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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

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1684 1229.10 Radiation Sterilization / General Information USP 39

minimum sterilization dose as well as the maximum dose the material can withstand without negative effects. With this infor-
mation analysts can set the dose for a specific radiation sterilization application.
Dose setting or dose establishment typically is achieved by following one of the ISO methods. These are Method 1, Method
2A, Method 2B, and Method VDmax. The choice of the most appropriate method depends chiefly on production batch size,
knowledge of the normal bioburden, and the material's sensitivity to radiation. Dose mapping plays an important role through
the cycle development and dose-setting exercise.

Dose Setting

Method 1 is based upon the assignment and verification of a sterilization dose based on a microbial population. The resist-
ance of the microbial population is not determined, and dose setting is based on a standard radiation resistance assigned to
the microbial population, derived from data obtained from medical device manufacturers and from the literature. This analysis
assumes that the distribution of standard resistance represents a more severe challenge than the natural microbial population
on the material to be sterilized. A verification dose study should confirm the relative resistance assumption. The VDmax method
is similar to Method 1 (it requires both bioburden and dose verification testing) but relies on bioburden ranges (e.g., <1000
CFUs per item for a 25-kGy sterilization dose and, for example, 0.11.5 CFUs per item for a 15-kGy sterilization dose).
The more complex Method 2 does not require the enumeration of the microbial population for the purpose of setting the
sterilization dose (although it is required for routine monitoring and control) but uses a series of incremental dose exposures to
establish a dose at which approximately 1 out of 100 samples irradiated at that dose will be nonsterile. This is not the steriliza-
tion dose, but it provides the basis to determine the sterilization dose by extrapolation from this information.
General Chapters

Material Compatibility

Once the required dose level has been established, the maximum dose should be established. Analysts typically establish the
maximum dose by evaluating the highest likely dose that might be seen during the sterilization process, adding a safety factor,
and evaluating the item for immediate and long-term effects of the radiation exposure. Some materials may appear un-
changed initially, and the effects may become evident only over time. The evaluation should consider all of the materials ex-
posed to the radiation processing, especially the drug product and its primary container. Product stability, safety, and func-
tionality should be confirmed over the product's intended use period.

Dose Verification

The methods for cycle development and dose setting rely on the bioburden approach. Analysts use defined presterilization
bioburden controls and periodic evaluation of the process effects on the bioburden to maintain cycle efficacy. Establishing the
required dose for microbial destruction during cycle development uses the bioburden's natural resistance; analysts then extrap-
olate the dose-setting algorithms to establish a dose that is capable of delivering a probability of a nonsterile unit (PNSU, a
standard measurement) of 1 106. The results of the dose-setting approaches for initial bioburden with different populations
and resistance to radiation sterilization are depicted in Figure 1.

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USP 39 General Information / 1229.10 Radiation Sterilization 1685

General Chapters
Figure 1. Results of radiation dose setting using VDmax. (top) Higher bioburden population, higher resistance to radiation
sterilization. (bottom) Lower bioburden population, lower resistance to radiation sterilization.

Validation Activities

Confirmation of appropriate dose delivery when using the sterilization dose requires a number of supportive activities.

EQUIPMENT QUALIFICATION

The use of gamma sterilization requires initial and periodic assessment of equipment controls and parameters necessary to
establish the system's capability. Sterilization systems that deliver directed beams or rays have controls for scan speed, source
intensity, and system timers. The other elements of radiation sterilization equipment largely are related to material transport
and are easily qualified. Qualification of safety controls, devices, and software is required.

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1686 1229.10 Radiation Sterilization / General Information USP 39

EMPTY CHAMBER DOSE MAPPING

This optional exercise entails mapping the target area for radiation dose in the absence of a load and is a possible means to
evaluate a focused beam or ray system. It provides a baseline of performance that may be useful over time.

LOAD DOSE MAPPING

The arrangement of items in irradiation containers, carriers, or pallets is an essential part of the initial validation exercise. The
goal of the mapping is to define the distribution of a dose throughout the load items and establish a configuration that mini-
mizes dose variation across the materials. The items are mapped using multiple dosimeters positioned internally and externally.
Identification of maximum dose location is important in evaluating the effects of the radiation on the load items. The location
of minimum and maximum dose can be identified from the dose mapping data for monitoring in routine sterilization of mate-
rials.

BIOLOGICAL INDICATORS

The use of biological indicators for radiation sterilization is not indicated because the physical and dosimetric measurements
employed are more reliable, reproducible, and robust than biological systems.

DOSIMETRY
General Chapters

Process control for radiation sterilization relies heavily on dosimetry for both initial development and ongoing verification.
For guidance on the selection and use of a dosimetry system for use in radiation sterilization refer to ASTM E2628 Practice for
Dosimetry in Radiation Processing. The dosimeters and the instruments used with them should be calibrated according to ISO/
ASTM 51261 Practice for Calibration of Routine Dosimetry Systems for Radiation Processing.

PROCESS CONFIRMATION

The core of the validation activity is the confirmation of acceptable lethality using dosimeters that are positioned across the
material as it is processed through the radiation-sterilizing equipment. Proof of sterilization cycle efficacy is provided in repli-
cate studies in which the dosimetry results correspond to the required minimum value for sterility assurance and demonstrate
that the maximum value has not been exceeded.

Routine Process Control

Radiation sterilization should be subject to formal controls that maintain the validated status. The practices outlined in Gen-
eral Principles of Sterilization of Compendial Articles 1229 provide the general requirements appropriate for all sterilization sys-
tems. This is accomplished by a number of related practices that are essential for the continued use of the process over an
extended period of time. The practices that are essential to maintain validated status for radiation include training, calibration,
physical measurements, bioburden monitoring, change control, preventive maintenance, and periodic dose audits.

