TEORI DAN PRINSIP DASAR

TEHNIK ASEPTIK

I Gusti Ngurah Jemmy Anton Prasetia

All drugs must be manufactured in accordance with the current Good
Manufacturing Practice (cGMP) regulations.
The products are manufactured in a controlled environment.
Cleanrooms are employed to reduce the variability of potential
production environments and as controlled environments can be
regulated to meet specific standards. GMP regulations require that
these environments be rigorously monitored to ensure that there is
full and constant awareness of current environmental conditions, both
for viable and non-viable contamination.

Cleanroom :
“a room in which air filtration, air distribution, utilities, materials of
construction, and equipment are maintained in a controlled manner.”
Aseptic filling strategy seems simple:

“sterilize the containers and everything that

enters them and fill them in a virtually
sterile environment”
Cleanrooms / aseptic fill areas
Aseptic process
Aseptic processing, which includes as appropriate:
(i) Floors, walls, and ceilings of smooth, hard surfaces that
are easily cleanable;
(ii) Temperature and humidity controls;
(iii) An air supply filtered through high-efficiency
particulate air filters under positive pressure,
regardless of whether flow is laminar or nonlaminar;
(iv) A system for monitoring environmental conditions;
(v) A system for cleaning and disinfecting the room and
equipment to produce aseptic conditions;
(vi) A system for maintaining any equipment used to
control the aseptic conditions.”
Personel yang bekerja dalam ruang
steril dipersyaratkan :

 Jumlah sesedikit mungkin

 Memiliki integritas dan motivasi yang tinggi
 Qualify = terlatih
 Sehat
 Memakai pakaian pelindung yang terdiri
dari baju, sarung tangan, penutup kepala,
masker dan penutup sepatu
Pakaian pelindung :
 Dibuat dari bahan yang tidak melepaskan
partikel dan serat, nyaman dipakai, mudah
dibersihkan dan dapat disterilkan berulang
Standar pakaian untuk ruang bersih :
 Bebas serat dan bebas partikel
 Bahan taffeta pliester ditenun tanpa
sambungan
 Tidak dapat ditembus oleh bakteri
 Bagian luar dan dalam dilapisi film plastik
 Aturan Dasar Bekerja Aseptik

1. No touch technique (hindari sentuan jika

memungkinkan)
2. Hambatan terhadap aliran udara bersih
harus seminimal mungkin
3. Pengaturan tata letak alat yang tepat
4. Semua proses aseptik dikerjakan pada
jarak minimal 6 inci dari tepi bagian luar
LAFC
LAFC Laminar Air Flow Cabinet (LAFC)
HEPA (High Efficiency Particulate Air)
Metode ventilasi pada Clean Room
1. Aliran tidak searah (Non unidirectional
type) / Conventional Flow
2. Aliran searah (Unidirectional type) /
Laminar Flow
Air Filtration
Membrane
A compressed gas should be of appropriate purity (e.g., free
from oil) and its microbiological and particle quality after
filtration should be equal to or better than that of the air in
the environment into which the gas is introduced.
Often used to produce a sterile compressed gas to conduct
operations involving sterile materials, such as components and
equipment.

High-Efficiency Particulate Air (HEPA)

