45

Part 5: Advanced pharmaceutical protection schemes

5.1 Maximizing sterility assurance

A pharmaceutical product is considered sterile if not more than one contaminated

unit is found among a million dosage units. This requirement - i.e. a sterility assur-
ance level SAL of at least 10-6 - has of course to be met by terminally sterilized
products.

In aseptic processing, sterility assurance is assessed through a process simulation

procedure, i.e. a filling operation using nutrient media - the so-called media fill.
What sterility assurance level can be achieved in this case?

For many years, a SAL value of 10-3, i.e. one contaminated dosage unit in 1 000 -
proven with an upper confidence limit of 95 % - has been considered acceptable
[5.01-5.02]. FDA has now pushed this limit, in its current Guidance for Industry for
aseptic processing [5.03], to almost an SAL value of 10-4 (Fig. 5.03), and Europe has
recently followed suit [5.04]. This is an improvement, but still two orders of magni-
tudes away from the sterility assurance level of terminally sterilized products. Thus,
FDA continues to require that products should be submitted to terminal sterilization
whenever possible [5.03]. However, the percentage of parenterals not permitting
terminal sterilization is steadily increasing: this is due, to a large extent, to the in-
creasing number of protein-based active substances. Therefore, there is a continu-
ing challenge to improve the protection schemes for aseptic processing operations
towards still higher levels of sterility assurance. Long term, the goal to be met is ob-
vious: to achieve the same level of sterility assurance as that applying to terminally
sterilized products: a SAL value of 10-6.

Blow-fill-seal technology, restricted access barrier systems (RABS systems) and iso-
lator technology are the predominant protection concepts committed to the goal of
improving the Sterility Assurance Level.

5.2 Blow-fill-seal technology – basic principles

In the blow-fill-seal process, containers are formed from a thermoplastic granulate,

filled and then sealed: all within one single automatic machine and in one continuous
operation. As a first step, the thermoplastic granulate is heated above melting point
in order to permit the primary container first to be extruded and then blown into its
final shape. After cooling, it is immediately filled with product and subsequently
thermally sealed. The immediate vicinity of the blowing, filling and sealing module
within the machine is flushed with HEPA filtered air, thus maintaining grade A condi-
tions there, and grade B conditions in the area surrounding the grade A core (Fig.
5.04). The process being fully automatic, no persons are normally expected to be
present in the air cleanliness controlled area (Fig. 5.05). Thus, a number of safety
features act in combination in order to guarantee the high level of sterility assurance
 46

distinguishing this technology [5.05-5.08]. That only containers blown from thermo-
plastic raw materials are feasible with this technology is, however, a serious draw-
back: many drug products do not remain stable in this kind of container.

For GMP stipulations regarding blow-fill-seal technology and its environmental re-
quirements please refer to Annex 1 of the GMP guideline of the European Union and
PIC/S [5.01-5.02] and to Appendix B of the FDA Guidance for Industry compendium
for aseptic processing [5.03].

5.3 Pharmaceutical isolator technology

5.3.1 Conceptual features

Particularly efficient protection of critical pharmaceutical work areas can be provided

by means of isolator technology [5.09]. Isolators offer a highly enhanced level of ste-
rility assurance for aseptic processing and are particularly indicated when combined
product and operator protection is required - as during handling of highly active sub-
stances such as cytotoxics.

Isolators combine the traditional safety features of the conventional cleanroom con-
cepts - i.e. establishing contamination control by means of HEPA filtered air and ap-
propriate airflow patterns - with four additional safety features (Fig. 5.06):
- consequent separation of process and operator;
- isolator-internal air circulation independent from the room air circulation system;
- the feasibility of effective biodecontamination of the internal surfaces of the isola-
tor, as a consequence of the isolator's independent air circulation system;
- sterile transfer of materials and objects into, and out of, the isolator.

Glove/sleeve units or half-suits integrated into the isolator contour permit operator
manipulations without breaking sterility (Fig. 5.07). Special transfer modules such
as the double door airtight transfer port (Fig. 5.08) permit the sterile transfer of mate-
rials to and from the isolator. For ventilation purposes, both turbulent mixing or unidi-
rectional airflow may be employed according to circumstances. For isolators protect-
ing batch processes, turbulent airflow with its modest air flow rate requirements may
well be adequate, whereas unidirectional airflow is, as a rule, more appropriate for
the protection of continuous processes.

