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Objective:The aim of this study was to examine the influence of the partial hydrolysis of virgin coconut oil (VCO) on it's antibacterial activity. Methods:The VCO used in this study was the productof UD SinarNias. Hydrolysis was carried out by enzyme and sodium hydroxide. Enzymatic hydrolysis using lipozyme was conducted in four different incubation time namely, 3 hours, 6 hours, 9 hours and 12 hours. Alkaline hydrolysis preformed with 25%, 50% and 75% NaOH calculated from the saponification valueof coconut oil. Acidified hydrolyzed VCO was extracted with n-hexane. Recovered hydrolyzed products were mixed with water (5 g in water to make 10 ml) to form water in oil emulsion (w/o). Antibacterial activity test was conducted against bacteria Pseudomonasaeruginosa (ATCC 25619), Staphylococcusaureus (ATCC 29737), Staphylococcus epidermidis (ATCC 12228) and Propionibacterium acnes (ATCC 6918) by diffusion agar method using the paper disc of 6 mm in diameter. Antibacterial activity of hydrolyzed VCO was compared with tetracycline and ampicillin. Results: Un-hydrolyzed VCO did not show antibacterial activity but hydrolyzed oil did. The longer the incubation time and the higher the amount of NaOH used in the hydrolysis increased antibacterial activity. VCO hydrolyzed by enzyme was more effective than those hydrolyzed by sodium hydroxide. Hydrolyzed VCO were more effective against Pseudomonas aeruginosa than other bacteria. Conclusions: Un-hydrolyzed VCO did not inhibit bacterial growth, while VCO after hydrolysis was found to have antibacterial activity. Hydrolyzed VCO by enzyme is more active asantibacterial than VCOhydrolyzed by NaOH. Tetracyclin and ampicillin were more active than those of hydrolyzed VCO.
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Research Article
Faculty of PharmacyUniversity of Sumatra Utara, MedanIndonesia 20155
Received: 24January 2014, Revised and Accepted: 20February 2014
Objective:The aim of this study was to examine the influence of the partial hydrolysis of virgin coconut oil (VCO) on it’s antibacterial activity.
Methods:The VCO used in this study was the productof UD SinarNias. Hydrolysis was carried out by enzyme and sodium hydroxide. Enzymatic
hydrolysis using lipozyme was conducted in four different incubation time namely, 3 hours, 6 hours, 9 hours and 12 hours. Alkaline hydrolysis
preformed with 25%, 50% and 75% NaOH calculated from the saponification valueof coconut oil. Acidified hydrolyzed VCO was extracted with n-
hexane. Recovered hydrolyzed products were mixed with water (5 g in water to make 10 ml) to form water in oil emulsion ( w/o). Antibacterial
activity test was conducted against bacteria Pseudomonasaeruginosa (ATCC 25619), Staphylococcusaureus (ATCC 29737), Staphylococcus
epidermidis (ATCC 12228) and Propionibacterium acnes (ATCC 6918) by diffusion agar method using the paper disc of 6 mm in diameter.
Antibacterial activity of hydrolyzed VCO was compared with tetracycline and ampicillin.
Results: Un-hydrolyzed VCO did not show antibacterial activity but hydrolyzed oil did. The longer the incubation time and the higher the amount of
NaOH used in the hydrolysis increased antibacterial activity. VCO hydrolyzed by enzyme was more effective than those hydrolyz ed by sodium
hydroxide. Hydrolyzed VCO were more effective against Pseudomonas aeruginosa than other bacteria.
Conclusions: Un-hydrolyzed VCO did not inhibit bacterial growth, while VCO after hydrolysis was found to have antibacterial activity. Hydrolyzed
VCO by enzyme is more active asantibacterial than VCOhydrolyzed by NaOH. Tetracyclin and ampicillin were more active than those of hydrolyzed
Keywords:VCO, MCT, MCFA, lauric acid, monolaurin,partial hydrolysis, antibacterial
Two kinds of oils can be obtained from coconut tree (Cocosnucifera)
they are coconut oil (copra oil) and virgin coconut oil (VCO). Coconut
oil is extracted from copra by heating process, while VCO is from
coconut milk prepared from fresh and mature coconut meat of
coconut fruit and processed at low temperature. Coconut oil and
VCO are different from most of the other common oils which are
usually composed of long chain fatty acids, while coconut oil is
composed of short and medium chain fatty acids, and therefore
classified as medium chain triglyceride (MCT). Coconut oil has been
used in health promotion and also in ailments prevention and
medication [1,2]. The quality of VCO is determined by medium chain
fatty acid (MCFA) content, especially lauric acid which is influenced
by variety and oil extraction process [3].
Antibiotic resistance is a consequence of the evolutionary adaptation
of bacteria due to the indiscriminate use of antibiotics. In addition, a
high cost and adverse effects are generally associated with synthetic
antibiotics. The emergence of antibiotic resistance in
microorganisms becomes a threat among medical community. There
is a continuous need to discover new antimicrobial compounds with
novel mechanisms of action for new infection diseases. Therefore,
researchers are turning their attention to antimicrobial of plant
origin. Antibacterial activity of ethanolic extract of citrus leaves on
Escherichia coli and Pseudomonas aeruginosa was studied and
found to be active [4]. Essential oils of some selected plants was
evaluated for antibacterial activity on methicillin resistant
staphylococcus aureus, and found that essential oils of Clove and
Cinnamon to be more active against tested bacteria [5]. Evaluation of
antimicrobial activity of Pithecellobiumdulce pod pulp was
conducted and found to be potential bactericidal and fungicidal [6].
Potential of medication of coconut oil and coconut products was
discovered by Jon Kabara in the year of 1970s, who found that
coconut oil has antibacterial, antiviral and antifungal activities
exerted by free MCFAs and mainly by their monoglycerides
molecule, especially monolaurin[7,8].
