Jurnal praktikum teknologi farmasi steril

VALIDASI
( VALIDATION )
Edward Andre Meide Saputra Pardede, Jella Iranda, Reza Amelia Octaviani,
Yasrina
Pharmacy Study Program Faculty of Mathematics and Natural Sciences Sriwijaya
University Indralaya

ABSTRAK
Validasi merupakan sebuah kegiatan untuk melakukan pembuktian, artinya validasi adalah
sebuah pekerjaan atau bisa juga disebut sebagai dokumentasi. CPOB mensyaratkan industri
farmasi untuk mengidentifikasi validasi yang perlu dilakukan sebagai bukti pengendalian
terhadap aspek kritis dari kegiatan yang dilakukan. Perubahan signifikan terhadap fasilitas,
peralatan dan proses yang dapat mempengaruhi mutu produk hendaklah divalidasi.Validasi
adalah tindakan pembuktian dengan cara yang sesuai bahwa tiap bahan, proses, prosedur,
kegiatan, sistem, perlengkapan atau mekanisme yang digunakan dalam produksi maupun
pengawasan mutu akan senantiasa mencapai hasil yang di inginkan. Percobaan yang
dilakukan berupa validasi autoklaf, validasi oven, validasi kimiawi dan validasi timbangan.
Dari percobaan ini, dapat diketahui bahwa pada validasi oven pada suhu 200C selama 30
menit valid dapat mensterilkan alat gelas dan logam.
Kata Kunci : validasi, oven, autoklaf, timbangan analitik

ABSTRACT

Validation is an activity to perform proof, meaning validation is a job or can also be

referred to as documentation. CPOB requires the pharmaceutical industry to identify the
validation that needs to be done as a proof of control over the critical aspects of the
activities undertaken. Significant changes to facilities, equipment and processes that may
affect the quality of the product shall be validated.Validation is an evidentiary act in an
appropriate manner that any material, process, procedure, activity, system, equipment or
mechanism used in production or quality control will always achieve results which are
desired. Experiments conducted in the form of autoclave validation, oven validation,
chemical validation and scalability validation. From this experiment, it can be seen that in
oven validation at 200 C for 30 minutes valid can sterilize glass and metal tools. From
autoclave validation at 121 C for 20 minutes, it is known that this method can validly
sterilize the existing glass container.