Regarding the isolator enclosure, two options are available:

- the soft wall concept, where the enclosure is made up of flexible, transparent
PVC sheets, stabilised by a simple framework structure;
- the hard wall concept, a combination of a solid metallic framework with hard
panel elements, into which transparent glass panels are integrated to the largest
feasible extent.
 47

The use of the soft wall principle is now largely limited to laboratory applications. In
isolators protecting production machines, the hard wall principle predominates almost
without exceptions.

5.3.2 Isolator designs for batch and continuous processes

When protecting batch processes, the key features of the isolator can be converted
almost ideally into technical reality. An example of a batch process is the sterility test
of ampoules and vials after filling (Fig. 5.09): in comparison with sterility testing in
clean work stations, the risk of false positives can be eliminated almost completely
when performing the test in an isolator [5.10].

When protecting continuous processes such as high-productivity filling operations,

deviations from the ideal isolator principles are inevitable: the isolator then provides
no longer an absolute barrier against contamination, but rather partial barrier pro-
tection. In the case of ampoule filling, for example (Fig. 5.10), ampoules enter and
leave the isolator in a continuous stream through so-called mouseholes. As the
ampoules are closed by means of a gas flame, also process air has to be extracted
continuously. The strict principle of separating the processing area from its environ-
ment by means of impervious physical barriers - i.e. walls - is thus broken locally at
these spots: aerodynamic barriers - driven by overpressure in this schematic draw-
ing - have to assume the function of efficiently isolating the inside against the outside
environment. Figs. 5.11-5.12 present an ampoule filling machine with isolator-
protected filling and closing unit, and Fig. 5.13 an isolator protecting a filling line for
syringes.

5.3.3 Regulatory and normative guidance

The regulatory authorities of the European Union and PIC/S were first in incorporat-
ing isolator-specific determinations into their GMP guidance [5.01-5.02]: in Annex 1,
a short chapter addresses this topic, with particular attention to (Fig. 5.14):
- the cleanliness of the air inside the isolator and in its background (requirements
being grade A (FDA class 100 and ISO Class 5 in operation) for the aseptic core
area, with a grade D background);
- biodecontamination of the isolator's inner surfaces;
- material transfer processes;
- isolator integrity (shell and glove/sleeve systems);
- monitoring, which should be carried out routinely and include frequent leak testing
of the isolator shell and of its glove/sleeve systems.

FDA, on the other hand, had for many years been reluctant in pronouncing them-
 48

selves on isolator technology. This has changed, though: in their present Guidance
for Industry document on aseptic processing [5.03] - see also Friedman and Sausville
[5.11] - a complete 5-page Annex is devoted to this technology. Regarding air
cleanliness requirements, for the process area class 100 (ISO Class 5) is stipulated
in the occupancy state in operation, and class 100 000 (ISO Class 8) for the isolator's
background, again in the occupancy state in operation. This background classifica-
tion requirement, corresponding with the European room grade C, is thus somewhat
more exacting than its European grade D equivalent.

Very detailed regulatory stipulations are to be found in the recommendation PIC/S PI

014-3 [5.12]: devoted exclusively to isolators for aseptic processing and sterility test-
ing, it addresses in detail the many aspects to be assessed by inspectors during au-
dits of isolators employed for these purposes. A very cautious attitude is apparent:
although the isolator's potential is appreciated, a clear warning is issued against
over-confidence and blind trust in this technology.

Most comprehensive guidance on design and qualification of isolators can be found

in the Technical Report no. 34 [5.13] issued by PDA - an International Association for
Pharmaceutical Science and Technology (formerly Parenteral Drug Association) -
with head office in the USA. It addresses all possible aspects of pharmaceutical iso-
lator technology in a both scientifically sound and practice-oriented attitude.

In the context of the ISO series of contamination control standards, isolator technol-
ogy is addressed in ISO 14644-7 [5.14], and in the context of the ISO series on asep-
tic processing, in ISO 13408-6 [5.15].

5.3.4 Biodecontamination of the isolator's internal surfaces

Biodecontamination of the isolator's internal surfaces by means of powerful agents in

the vapour phase is a particularly distinctive feature of pharmaceutical isolators. The
feasibility of the procedure is a direct consequence of the isolator's capability of
closed loop internal air circulation independent from the air circulation system serving
the isolator’s background, and of the very low leakage level of the isolator's contour.
Mouseholes and other openings must be sealed before starting biodecontamination.