VCO contains high lauric acid content (46-50%) attached to glycerol
backbone to form a triglyceride. In the human gastrointestinal
tract,triglycerides in VCO can be converted into free fatty acids
mainly lauric acid and monolurin which are active as antibacterial,
antiprotozoal, and antiviral components. Moreover, MCFAsare easily
absorbed into cells and then to mitochondria, increase metabolism,
and hence the cells work more efficiently to form new cells and
hence substitute damaged cells faster [3,8,9].
Antimicrobial activity is due to free fatty acids of medium chain and
their monoglycerides. Triglyceride and diglyceride are not effective
as antibacterial. Of the free fatty acids present in coconut oil, lauric
acid (C:12:0) is proven to be more active as antibacterial agent
compared to caprilic acid (C8:0), capric acid (C10:0), and myristic
acid (C14:0). Free fatty acids and their monoglycerides inactivate
bacteria by disrupting plasma membrane of lipid bilayer[7,10].
Antibacterial activity of free fatty acid or its monoglyceride has been
tested separately [7]. Combination or mixture of free fatty acidsand
their monoglycerides generated from coconut oil could be
synergistic in bacterial inactivation. To generate monoglyceride
from VCO can be done by enzymatic hydrolysis which is effective
specifically on sn-1 and sn-3 position. This specific enzyme can be
obtained from pancreas, Thermomyces lanuginose and
Mucormiehei[11]. Hydrolysis can also be done by saponification
reaction with alcoholic sodium or potassium hydroxide solution.
Saponification byNaOH with or above saponification value will
hydrolyze all triglycerides completely in to glycerol and free fatty
acids as soap [12,13]. However, hydrolysis using NaOH lower than
NaOHneeded for total hydrolysis (saponification value) would
partially hydrolyze oil into mixture of free fatty acids and their
diglyceride or monoglyceride derivatives depending on the amount
NaOH used. The aim of this study was to compare the influence of
Vol 7, Suppl 2, 2014 ISSN - 0974-2441
Jansenet al.
Asian J Pharm Clin Res, Vol 7, Suppl 2, 2014, 90-94
partial hydrolysis of VCO by enzymeand NaOH on their antimicrobial
Apparatus used including vortex (Bender,Germany), analytical
balance (Sartorius, Japan), hotplate (Heidelberg, Germany),
autoclave, oven, spectrophotometer (Shimadzu, Japan), incubator,
reflux condenser, water bath, burette, and glass wares. All chemicals
were pro analysis grade product of E. Merck (Germany) including
potassium and sodium hydroxide, n-hexane, methanol, ethanol, tris-
hydroxymethylaminomethane, hydrochloric acid, calcium chloride,
anhydrous sodium sulfate, phenolphthalein (1% in ethanol) and
Lipozyme TL IM.
VCO used in this study was product of UD SinarNias. Culture media
used were Nutrient Agar(NA), Nutrien Broth (NB), and Mueller
Hinton Agar (MHA). Bacteria tested were Pseudomonas aeruginosa
(ATCC 25619), Staphylococcus aureus (ATCC 29737), Staphylococcus
epidermidis (ATCC 12228) and Propionibacterium acnes (ATCC
6918). Paper disc used was of Machereynagel with 6 mm in
diameter. Antibacterial activity of hydrolyzed VCO was compared
with those of tetracycline and ampicillin.
Reagents used were calcium chloride solution of 0.063 M, Tris-HCl
buffer solution with the pH of 8, HCl solution of 0.5 N, KOH of 0.5 N,
NaOHof 0.5 N, 1% phenolphthalein indicator solution. These
solutions prepared according to procedure described in Indonesian
Pharmacopeia [14]. Medium used were Nutrient Agar(NA), Nutrien
Broth (NB), and Mueller Hinton Agar (MHA). Preparation of these
media used as described in Difco Laboratory Manual [15].
Enzymatic Hydrolysis
Fifty (50) g of oil placed in an erlenmeyer of 250 ml to which 50 ml
water, 12.5 ml CaCl2of 0.063 M, 25 ml buffer solution Tris-HCl and
500 mg lipozyme were added. This mixture was stirred with
magnetic stirrer for 10 minutes to homogenize. Then it was allowed
to stand (incubated) for various length of time; 3, 6, 9, and 12 hrs at
temperature of 40 ± 0.5oC, and shaken the mixture for 10 minutes in
every one hour during incubation. After the hydrolysis was
completed, the mixture was transferred into separating funnel,
acidified with dilute HCl, extracted with 50 ml n-hexane resulted in
two separated layers [13,16]. The upper layer (n-hexane fraction)
was separated and called as filtrate I. The bottom layer was
extracted again with 50 ml n-hexane and separated as filtrate II.
These two filtrates were combined to which then 50 mg anhydrous
Na2SO4 added and allowed to stand for 15 minutes. It was then
evaporated on a water bath to dryness. The recovered hydrolyzed oil
was used in the antibacterial experiment after acid value was
Hydrolysis with Sodium hydroxide
Ten (10) g of oil placed in a 250 ml conical flask and 100 ml
methanolicNaOHof 0.5 N was added in to it. The flask was attached
with reflux condenser and heated. As the ethanol boiled, the flask
occasionally shaken till the fat completely saponified (~3 hours).
Solution allowed to cool and added 1 ml solution of 1%
phenolphthalein indicator, titrated with HClof 0.5 N till the red color
disappeared. Saponification value was calculated as the amount in
mg of NaOH needed for the saponification [12,13].
Partial hydrolysis of oil was performed with the same procedure as
described in saponification procedure but the amount of NaOHused
was lowerthan the amount of NaOHused in the total saponification
value. Fifty (50) g oil was weighed then added methanolicNaOH with
the amount of 25%, 50% and 75% from saponification value, and
hydrolysis procedure conducted as already described for 3 hrs. After
hydrolysis, then the mixture acidified with dilute HCl in order to
convert soap (sodium salt of fatty acids) into free fatty acids.