Keywords: validation, oven, autoclave, analytical scales

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Jurnal praktikum teknologi farmasi steril
1. INTRODUCTION
Validation is an evidentiary act in an
appropriate manner that any material, process,
procedure, activity, system, equipment or
mechanism used in production and control will
always achieve the desired result (Voight,
1995).
Validation is an action that proves that a
process or method can provide consistent
results in accordance with established
specifications and well documented. Validation
is done when there are changes that affect the
product directly, new product or old product
with new method, exiting and legacy product.
Where the validation of the production process
is an action that proves that the process
performed can provide consistent results in
accordance with the specified specifications.
While the packaging process validation is an
action that proves that the process performed
can provide consistent results in accordance
with the specified specifications. Calibration is
an action to compare between the value
indicated by a tool or instrument with a value
already known by its standard or calibrator.
Calibration aims to prove that the tools used
can provide true value (Liana, 2008).
Oven is a tool used for sterilization by
using dry air. This sterilizer is used to sterilize
glassware such as Erlenmeyer, Petridisk (petri
dish), test tube and other glasses. Materials
such as cotton, cloth and paper can also be
sterilized in the oven but in certain
Farmasi, Fakultas Matematika dan Ilmu Pengetahuan Alam Universitas Sriwijaya
temperatures, generally the temperature used in
dry-way sterilization is about 140-170 C for at
least 2 hours. It should be noted that the length
of sterilization depends on the amount of
sterilized equipment and the resistance of the
device to heat. Sterilization with oven can not
be used for glass tools requiring accuracy (eg
measuring instrument) and rubber or plastic
cover (Lachman, 1989)
An autoclave is a closed heating device
used to sterilize an object using high pressure
and pressure vapor (121 C., 15 lbs) for about
15 minutes. The pressure drop in the autoclave
is not intended to kill microorganisms, but
rather increases the temperature in the
autoclave. This high temperature will kill
microorganisms. The autoclave is primarily
aimed at killing endospores, which are resistant
cells produced by bacteria, they are resistant to
heating, dryness, and antibiotics. In the same
species, endospores can withstand the
environmental conditions that can kill the
bacterial vegetative cells. Endospores can be
killed at 100 C, which is the boiling point of
water at normal atmospheric pressure. At 121
C, endospores can be killed within 4-5 minutes,
in which bacterial vegetative cells can be killed
in just 6-30 seconds at 65 C (Lachman 1989)
The autoclave sterilization time
calculation begins when the temperature inside
the autoclave reaches 121 C. If the sterilized
object is thick or large enough, the heat transfer
on the inside of the autoclave will slow down,
Jurnal praktikum teknologi farmasi steril
resulting in an extension of the total heating
time to ensure that all objects are at 121 C for
10-15 min. Extension of time is also required
when large volume fluids will be autoclaved
because large volumes take longer to reach
sterilization temperatures. Performance
autoclave tested with biological indicators, such
as Bacillus stearothermophilus (Lachman,
1989)
There are three types of autoclaves,
namely gravity displacement, prevacuum or
high vacuum, and steam-flush pressure-pulse.
The difference between these three types of
autoclaves lies in how air is removed from the
autoclave during the sterilization process (Harr,
1994).
Gravity Displacement Autoclave. The air
in the autoclave space is moved only by
gravity. The principle is to utilize vapor relief in
comparison with air, so that air lies beneath the
vapor. How it works begins by inserting the
steam through the top of the autoclave so that
the air is pressed down. Slowly, the steam
began to accumulate more and more air down
and out through the channel at the bottom of the
autoclave, then the temperature increases and
sterilization occurs. The autoclave can work
with a temperature range of 121-134 C with a
time of 10-30 minutes (Harr, 1994).
Prevacuum or High Vacuum Autoclave.
The autoclave is equipped with a pump that
evacuates almost any air from the autoclave.
The way it works begins with air expenditure.
Farmasi, Fakultas Matematika dan Ilmu Pengetahuan Alam Universitas Sriwijaya
This process lasts for 8-10 minutes. When a
vacuum state is created, steam is inserted into
the autoclave. As a result of vacuum air, vapor
immediately associated with the entire surface
of the object, then there is an increase in
temperature so that the sterilization process
takes place. The autoclave works with a
temperature of 132-135 C with a time of 3-4
minutes (Harr, 1994).
Steam-Flush Pressure-Pulse Autoclave.
This autoclave uses steam flow and pressure
impulse above atmospheric pressure with
repeating circuit. The cycle time in this
autoclave depends on the sterilized object
(Harr, 1994).
The analytic balance (often called the
"laboratory balance") is a type of balance sheet
designed to measure small masses within the
sub-milligram range. Analytical balance sheets
(0.1 mg or better) are in transparent boxes so
that they are not dusty and the wind indoors
does not affect the weighing operation. This
enclosed space is often referred to as a
protective angina (AOAC, 1984).
The sample to be weighed shall be at
room temperature to prevent natural convection
from forming airflow within the balance sheet
space which may cause the reading error. The
electronic analytical balance measures the
pressure required to calculate the mass to be
measured, rather than measuring its real mass.
Therefore, this tool must be calibrated to
compensate for the difference in gravity. It uses
Jurnal praktikum teknologi farmasi steril
an electromagnet to generate a repulsive force
against the sample to be measured and outputs
the result by measuring the forces required to
achieve equilibrium. Such a measuring
instrument is called an electromagnetic force
restoration sensor (AOAC, 1984).
The weighing or weighing scales we
know may be weights that are used for trading
in markets. Usually weigh the child is used as a
single device weighing on the table scales or cb
scales. The child weighing class used in the
trading process is a child weighing with M3
class diamana at this time usually weigh the
child is only made of cast iron (Ansel, 2005).
Outside of the weighing scales used for
the process of trading with the M3 class is still
