zone of 14.5 mm. The chloroform fraction only inhibited S. aureus with a zone of 9.9 mm at 150 mg/ml. The methanol extract demonstrated higher antibacterial activity compared to the chloroform fraction against both bacterial strains.
Lipase production and purification Likhith KLIKHITHK1
Lipase (tri acyl glycerol acyl hydrolase, EC 3.1.1.3) catalyzes the hydrolysis of the carboxyl ester bonds in tri acyl glycerols to produce di acyl glycerols, mono acyl glycerols, fatty acids and glycerol under aqueous conditions and the synthesis of esters in organic solvents.
Under the controlled conditions, lipases are able to catalyze a large number of reactions. Lipases of microbial origin are of considerable commercial importance, because of the high versatility and high stability, moreover, the advantage of being readily produced in high yields.
Many microbial lipases have been commercially available in free or immobilized form. Numerous species of bacteria (Bacillus, Pseudomonas, and Burkholderia), yeasts (Candida rugosa, Yarrowia lipolytica, and Candida antarctica) and molds (Aspergillus, Trichoderma viride) produce lipases with different enzymological properties and specificities but microbes are known to be more potent lipase producer.
Download luận văn đồ án tốt nghiệp ngành công nghệ thực phẩm với đề tài: Sử dụng enzym pectinase trong sản xuất nước rong nho (Caulerpa lentillifera J. Agardh) đóng chai, cho các bạn tham khảo
Antimicrobial activity of annona muricata L. (Soursop Leaves)Fattah Fazel
Testing antimicrobial activity of Annona Muricata L. (Soursop) using Disk diffusion method and Well diffusion method.
Bacteria used: MRSA, S.pyogenes, B. fragilis, C. perfringens
Note: No results. Reasons in the description section.
Lipase production and purification Likhith KLIKHITHK1
Lipase (tri acyl glycerol acyl hydrolase, EC 3.1.1.3) catalyzes the hydrolysis of the carboxyl ester bonds in tri acyl glycerols to produce di acyl glycerols, mono acyl glycerols, fatty acids and glycerol under aqueous conditions and the synthesis of esters in organic solvents.
Under the controlled conditions, lipases are able to catalyze a large number of reactions. Lipases of microbial origin are of considerable commercial importance, because of the high versatility and high stability, moreover, the advantage of being readily produced in high yields.
Many microbial lipases have been commercially available in free or immobilized form. Numerous species of bacteria (Bacillus, Pseudomonas, and Burkholderia), yeasts (Candida rugosa, Yarrowia lipolytica, and Candida antarctica) and molds (Aspergillus, Trichoderma viride) produce lipases with different enzymological properties and specificities but microbes are known to be more potent lipase producer.
Download luận văn đồ án tốt nghiệp ngành công nghệ thực phẩm với đề tài: Sử dụng enzym pectinase trong sản xuất nước rong nho (Caulerpa lentillifera J. Agardh) đóng chai, cho các bạn tham khảo
Antimicrobial activity of annona muricata L. (Soursop Leaves)Fattah Fazel
Testing antimicrobial activity of Annona Muricata L. (Soursop) using Disk diffusion method and Well diffusion method.
Bacteria used: MRSA, S.pyogenes, B. fragilis, C. perfringens
Note: No results. Reasons in the description section.
Bài giảng được xuất bản mong nhận được ý kiến đóng góp từ kinh nghiệm sản xuất thực tế như số liệu, các thông số nhà máy ... mà hoàn thiện hơn và là sổ tay cho mọi tân sinh viên mới ra trường bốt bỡ ngỡ trong công việc
Mọi ý kiến đóng góp gửi về ngconghoan2881985@gmail.com, cong6hoan@gmail.com
Số Điện thoại 0918001595
Moringa is used for “tired blood” (anemia); arthritis and other joint pain
(rheumatism); asthma; cancer; constipation; diabetes; diarrhea; epilepsy;
stomach pain; stomach and intestinal ulcers; intestinal spasms; headache; heart
problems; high blood pressure; kidney stones; fluid retention; thyroid disorders;
and bacterial, fungal, viral, and parasitic infections.
Oil from moringa seeds is used in foods, perfume, and hair care products, and as a
machine.