As biodecontamination agent, vapour phase hydrogen peroxide VPHP (H2O2) is

presently clearly predominant. The sequence of steps during the biodecontamination
procedure is shown in Fig. 5.15. The outside and extract air dampers of the isolator
as well as all mouseholes are hermetically closed and air circulation is switched to
the closed loop mode, i.e. to 100 % air recirculation. The recirculated air is then first
dried to a relative humidity below 30 %. Now, the hydrogen peroxide vapour, fre-
quently produced on-site by a special generator unit, is added to the recirculating air,
either at a concentration slightly below, or slightly above, saturation. After a prede-
termined period shown to be sufficient for effective biodecontamination of the isola-
tor's interior, a purging cycle is initiated: the hydrogen peroxide vapour is catalytically
decomposed into its constituents water and oxigen - both, of course, natural compo-
 49

nents of the atmosphere. After removing most of the sterilizing agent by this proce-
dure, the outside and extract air dampers of the isolator can be opened again and the
remaining traces of hydrogen peroxide vented off into the atmosphere. Details re-
garding the optimization of the VPHP process are discussed in Unger-Bimczok et al.
[5.16-5.17] and regarding the physical chemistry in Hultman et al. [5.18].

The duration of the biodecontamination cycle is established experimentally with the

help of test strips carrying spores of geobacillus stearothermophilus or bacillus atro-
phaeus (previously denominated bacillus subtilis), spores which are particularly diffi-
cult to kill. If even on the most remotely positioned spore strips no growth is found
after due incubation and assessment, then the cycle can be considered as qualified.

The merits of the VPHP process are summed-up in Fig. 5.16, whereas aspects mer-
iting specific attention are listed in Fig. 5.17. Special care must be taken that all ma-
terials entering into contact with VPHP must be compatible with this biodecontamina-
tion agent. Copper alloys, for example, are unsuitable, as they tend to decompose
the hydrogen peroxide catalytically, and also some rubbers are adversely affected by
this product. Easy and direct access of the sterilizing vapour to all surfaces must be
ascertained, and long diffusion paths for the agent have to be avoided. Gloves/
sleeves should therefore be tucked into the inside of the isolator for their effective
sterilization.

The VPHP process is increasingly also utilized for the biodecontamination of entire
rooms [5.19].

5.4 Restricted access barrier systems (RABS technology)

5.4.1 Point of departure

Regarding the protection of aseptic filling operations, an alternative barrier isolation

concept has come into the limelight lately: the Restricted Access Barrier System -
RABS in short [5.20-5.23]. A prerequisite for the feasibility of technologies of this
kind was the impressive increase in reliability of filling processes - a development
strongly driven by isolator technology.

Point of departure for the development of RABS concepts were the two cornerstones
of protecting aseptic filling operations:
- conventional cleanroom technology;
- isolator technology.

In conventional cleanroom technology (Fig. 5.18, from Rauschnabel [5.20]), the

aseptic core is protected with unidirectional downward airflow of HEPA-filtered air.
From the surrounding room it is separated by curtains or solid partitions, with air spill-
over from the aseptic core into the surrounding environment. There, room grade B,
corresponding to ISO class 7 in the occupancy state in operation, has to be main-
 50

tained. Operators interfere, when required, into the aseptic core with their sterilely
gloved hands, and sanitization of the internal surfaces of the processing area is by
means of periodical disinfection: as a rule, by alcohol spray and wipe.

Isolator protection of the aseptic core (Fig. 5.19, from [5.20]), on the other hand, is
distinguished by:
- strict separation of process and personnel;
- isolator-internal air circulation independent from the room air circulation system.

Operators interfere into the aseptic core by means of glove/sleeve systems, and bio-
decontamination is effected by means of VPHP, a very effective, but time-consuming
process: hours may be required for biocontamination of a voluminous isolator. As a
consequence of the high level of airtightness of the isolator shell, room grade D is
acceptable according to the GMP Guide of the European Community and PIC/S.
FDA, on the other hand, expects the environment to be maintained at ISO class 8 in
the occupancy state in operation; this corresponds with room grade C.

5.4.2 RABS concepts

The RABS philosophy requires limiting interference into the critical area through hu-
man interventions to the absolute minimum. If such access is inevitable, it is per-
formed as far as possible through glove/sleeve units incorporated into the RABS en-
closure. Two design alternatives are distinguished:
- passive RABS units (Fig. 5.20, from [5.20]), connected to the central HVAC sys-
tem of the facility (an example is shown in Fig. 5.21);
- active RABS units (Fig. 5.22, from [5.20]), i.e. stand-alone units equipped with
an integrated air handling unit for maintaining the required conditions in the asep-
tic core and drawing its supply air from the surrounding room.

