ASTM F2103-01(2007)e2
(Guide)Standard Guide for Characterization and Testing of Chitosan Salts as Starting Materials Intended for Use in Biomedical and Tissue-Engineered Medical Product Applications
Standard Guide for Characterization and Testing of Chitosan Salts as Starting Materials Intended for Use in Biomedical and Tissue-Engineered Medical Product Applications
SIGNIFICANCE AND USE
This guide contains a listing of those characterization parameters that are directly related to the functionality of chitosan. This guide can be used as an aid in the selection and characterization of the appropriate chitosan or chitosan salt for a particular application. This standard is intended to give guidance in the methods and types of testing necessary to properly characterize, assess, and ensure consistency in the performance of a particular chitosan. It may have use in the regulation of devices containing chitosan by appropriate authorities.
The chitosan salts covered by this guide may be gelled, extruded, or otherwise formulated into biomedical devices for use as tissue-engineered medical products or drug delivery devices for implantation as determined to be appropriate, based on supporting biocompatibility and physical test data. Recommendations in this guide should not be interpreted as a guarantee of clinical success in any tissue-engineered medical product or drug delivery application.
To ensure that the material supplied satisfies requirements for use in TEMPs, several general areas of characterization should be considered. These include identity of chitosan, physical and chemical characterization and testing, impurities profile, and performance-related tests.
SCOPE
1.1 This guide covers the evaluation of chitosan salts suitable for use in biomedical or pharmaceutical applications, or both, including, but not limited to, tissue-engineered medical products (TEMPS).
1.2 This guide addresses key parameters relevant for the functionality, characterization, and purity of chitosan salts.
1.3 As with any material, some characteristics of chitosan may be altered by processing techniques (such as molding, extrusion, machining, assembly, sterilization, and so forth) required for the production of a specific part or device. Therefore, properties of fabricated forms of this polymer should be evaluated using test methods that are appropriate to ensure safety and efficacy.
1.4 Warning—Mercury has been designated by EPA and many state agencies as a hazardous material that can cause central nervous system, kidney, and liver damage. Mercury, or its vapor, may be hazardous to health and corrosive to materials. Caution should be taken when handling mercury and mercury-containing products. See the applicable product Material Safety Data Sheet (MSDS) for details and EPA’s website (http://www.epa.gov/mercury/faq.htm) for additional information. Users should be aware that selling mercury or mercury-containing products, or both, in your state may be prohibited by state law.
1.5 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use.
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Designation:F2103–01 (Reapproved 2007)
Standard Guide for
Characterization and Testing of Chitosan Salts as Starting
Materials Intended for Use in Biomedical and Tissue-
Engineered Medical Product Applications
This standard is issued under the fixed designation F2103; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
´ NOTE—Formatting and grammar were corrected editorially throughout in April 2007.
´ NOTE—Mercury warning was editorially added in April 2008.
INTRODUCTION
Biopolymers from marine sources have been studied and used in commercial applications and
product development for a number of years. Chitosan, a linear polysaccharide consisting of
glucosamine and N-acetyl glucosamine derived mainly from crustacean shells, has been used in many
technical applications such as water purification (as a flocculant), in cosmetics, and recently as a
proposed fat-binding weight control product. In solution, the cationic nature of chitosan gives this
polymer a mucoadhesive property. Chitosan salts can be used as a matrix or scaffold material as well
as in non-parenteral delivery systems for challenging drugs. Chitosan salts have been shown to
increase the transport of polar drugs across the nasal epithelial surface. The purpose of this guide is
to identify key parameters relevant for the functionality and characterization of chitosan salts for the
development of new commercial applications of chitosan salts for the biomedical and pharmaceutical
industries.
1. Scope materials.Cautionshouldbetakenwhenhandlingmercuryand
mercury-containing products. See the applicable product Ma-
1.1 This guide covers the evaluation of chitosan salts
terial Safety Data Sheet (MSDS) for details and EPA’s website
suitable for use in biomedical or pharmaceutical applications,
(http://www.epa.gov/mercury/faq.htm) for additional informa-
or both, including, but not limited to, tissue-engineered medi-
tion. Users should be aware that selling mercury or mercury-
cal products (TEMPS).
containingproducts,orboth,inyourstatemaybeprohibitedby
1.2 This guide addresses key parameters relevant for the
state law.
functionality, characterization, and purity of chitosan salts.
1.5 This standard does not purport to address all of the
1.3 As with any material, some characteristics of chitosan
safety concerns, if any, associated with its use. It is the
may be altered by processing techniques (such as molding,
responsibility of the user of this standard to establish appro-
extrusion, machining, assembly, sterilization, and so forth)
priate safety and health practices and determine the applica-
required for the production of a specific part or device.
bility of regulatory limitations prior to use.
Therefore, properties of fabricated forms of this polymer
should be evaluated using test methods that are appropriate to
2. Referenced Documents
ensure safety and efficacy.
2.1 ASTM Standards:
1.4 Warning—Mercury has been designated by EPA and
D2196 Test Methods for Rheological Properties of Non-
many state agencies as a hazardous material that can cause
Newtonian Materials by Rotational (Brookfield type) Vis-
central nervous system, kidney, and liver damage. Mercury, or
cometer
its vapor, may be hazardous to health and corrosive to
F619 Practice for Extraction of Medical Plastics
This guide is under the jurisdiction of ASTM Committee F04 on Medical and
Surgical Materials and Devices and is the direct responsibility of Subcommittee
F04.42 on Biomaterials and Biomolecules for TEMPs.
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
Current edition approved Feb. 1, 2007. Published February 2007. Originally
contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM
approved in 2001. Last previous edition approved in 2001 as F2103 – 01. DOI:
Standards volume information, refer to the standard’s Document Summary page on
10.1520/F2103-01R07E02. the ASTM website.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959, United States.