Bài giảng được xuất bản mong nhận được ý kiến đóng góp từ kinh nghiệm sản xuất thực tế như số liệu, các thông số nhà máy ... mà hoàn thiện hơn và là sổ tay cho mọi tân sinh viên mới ra trường bốt bỡ ngỡ trong công việc
Mọi ý kiến đóng góp gửi về ngconghoan2881985@gmail.com, cong6hoan@gmail.com
Số Điện thoại 0918001595
Bioassays are assays or biological techniques to measure strength, potency, concentration or efficacy of any substance by its effect on biological substance like tissues, cells, animals or enzymes etc
Bài giảng được xuất bản mong nhận được ý kiến đóng góp từ kinh nghiệm sản xuất thực tế như số liệu, các thông số nhà máy ... mà hoàn thiện hơn và là sổ tay cho mọi tân sinh viên mới ra trường bốt bỡ ngỡ trong công việc
Mọi ý kiến đóng góp gửi về ngconghoan2881985@gmail.com, cong6hoan@gmail.com
Số Điện thoại 0918001595
Moringa is used for “tired blood” (anemia); arthritis and other joint pain
(rheumatism); asthma; cancer; constipation; diabetes; diarrhea; epilepsy;
stomach pain; stomach and intestinal ulcers; intestinal spasms; headache; heart
problems; high blood pressure; kidney stones; fluid retention; thyroid disorders;
and bacterial, fungal, viral, and parasitic infections.
Oil from moringa seeds is used in foods, perfume, and hair care products, and as a
machine.
Bài giảng được xuất bản mong nhận được ý kiến đóng góp từ kinh nghiệm sản xuất thực tế như số liệu, các thông số nhà máy ... mà hoàn thiện hơn và là sổ tay cho mọi tân sinh viên mới ra trường bốt bỡ ngỡ trong công việc
Mọi ý kiến đóng góp gửi về ngconghoan2881985@gmail.com, cong6hoan@gmail.com
Số Điện thoại 0918001595
Bioassays are assays or biological techniques to measure strength, potency, concentration or efficacy of any substance by its effect on biological substance like tissues, cells, animals or enzymes etc
This is an Engg Biotechnology project based on medicinal plant i.e singapore cherry or jamaican cherry tree (scientific name Muntingia calabure ), we did in 2013 in GMIT college Davangere, karanataka, India. i have complete project detail what we did..,
Introduction to Diabetes & anti diabetic drug screening methodsAnurag Raghuvanshi
Diabetes is one of the most common life long disese now days & there are various works done in pharma. how dugs are developed & the animals used in this methodolgy is well brief in these slides.
Anti Microbial Activities and Phytochemical Screening of the Premna Odorata B...ijtsrd
The researcher focused mainly to determine the antimicrobial properties and phytochemical screening of the Alagaw leaf extract. Specifically, this study was conducted to determine the percent yield, antimicrobial activities and the secondary metabolites of Alagaw leaf extract which was analyzed and it include alkaloid, anthraquinone, saponins and steroid. Findings of the study showed that the Alagaw leaf extract has a percent yield of 11.5 . Anti Microbial Activity was tested by petri disk on a plate Nutrient Agar streaked with the E. coli bacteria, the plates were incubated for 24hrs and 37oC. Results were observed for the presence of zone inhibition clear area around the test disk. Results from this study showed that the antimicrobial activity on E. coli as indicated as negative by the presence of Alagaw leaf extract. Furthermore, the result suggests that the Alagaw leaf extract did not suppress the growth of the E. coli bacteria, hence it indicates that it has no anti microbial effect to the test organism. While the secondary metabolites such as alkaloid, anthraquinone, saponins and steroid is found negative. It is therefore recommended that further study of the chemical properties of alagaw leaf extract, barks and roots should be conducted Bernadette C. Mollejon | Charito V. Mollejon ""Anti-Microbial Activities and Phytochemical Screening of the Premna Odorata Blanco (Alagaw) Leaf Extract"" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-3 | Issue-4 , June 2019, URL: https://www.ijtsrd.com/papers/ijtsrd23751.pdf
Paper URL: https://www.ijtsrd.com/other-scientific-research-area/other/23751/anti-microbial-activities-and-phytochemical-screening-of-the-premna-odorata-blanco-alagaw-leaf-extract/bernadette-c-mollejon
Phytochemical Potential and Antimicrobial Activity of Andrographispaniculataiosrjce
The Herbal medicine today ensures safety in contrast to the synthetic preparations. Herbs the Nature’s
Physician, have been reported as an important source of medicine for years and years. Using of herbs for
curing diseases dated back to prehistory and people of all continents have this old tradition.Recently, wide
research proposals highlight the property of medico potential from phytalsources. My herb of interest is also the
above said, ofcourseAndrographispaniculata (Acanthaceae) is a medicinal plant used for the treatment of
various ailments, which has been documented in history of all civilizations. The present study is to learn the
phytochemical properties and the antimicrobial activity of the above using disc diffusion method
Anti-Oxidant and Antimicrobial Studies of Tinospora cordifolia (Guduchi/Giloy...SUS GROUP OF INSTITUTIONS
Plants produce a diverse range of bioactive molecules, making them a rich source of
different types of medicines and healing properties. The present study was aimed to
evaluate the anti-oxidant and antimicrobial properties of stem and root of T. cordifolia.