Acidified mixture was then shaken and extracted with 50 ml n-
hexane resulted in two separated layers. The upper layer (n-hexane
fraction) separated called as fraction I. The bottom layer shaken
with 50 ml n-hexane and allowed to stand for a while then hexane
fraction was taken as fraction II. The two fractions were combined
and dried by adding 50 mg anhydrous Na2SO4, allowed to stand for
15 minutes. Dehydrated hexane fraction was then heated on a water
bath to evaporate hexane, and dried hydrolyzed oil was then used
for acid value determination prior to antimicrobial test.
Acid Value Determination
Acid value determination was carried out for un-hydrolyzed VCO
and hydrolyzed oil. Five gram oil was transferred in to 200 ml
erlenmeyer, added 25 ml neutralized ethanol of 95%, then heated
for ten minutes on a water bath and occasionally shaken. This
solution then titrated with KOH of 0.1 N using phenolphthalein
indicator solution. Acid value of the oil was calculated [12].
A= the amount of ml KOH for titration
N = normality of KOH solution
W= weight of oil (g)
Antibacterial Activity Test
Bacterial inoculum was prepared by suspending bacterial colony in
Nutrient Broth Media solution and turbidity was measured at 580
nm to have transmittance of 25% (bacterial concentration is
106cfu/ml). Antibacterial activity test of VCO and hydrolyzed
VCOwas conducted and the results compared with tetracyclineand
ampicillin.The volume of 0.1 ml bacterial inoculum was mixed with
15 ml MHA in a petri dish, allowed to stand until the media
solidified. Tested material was prepared as an emulsion by mixing
VCO and hydrolyzed VCO in water at the same amount of sterile
distilled water (5 g oil mixed with water to 10 ml, concentration was
500 mg/ml). Paper disc was then dipped in the emulsion for 15
minutes and then incubated in prepared media at 36 - 37oC for 24
hours. Antibacterial activity was determined by measuring diameter
of transparent area around the paper disc (zone of inhibition).The
concentration of the tetracyclin and ampicillin tested were prepared
in 5 mg/ml, 1 mg/ml and 0.1 mg/ml. The test was conducted in
three replicates [14,17].
Acid value of VCO and hydrolyzed VCO
Partial hydrolysis of VCO resulted in the generation of free fatty
acids in hydrolyzed VCO, which was measured by acid value. Acid
value of VCO and VCO partially hydrolyzed by NaOH and enzyme is
presented in Table 1.
Tabel 1: Acid value of hydrolyzed virgin coconut oil
time and
degree of
Acid values (n = 3)
(mg KOH/g oil)
0.74 ± 0.153
3 hour
72.02 ± 0.517
6 hour
79.05 ± 3.405
9 hour
108.08 ± 0.845
12 hour
150.88 ± 0.818
The percentage
25 %
68.15 ± 0.483
50 %
133.87 ± 0.796
75 %
199.77 ± 2.575
From Table 1 can be seen that the acid value of VCO increased after
hydrolysis by enzyme and NaOH. The longer the incubation period in
enzymatic hydrolysis and the higher the amount of NaOH used in the
hydrolysis the higher the acid value.Acid value exerted by NaOH75%
Jansenet al.
Asian J Pharm Clin Res, Vol 7, Suppl 2, 2014, 90-94
was higher than that by enzymatic hydrolysis with incubation for 12
h. Enzymatic hydrolysis of a triglyceride molecule resulted in 2 fatty
acid molecules and 1 molecule of 2-monoglyceride, while partial
hydrolysis by alkaline was difficult to predict [7,13,19].
Zonesof Inhibition by VCO and Partially Hydrolyzed VCO on
Tested Bacteria
Typical zonesof inhibition to evaluate the antibacterial activities by
measuring diameter of paper disc in agar media of different
hydrolyzed products are presented in Fig. 1.
Fig. 1:Antibacterial activities shown by zones of inhibition by
VCO, hydrolyzed VCO compared with tetracyclin and
ampicillin against Pseudomonas aeruginosa
Note: (A) Zone of inhibition by VCO and hydrolyzed VCO by NaOH;
(B) zone of inhibition by VCO and hydrolyzed VCO by enzyme; (C)
zone inhibition by ampicillin; (D) zone of inhibition by tetracycline.
Zones of Inhibition of VCO hydrolyzed by enzyme and alkaline
(NaOH) are presented in Table 2 and Table 3.Bacterial inactivation
by enzymatic hydrolysis for 12 hours and that by alkaline hydrolysis
(75%) were compared with those of tetracycline and ampicillin
(Table 4).
Table 2: Antibacterial activityof VCO hydrolyzed by enzyme
Antibacterial activity of Hydrolyzed VCO by
enzyme at different incubation time shown by
zones of inhibition (mm)
9 hours
12 hours
S. aureus
Notea)Zonesof inhibition is significantly difference (P<0.05)
compared with hydrolyzed by enzyme for 3 hours of incubation.
Table 3: Antibacterial activityof VCO hydrolyzed by NaOH
Antibacterial activity of hydrolyzed VCO
byNaOHat different percentage relative to total
saponification value shown by zonesof inhibition
S. aureus
Note:b)Zone of inhibition is significantly difference (P<0.05)
compared with hydrolyzed by NaOH at 25% of saponification
value.Negative antibacterial activity indicated by 6 mm in diameter.
Table 4: Bacterial inhibition of hydrolyzed VCO and antibiotic
tetracycline and ampicillin against tested bacteria
Zones of Inhibition (mm)
Hydrolyzed VCO
(500 mg/ml)
on (75%)
S. aureus
Note:(-)zone of inhibition is zero (if diameter is 6 mm)
Hydrolysis of VCO, either by enzyme or NaOH induced antibacterial
activity, but enzymatic hydrolysis was more inductive than by
alkaline hydrolysis. Enzymatic hydrolysis resulted in the formation
of a mixture containing free fatty acids, monolaurin, and
triglycerides depending on the incubation time. The composition
ofoil after alkaline hydrolysis (partial hydrolysis) would be
composed of free fatty acids, monoglycerides, diglyceridesand/or
un-hydrolyzed triglycerides depending on the amount of NaOH used.
