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formula 1 of 28.814 ± 0.431; formula 2 is 32.299 ±
1.893 and formula 3 is 30.234 ± 0.531 which shows
that the three formulas have very strong antioxidant
activity (Kasuan N,et all , 2009).
2 METHOD
To determine the inhibition of citrus hystrix leaf
extract against the fungus Pityrosporum ovale, the
sensitivity test method (disc diffusion) was used. In
the experiment, 5 treatment groups included negative
control using aquadest, positive control using
ketoconazole ointment 2%, concentrations of 25%,
50%, and 75%. The medium used was Potato
dextrose agar. The research was conducted at the
Microbiology Laboratory of the Faculty of Medicine,
Prima Indonesia University.
The tools and materials used in the experiment
were petri dishes (7 pieces), test tubes, filter paper,
measuring cups, autoclaves, incubators, disc paper,
cotton swabs, tweezers, sterile cotton, calipers, drop
pipettes, erlenmeyer flasks, analytical scales. , test
tube racks, 5 kg kaffir lime leaves, 70% ethanol, 96%
ethanol, Pityrosporum ovale mushrooms, 2%
ketoconazole ointment, alcohol, aquabidest,
physiological NaCl, spiritus, potato dextrose agar
(PDA) media, plastic wrap, and aluminum foil.
Preparation stages, namely: The tools and
materials used are sterilized in an oven at a
temperature of 40-70º C for ± 2 hours, At this stage
of making citrus hystrix leaf water, first clean the
citrus hystrix leaves with water until clean then the
citrus hystrix leaf water is sliced small and blended
(Figures 1 and 2.
The citrus hystrix leaf extract which has been
evaporated with ethanol was divided into 5 treatment
groups with a concentration of 25%, 50%, 75%,
positive control, and negative control by dissolving
with 96% ethanol technique. Dissolving with ethanol
aims to remove the essential oil content in the extract.
The extract was made by macerating the citrus hystrix
leaves which had been cleaned with running water
before drying. The dried citrus hystrix leaves were
mashed using a blender to form simplicia powder.
From each simplicia, 1 kg of sample was taken and
then dissolved or soaked using 10 liters of absolute
ethanol (96% ethanol) solvent per each simplicia
(Figure 1). Then stirring per day for 1 hour. This
maceration process was carried out 3 times.
The citrus hystrix leaves are washed with running
water then drained until dry and put in a drying
cabinet for 3 days before mashed using a blender. The
powder was sifted to a fine powder of the citrus
hystrix leaves.
For the experiment, several tools such as petri
dishes (7 pieces), test tubes, measuring cups, are
sterilized first using an autocalved at a temperature of
121ºC for about 15 minutes. Meanwhile, the tools
made of metal are sterilized on an incandescent fire
for about 1 minute.
Preparation of Potato Dextrose Agar (PDA) as
much as 65 grams of medium is suspended in 1 liter
of distilled water into an erlenmayer flask and then
stirred using a hotplate strirer for 1 minute or until
dissolved. The sterilization was then carried out by
autoclave for 15 minutes at 121 ° C and allowed to
cool until the temperature has decreased to 40-45 ° C.
The results obtained were in the form of jelly poured
into a petri cloud and cooled until frozen.
The preparation of the Pityrosporum Ovale
Mushroom suspension used in this study was
obtained from the USU Pharmacy Laboratory. The
Pityrosporum Ovale used was made by taking a loop
of germs from the culture so that it is tilted and then
put into a tube containing physiological NaCl then
stirring until all fungal colonies dissolve in NaCl.
The diffusion test of pityrosporum ovale was
carried out by: PDA media that had hardened, evenly
scratched the entire surface of the media using a
cotton swab containing the suspension of
Pityrosporum Ovale fungus; a sterile blank disk is
taken; dipped in 50% concentration, and finally
placed on the surface of the media that has been
scratched by the fungus Pityrosporum Ovale. The
treatment is carried out for all concentrations then
spaced apart to prevent the inhibition zones from
forming. The same experiment was repeated 3 times
for data collection needs. All test isolates were
incubated for 36-48 hours at 37ºC in an incubator.
After 36-48 hours, the zone of inhibition is measured
using a caliper. The classification of the response to
fungal growth inhibition is given in Table 1
(Maryanti, Marta, Della, and Hamidy, 2017).
Table 1 Classification of fungal growth inhibition
responses.
Inhibition Growth
Rresponse
From Table 1, it is obtained that the larger the
diameter of the clear inhibition zone was > 2 cm, then
the response to fungal growth will be stronger and on
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