HEPA filter: should be capable of retaining at least 99.97 % of
particulates greater than 0.3 μm in diameter.
 HEPA-filtered air should be supplied in
critical areas at a velocity sufficient to sweep
particles away from the filling/closing area
and maintain unidirectional airflow during
operations.
 Laminar Air Flow Cabinet (LAFC)
 Merupakan suatu cabinet yang dapat menjamin
ruangan dalam cabinet tersebut memenuhi
persyaratan ruang kelas A
 LAFC Horisontal
◦ Udara mengalir ke arah operator
◦ Digunakan untuk sediaan non kemoterapi
◦ Kecepatan aliran udara 0,45 m/detik
 LAFC Vertical
◦ Udara mengalir dari atas ke bawah untuk menjaga
sterilitas dan melindungi operator
◦ Digunakan untuk penyiapan sediaan
kemoterapi/sitotoksik
◦ Kecepatan aliran udara 0,3 m/detik
CLEANING THE ASEPTIC FILL AREA
Cleaning the aseptic fill area
Cleaning the aseptic fill area is made more
difficult by the variety of surfaces that require
attention: barrier curtains, walls, windows,
floors, ceilings, table-tops and machinery with
complicated inner and outer surfaces.
Cleaning the aseptic fill area
Cleaning efforts often centre on viable and
non-viable particles. Particles can be hard to
remove because of capillary forces,
electrostatic forces, hardened chemical bridges,
thermal solidification, tackiness, Van der Waals
forces, deformation and imbedding, surface
roughness and gravity.Viable particles are often
on, or under, non-viable particles on surfaces
and cannot be killed until the covering
particles are removed.
Cleaning materials
Wipers for cleaning aseptic fill areas are typically knitted
from continuous filament polyester and may have sealed
edges to further reduce particle release. Pre-sterilized by
gamma radiation at a dose level adequate to assure a less
than one in a million chance of a viable organism
liquid cleaners
reduced surface tension to wet surfaces; quick evaporation
and minimal residue. They should possess an aqueous
component; a hydrocarbon solvent; an alkaline component
to help dissolve oils and greases; and should be non-toxic,
non-flammable and not ozone-depleting.
Disinfectants
 Suitablility, efficacy, limitations of disinfectants and procedures
should be assessed minimum contact time established
 Disinfectants in Grade A/B areas should be sterile, supplied in
sterile containers and used for a defined period
 Should be shown to be effective against facility microbial flora
 Should be sporicidal (if spores found in the environment) and
for “spraying in” of components and equipment
 Disinfection SOPs should include sufficient detail to enable
reproducibility preparation, work sequence, contact time
 Organisms identified from adverse trends should be tested for
their sensitivity to the disinfectants used
Cleaning Method
Wet wiping is used not only to disinfect but also to dissolve
contaminating materials and to weaken bonds between the particles
and the surface.
Procedures:
1. Fold the wiper in quarters, obtaining several clean wiper surfaces.
Folding helps to make the pressure of the hand and fingers more
uniform (Figure 1).
2. Wipe in a pattern of parallel strokes with some overlap, changing
the surface of the wiper or the wiper itself at the beginning of each
stroke (Figures 2 and 3).
3. Start each stroke at the cleaner end of the planned path.
4. Change the wiper surface approximately every 10 wiper lengths of
wiping (more often if the surface is visibly dirty).
5. To disinfect, use a nearly saturated wiper and leave a visible
disinfectant film.
Standar prosedur cleaning tertuang dalam:
PETUNJUK OPERASIONAL PENERAPAN
PEDOMAN CARA PEMBUATAN OBAT
YANG BAIK (CPOB)
Monitoring
Manufacturing Environment
Four grades of clean areas:
 Grade D (equivalent to Class 100,000, ISO 8):
◦ Clean area for carrying out less critical stages in manufacture of
aseptically prepared products eg. handling of components after
washing.
 Grade C (equivalent to Class 10,000, ISO 7):
◦ Clean area for carrying out less critical stages in manufacture of
aseptically prepared products eg. preparation of solutions to be
filtered.
 Grade B (equivalent to Class 100, ISO 5):
◦ Background environment for Grade A zone, eg. cleanroom in
which laminar flow workstation is housed.
Manufacturing Environment
 Grade A (equivalent to Class 100 (US Federal Standard
209E), ISO 5 (ISO 14644-1):
◦ Local zone for high risk operations eg. product filling, stopper bowls,
open vials, handling sterile materials, aseptic connections, transfer of
partially stoppered containers to be lyophilized.
◦ Conditions usually provided by laminar air flow workstation.
 Each grade of cleanroom has specifications for viable and
non-viable particles
◦ Non-viable particles are defined by the air classification
Manufacturing Environment
Environmental Monitoring - Physical
 Particulate matter
◦ Particles significant because they can contaminate and also carry
organisms
◦ Critical environment should be measured not more than 30cm from
worksite, within airflow and during filling/closing operations
◦ Preferably a remote probe that monitors continuously
◦ Difficulties when process itself generates particles (e.g. powder
filling)
◦ Appropriate alert and action limits should be set and corrective
actions defined if limits exceeded
Manufacturing Environment
Environmental Monitoring - Physical
 Differential pressures
◦ Positive pressure differential of 10-15 Pascals should be maintained
between adjacent rooms of different classification (with door closed)
◦ Most critical area should have the highest pressure
◦ Pressures should be continuously monitored and frequently
recorded
◦ Alarms should sound if pressures deviate
◦ Any deviations should be investigated and effect on environmental
quality determined
Manufacturing Environment
Environmental Monitoring - Physical
 Air Changes/Airflow patterns
◦ Air flow over critical areas should be uni-directional (laminar
flow) at a velocity sufficient to sweep particles away from
filling/closing area
◦ for B, C and D rooms at least 20 changes per hour are ususally
required
 Clean up time/recovery
◦ Particulate levels for the Grade A “at rest” state should be
achieved after a short “clean-up” period of 20 minutes after
completion of operations (guidance value)
◦ Particle counts for Grade A “in operation” state should be
maintained whenever product or open container is exposed
Manufacturing Environment
Environmental Monitoring - Physical
 Temperature and Relative Humidity
◦ Ambient temperature and humidity should not be
uncomfortably high (could cause operators to generate
particles) (18°C)
 Airflow velocity
◦ Laminar airflow workstation air speed of approx 0.45m/s ±
20% at working position (guidance value)
Particle Count Room Classification
There are three measurement phases involving particle
counting in cleanrooms:
 As-Built: a completed room with all services connected and
functional, but without production equipment or personnel
within the facility.
 At Rest: all the services are connected, all the equipment is
installed and operating to an agreed manner, but no
personnel are present.
 Operational: all equipment is installed and is functioning to
an agreed format, and a specified number of personnel are
present, working to an agreed procedure.
Monitoring Non-Viable Particle Levels in
Cleanrooms
The cGMP guides for monitoring Particle:
Portable Particle Counter : A compact
approach to maintaining cleanliness is to build
data storage into a portable particle counter.
Facility Monitoring System : Single continuous
particle counter installed into a critical location,
or an arrangement of instruments linked to a
central monitoring computer suitable for making
the measurements required
Particles in Pharmaceutical
Cleanrooms
 Pharmaceutical cleanrooms are classified
according to the particle concentration of the
air that is required to meet the cleanliness
criteria for the manufacturing process being
performed.
 Using the ISO standards, the higher the
classification number, the lower the particle
concentration. Originally cleanrooms were
classified according the number of particles per
cubic foot at 0.5 microns (μm).
There are three basic approaches to obtaining
automated particle counts:
 Manifold system (using multiple tubes via
multi-port scanning) linked to a particle
counter
 Individual particle sensors
 Combination of manifolds and particle
sensors
Manifold system
Individual Particle Sensors
Increasingly, to ensure that continuous monitoring is preserved,
dedicated, locally-mounted particle sensors are being used.A sensor
consists of a small enclosure, housing an optical system, a light source
(laser diode), and signal generation electronics. The sensors often require
an external vacuum source and signal communication cable to transmit
data to the central monitoring computer.
Advantages of individual sensors:
 Automated, so lower personnel costs
 Continuous monitoring and reporting of data, therefore detecting short-lived particle
bursts
 Simple, low-cost installation
 Ease of relocation to alternative positions; easy to rotate out for servicing
 Highest level of confidence
Disadvantages of individual sensors:
 More instruments
 Higher initial equipment cost
 Higher maintenance cost
Combination System
 An alternative to the above choices
(manifold or individual sensors) is a
combination which uses the advantages of
both systems. In this solution, the majority
of sampling is monitored using the manifold
system, while specific critical locations are
continuously sampled by individual sensors.
Particle Measuring
Systems recommends
that measurements to
confirm air cleanliness
in critical areas be
taken at sites where
there is the most
potential risk to the
exposed sterilized
product, containers,
and closures.
Manufacturing Environment
Classification of Clean Areas
Monitoring Viable Particle Levels in
Cleanrooms
Methods
 Surface monitoring
◦ Product contact surfaces, floors, walls, and equipment should be tested on a regular
basis
◦ Touch plates - used for flat surfaces
 sample area of 25cm2
 medium protrudes above sides
 medium contains neutralisers