The most potential antibacterial activity of MCFA exerted by free
fatty acid and monoglycerides which may inactivate bacteriaby
disrupting microbial plasma membraneof lipid bilayer. Of the many
saturated fatty acids, lauric acid (C:12) shown to be the most active
as antibacterial compared to caprilic(C8:0), carpric(C10:0), and
myristicacid (C14:0) [7,10,18].
In this study, VCO did not show to have antibacterial activity on
tested bacteria, because it contained small amount of free fatty acid
and there was no monolaurin present.On the other hand, a study
showed that VCO without hydrolysis was effective on Pseudomonas
aeruginosaandStaphylococcus aureus, using glycerin as solvent
[20].Bacterial growth inhibition by hydrolyzed VCO was found to be
more active against gram negative Pseudomonas aeruginosathan
gram positive Staphylococcus aureus. The inhibition
ofStaphylococcus epidermidis, was found to be higher by VCO
hydrolyzed by alkaline than that by enzyme, but antibacterial
activity was very low against Propionibacterium acnes.
The evaluation of inhibition can be classified into three categories
based on the diameter of zones of inhibition; very active (above 11
mm), medium activity (active) (between 6-11 mm), while non-active
(6 mm).According to this criterion, un-hydrolyzed VCO was not
active as antimicrobial, where as hydrolyzed VCO by enzyme for 12
hours and by alkaline of 75% were very active since the diameter of
zones of inhibition were above 11 mm (13.43 mm) and 11.35 mm
respectively [21]. The antibacterial activity of synthetic
Jansenet al.
Asian J Pharm Clin Res, Vol 7, Suppl 2, 2014, 90-94
monolaurinagainst Staphylococcus aureuswas previously conducted
[18] and reported that zone of inhibition was 13 mm (500 mg/ml)
was better than hydrolyzed VCO in this study with inhibition zone
ranged from 10-11 mm (500 mg/ml) on the same species of
bacteria.This difference could be due to lower content of monolaurin
in the partially hydrolyzed VCO in the present study.
Bacterial inhibition was more effective on gram negative than gram
positive bacteria. It is probably due to the components of hydrolyzed
VCO are non-polar molecules, and therefore they easily interact with
cell membrane and disrupting lipid layer present in the outer part of
cell membrane of gram negative bacteria, while the cell membrane
of gram positive bacteria composed of more peptidoglucan layer
compared with that in gram negative bacteria.The peptidoglucan
layer in gram positive bacteria is rigid and resistant to osmotic lysis
Pseudomonas aerugiosa is an opportunistic bacteriacausing infection
when the immunity system of the host is getting
weaker.Pseudomonas aeruginosacould survive from host immunity
system because this bacteria has lipidpolysacharide as a
protectingcomponent [25,26]. It is postulated that the mechanism of
how lauric acid and monolaurin may inactivate bacteria is that by
dissolving lipid component present in bacterial cell membrane [27].
Lipidpolysacharide present in Pseudomonas aeruginosamembrane
through which lauric acid and monolaurin may interact and disrupt
bacterial cell membrane.
Propionibacterium acnes is a gram positive bacteria, itcan not be
inhibited by hydrolyzed VCO. This bacteria may cause skin acnes, a
local inflammation on hair follicle resulted from two stages. In the
first stage is that the excessive sebaceous secretion accumulates in
the hair follicle that is previously blocked by ceratine cells (komedo).
On the second stage is the formation of acne, the excessive sebum
converted into fatty acid by lipase enzymereleased by skin normal
flora Propionibacterium acnes, resulting in inflammation on the
follicle. Acne medication can be done by reducing sebum
productionwith retinoic acidor by lifting off komedoand decreasing
fatty acid content or lipid on the skinwith benzoyl peroxide [24,27].
It is still not clear by which mechanism the fatty acids acting as
antimicrobial agent. But the main target is cell membrane of bacteria
and other mechanisms may involve on the membrane. Retarding
growth effect is related to amphiphilic property of fatty acids
enabling them to interact with cell membrane generating
temporarily or permanent pores of various sizes. With the high
concentration, detergent such as free fatty acids being able to
dissolve cell membrane and hence releasing or disrupting larger
portion.Free fatty acids also influence energy production in cell
membrane by disturbing electron transport chain and oxidative
phosphorilation [28].Probable other processes are cell lysis,
impairing enzyme activities, inactivating macromolecular synthesis,
disturbing nutrient absorption or protein DNA denaturation.
Monolaurinmay act as antimicrobial agent by this mechanism
From Table 4 can be seen that antibacterial activity of enzymatic
hydrolysis is greater than that of alkaline hydrolysis agai nst P.
aeruginosa and S. aureus, but similar toward S. epidermis and P.
acnes. Hydrolyzed VCO indicates much lower antibacterial activity
compared with tetracyclin and ampicillin at very low concentration.
Tetracyclin and ampicillin show different activity against tested
bacteria. Tetracycline is most active toward S. epidermis and the
lowest on S. aureus. On the other hand ampicillin is active against P.
aeruginosa and it is most active against P. acnes. It is reported that
monolaurin and lauric acid derived from coconut oil inactivate
pathogenic bacteria but not the beneficial microorganismor
probiotic. In addition, lauric acid and monolaurin do not develop
microbial resistance while the antibiotic would do [30,31].
Un-hydrolyzedVCO is not active as antimicrobial, but partial
hydrolysis will increase antibacterial activity. The longer incubation
time in enzymatic hydrolysis and the higher the percentage of NaOH
relative to total saponification during alkaline hydrolysis resulted in
the more effective in antimicrobial activity of hydrolyzed
VCO.Hydrolyzed VCO is more effective against Pseudomonas
aeruginosa (gram negative) compared to other tested bacteria.
Hydrolyzed VCO is not as effective as tetracycline and ampicillin.
Ampicillin is not effective againstPseudomonas aureginosa. The
benefit of VCO used orally as antibacterial is that VCO does not cause
any side effectsince it is a common food component which will be
hydrolyzed by lipase in the gastrointestinal tract. Antibacterial
activity of hydrolyzed VCO is necessary evaluated by in vivo
experiment in order to determine the effective dosage of VCO.