a lot of other children weighing scales from the
children of the M2 to the most accurate scale is
the child E1 scales. What distinguishes the
grade of the weights is the accuracy of the
weighing child, this is the main differentiator of
the weight class of the weighing child. For
example a child of 20 kg with a M3 class
according to the OIML standard can have a
deviation of +/- 10,000mg whereas a child with
a higher grade such as a F1 20 kg weight can
have a deviation of +/- 100mg.
Material weighing scales, The higher the grade
of the child weigh then the materials used
should also be good semakain. For children
weigh M3 to M1 can still use iron raw
materials. But the higher the grade of the weigh
scale for example starting M1 better if using
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stainless steel. Materials for the manufacture of
weighing scales affect the resistance of the
child's scales and also the magnetization level.
The function of the weighing child with a high
grade is as a child of the calibration scales. The
child of the calibration scales is the child of the
scales that will be used as a reference to the
adjustment of the scales. Both on mechanical
scales and on digital scales (Ansel, 2005)
High-grade scales are also a standard
completeness requirement in a lab or a
company that wants to get an iso certification.
With the existence of a standard weighing
scales with good quality then the company or
institution can check the instrument weighing
every time. Thus, the quality control also
becomes more awake (Ansel, 2005)
2. METHODOLOGY
2.1 Time and Place
The research is done in laboratorium
Pharmacocinetic of Pharmacy Math and
Science Faculty Sriwijaya University, on
Wednesday, August 16th at 08.00 to 13.00 pm.
2.2 Tools and Materials
The tools used in this research are validation
method oven, autoclaf, vials, infusion
container, syiringe injection, scales analytic.
Materials used in this research are aquadest,
Water for Injection, alcohol 70 %, and NaOH
0,1 N, .
Jurnal praktikum teknologi farmasi steril
2.3 Procedure
Calibration
Calibration Scales analytic
Do it Weighing child Scales standard, then
Record results weighing, repeat Weighing up to
5 times the count value - average, tolerance
difference weight which is still could be
accepted that is for weight of 1-50 mg:
0,014mg, 100-500 mg: 0.025 mg, 1-5 mg:
0,054 mg. Then count range of values tolerance
on weight child Scales and Count difference
value - average weighting with weight child
scales.
Calibration Autoclave
Do it Testing microbes that are Thermophilic
and have Endospores that is Bacillus
stearothermophilus. Media for inclusion to in
Autoclave and sterilize, incubation 24 hours
ago Observe growth Bakter when the media
remains clear then Autoclave work with good.
Calibration Oven
Prepare the media to be put into the oven and
incubate for 24 hours, then observe the growth
of bacteria on the media.
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VALIDATION OF STERILIZATION
METHODS
Validation Method Sterilization with
Autoclave
Set it up container Preparations Form bottle
infusion, vial, and ampoule, then Preparations
Inserted Aquadest and Inserted to in Autoclave
with a temperature of 121 * C for 15 minutes,
preparation do on box aseptic, remove
Preparations from Autoclave and Make sure
still Spared from contact air Outside to Sign in
into the box aseptic, Sterilization Aquadets in
each container in check with the media in order,
with dripping sufficiently and flattening,
incubation agar medium containing Sample
preparations 24 hours, check there is at least
growth Bacteria on an agar medium.
Validation Chemical
Rinse cap vial with alcohol 70%, then Rinse
with distilled water, then sterilization Aquadest
checked with the media in order, with dripping
sufficiently on surface and leveling her, then
Incubated 24 hours, check there is at least
growth bacteria on an agar medium.
Validation Method Sterilization with Oven
Container vials, ampoules, and glass alarm
respectively - Respectively two fruit Wrapped
with Aluminum voil, Sterilize in oven with a
temperature of 180 * C for 30 minutes, each
container rinsed with Water for Injection in part
Jurnal praktikum teknologi farmasi steril
dalamya, results rinse Inserted to in an agar grams. These two weighing results do not enter
medium, and WFI are not inserted to in the allowed range, so the analytical scales are
container Used as a control, incubation for 24 not eligible and need to be re-calibrated.
hours on temperature of 37 * C and then Do it In the validation of dry heat sterilization
checking sterilization media in order. methods, use the oven. This method of dry heat
sterilization is done for the tools made of glass.
3. RESSULT AND DISSCUSION
In the experiment, used glass beaker, vial, and
This experiment, calibration of analytical
ampoule wrapped with two layers of aluminum
scales and validation of sterilization method
foil, to prevent contamination after sterilization.
using autoclave and oven, and validation of
The critical parameters in the oven are
chemical sterilization. Calibration is an action
temperature and time. Temperature used 180
to compare between the values indicated by a
C for 30 minutes. The sterility test is done by
device with a known standard value. While
rinsing the container with WFI then rinsing
validation is an evidentiary act in an appropriate
water is fed into agar medium. The use of WFI
manner that every material, process, procedure,
aims to prevent the false results or
activity, system, equipment, or mechanism used
contamination of water.
in production and control always achieves the
desired result. Validation of this sterilization There are two types of bacterial growth
method needs to be done to identify the critical media used, namely media for chicken broth
parameters of sterilization as well as to know and media for potato broth. Both types of media
the limits of tolerance. have different functions in their microbial
growth, where the media using chicken broth is
Group average weight
intended for bacterial growth, while the
Team A 1,966
medium using potato broth is intended for
Team B 1,968
fungal growth.
On calibration of the analytical scale, the
tolerance of acceptable weight difference is *D value A = Team Blue 1
0.014 mg for weight 1 - 50 mg, 0.025 mg for *D value B = Team Blue 2
weight 100- 500 mg, and 0.054 mg for weight D-value potato oven broth
of 1-5 g. In this laboratory weighing weighing 2 Tools D value A D value B
grams, therefore used range 0.054 mg, ie Ampule 29,002 37,0074
Vials 31,044 42,7640
1999.946 mg - 2000.054 mg. In the blue group
Beaker glass 28,006 29,0987
1, the average gain of five weights was 1.966
grams, whereas in the blue group 2 was 1.968