◦ Surface Swabs - used for irregular surfaces

 area approx 25cm2 is swabbed
 qualitative or quantitative

Surface monitoring should be performed at conclusion of aseptic processing (to

minimise risk of contaminating critical surfaces during production)
Environmental Monitoring
Methods
◦ Active Air Monitoring
 impaction, centrifugal and membrane (or gelatin) samplers
 a certain volume of air is sampled (volume and location should be
meaningful)
 instruments should be calibrated
◦ Passive Air Monitoring
 Settle plates exposed for 30-60 minutes (longer may result in agar drying
out) and replaced for duration of filling
 Media should be capable of growing a range of bacteria and moulds (e.g.
Soybean Casein Digest Agar (SCDA)/Trypticase Soy Agar (TSA)
 Should consider use of medium specific for moulds if shown to be a
problem in the environment
 Only give qualitative or semi-quantitative results
 Data generated considered in combination with active air sampling results
Environmental Monitoring
Sampling Locations
◦ Should be based on risk of microbiolgical contamination
◦ Should be clustered around areas where product or components are
exposed e.g.
 at filling heads on filling lines
 loading of product into lyophilizers
 stopper bowls
 where aseptic connections are made
 where there are high levels of operator activity (but without impacting on
production)
◦ Lower grade areas are monitored less frequently and trends monitored
 Manufacturing Environment
 Limits for viable particles (microbiological
contamination)
Grade Air sample Settle plates (90mm Contact plates Glove print
(CFU/m3) diameter) (55mm (5 fingers)
(CFU/4hours) diameter) (CFU/glove)
(CFU/plate)
A <3 <3 <3 <3
B 10 5 5 5
C 100 50 25 -
D 200 100 50 -

These are average values

– Individual settle plates may be exposed for less than 4 hours
• Values are for guidance only - not intended to represent
specifications
• Levels (limits) of detection of microbiological contamination should be
established for alert and action purposes and for monitoring trends of air
quality in the facility
Terima Kasih