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... [11,12] Silalahi's research et al. proved that hydrolyzed VCO (HVCO) was active against bacteria. [13] HVCO can inhibit Clostridium difficile of 99.9% whereas 0.15% HVCO gave 50% inhibition. [14] Other researchers tried using papaya enzyme. ...
... [19] Hydrolyzed virgin coconut oil Hydrolysis of VCO was conducted using Rhizomucor miehei lipase enzyme as previous. [13] Physical characteristics and IR evaluation Characteristics of VCO and HVCO test were done including organoleptic, density, and content of lauric acid which compared to the reference. [7] Infrared spectra was compared to the reference. ...
... This study compared VCO and HVCO which according to the literature can increase antibacterial activity. [13] Density and fatty acid content A c c o r d i n g t o t h e A P C C , r e s u l t i n g V C O o f 0.920 ± 0.002 g/cm 3 met the standard. Results of fatty acid content are shown in Table 2. Lauric acid content was lower than the APCC, which is 43.0%-53.0%. ...
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Virgin Coconut Oil (VCO) with potential fatty acid content has a strong antibacterial effect for drug and cosmetics. VCO was hydrolyzed as hydrolyzed VCO (HVCO) to increase its activity. This study aims to optimize VCO and HVCO microspheres. Examination was included fatty acid content, density, and organoleptic. VCO and HVCO microspheres were characterized by yield, moisture content (MC), morphology, and size. Fatty acid content was done through gas chromatography mass spectrometry whereas for microspheres, effect of alginate concentration (1%-2%), and addition of poly ethylene glycol (PEG) were studied. VCO and HVCO microspheres formulas showed that an increase in polymer concentration, increasing yield, and improving MC. The most optimal 2% alginate with the addition of PEG increased yield up to 59% and reduced size. F4 microspheres resulted the highest yield, low MC, and high fatty acids. Morphology was spherical and smooth with small size. Optimization of HVCO microspheres showed its potential which recommend for antibacterial and antifungal activity.
... The distribution of lauric acid on the position of sn-2 also influences the effectiveness of the antibacterial activity of monolaurin [12]. As an emulsifier, 2-monoglycerides (monoglycerides with fatty acid chains at the positions of sn-2) have only 1 form of β polymorphic [13]. ...
... Furthermore, palm olein contains about 87 % unsaturated fatty acids (oleic acid and linoleic acid) and 7 -11 % palmitic acid at the position of sn-2 [37]. The distribution of lauric acid on the position of sn-2 influences the effectiveness of the antibacterial activity and capability emulsion of monolaurin [12,13]. ...
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This study aims to analyze the physicochemical properties of fat blends between palm kernel olein and stearin in different ratios. Furthermore, it determines the fat blends, which have more than 50 % lauric acid at the sn-2 position. The liquefied RBDPKOo (refined bleached deodorized palm kernel olein) and RBDPKOs (refined bleached deodorized palm kernel stearin) were mixed in proportions of 100:0; 80:20; 60:40; 40:60; 20:80; and 0:100 (w/w), and homogenized at 70 °C for 15 min. Also, the samples analyzed were fatty acid composition, triglyceride profile and physicochemical properties. The results showed the blending of RBDPKOo with RBDPKOs resulted in a change in melting point, iodine value, and behavior. Increased RBDPKOs resulted to a higher melting point, and a lower iodine value of fat blends. A blending with higher RBDPKOs (more than 60 %) resulted in a sharp endothermic peak. Meanwhile, blending of RBDPKOo with RBDPKOs significantly changes the fatty acid composition and triglycerides profiles. The RBDPKOs in the fat blends increased the trilaurin, and the composition as well as the amount of lauric acid at the position of sn-2. In addition, the 2 fat blends obtained with more than 50 % lauric acid at the position of sn-2 were RBDPKOo: RBDPKOs ratios of 40:60 and 20:80, respectively. HIGHLIGHTS Palm kernel olein and stearin blend Improving the Physicochemical Properties of Palm Kernel Oil Raw material of monolaurin synthesis
... The VCO was diluted in dimethyl sulphoxide as the negative control in the dilution factors of 2X, 4X, and 8X, so the samples are denoted VCO-2X, VCO-4X, and VCO-8X, respectively, whereas VCO sample is the pure sample. Referring to the inhibition zones in S. aureus, P. acne, and P. aeruginosa bacteria test, the antibacterial activity of VCO suggests a higher activity compared to hydrolyzed VCO [24]. The inhibition zones are higher, even though the dilution factors are also higher in this research. ...
... was diluted in dimethyl sulphoxide as the negative control in the dilution factors of 2X, 4X, and 8X, so the samples are denoted VCO-2X, VCO-4X, and VCO-8X, respectively, whereas VCO sample is the pure sample. Referring to the inhibition zones in S. aureus, P.acne, and P. aeruginosa bacteria test, the antibacterial activity of VCO suggests a higher activity compared to hydrolyzed VCO [24]. The inhibition zones are higher, even though the dilution factors are also higher in this research. ...
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This study concerns papain enzyme assisted synthesis of virgin coconut oil (VCO) as a candidate in-house reference material. The study was conducted to obtain optimization of the VCO preparation with green processes as the standardized product which high stability and homogeneity. The method is expected to produce the candidate of in-house reference material to ensure the standards and quality of the VCO product. Based on the results of this study, the preparation of VCO was carried out using the papain. An optimum yield of 24.30%. was achieved under the following conditions: enzyme to coconut milk with a mass ratio of 0.6 g/L, under 500 mL water/g of coconut powder, at the temperature of 70 °C by five stages of extraction. The physicochemical properties as well as organoleptic feature of VCO which consist water content, peroxide number, free fatty acids, and iodine numbers are fit with the standard. The parameters exhibited the homogeneity and stability which be able recommended as candidate in-house reference material and have potentially as antibacterial agent. Antibacterial activity test showed that VCO has potential against Escherichia coli, Staphylococcus aureus, Propionibacterium acnes and Pseudomonas aeruginosa as shown by the inhibition zone in the testing.
