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1680 1229.7 Gaseous Sterilization / General Information USP 39

Component and Load Mapping

Component and load mapping using invasive sampling are not a part of gas sterilization because sampling systems placed
within the load items would alter gas and humidity penetration. Evaluation of lethal conditions with individual items and
across loading patterns is best provided by biological indicators or process challenge devices placed within the load items and
distributed within the load. Indicators or process control devices are placed within the items and load at locations believed to
be hardest for the gas and humidity to penetrate.

Biological Indicators

The biological indicator of choice for gas sterilization varies, as noted above. B. atrophaeus (ATCC 9372) is used with EO and
chlorine dioxide, and ozone sterilization is monitored with G. stearothermophilus (ATCC 12980 or 7953). D-values for the bio-
logical indicator can be used to establish exposure periods for the sterilization process to ensure adequate process efficacy.
When positioning biological indicators within items it is important to ensure that the placement of the BI does not occlude gas
passage or otherwise interfere with the distribution/penetration of the sterilant within the item.

Process Confirmation and Microbiological Challenge

The core of the validation activity is the confirmation of acceptable process parameters with simultaneous physical and
chemical measurement and microbial challenge. Sensors are placed in the chamber, or biological indicators are positioned
General Chapters

within the load items. Proof of cycle efficacy is provided in replicate studies in which the biological indicators are killed and the
physical measurements correspond to the expected values.

ROUTINE PROCESS CONTROL

Gas sterilization is subject to formal controls that maintain a validated state over time. The practices outlined in 1229 in-
clude the general requirements appropriate for all sterilization systems. Sterilization is accomplished by a number of related
practices that are essential for continued use of the process over an extended period of time. The essential practices to main-
tain validated status include calibration, physical measurements, ongoing process control, change control, preventive mainte-
nance, periodic reassessment, and training. When parametric release has not been established, biological indicators positioned
within the load are used for routine release of each sterilization load, along with a review of documentation from the sterilizer
control system.

REFERENCES

1. USP General ChaptersMicrobiology Expert Committee. An outline of planned changes to USP 1211 Sterilization and
Sterility Assurance of Compendial Articles. Pharmacopeial Forum. 2012; 38(2).
2. ISO 11135-1:2007 Sterilization of Health Care ProductsEthylene OxidePart 1: Requirements for Development, Valida-
tion, and Routine Control of a Sterilization Process for Medical Devices. Geneva: International Organization for Standards
(ISO); 2007.
3. ISO 11135-2:2008 Sterilization of Health Care ProductsEthylene OxidePart 2: Guidance on the Application of ISO
11135-1. Geneva: International Organization for Standards (ISO); 2008.
4. ISO 10993-7 Biological Evaluation of Medical Devices, Part 7: Ethylene Oxide Sterilization Residuals. Geneva: International
Organization for Standards (ISO); 2008.
5. Ethylene oxide, ethylene chlorohydrin, and ethylene glycol proposed maximum residue limits and maximum levels of ex-
posure. Fed Regist. 1978; 43(122):2747427483.
6. Gillis J, Mosley G. Validation of ethylene oxide sterilization processes. In: Agalloco J, Carleton FJ, eds. Validation of Pharma-
ceutical Processes. 3rd ed. New York: InformaUSA; 2007.

1229.8 DRY HEAT STERILIZATION

Dry heat sterilization is a process utilized for heat-stable items (glass, stainless steel, nonaqueous liquids, powders, etc.) that
are unsuited for steam sterilization because of either an absence of water (nonaqueous liquids and powders) or requirements
for absolute dryness following processing (product contact parts for nonaqueous products). Because dry heat relies on air for
the transfer of heat to and from the load items, the process takes longer than a steam process for a comparable size item or
load. Lengthy heating and cooling periods require that the load items be unaffected by heat over a long period of time and
also require the use of the overkill method for cycle development and validation.