Total phenolic contents of different solvent extracts were determined and found that ethanol
extract had the highest phenolic content of 0.3213 mg g-1. Antioxidant assays were also
carried out by using different in vitro models such as total reducing power, hydrogen
peroxide scavenging activity assay and hydroxyl redical scavenging activity. The Ethanol
extract showed the highest total antioxidant activity. The H2O2 scavenging and hydroxyl
free radical scavenging activity was maximum 87.2 % and 91.0% found in case of ethanolic
steam extract respectively. The antimicrobial activity of ethanolic and methanolic extract of
root and stem of T. cordifolia were also evaluated against some pathogenic microorganisms
viz. E. coli, B. subtilis, A. niger and Candida sp. it was found that the various concentration
of extract viz. 50, 100, 150 and 200 mg ml-1 were tested. It was observed that the
increasing in concentration there was also increasing in antimicrobial activity reveled by
increase in size of zone of inhibition. The methanolic stem extract exhibits highest
antimicrobial activity against all four pathogens. The study shown that the extract of T.
cordifolia has a wide range of anti-oxidant as well as antimicrobial activity against bacterial
as well as fungal pathogens.
In Vitro Antibacterial Activities of Cochlospermum planchonii Roots Crude Ext...iosrjce
The antibacterial activities of the methanolic, hot water, chloroform and petroleum ether of
Cochlospermum planchonii root extracts on some clinical bacterial isolates and reference organisms were
investigated using conventional microbiological and microdilution indicator technique. Phytochemical
screenings were also carried on the extracts. The root extracts of the plant exhibited antibacterial activities
against reference strains and clinical isolates of Escherichia coli, Pseudomonas aeruginosa, Staphylococcus
aureus, Shigella flexneri, and Salmonella typhii. However, the susceptibility pattern of the bacteria did not
differ significantly from each other (p>0.05). The methanolic root extracts exhibited the highest antibacterial
activity, its minimum inhibitory concentration (MIC) ranging between 1.25 mg/ml and 5.00mg/ml; and its zones
of inhibition diameter on the various test microorganisms ranging between 8mm and 12mm. The petroleum
ether extracts had the weakest antibacterial activity, with minimum inhibitory concentration of 5.00mg/ml and
its zones of inhibition diameter ranging between 4mm and 7mm. The bioactive constituents in the plant were
alkaloids, tannins, saponins, cardiac glycosides, and sterols. The methanolic extracts of root appeared to be
more biologically active than other extracts and may be more useful in treating human infections caused by
these pathogens.