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Jurnal praktikum teknologi farmasi steril
Based on the results obtained by D value in
group A group potion broth for amps and vials
smaller than D group B. This is due to different
tools used so that the sterilization process with
autoclave made possible group A more sterile
than group B. And for a larger glass of D value
of group A then the sterilization process of
beaker glass is less sterile than group B.
D-value Chicken oven brooth
Tools D value A D value B
Ampule 20,21 18,987
amount on ampoules and vials that tested its
microbial growth with the medium for potato
broth. This means that sterilization method with
oven at 180 C for 30 minutes is not yet valid
for sterilization of glass container. In addition,
it can be seen that the blue group D-value 2 is
better than the blue group 1, where in the blue
group 1 there is more D-value deviation than
the blue group 2. In the validation of hot steam
sterilization method using an autoclave at 121
C for 15 minutes. The critical parameters of the
autoclave are temperature, time, and pressure.
The difference is in autoclave, sterilized glass

Vials 25,11 28,905 container filled first with new aquades

continued sterilization. This water is included
Beaker 34,92 28,991
in the agar medium and then incubated for 24
glass
hours at 37 C.

D-value Potato stock autoclave

Tools D value A D value B
Based on the results obtained by D value in
Ampule 18,532 20,876
chicken broth medium by sterilization process
Vials 17,987 19,112
using oven group A is greater than group B Infusion 16,098 17,098
value of group B. This is because sterilization
process of group B is more sterile than in group Based on the result obtained D value in potato

A. The result of sterilization test of gray 1 broth medium in all group A tool is smaller

group shows that the infusion sterilization than Group B value D. Because the sterilization

tested with the media for chicken broth, as well process with autoclave by group A is more

as sterilization of all the tools tested with the sterile than group B.

media to make potato broth shows D-value

D-value Chicken stock autoclave
more than 30. That is, there has not been a Tools D value A D value B
reduction of one log of the number of microbes Ampule 17,987 15,598
in 30 minutes. While in the gray 2 group, there Vials 16,890 13,908
has been no reduction of one log of microbial Infusion 19,870 16,876