... Virgin Coconut Oil (VCO) adalah suplemen nutrisi sebagai minyak kesehatan yang sangat popular, dibuat dari daging buah kelapa tanpa pemanasan (Agarwal and Bosco 2017 ;Sant'Anna et al. 2003). Kualitas VCO ditentukan oleh kandungan asam lemak rantai medium (MCFA) terutama asam laurat dan bentuk monogliseridanya, monolaurin yang membuat VCO sangat aktif sebagai antibakteri, anti protozoa, dan anti virus ( Maria et al, 2005 ;Silalahi, et al, 2014). Berbagai cara dilakukan untuk mendapatkan VCO dari buah kelapa seperti: ekstraksi pelarut, pengepresan, dan ekstraksi enzimatis (Subroto et al. 2020;Agarwal and Bosco. ...
ABSTRAK Telah dilakukan pembuatan virgin coconut oil (VCO) dengan ekstrak jamur Aspergillus niger serta uji antibakteri VCO dengan bakteri Staphylococcus aureus. Kualitas VCO ditentukan dengan uji kadar air, angka asam, angka penyabunan, angka iod, uji organoleptik, dan analisis GC-MS. Ekstraksi VCO tanpa menggunakan ekstrak jamur hanya mendapatkan VCO sebanyak 5,859 g. Penambahan ekstrak jamur A. niger 0,5% b/v menghasilkan VCO sebanyak 8,832 g, menunjukkan terjadi kenaikan yang sangat signifikan (p<0,05). Hasil uji kadar air, angka asam, angka penyabunan, dan angka iod masing-masing diperoleh: 0,1958; 0,2929; 5,0487; dan 0,2781, hasil ini sesuai dengan baku mutu VCO yang ditetapkan. Hasil uji organoleptik memberikan VCO yang tidak berwarna dan tidak berbau, dan hasil analisis GC-MS diperoleh kandungan asam lemak rantai medium dengan kandungan asam laurat sebagai komponen terbanyak. Hasil uji antibakteri terhadap bakteri Staphylococcus aureus menunjukkan kemampuan VCO dalam menghambat pertumbuhan bakteri dengan zona hambat 13,5 mm. Kata kunci: antibakteri, Aspergillus niger, Staphylococcus aureus, virgin coconut oil. ABSTRACT Preparation of virgin coconut oil (VCO) with Aspergillus niger fungi extract and the antibacterial test of the VCO with Staphylococcus aureus has been carried out. The quality test of the VCO included water content, acid number, saponification number, iodine value, organoleptic test, and GC-MS analysis. VCO extraction without using fungi extract only got 5.859 g of VCO. The addition of 0.5% w/v of A. niger fungi extracts produced 8.832 g of VCO, indicating a very significant increase (p <0.05). The water content, acid number, saponification number, and iodine value obtained were 0.158; 0.2929; 5.0487; and 0.2781 respectively, which met the VCO quality standard. The organoleptic test proved that the VCO was colourless and odourless. Meanwhile, the GC-MS analysis showed the content of medium-chain fatty acids with lauric acid as the largest component. The antibacterial test against Staphylococcus aureus indicated the ability of VCO to inhibit the growth of bacteria with an inhibition zone of 13.5 mm. Keywords: antibacterial, Aspergillus niger, Staphylococcus aureus, virgin coconut oil.
... VCO contains higher antioxidant components such as tocopherol content and total phenolic compounds compared with coconut oil due to different extraction methods, but not fatty acid profile. [26,27,28]. ...
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A ketogenic diet consists of high fat, with moderate to low protein content, and very low carbohydrates, which forces the body to use fat instead of glucose to produce energy which is called ketosis. Ketogenic diets are commonly used in patients suffering from neurological disorders, and mostly epilepsy. Fat is a mixture of different triacylglycerol molecules formed by esterification of glycerol with three fatty acids. Based on the length of fatty acids in triacylglycerols, fats and oils can be classified into two groups; medium chain triglycerides composed of short and medium chain fatty acids containing 4 to 12 carbon atom, and long chain triglycerides composed of long chain fatty acids containing 14 to 22 carbon. Metabolism of medium chain triglycerides is different from that of long chain triglycerides. Classic or old ketogenic diet use long chain triglycerides as fat component may cause side effects such as dislipidemia and hence increase cardiovascular disease risk.This drawback may be reduced by replacing long chain triglycerides with medium chain triglycerides. Coconut oil belongs to medium chain triglyceride fats because it’s fatty acids consist mostly of medium chain fatty acids. There are two kinds of coconut oil obtained from coconut meat namely coconut oil used for frying, and virgin coconut oil is directly consumable and more suitable in Ketogenic diets
... Saponification was performed on commercially available KLF Nirmal cold-pressed VCO (produced in India). 4 All of the fatty acids found in VCO, including lauric acid, caprylic acid, capric acid, myristic acid, palmitic acid, stearic acid, oleic acid, and linoleic acid, were produced as potassium salts. ...
... Alkaline hydrolysis was performed using NaOH concentrations of 25%, 50%, and 75% based on the saponification value of coconut oil. [11] The formulation, characterization, and penetration of VCO-solid lipid particles have been investigated by Noor. [12] Ibrahim [13] examined the effect of fermented VCO on wound healing in Sprague-Dawley rats. ...
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Background: Coconut oil is of two varieties: virgin and refined oil. Virgin coconut oil (VCO) is made by cold-pressing the liquid from the fresh part of coconut meat. It has a milky appearance. This oil extraction method prevents the loss of vitamin E, pro-vitamin A, and polyphenols. It has various properties such as analgesic, anti-inflammatory, and anti-cancer. Skin is the general structure of the body. It is the first line of protection against traumatic injuries and microorganisms. Aim: This review is focussed on the existing data on the effect of VCO on the skin. Materials and Methods: PubMed and Google Scholar were searched for citations for keywords "virgin coconut oil and dermatology" and "virgin coconut oil and skin." In search of the various databases, 13 articles were found on VCO related to skin. Result: Virgin coconut oil is used as antioxidant, anti-inflammatory, as skin protector, in Alzheimer's disease, in wound healing and as moisturizer. Conclusion: From this review, it can be concluded that VCO is beneficial for various dermatological disorders. It is antifungal and antibacterial and also acts as an immunomodulator. It also has anti-inflammatory, angiogenic, wound-healing, and skin protective properties.