A Preliminary Study on Phytochemical Screening of Boerhaavia Diffusa, Euphorb...ijtsrd
Medicinal plants are of great importance to the health of individuals and communities. The medicinal value of these plants lies in some chemical substances that produce a definite physiological action on the human body. The objective of the present study was to evaluate the phytochemical constitution and antioxidant activity of Aqueous extracts of three selected plant Boerhaavia diffusa, Euphorbia hirta and Amaranthus polygonoides. Preliminary phytochemical screening revealed the presence of phytochemicals like alkaloids, flavonoids, Steroids, phenols, tannin and carbohydrates in Boerhaavia diffusa and Euphorbia hirta where as in Amaranthus polygon ides many phytoconstituents like alkaloids, flavonoids, Steroids, terpenoids, phenols, saponin, tannin and carbohydrates were present. Antioxidants are the compounds which terminate the attack of reactive species and reduce the risk of diseases. The free radicals oxidants are species with very short half life, high reactivity and damaging activity towards macromolecules like proteins, DNA and lipids. The results of antioxidant activity of three aqueous extract showed maximum activity in different concentration of 50, 250, 500, 750 and 1000 µg ml. The percent inhibition of aqueous extract of Boerhavia diffusa , Euphorbia hirta , Amaranthus polygonoides was 176.15, 404.78 and 413.06 respectively. In the present work potent anti oxidant activity of aqueous extract of Boerhaavia diffusa was higher when compared to other two extracts. The present study revealed that the plant extract possessed good antioxidant activity and less quantity of toxic metals, which therefore can be used as a natural source of free radical scavenger. However, further study needs to be carried out to know its mode of action. R. Ezhilarasi | Dr. B. Senthilkumar | Dr. K. Devi "A Preliminary Study on Phytochemical Screening of Boerhaavia Diffusa, Euphorbia Hirta and Amaranthus Polygonoides" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-4 | Issue-1 , December 2019, URL: https://www.ijtsrd.com/papers/ijtsrd29435.pdfPaper URL: https://www.ijtsrd.com/biological-science/zoology/29435/a-preliminary-study-on-phytochemical-screening-of-boerhaavia-diffusa-euphorbia-hirta-and-amaranthus-polygonoides/r-ezhilarasi
Membrane Stabilizing And Antimicrobial Activities Of Caladium Bicolor And Che...IOSR Journals
The crude methanol extracts of whole plant of Caladium bicolor (Aiton) Vent. and leaf of Chenopodium album L. as well as their pet-ether, carbon tetrachloride, chloroform and aqueous soluble fractions were evaluated for membrane stabilizing and antimicrobial activities. At concentration 1.0 mg/ml, the carbon tetrachloride soluble fraction of C. bicolor inhibited 43.92±1.63% and 38.08±0.83 % hypotonic solution and heat induced haemolysis of RBCs, respectively. Among the extractives of C. album, the aqueous soluble fraction inhibited 47.11±0.49 % and 36.73±0.76 % hypotonic solution and heat induced haemolysis of RBCs as compared to 72.79 % and 42.12 % by acetyl salicylic acid (0.10 mg/ml), respectively. C. bicolor test samples demonstrated zone of inhibition ranging from 6.0 to 20.0 mm. The chloroform soluble fraction showed the highest zone of inhibition (20.0 mm) against Staphylococcus aureus. The test samples of C. album displayed zone of inhibition ranging from 7.0 to 13.0 mm. The highest zone of inhibition (13.0 mm) was showed by the chloroform soluble fraction against Salmonella paratyphi
Antibacterial property of Atuna racemosa Rafin. Chrysobalanaceae shell and ke...Innspub Net
This research evaluated the antibacterial potential of the aqueous, ethyl acetate, methanol, and decocted extracts of the shell and kernel of Atuna racemosa Rafin. Chrysobalanaceae (tabon-tabon). The antimicrobial screening was done against Escherichia coli and Staphylococcus aureus by paper disc diffusion method. The A. racemosa shell and kernel showed resistant to intermediate antimicrobial activity against E. coli and S. aureus in aqueous extracts with mean zone of inhibition of 7.7 mm and 9.8 mm, ethyl acetate extracts with 9.2 mm and 12.8 mm, methanol extracts with 9.5 mm and 13.2 mm, and decoction extracts with 7.3 mm and 11.0 mm, respectively. Ethyl acetate extracts with the highest antibacterial activity against E. coli obtained minimum inhibitory concentration values of 0.11375 mg/mL in shell and 2.92 mg/mL in kernel for both bacterial strains. Methanol extracts with the highest antibacterial activity against S. aureus obtained minimum inhibitory concentration values of 0.81375 mg/mL in shell for both test organisms, and 8.57 mg/mL for E. coli and 2.138 mg/mL for S. aureus in kernel. Overall, the ethyl acetate and methanol extracts of A. racemosa kernel showed good antibacterial potential against bacterial strains. Further investigation is needed to determine the bioactive components present in these extracts.
The IOSR Journal of Pharmacy (IOSRPHR) is an open access online & offline peer reviewed international journal, which publishes innovative research papers, reviews, mini-reviews, short communications and notes dealing with Pharmaceutical Sciences( Pharmaceutical Technology, Pharmaceutics, Biopharmaceutics, Pharmacokinetics, Pharmaceutical/Medicinal Chemistry, Computational Chemistry and Molecular Drug Design, Pharmacognosy & Phytochemistry, Pharmacology, Pharmaceutical Analysis, Pharmacy Practice, Clinical and Hospital Pharmacy, Cell Biology, Genomics and Proteomics, Pharmacogenomics, Bioinformatics and Biotechnology of Pharmaceutical Interest........more details on Aim & Scope).