In both cases, after having provided protection to the aseptic core, the air spills over
into the surrounding room - in the same way as in conventional cleanroom technol-
ogy. As a consequence of this air spill-over into the surroundings the same stipula-
tions for the background of the core area as in conventional filling operations have to
apply: room grade B corresponding to ISO Class 7 in the occupancy state opera-
tional. Sanitization of the surfaces in the processing area is performed in the same
way as in conventional cleanroom technology, i.e. by spraying and/or wiping proce-
dures.

A modification of the basic RABS concept is the closed RABS, also denominated
cRABS. Here, the airflow protecting the aseptic core is recirculated internally - just
as in an isolator. Again, distinction is made between a passive cRABS (Fig. 5.23,
from [5.20]), where the central HVAC system of the facility is responsible for supply-
ing the aseptic core with HEPA-filtered air, and the active cRABS (Fig. 5.24, from
[5.20]), equipped with its own stand-alone air handling unit.
 51

For sanitization of the cRABS, the same procedures apply as for "normal" RABS
units; therefore a high degree of airtightness of the cRABS shell is neither necessary
nor attempted. The lower requirements regarding airtightness of the enclosure are
the most important distinctive feature in comparison with the isolator.

RABS technology has not yet been specifically addressed in regulatory documents;
the present FDA position has been outlined by Friedman and Sausville [5.24].

5.5 RABS technology versus isolators

Operated correctly and with a high degree of discipline, i.e. with a minimum of inter-
ference into the aseptic core, RABS- and cRABS-protected filling lines achieve al-
most the same Sterility Assurance Level as isolator-protected ones. As a conse-
quence of the relatively modest air leakage requirements of RABS and cRABS units,
costs are below those of the isolator. From the rapid sanitization procedure, a faster
start-up results in comparison with the isolator; in addition, format changes and
change-over from one product to the next are easier and quicker. RABS and cRABS
concepts are equivalent in these respects. Contract manufacturers therefore tend to
gravitate towards RABS technology because of its operational flexibility [5.25].

Due to the internal recirculation of the air protecting the aseptic core, cRABS pro-
tected filling lines are said to be capable of also handling highly active products ade-
quately [5.20], even if operated - as required by the GMP authorities - under over-
pressure.

Due to the very low air leakage level of the isolator's contour providing a highly effec-
tive barrier between the aseptic core and the environment, the isolator is to be given
preference where both the product and the operator require reliable protection.
Therefore, the isolator is indeed the technology of choice where highly active or toxic
products require to be handled - especially in the case of crystalline products where
the filling process is distinguished by a high degree of particle liberation. As there is
a clear trend towards highly active pharmaceutical agents, the future of isolator-
protected filling lines seems assured. Additional safety results from the high degree
of microbiological cleanliness of the isolator's internal surfaces as a result of biode-
contamination by means of VPHP - a time-consuming procedure though. Pros and
cons must therefore be balanced against the comparative rapidity of the traditional
sanitization procedures.

An exhaustive comparative assessment of RABS concepts and isolator technology

versus conventional cleanroom technology has been compiled by Agalloco et al.
[5.26]. They considered a total of 22 project criteria, and there were, of course, pros
and cons for each of the assessed technologies. For process and product protection,
both technologies are appropriate; however, the isolator should be favoured where
also personnel protection is required. Therefore, case by case assessment is indi-
cated for deciding how best to meet a specific protection requirement. However, a
 52

clear trend away from conventional cleanroom technology is evident, and this is
bluntly highlighted by the authors with their question: Will conventional cleanroom
technology still be GMP compatible in 10 years' time for protecting aseptic filling
lines?

5.6 References

[5.01] Eudralex, the rules governing medicinal products in the European Union - Vol.
4: EU guidelines to Good Manufacturing Practice for medicinal products for
human and veterinary use. European Commission, Brussels (frequently up-
dated).

[5.02] PIC/S PE 009-9: Guide to Good Manufacturing Practice for medicinal prod-
ucts (divided into four parts: Introduction, Part I, Part II, Annexes). Pharma-
ceutical Inspection Convention PIC, Pharmaceutical Inspection Co-operation
Scheme PIC/S, Geneva (1 September 2009).

[5.03] Guidance for industry: Sterile drug products produced by aseptic processing
- current Good Manufacturing Practice. U.S. Department of Health and Hu-
man Services, Food and Drug Administration (September 2004).