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USP 39 General Information / 1229.8 Dry Heat Sterilization 1681

Dry heat sterilization is typically performed in the range of 160190 where the objective is sterilization rather than depyro-
genation. (Depyrogenation will be covered separately in Dry Heat Depyrogenation 1228.1). In dry heat sterilization, hot air is
in direct contact with the load items (whether wrapped or unwrapped) and transfers some of its thermal energy. Unlike steam
sterilization, in dry heat sterilization there is no phase change of the heating medium, and thus heat transfer is less efficient.
The items can be stainless steel, glass, ceramic, or other heat-stable materials and may be wrapped or covered with aluminum
foil to protect them during pre- and postprocess handling. Dry heat sterilization is commonly used for heat-stable materials
(e.g., petrolatum or powders).
The limited heat transfer capacity of air requires that items in the oven be placed in locations that were confirmed to be
acceptable during the validation effort. Manufacturers should exercise caution with varying load sizes because in some instan-
ces (resulting from system design and control probe positioning) minimum load sizes may present a worst case.

STERILIZATION CYCLE CONTROL

Process equipment for dry heat sterilization is controlled by calibrated temperature sensors. During the exposure portions of
the cycle, attainment of a minimum dwell time at a predefined temperature is used to document process lethality. Cycle effica-
cy for dry heat sterilization customarily is measured using FH, which typically is defined as the amount of time the load receives
the equivalent of exposure at 170. The FH approach is used to compare sterilization processes that operate at varying temper-
ature conditions to a single standard. The process lethality at temperatures other than 170 can be calculated to determine
lethality equivalent to that provided at 170. Sterilizer control systems must deliver conditions within a predefined timetem-
perature or FH range. Simple mathematics can be used to calculate the total lethality over the course of the process. For the

General Chapters
specific reference temperature of 170 and a z-value (for definitions see Steam Sterilization by Direct Contact 1229.1) of 20,
the FH calculation can be determined by the following equation:

FH = accumulated lethality
t2 = end time
t1 = start time
T = temperature
Accumulation of the lethality (FH) for the sterilization process across the entire cycle (heat-up and cool-down segments inclu-
ded) includes the contribution of those segments and allows the cycle to be defined by a targeted lethality rather than by a
time at a defined minimum temperature.

VALIDATION OF DRY HEAT STERILIZATION

Because dry air has limited heat capacity and dry heat conditions are more variable than those encountered with other ther-
mal sterilization methods, analysts routinely validate their dry heat sterilization procedures using the overkill method as defined
in Sterilization of Compendial Articles 1229.
Overkill sterilization can be defined as a method in which the destruction of a high concentration of a resistant microorgan-
ism supports the destruction of reasonably anticipated bioburden that could be present during routine processing. That objec-
tive can be demonstrated by attaining any of the following: a defined minimum lethality; a defined set of method conditions;
or confirmation of minimum log reduction of a resistant biological indicator.
The validation requirements for the overkill method are less onerous than those of the other sterilization approaches. Be-
cause the load items can withstand substantial amounts of heat without adverse consequence, the greater lethality provided
by the overkill method clearly is justifiable.

Equipment Qualification

Equipment qualification is a predefined program that focuses on the sterilizing equipment to confirm that it has been prop-
erly installed and operates as intended before evaluation of the sterilization process. In some companies, equipment qualifica-
tion is separated into installation qualification and operational qualification or is lumped together under a joint terminology of
installation/operational qualification. The major use of qualification of the sterilizing equipment is to provide a baseline for pre-
ventive maintenance and change control, ensuring reproducibility of operation over time and assurance that the sterilization
process is constantly and accurately performed.

Empty Chamber Temperature Distribution

The equipment should be evaluated for empty chamber temperature distribution. The oven or tunnel should be evaluated
to determine the range of temperatures within the system, and the cycle parameters should be determined to ensure ade-
quate lethality across the expected load. The acceptance criteria for empty chambers can vary with the equipment capabilities

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1682 1229.8 Dry Heat Sterilization / General Information USP 39

and customary use, but it is typically less uniform than observed in steam sterilizers. Biological indicators are not required dur-
ing the evaluation of empty chamber temperature distribution.