All manuscripts are subject to rapid peer review. Those of high quality (not previously published and not under consideration for publication in another journal) will be published without delay.
The IOSR Journal of Pharmacy (IOSRPHR) is an open access online & offline peer reviewed international journal, which publishes innovative research papers, reviews, mini-reviews, short communications and notes dealing with Pharmaceutical Sciences( Pharmaceutical Technology, Pharmaceutics, Biopharmaceutics, Pharmacokinetics, Pharmaceutical/Medicinal Chemistry, Computational Chemistry and Molecular Drug Design, Pharmacognosy & Phytochemistry, Pharmacology, Pharmaceutical Analysis, Pharmacy Practice, Clinical and Hospital Pharmacy, Cell Biology, Genomics and Proteomics, Pharmacogenomics, Bioinformatics and Biotechnology of Pharmaceutical Interest........more details on Aim & Scope).
All manuscripts are subject to rapid peer review. Those of high quality (not previously published and not under consideration for publication in another journal) will be published without delay.
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Study of the antibacterial activities of soursop (annona muricata l.) leaves
1. International Journal of PharmTech
Research
CODEN (USA): IJPRIF
ISSN : 0974-
4304
Vol.6, No.2, pp 575-581,
April-June
2014
Study Of The Antibacterial Activities Of
Soursop (Annona muricata L.)
Leaves
Ginda Haro*, Niky Puji Utami, and Erly Sitompul
Faculty of Pharmacy, University of Sumatera Utara,
Jl. Tri Dharma No. 5, Pintu 4, Kampus USU, Medan, Indonesia, 20155
*Corres. Author :
gindaharo@yahoo.com Phone
number : +62 812 6477 1909
Abstract : Background: The use of natural product medicines has emerged from traditional to modern
therapy in order to increase the quality of health worldwide. Nature-derived medicines are considered safer. The
leaves of soursop (Annona muricata L.) has long been used by certain local communities in Indonesia as an
alternative treatment of bacterial diseases.
Objective: Objective of this study was the investigation of antibacterial activities of methanol extract and
chloroform fraction of the leaves of soursop (Annona muricata L.) of the family of Annonaceae against
Escherichia coli and Staphylococcus aureus
Methods: The methanol extract and chloroform fraction were obtained by means of maceration method using
methanol as solvent and then fractionated, with chloroform. The antibacterial activities were measured in vitro
by means of agar diffusion method using paperdisc.
Results: Methanol extract of soursop leaves at concentration of 150 mg/ml could inhibit the growth of
Staphylococcus aureus with inhibition zone diameter of 14.1 mm and of 13.1 for Escherichia coli. But the
methanol extract of soursop leaves at concentration of 250 mg/ml could inhibit the growth of Escherichia coli
with inhibition zone diameter of 14.5 mm. It fulfilled the requirement of Farmakope Indonesia. Chloroform
fraction of soursop leaves at concentration of 150 mg/ml could only inhibit the growth of Staphylococcus
aureus with inhibition zone diameter of 9.9 mm and of 9.2 mm for Escherichia coli.
Conclusion: The methanol extract indicated a higher inhibition zone diameter than chloroform fraction against
Staphylococcus aureus and Escherichia coli.
2. Key words: soursop leaves, antibacterial activities, Escherichia coli and Staphylococcus aureus.
Introduction
We previously have reported our investigation about the characterization and the phytochemical screening of
the leaves of soursop (Annona muricata L.). The alkaloids, flavonoids, tannins, saponins, glycosides and
steroid/triterpenoids were present on the leaves. The antibacterial activities against Staphylococcus aureus,
Stapylococcus epidermidis, Propionibacterium acne and Pseudomonas aeruginosa were also positively found1.
Soursop (Annona muricata L.) is a tropical plant and familiar to the Indonesian certain local communities. This
plant has great benefits for human life which is full with nutrition. In the food industry soursop can be
Ginda Haro et al /Int.J.PharmTech Res.2014,6(2),pp 575-581. 576
processed into jam, fruit juice, syrup2. Soursop leaves contain flavonoid, tannin, alcaloid, saponin, calcium,
phosphor, carbohydrate, vitamine A, B and C, phytosterol, calcium oxalate3.