[5.04] Eudralex: The rules governing medicinal products in the European Union,
vol. 4: EU guidelines to Good Manufacturing Practice - Medicinal products for
human and veterinary use: Annex 1: Manufacture of sterile medicinal prod-
ucts. Brussels, 14 February 2008.

[5.05] Whyte W. et al.: Airborne contamination during blow-fill-seal pharmaceutical

production. PDA Journal of Pharmaceutical Science & Technology 52 (1998)
3, 89-99.

[5.06] Haerer M., Lichtenstein U.: Blow-fill-seal technology for aseptic production.
European Journal of Parenteral Sciences 2 (1997) 4, 119-121.

[5.07] Ljungqvist B. et al.: Current practice in the operation and validation of aseptic
blow-fill-seal processes. PDA Journal of Pharmaceutical Science and Tech-
nology 60 (2006) 4, 254-258.

[5.08] Sundström St. et al.: Some observations on airborne particles in blow-fill-seal

filling rooms. PDA Journal of Pharmaceutical Science and Technology 61
(2007) 3, 147-153.

[5.09] Coles T.: Isolation technology - a practical guide, 2nd ed. Taylor & Francis
CRC Press, London (2004).

[5.10] Akers J.E., Agalloco J.P., Kennedy C.M.: Experience in the design and use
of isolator systems for sterility testing. PDA Journal of Pharmaceutical
 53

Science and Technology 49 (1995) 3, 140-144.

[5.11] Friedman R., Sausville R.: FDA response to industry questions on filling-line
isolators. Pharmaceutical Engineering 24 (2004) 1, 50, 72-76.

[5.12] PIC/S PI 014-3: Isolators used for aseptic processing and sterility testing.
Pharmaceutical Inspection Co-operation Scheme PIC/S, Pharmaceutical In-
spection Convention PIC, Geneva (25 September 2007).

[5.13] Design and validation of isolator systems for the manufacturing and testing of
health care products. PDA Journal of Pharmaceutical Science and Technol-
ogy 55 (2001) 5, supplement TR 34.

[5.14] ISO 14644-7: Cleanrooms and associated controlled environments - Part 7:

Separative devices (clean air hoods, gloveboxes, isolators, mini-environ-
ments). International Organization for Standardization ISO, Geneva (October
2004).

[5.15] ISO 13408-6: Aseptic processing of health care products - Part 6: Isolator
systems. Ibid. (June 2005).

[5.16] Unger-Bimszok Beatriz et al.: The influence of humidity, hydrogen peroxide

concentration, and condensation on the inactivation of geobacillus
stearothermophilus spores with hydrogen peroxide vapour. Journal of Phar-
maceutical Innovation 3 (2008) 2, 123-133.

[5.17] Unger-Bimczok Beatriz et al.: Suitability of different construction materials for

use in aseptic processing environments decontaminated with gaseous hydro-
gen peroxide. PDA Journal of Pharmaceutical Science and Technology 61
(2007) 4, 255-275.

[5.18] Hultman C., Hill A., McDonnell G.: The physical chemistry of decontamina-
tion with gaseous hydrogen peroxide. Pharmaceutical Engineering 27 (2007)
1, 22-32.

[5.19] Otter J.: Blazing a vapour trail. Cleanroom Technology 12 (2006) 11, 15-17.

[5.20] Rauschnabel J.: Zwischen Isolator und Sterilraum (Between isolator and
sterile room) - Restricted Access Barrier System (RABS). Pharm. Ind. 68
(2006) 6, 767-773.

[5.21] Isberg E.A.: Advanced aseptic processing: RABS and isolator operations.
Pharmaceutical Engineering 27 (2007) 1, 18-21.

[5.22] Lysfjord J.: The ISPE RABS definition - an introduction. Pharmaceutical En-
gineering 25 (2005) 11/12, 116-117.
 54

[5.23] Drinkwater J.: RABS - performance levels defined (part 1); RABS - in opera-
tion with isolators (part 2). Cleanroom Technology 11 (2005) 9, 22-23 and 10,
27-29.

[5.24] Friedman R., Sausville R.: The FDA answers your questions on barrier isola-
tion technology. Pharmaceutical Engineering 27 (2007) 2, 102-106.

[5.25] Lysfjord J.: Using RABS and isolator in pharmaceutical applications. Clean-
Rooms 21 (2008) 1, 24-26.

[5.26] Agalloco J., Akers J., Madsen R.: Choosing technologies for aseptic filling:
"Back to the future, forward to the past"? Pharmaceutical Engineering 27
(2007) 1, 8-16.