Component Mapping

Load items that are complex and feature enclosed volumes and product contact surfaces should be subjected to component
mapping to determine internal cold spots. This is particularly important in powder sterilization. For each load item, manufac-
turers should establish the ability of heat to penetrate the items or containers and to bring them to the required temperature.
These studies can be performed in a laboratory setting and need not be repeated when the same item is sterilized in other
equipment. Thermocouples should be placed into direct contact with the item(s) being evaluated. During component map-
ping load items should be prepared and oriented in a manner that is consistent with how they will be processed.

Load Mapping

Fixed loading patterns for dry heat sterilization in batch ovens are preferable because the limited heat capacity of the air
allows substantial temperature differences across the load. It may be possible to validate maximum and minimum loads as de-
termined by either the number of items or their mass within the oven. Loading in a continuous tunnel process is typically well
defined by the limitations of the conveying system. Load and component mapping ensures that all load items attain the re-
quired temperature. Information from the load mapping is used to adjust cycle timing to ensure appropriate lethality. System
control must consider the relationship between load position and size relative to temperature control locations.
General Chapters

Biological Indicators

The biological indicator (BI) for dry heat sterilization is Bacillus atrophaeus (ATCC 9372), a thermophilic spore-former with
high resistance to dry heat. The spore challenge is placed on a substrate positioned within the load or on a load item. If spores
are used as intended by the BI manufacturer, the population and resistance information provided by the vendor can be used.
End users should determine the population and resistance of their biological indicator used when inoculating their own items.

Heat Penetration and Microbiological Challenge

The core of the validation activity is the confirmation of acceptable heat penetration using temperature measurements and
microbial challenges. Thermocouples and BIs are placed within the load items at the locations determined during the compo-
nent and load mapping to present the worst case. Thermocouples should be placed into direct contact with the item(s) being
monitored. Proof of cycle efficacy is provided by replicate studies in which the BIs are killed and the physical measurements
correspond to the expected values of timetemperature or FH. If the microbial and physical measurements do not correlate,
manufacturers should conduct an investigation and should take corrective action to rectify the discrepancy. This study custom-
arily is performed slightly subminimal to the lower specification limits for time, temperature, and/or cumulative lethality.

ROUTINE PROCESS CONTROL

As with all sterilization processes, after the dry heat sterilization process has been validated, it must be subject to formalized
controls that keep it in a validated state over time. General chapter Sterilization of Compendial Articles 1229 details the general
practices that are appropriate for all sterilization systems. This is accomplished by a number of related practices that are essen-
tial for the continued use of the process over an extended period of time. The essential practices to maintain validated status
include calibration, physical measurements, physical integrators and indicators, ongoing process control, change control, pre-
ventive maintenance, and periodic reassessment and training.

REFERENCES

1. USP General ChaptersMicrobiology Expert Committee. An outline of planned changes to USP 1211 Sterilization and
Sterility Assurance of Compendial Articles. Pharm Forum. 2012; 38(2).

Official from December 1, 2016

Copyright (c) 2017 The United States Pharmacopeial Convention. All rights reserved.
Accessed from 10.6.1.1 by mvpstn3kts on Thu Apr 06 04:42:13 EDT 2017
USP 39 General Information / 1229.10 Radiation Sterilization 1683