These leaves are traditionally used to prevent and treat arthritis, asthma, bronchitis biliary disorder, diabetic,
heart diseases, hypertension, worm disease, liver disorder, malaria, rheumatism, sedative, tumor, and cancer 4.
The leaves are also used for the treatment of several types of diseases caused by bacteria such as pneumonia,
diarrhea, urinary tract infection, and other kinds of skin diseases5.
Staphylococcus aureus is a gram positive bacteria that is commonly found on human skin and mucous
membranes. This bacteria can cause skin infection6. Escherichia coli is a gram negative bacteria that is
commonly found in the human colon. This bacteria is one of the most common pathogenic bacteria in food
causing the primary infection of the intestine such as diarrhea7.
Based on those facts, in order to continue our investigation we report here our study about the antibacterial
activities of soursop (Annona muricata L.) leaves against Escherichia coli and Staphylococcus aureus.
Experimental Methods
Materials
Materials used in this study were soursop (daun sirsak, Annona muricata L.) leaves, agar nutrient media, broth
nutrient media, Staphylococcus aureus (ATCC No. 6538) and Escherichia coli (ATCC No. 25922), and
aquadest.
Apparatus
Apparatus used in this study were glasses, autoclave (Fisons), blender (Philips), freeze dryer (Modulio),
incubator (Fiber Scientific), filter paper, Laminar Air Flow Cabinet (Astec HLF 1200L), refrigerator (Toshiba),
oven (Memmert), heater, micro pipet (Eppendorf), rotary evaporator (Haake D), visible spectrophotometer
(Dynamic), digital balance (Mettler Toledo).
The Collection and The Treatment of Soursop Leaves
The leaves of soursop (Annona muricata L.) were purposively collected from Kelurahan Tanjung Mulia,
Kecamatan Medan Deli, Medan city, Indonesia in January 2013. The fourth and the fifth leaves were picked out
of the point of a young leaf.
The Identification of Soursop Leaves
It was conducted by the Herbarium Medanense, University of Sumatera Utara.
The Preparation of Symplex
The leaves of soursop was weighed after washing and drying in open air. They were blended to obtain a powder
3. mass and kept in a closed pocket of plastics.
The Preparation of Extract of Soursop (Annona muricata L.) Leaves
500 g of symplex powder of soursop leaves were macerated for 5 days out of light, occasionally stirred, by
maceration method using methanol as solvent. After 5 days, the mixture was filtered and the residue washed by
using methanol and treated for 2 days of the same treatment as before. The macerates were then mixed together
and concentrated by means of rotary evaporator with the temperature not exceeding 40°C untill obtained
spissum extract by means of freeze dryer8.
Ginda Haro et al /Int.J.PharmTech Res.2014,6(2),pp 575-581. 577
The Preparation of Media
Agar nutrient media
Composition: Lab-lemco powder
1
g
Yeast extract
2
g
Peptone
5
g
Sodium chloride 5 g
Agar
1
5
g
The preparation9: 28 g of agar nutrient media was suspended in 1000 ml of distilled water and heated to
completely dissolved. The media was then put into flask and sterillized for 15 minutes at 121°C.
Broth nutrient media
Composition: Lab-lemco powder
1
g
Yeast extract
2
g
Peptone
5
g
Sodium chloride
5
g
The preparation9 : 13 g of broth nutrient media was suspended in 1000 ml of distilled water and heated to
completely dissolved. The media was then put into flask and sterillized for 15 minutes at 121°C.
The Sterilization of Apparatus
The glasses used in this antibacterial activities test was sterillized in oven for 1 hour at the temperature of
170°C. The media was sterillized in autoclave for 1 5 minutes at121°C. Alat-alat yang digunakan dalam uji
aktivitas antibakteri ini, disterilkan terlebih dahulu sebelum dipakai. The ose syringe and pincet were sterillized
by means of Bunsen lamp10.
The Preparation of Chloroform Fraction of Soursop (Annona muricata L.) Leaves
10 ml of methanol solvent was added into 10 g of methanol extract, then homogenously stirred and removed
into separating funnel and 20 ml of distilled water and 40 ml of n-hexane were added, stirred and let it separated
and fractionated completely by n-hexane, the residue of methanol extract was then fractionated by 40 ml of
chloroform. The result of chloroform fractionation was then evaporated on a waterbath to obtain a dry extract of
chloroform fraction.