... Many types of lipase have been currently used to obtain the FFAs from coconut oil such as lipozyme TL IM, lipozyme TL 100 L, CRL, and PPL (Silalahi et al. 2014;Sun et al. 2012;Lan and Hoa 2015). Most studies have been carried out to evaluate the antibacterial efficacy of extracted FFAs, which consist of a mixture of different chain-length fatty acids (C8-C18) (Nguyen et al. 2017(Nguyen et al. , 2018Silalahi et al. 2014). The efficacy of each specific fatty acid in VCO against food pathogens has been yet indicated. ...
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Free fatty acids (FFA) have been previously shown to be an effective bactericidal agent. In this study, the FFA mixture was attempted to isolate from virgin coconut oil (VCO) using lipases and was fractionated to obtain the enriched medium-chain fatty acids (MCFA) to compare its antibacterial efficacy against food pathogens. Three types of lipases-namely, lipozyme TL 100L, Candida rugosa lipase (CRL), and porcine pancreas lipase (PPL)-were used to compare their efficacy in VCO hydrolysis, in which CRL was found to be the most efficient lipase for the process of VCO hydrolysis at the water: buffer ratio of 1:5, lipase concentration of 5310 U/g, pH 7, and temperature of 40 °C. The vacuum-distillation process was successfully found to fractionate the initial FFA into three fractions: Fraction I containing MCFA (C8-C12) plus Fraction II (C12-C14) and Fraction III containing LCFA (C14-C20). Among these fractions, Fraction I was observed to be the most effective anti-agent against four out of five food pathogens (negative result against Salmonella typhimurium), followed by Fraction II and initial FFA. Antimicrobial activities of study fractions were found effective on both Gram-positive strains and Gram-negative strains.
... The antimicrobial effect of VCO has been shown to inhibit Pseudomonas aeruginosa (ATCC 25619), Staphylococcu saureus (ATCC 29737), Staphylococcus epidermidis (ATCC 12228) and Propionibacterium acnes (ATCC 6918) (Silalahi et al., 2014). Apart from being an antibacterial, VCO is also effective against several fungal species, such as Candida albicans, Candida glabrata, Candida tropicalis, Candida parapsilosis, Candida stellateoidea, and Candida krusei (Ogbolu et al., 2007). ...
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This study aims to describe the pregnancy care of the Muna tribe in Muna Regency. The research method used is descriptive qualitative, to determine the informants using purposive sampling. The research was conducted in Muna Regency, Southeast Sulawesi. Data was collected through interviews, observation, and documentation. The results of this study indicate that the treatment carried out by the people of Muna Regency is very beneficial. Pregnancy treatment, which is commonly known as doforoghu mina or drinking oil, has an association with medical treatment because the oil used in the treatment of doforoghu mina is real coconut oil which has extraordinary benefits in health sciences such as anti-inflammatory. The diforoghu mina ritual is usually guided by sando, a person who is trusted to take care of pregnant women until the delivery process, sando is in charge of supervising the doforoghu mina ritual performed by pregnant women. The results of this study are expected to provide implications for the need for education from health workers about health for pregnant women with a special target, namely the Muna tribe. This research on prenatal care requires tripartite cooperation between the government, health workers and the community which must be improved and realized in all aspects. With the responsibility of all parties, pregnant women will get the special attention needed.
In the present research work, medium-chain triglyceride (MCT) is used in the preparation of puran poli. Effect of MCT on various attributes likes textural, microbiological, sensory and oxidative stability of puran poli was studied. Use of MCT showed a positive effect on the texture of puran poli without use of hydrocolloids. Texture of puran poli became soft after storage of 15 and 25 days at 25 ± 2 °C and 4 ± 2 °C respectively. Puran poli showed no bacterial growth at both the storage conditions, however, there was yeast and mould growth on Puran poli stored at 25 ± 2 °C after 25 days i.e., 3 × 101 CFU/gm sample, which was safe for consumption as per WHO guidelines. pH showed a marginal change from 6.56 to 6.11 for puran poli stored at 25 ± 2 °C and from 6.62 to 6.33 for puran poli stored at 4 ± 2 °C. Sensory attributes like colour, taste, texture was not affected by the use of medium-chain triglyceride. Overall acceptability of puran Poli was satisfactory for the storage period of 30 days at 4 ± 2 °C.
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Objective-Emergence of Multi Drug Resistance indicates a dire to understand the bacterial involvement in infections and find out new alternative approaches in its therapeutics and prevention. The present study was undertaken to study the antimicrobial resistance patterns of S. aureus isolated from various samples collected from Hospitals of Gwalior. During the present study an effort was made to find out the information about mecA protein of Staphylococcus and their conserved regions were analyzed in order to assess their antigenic potential. Methods-In the present study, a total of 872 samples were collected and processed for MRSA screening. Conventional methods were used for the isolation and identification of bacteria. Thereafter, antibacterial property of 20 various drugs as well as aromatic compounds of 18 herbal plants was performed against multiple resistant Staphylococcus aureus (MRSA) isolates according to the guidelines of National Committee for Clinical Laboratory Standards (NCCLS). In silico prediction of vaccine candidates in mecA through bioinformatics approach was also performed. Results-The study revealed that drug resistance pattern of MRSA isolates is increasing. But the major concern is the development of resistance against Vancomycin which is thought be the most effective drug against Staphylococcus. In comparison to antibiotics, essential oils showed very good activity against the test bacteria with few exceptions. Conclusion-The essential oils of Clove and Cinnamon were found to be more active against the test organism. We predicted multi-epitope peptide which was having very good potential to induce B cell response and a very good candidate for binding to MHC II molecule and thus can act as a suitable vaccine target against S. aureus.