1229.10 RADIATION STERILIZATION

INTRODUCTION

Radiation sterilization utilizes the lethal effect of various forms of radiation as a means of microbial destruction. Ionizing radi-
ation (gamma, x-ray, or beam) sterilization is used extensively for the sterilization of medical devices and for a variety of other
materials and products. Nonionizing sterilization methods such as microwave, infrared, x-ray, and ultraviolet light may be use-
ful but have more restricted application, and are outside the scope of this chapter. This chapter provides an overview of sterili-
zation using ionizing radiation and its validation, including dose setting, material compatibility, and dose verification.
The effects of radiation on materials can be substantial and are a major consideration when manufacturers select radiation as
a processing method. The advantages of sterilization by irradiation include simplicity, absence of mechanical complexity, re-
producibility, and overall efficiency. In fact, radiation sterilization is unique because the basis of control essentially is the absor-
bed radiation dose, which can be precisely measured. Methods used to establish appropriate radiation doses to achieve the
desired sterility assurance level are defined in ISO 11137-1 Sterilization of Health Care Products; ISO 11137-2 Sterilization of
Health Care ProductsRadiationPart 2: Establishing the Sterilization Dose; and ISO TS 13004: 2013 Sterilization of Health
Care ProductsRadiationSubstantiation of Selected Sterilization Dose: Method VDmaxSD. These methods include Method
1, Method 2A, Method 2B, and Method VDmax, which differ in the specific testing scheme and the number of articles that are
needed for testing and are based on certain assumptions about bioburden. The use of a biological indicator is inappropriate
during radiation sterilization validation because (a) there are accurate correlations between dose measurement and microbial

General Chapters
destruction for a wide range of microorganisms and (b) the established dose setting methods are based on the material's bio-
burden in its natural state. These correlations have been developed by the medical device industry and provide a direct meth-
odology for process control. Dosimetry plays a central role in radiation sterilization and serves as a direct means for affirming
process lethality. The radiation dose measured in kGy (formerly MRads) is directly related to the lethal effects of the radiation
on microorganisms. The measured dose has the same utility as F0 in steam sterilization. Routine process control for radiation
sterilization is provided by one or more reference dosimeters on the exterior of the packages (after dosimeters have been cor-
related during validation with dose measurement inside the package). The robustness and reliability of the absorbed dose of
the article to be sterilized can support parametric release, as described in Terminally Sterilized Pharmaceutical ProductsPara-
metric Release 1222, for many items.

GAMMA STERILIZATION

Gamma sterilization entails the use of a specifically designed facility where items to be sterilized are exposed to a Co60 radia-
tion source in a manner that ensures uniform dosing. Highly penetrating photons (gamma rays) are emitted from Co60 as it
decays to Ni60. The half-life for this isotope is 5.27 years, which means that over the course of each year the source loses about
12% of its radioactivity. This steady reduction in radioactivity requires that radiation process operators adjust their process con-
trols (typically exposure time) to maintain the established dose required. Periodically, additional Co60 is required to maintain
practical throughput.

X-RAY STERILIZATION

X-ray sterilizers generate highly penetrative photons similar to the gamma photons from Co60 irradiators. X-ray photons are
generated when accelerated electrons impact a target such as tantalum. These systems rely on scanning of materials with x-ray
photons in order to sterilize them. Properly maintained, these systems are able to deliver a constant dose over time. No local
radioactive source is required for x-ray sterilization systems.

E-BEAM STERILIZATION

Electron beam systems rely on scanning of objects with focused electrons to sterilize the items within a defined radiation
field. Properly maintained and controlled, these systems deliver a constant dose, so there is no change in dose with respect to
time. The principal advantages of electron beam sterilization are a much higher dose rate and the absence of a localized radio-
active source. These systems can be installed and operated by the end user. Electron beam penetration is substantially less
than that obtained with photons, and therefore dose mapping is critical to ensure that items of varying density and complexity
are properly sterilized. Because of the high dose rates used with electron beam sterilization, some materials can experience
significantly higher temperatures than the same materials would experience in Co60 irradiation.

VALIDATION OF RADIATION STERILIZATION

Cycle development for radiation requires the identification of an appropriate radiation dose for the objects and confirmation
that the dose does not adversely affect the material's essential quality attributes. In other words, analysts should identify the

Official from December 1, 2016

Copyright (c) 2017 The United States Pharmacopeial Convention. All rights reserved.