4. The Preparation of Bacteria Culture Stock11
The colony of bacteria was taken by sterillized ose syringe, then planted into sloping agar nutrient media by
scratching it. It was then incubated in a incubator for 18 – 24 hours at the temperature of 36-37°C.
The Preparation of Bacteria Inoculumn
The colony of bacteria was taken from culture stock by sterillized ose syringe, then suspended into test tube of
broth nutrient media of 10 ml. The turbidity of the solution was then measured at the wavelength of 580 nm
untill obtained the transmittance of 25% equivalent with106 CFU (Colony Forming Units)11.
The Preparation of Solution Tests of Methanol Extract and Chloroform Fraction with Various
Concentrations
3 g of methanol extract was dissolved into DMSO untill 10 ml of volume in order to obtain the concentration of
the extract of 300 mg/ml. The dilution was then made in order to obtain extracts with the concentrations of 250
mg/ml; 200 mg/ml; 150 mg/ml; 100 mg/ml; 50 mg/ml; 25 mg/ml;10 mg/ml; 5 mg/ml. The same procedure was
done to chloroform fraction.
Ginda Haro et al /Int.J.PharmTech Res.2014,6(2),pp 575-581. 578
The In Vitro Antibacterial Activities Tests
0.1 ml of inokulum was put into petridisc was then added 20 ml of sterillized liquid agar nutrient media heated
gently to 45°C, homogenized and let it stand in ord er to obtain a solid media. The paper disc with diameter of 6
mm was soaked into solution tests of various concentrations, dried and let them on the surface of agar nutrient
media. It was then incubated for 18-24 hours at the temperature of 36-37°C. The same procedure was don e to
chloroform fraction. The inhibition zone diameter around the paperdisc was then measured and recorded by
what so called jangka sorong. The tests were respectively done 3 times11. The tests of antibacterial activities of
methanol extract and chloroform fraction of soursop (Annona muricata L.) leaves against Staphylococcus
aureus and Escherichia coli are depicted in the next Figure 1, 2, 3 and 4.
Fig. 1. The test of
antibacterial activities
of methanol extract of
soursop (Annona
muricata L.) leaves
against
Staphylococcus
aureus
5.
6.
7. Fig 2. The test of
antibacterial activities
of methanol extract of
soursop (Annona
muricata L.) leaves
against Escherichia
coli
Fig. 3. The test of
antibacterial
activities of
chloroform
fraction of
soursop
(Annona
muricata L.)
leaves
against
Staphylococ
cus aureus
Fig. 3. The test of
antibacterial
activities of
chloroform
fraction of
soursop
(Annona
muricata L.)
leaves
against
Escherichia
coli
Results And Discussion
The antibacterial activities
test was conducted in various
concentrations to investigate
their relationship. The results
of antibacterial activities
exami
nation
showe
d that
metha
nol
extract
and
chloro
form
fractio
n gave
antiba
cterial
effect
againt
s
Esche
richia
coli
and
Staphy
lococc
us
aureus
. They
were
denote
d by
the
existen
ce of
inhibiti
on
zone
around
the paperdisc. The results of
the average of inhibition
zone diameter of methanol
extract and chloroform
fractions of soursop leaves
for Staphylococcus aureus
and Escherichia coli can
completely be seen on Tabel
1 and Tabel 2 below.
8. Ginda Haro et al /Int.J.PharmTech Res.2014,6(2),pp 575-581. 579
Tabel 1. The results of the average of inhibition zone diameter of the growth of Staphylococcus
aureus and Escherichia coli of methanol extract of soursop leaves
Methanol extract Inhibition Zone Diameter
concentration (mm)*
(mg/ml) Staphylococcus aureus Escherichia coli
300
16.
4
15.
3
250
15.
4
14.
5
200
14.
7
13.
8
150
14.
1
13.
1
100
13.
1
12.
3
50
12.
2
11.
5
25
11.
2
10.
7
10 9.8 8.7
5 8.6 8.0
Blank - -
Informations: (*) = The average results of triple measurements, (-) = no inhibition
On the Tabel 1 above it is obvious that the increase of extract concentrations will enhance their antibacterial
activities caused by the increase of bioactive contents of the extract.