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Objective: The objective of the study was to evaluate the antibacterial activity of the ethanolic extracts of leaves of Citrus maxima (Burm.) Merr. (EECM) on Escherichia coli and Pseudomonas aeruginosa. Methods: The ethanolic extract of leaves of Citrus maxima (Burm.)Merr.(EECM) was prepared by percolation method. Pathological isolates Escherichia coli and Pseudomonas aeruginosa were obtained from the Department of Microbiology, Assam Medical College & Hospital. Disc diffusion method for antimicrobial susceptibility testing was performed according to the Kirby-Bauer method. The whatman-1 filter paper discs of 6mm sizes impregnated with the plant extract were placed on Mueller-Hinton agar plates seeded with bacterial cultures of 0.5 Mc Farland standards. The antibacterial activities were assessed by the presence or absence of inhibition zones after incubating the plates at 37°c for 24 hours. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of EECM for the selected pathogens were determined by broth macrodilution method. Results: Maximum zone of inhibition in antibacterial susceptibility test was shown by Pseudomonas aeruginosa. MIC value of the extract for Pseudomonas aeruginosa (0.312mg/ml) was found to be lower than Escherichia coli but MBC value (1.25mg/ml) was found to be the same for both the bacteria. Conclusion: The plant extract of Citrus maxima (Burm.) Merr showed significant antibacterial activity against Escherichia coli and Pseudomonas aeruginosa.
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The monolaurin compound had been synthesized from lauric acid and glycerol by using sulfuric acid (H2SO4) as catalyst. The synthesis of monolaurin was done by batch esterification on the free solvent system. The esterification reaction was performed on the equivalent mol ratio between lauric acid and glycerol 1:1, in the presence of 5% H2SO4, at 130 °C, for 6 hours, produced ester compounds on 59.29%. The products of column chromatography on silica gel purification are monolaurin and dilaurin in amount of 31.05 and 4.48%, respectively. The monolaurin and dilaurin were identified by TLC, FTIR, GC-MS, and NMR spectrometer. The spectral data of monolaurin was compared to spectral data of standard monolaurin. The result of NMR identifications showed that synthesis products were a-monolaurin and a,a'-dilaurin. The antibacterial activity of synthesis products was tested against Staphylococcus aureus. The activity result showed that the antibacterial activity of monolaurin is more active than dilaurin. Keywords: monolaurin, esterification, identification, antibacterial activity.
Analytical information on fats and oils is required for trading, quality control, nutritional labeling and forensics. Development of analytical procedures was one of the historical reasons for the organization of the Society of Cotton Products Analysits; it continues as a major effort of the successor organization, the American Oil Chemists’ Society, through its Uniform Methods Committee and final publication of methods in the Society’s “Official and Tentative Methods.” A review of the current status of methods development will be followed by a glimpse of methodological research currently underway.
Objective: Diseases due to pathogenic bacteria and fungi represent a critical problem to human health and they are the major cause of morbidity and mortality worldwide. Plants based antimicrobials are effective in the treatment of infectious diseases while simultaneously mitigating many of the side effects that are often associated with synthetic antimicrobials. In the series of medicinal plants, one such medicinal plant which has been widely used in traditional medicine but lacks scientific scrutiny is Pithecellobium dulce. The present study was aimed to investigate the antimicrobial properties of P.dulce pod pulp extract against common pathogenic gram positive, gram negative bacteria and fungi. Methods: Ethanolic extract was used for the study. Phytochemical screening, total phenolic and flavonoid content were determined. The antibacterial and antifungal activity of ethanolic extract of pod pulp was tested against clinically important Gram positive, Gram negative bacteria and pathogenic fungal strains. The inhibitory effect was assessed by well diffusion method. The Minimum Inhibitory Concentration (MIC), Minimum Bactericidal Concentration (MBC) and Minimum Fungicidal Concentration (MFC) were also determined by serial dilution method. Results: Phytochemical analysis of the pulp extract revealed the presence of alkaloids, flavonoids, glycosides, saponins, phytosterols, and triterpenoids. The pulp extract was found to contain significant amounts of total phenols and flavonoids. The pulp extract showed significant zone of inhibition in a dose dependent manner. The MIC and MBC values of the pulp extract against both Gram positive and Gram negative bacterial strains varies from 1mg to 5mg and the results are comparable with chloramphenicol. The MIC and MFC values of pod pulp extract against fungal strains varies from 1 mg to 7 mg and the results are comparable with Amphotericin B. Conclusion: It can be concluded that the pulp extract possesses potent bactericidal and fungicidal activity which in turn may be due to the presence of biologically active ingredients with antimicrobial activity in the pod pulp.
Coconut oil has a unique role in the diet as an important physiologically functional food. The health and nutritional benefits that can be derived from consuming coconut oil have been recognized in many parts of the world for centuries. Although the advantage of regular consumption of coconut oil has been underappreciated by the consumer and producer alike for the recent two or three decades, its unique benefits should be compelling for the health minded consumer of today. A review of the diet/heart disease literature relevant to coconut oil clearly indicates that coconut oil is at worst neutral with respect to atherogenicity of fats and oils and, in fact, is likely to be a beneficial oil for prevention and treatment of some heart disease. Additionally, coconut oil provides a source of antimicrobial lipid for individuals with compromised immune systems and is a nonpromoting fat with respect to chemical carcinogenesis.
Enzymatic modification of fats and oils using lipases has recently come into limelight, giving rise to a breakthrough in modification technology. This paper describes the latest development and future trend of this new technology with respect to the enzyme lipases, lipase reactions, and bioreactor systems. Particular attention is paid to the utilization of this technology in industrial production of functional fats and oils. A new topic of the reasearch work of an enzymatic reaction of conversion from diglycerides to triglycerides in crude palm olein is presented in detail.