REFERENCES

1. Jacobs, G., Validation of the Radiation Sterilization of Pharmaceuticals, chapter in Agalloco, J., & Carleton, F. J. (eds.),
Validation of Pharmaceutical Processes: 3rd Edition, InformaUSA, New York, 2007.
2. Herring, C., & Saylor, M., Sterilization with Radioisotopes, chapter in Morrissey, R., & Phillips, G. B. (eds.), Sterilization
TechnologyA Practical Guide for Manufacturers and Users of Health Care Products, Van Nostrand Reinhold, New York,
1993.
3. Cleland, M., O'Neill, T., & Thompson, C., Sterilization with Accelerated Electrons, chapter in Morrissey, R., & Phillips, G.
B., Sterilization TechnologyA Practical Guide for Manufacturers and Users of Health Care Products, Van Nostrand Reinhold,
New York, 1993.

Official from December 1, 2016

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USP 39 General Information / 1229.11 Vapor Phase Sterilization 1687

1229.11 VAPOR PHASE STERILIZATION

INTRODUCTION

Sterilization can be accomplished using sporicidal agents suspended in air (i.e., vapor). Sterilizing agents that operate in this
fashion include hydrogen peroxide (H2O2), peracetic acid (CH3CO3CH), formaldehyde (CH2O), and glutaraldehyde
[CH2(CH2CHO)2] in aqueous solution. At room temperature these are liquids or solids that can be vaporized for introduction
into a vessel or chamber. They differ from sterilizing gases and liquids in that there are multiple phases within the vessel during
sterilization. Vapor sterilization systems are well suited for heat-sensitive materials and surface sterilization. Items exposed to
the process should have their surfaces exposed to the greatest extent possible. Vapor sterilization processes require appropriate
sterilant concentration, temperature, and relative humidity, all of which may be variable during the exposure period. Because
the agent is ordinarily supplied as an aqueous solution, moisture is introduced with the agent. The consequences of variation
in these parameters may be localized differences in relative humidity, agent concentration, and condensation rates on the sur-
faces to be treated, resulting in variations in process lethality. The parameters to be established include sterilant amount (usual-
ly derived from injection quantities), relative humidity, and temperature. There is no demonstrated correlation between gas
phase conditions, surface conditions, and microbial kill. For this reason, online monitoring of vapor phase concentration is not
widely utilized as a control parameter. Efforts to develop a standardized biological indicator for vapor systems have been ham-
pered by the multiphasic nature of these sterilants. Selection of the appropriate biological indicator (BI) and resistance should
be based on experimentation within the user's system. Only under well-defined, specific conditions (e.g., agent concentration,

General Chapters
humidity level, temperature, substrate, and phase) can a reliable D-value be established (for a definition of D-value, see Sterili-
zation of Compendial Articles 1229).
This chapter will briefly review hydrogen peroxide and peracetic acid sterilization systems, because they are more widely
used than other vapor phase sterilizing agents in the pharmaceutical and medical device industries.

HYDROGEN PEROXIDE

The efficacy of hydrogen peroxide as a liquid sterilant has been long established.1 There are several effective approaches to
hydrogen peroxide (H2O2) injection, including continuous, intermittent, or all at once. Some of the systems utilize an evacua-
tion or drying step prior to introduction of the hydrogen peroxide (H2O2) to allow for increased concentration without excess
condensation. Alternatively, hydrogen peroxide (H2O2) can be introduced as a liquid, followed by target heating. Following
the exposure period, the chamber or target is aerated to an acceptable level for further processing of materials and/or person-
nel exposure (whichever is lowest) prior to opening and removing the sterilized article.

PERACETIC ACID

Peracetic acid (CH3CO3CH), alone or mixed with hydrogen peroxide, is a sporicidal agent that has been proven effective.1
Peracetic acid is introduced as a liquid through an atomizer, resulting in the presence of liquid and vapor phases within the
chamber. After the process dwell period, it is removed by evaporation.

VALIDATION OF VAPOR STERILIZATION

Standard sterilizing conditions have not been defined due to the varying phase and multiphase nature of the sterilant during
sterilization processes. Therefore, no standardized BIs with D-values that may be used for conventional predictive analysis of kill
rates exist. In the absence of BI D-values, and due to the variation in vapor sterilization cycle parameters, an empirical ap-
proach must be used. The kill rates in the gas and liquid phases that constitute the vapor may be substantially different (liquid
kill rates are considered greater than gaseous kill rates). Destruction of BIs distributed across the system or load demonstrates
lethality regardless of which phase effects the kill. Sterilization process parameters (usually time) that do not kill the BIs may be
adjusted until a complete kill is achieved. This establishes the minimum conditions necessary for a complete kill. Vapor steriliza-
tion may be validated using a half-cycle, bracketing, or other suitable approach as defined in 1229 and Gaseous Sterilization
1229.7.
The half-cycle validation method requires the destruction of a suitable concentration of a resistant microorganism under de-
fined, minimum conditions for a complete kill. Then, in routine operation, the minimum lethal time period is arbitrarily dou-
bled, which supports a doubling of the spore log reduction of the BI, and is more than sufficient to inactivate the bioburden.
A bracketing approach, which better supports process operating ranges for the critical parameters than does the half-cycle
method, also can be employed. In the bracketing method, one evaluates conditions that bracket the defined process condition
in order to establish parameters for the minimum and maximum effects on the materials and bioburden. The minimum lethal

1 Block SS, editor. Disinfection, Sterilization, and Preservation. 5th ed. Philadelphia: Lippincott Williams & Wilkins; c2001. Chapter 9, Peroxygen compounds; pp.

185204.

Official from December 1, 2016

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