The results of antibacterial activities examination showed that methanol extract at concentration of 150 mg/ml
could inhibit the growth of Staphylococcus aureus with inhibition zone diameter of 14.1 mm and at
concentration of 250 mg/ml could inhibit the growth of Escherichia coli with inhibition zone diameter of 14.5
mm. The Minimum Inhibition Concentration (MIC) of methanol extract at concentration of 5 mg/ml could
inhibit Staphylococcus aureus with inhibition zone diameter of 8.6 mm and of 8.0 mm for Escherichia coli.
The inhibition zone diameter of bacterial growth of methanol extract of soursop was larger than of ethanol
extract1 caused by the strength of methanol compared with ethanol as a solvent of bioactive compound12.
Tabel 2. The results of the average of inhibition zone diameter of the growth of Staphylococcus
aureus and Escherichia coli of chloroform fractions of soursop leaves
Chloroform
fractions Inhibition Zone Diameter
concentration (mm)*
(mg/ml) Staphylococcus aureus Escherichia coli
300 12.1
11.
5
250 11.5
10.
7
200 10.5 9.8
150 9.9 9.2
100 9.3 8.8
50 8.8 8.3
25 - -
10 - -
9. 5 - -
Blank - -
Informations: (*) = The average results of triple measurements, (-) = no inhibition
The chloroform fractions at concentration of 300 mg/ml showed the inhibition zone diameter of 12.1 mm for
Staphylococcus aureus and of 11.5 mm for Escherichia coli. The Minimum Inhibition Concentration (MIC) at
concentration of 50 mg/ml showed the inhibition zone diameter of 8.8 mm for Staphylococcus aureus and of
8.3 mm for Escherichia coli.
A compound with inhibition zone diameter about 14 through 16 mm is said to have a satisfied inhibition zone
diameter11. The inhibition zone diameter of methanol extract fulfilled the requirement. It could be understood,
Ginda Haro et al /Int.J.PharmTech Res.2014,6(2),pp 575-581. 580
maybe because of methanol extract containing tannin and flavonoids, whereas chloroform fraction containing
only flavonoids in small amount. The mechanism of tannin with antibacterial activities on low concentration
was by destroying the cytoplasm membrane causing the leak of cell, on high concentration the tannin would
coagulate with cellular protein13. The flavonoids could damage the permeability of cell wall of bacteria14.
The result of this study indicated that Staphylococcus aureus as positive gram bacteria is more sensitive to
chemicals than Escherichia coli as negative gram bacteria. This thing could be caused by the difference in
composition and structure of cell wall of those bacteria. The structure of cell wall of positive gram bacteria
consists of lipid only (1 – 4 %), whereas of negati ve gram bacteria consists of multi-layers of lipoprotein,
liposaccharide and peptidoglican with a high lipid content (11 – 12 %).
Conclusion
This study indicated that methanol extract of soursop leaves at concentration of 300 mg/ml could inhibit the
growth of Staphylococcus aureus with inhibition zone diameter of 16.4 mm and of 15.3 mm for Escherichia
coli. Chloroform fraction of soursop leaves at concentration of 300 mg/ml could only inhibit the growth of
Staphylococcus aureus with inhibition zone diameter of 12.1 mm and of 11.5 mm for Escherichia coli.
Methanol extract of soursop leaves at concentration of 150 mg/ml could inhibit the growth of Staphylococcus
aureus with inhibition zone diameter of 14.1 mm and of 13.1 for Escherichia coli. But the methanol extract of
soursop leaves at concentration of 250 mg/ml could inhibit the growth of Escherichia coli with inhibition zone
diameter of 14.5 mm. It fulfilled the requirement of Farmakope Indonesia, they are of 14 through 16 mm. The
minimum inhibitory concentration (MIC) of methanol extract at concentration of 5 mg/ml could inhibit the
growth of Staphylococcus aureus with inhibition zone diameter of 8.6 mm and of 8.0 mm for Escherichia coli
respectively. The MIC of chloroform fraction at concentration of 5 mg/ml did not inhibit growth of
Staphylococcus aureus and Escherichia coli at all. Whereas the MIC of chloroform fraction at concentration of
50 mg/ml could only inhibit Staphylococcus aureus with inhibition zone diameter of 8.8 mm and of 8.3 mm for
Escherichia coli.
The methanol extract indicated a more effective bacterial inhibition than chloroform fraction of the same
concentration.
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