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Jurnal praktikum teknologi farmasi steril
A sterilization process is more sterile than
Based on the results obtained by D value in group B.
chicken broth medium with sterilization process
using autoclave group A is greater than group B D-value chemical chicken broth
Tool D value A D value B
value of group B. This is due to sterilization
Vial Tiip 8,123 5,098
process group B more sterile than in group A.
The results obtained in the blue group 1
Based on the results obtained by D value in
showed a D-value value above 15 in the
chicken broth medium chemically with group
infusion tested for bacterial growth on the
vial cap A is larger than group B, because
media for chicken stock after sterilization, and
group B sterilization process is more sterile
on ampoules and infusions tested with the
than in group A. In the validation of the
medium for potato broth. While in the blue
chemical method using alcohol 70% for 10
group 2, the above D-value values were
minutes, obtained D-value of both groups, both
obtained from infusion sterilization tested on
tested on the media for potato broth or chicken
the media for chicken stock, and sterilization of
chicken medium, has D-value below 10. This
all tools tested with potato broth medium. A D-
indicates that there has been a reduction of one
value value above 15 on sterilization using an
log from Number of early microbes after
autoclave indicates a no log reduction of the
sterilization. Therefore, chemical sterilization
initial microbial count after the sterilization
using 70% alcohol for 10 minutes can be
process. This shows that the method of
considered valid for vial lid sterilization
sterilization with autoclave at 121 C for 15
minutes is not yet valid for sterilization of glass 4. CONCLUSION
container. In addition, it can be seen that D- Validation is a method of sterilization needs to
value of blue group 1 is better than blue group be done to identify the critical parameters of
2, where in blue group 2 occurs more D-value sterilization as well as know the limits of
deviations from blue group 1. tolerance. Previously the implementation of the
validation process carried out calibration of the
D-value chemical potato broth
Tool D value A D value B equipment to be used, among others, the scales,
ovens and autoclaves. This calibration stage
Vials tip 4,89 6,90
aims to prove that the tool used can give value
Based on the results obtained by D value in
in accordance with the standard value or
chemical potato broth medium with group vial
calibrator. The three calibrated tools show good
closure B is larger than group A, because group
results and are in accordance with the standard

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Jurnal praktikum teknologi farmasi steril
values so they can proceed to the validation Voight, R., 1995, Textbook of Pharmaceutical
stage. Validation of sterilization methods Technology, Gadjah Mada University
conducted among others using an oven and Press, Yogyakarta
autoclave. In sterilization with oven, sterility
tests carried out by using water for injection
(WFI) sterile rinsing results sterilized tool
which drops to media agar and incubated for 24
hours. While the autoclave sterilization method
using water put into the equipment before the
sterilization process took place and dripped
onto the agar medium and incubated for 24
hours. The results obtained after observation
indicate that microbes are grown based on the
temperature and cleanliness of the tool.

BIBLIOGRAPHY

Ansel, Howard C., 2005, Introduction to

Pharmaceutical Dosage Forms, Issue IV,
UI Press, Jakarta
AOAC. 1984, Official Methods of
Analysis, Washington: Association of
Official Analytical Chemists.
Liana, 2008. General laboratory techniques.,
Vol 2, 11-12 Engineering
Laboratories. Jakarta. Erland.
Harr, Robert R., 1994, reviewer Labaratorium
Studies, Jakarta, Publisher ECG Johnson.
Lachman, L., 1989, Theory and Practice of
Pharmaceutical Industry II, Publisher
University of Indonesia, Jakarta
Lachman, L., 1989, Theory and Practice of
Pharmaceutical Industries III, UI Press,
Jakarta

Farmasi, Fakultas Matematika dan Ilmu Pengetahuan Alam Universitas Sriwijaya

Jurnal praktikum teknologi farmasi steril

LAMPIRAN

Pencucian alat gelas

Pencucian Alat Plastik Pencucian Alat logam

Farmasi, Fakultas Matematika dan Ilmu Pengetahuan Alam Universitas Sriwijaya

Jurnal praktikum teknologi farmasi steril

Pembungkusan alat sebelum masuk ke autoklaf

Farmasi, Fakultas Matematika dan Ilmu Pengetahuan Alam Universitas Sriwijaya

Jurnal praktikum teknologi farmasi steril

Pengovenan alat didalam oven

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