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F2103–01 (2007)
F748 Practice for Selecting Generic Biological Test Meth- ance for Industry S2B Genotoxicity: A Standard Battery
ods for Materials and Devices for Genotoxicity Testing of Pharmaceuticals 62 FR
F749 Practice for Evaluating Material Extracts by Intracu- 62472
taneous Injection in the Rabbit International Conference on Harmonization (1994) Guide-
F756 Practice for Assessment of Hemolytic Properties of line for Industry S5A Detection of Toxicity to Reproduc-
Materials tion for Medicinal Products 59 FR 48746
F763 Practice for Short-Term Screening of Implant Materi- International Conference on Harmonization (1996) Guid-
als ance for Industry S5B Detection of Toxicity to Reproduc-
F813 Practice for Direct Contact Cell Culture Evaluation of tion for Medicinal Products: Addendum on Toxicity to
Materials for Medical Devices Male Fertility 61 FR 15360
F895 Test Method for Agar Diffusion Cell Culture Screen- International Conference on Harmonization (1996) Guide-
ing for Cytotoxicity line for Industry S1A The Need for Long-term Rodent
F981 Practice forAssessment of Compatibility of Biomate- Carcinogenicity Studies of Pharmaceuticals 61 FR 8153
rials for Surgical Implants with Respect to Effect of International Conference on Harmonization (1998) Guid-
Materials on Muscle and Bone ance for Industry S1B Testing for Carcinogenicity of
F1251 Terminology Relating to Polymeric Biomaterials in Pharmaceuticals 63 FR 8983
Medical and Surgical Devices International Conference on Harmonization (1995) Guide-
F1439 Guide for Performance of Lifetime Bioassay for the line for Industry S1C Dose Selection for Carcinogenicity
Tumorigenic Potential of Implant Materials Studies of Pharmaceuticals 60 FR 11278
F1903 Practice for Testing For Biological Responses to International Conference on Harmonization (1997) S1C[R]
Particles in vitro Guidance for Industry Addendum to Dose Selection for
F1904 Practice for Testing the Biological Responses to Carcinogenicity Studies of Pharmaceuticals:Addition of a
Particles in vivo Limit Dose and Related Notes 62 FR 64259
F1905 Practice For Selecting Tests for Determining the International Conference on Harmonization (ICH) Q1AICH
Propensity of Materials to Cause Immunotoxicity Harmonized Tripartite Guidance for Stability Testing of
F1906 Practice for Evaluation of Immune Responses In New Drug Substances and Products (September 23,
Biocompatibility Testing Using ELISATests, Lymphocyte 1994)
Proliferation, and Cell Migration 2.5 FDA Documents:
2.2 Ph. Eur. Document: FDA Guideline on Validation of the Limulus Amebocyte
Ph. Eur. Monograph Chitosan Chloride, Nov. 2000 Test as an End-Product Endotoxin Test for Human and
2.3 ISO Documents: AnimalParenteralDrugs,BiologicalProductsandHealth-
4 6
ISO 10993 Biological Evaluation of Medical Devices care Products DHHS, December 1987
ISO 10993-1 Biological Evaluation of Medical Devices— FDA Interim Guidance for Human and Veterinary Drug
Part 1: Evaluation and Testing Products and Biologicals. Kinetic LAL Techniques-
ISO10993-3—Part3: TestsforGenotoxicity,Carcinogenic- DHHS, July 15, 1991
ity and Reproductive Toxicity 2.6 ANSI Documents:
ISO 10993-9—Part 9: Framework for Identification and ANSI/AAMI/ISO 11737-1: 1995 Sterilization of Medical
Quantification of Potential Degradation Products Devices—Microbiological Methods—Part 1: Estimation
ISO 10993-17—Part 17: Methods for Establishment of of Bioburden on Product
AllowableLimitsforLeachableSubstancesUsingHealth- ANSI/AAMI/ISO 11737-2: 1998 Sterilization of Medical
Based Risk Assessment Devices—Microbiological Methods—Part 2:Tests of Ste-
ISO 13408-1: 1998: Aseptic Processing of Health Care rility Performed in the Validation of a Sterilization Pro-
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Products—Part 1: General Requirements cess
2.4 ICH Documents: 2.7 AAMI Documents:
International Conference on Harmonization (1997) Guid- AAMI TIR No. 19—1998: Guidance for ANSI/AAMI/ISO
ance for Industry M3 Nonclinical Safety Studies for the 10993–7: 1995, Biological Evaluation of Medical
Conduct of Human ClinicalTrials for Pharmaceuticals 62 Devices—Part 7: Ethylene Oxide Sterilization Residuals
FR 62922 AAMI/ISO 14160—1998: Sterilization of Single-Use
International Conference on Harmonization (1996) Guide- Medical Devices Incorporating Materials of Animal
line for Industry S2A Specific Aspects of Regulatory Origin—Validation and Routine Control of Sterilization
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Genotoxicity Tests for Pharmaceuticals 61 FR 18199 by Liquid Chemical Sterilants
International Conference on Harmonization (1997) Guid- AAMI ST67/CDV-2: 1999: Sterilization of Medical
Devices—Requirements for Products Labeled “Sterile”
Available from EDQM, Publications and Services European Pharmacopoeia,
BP 907 226, avenue de Colmar, F-67029 Strasbourg Cedex 1, France.
4 6
Available fromAmerican National Standards Institute (ANSI), 25 W. 43rd St., Available from Food and Drug Administration (FDA), 5600 Fishers Ln.,
4th Floor, New York, NY 10036, http://www.ansi.org. Rockville, MD 20857, http://www.fda.gov.
5 7
Available from ICH Secretariat, c/o IFPMA, 30 rue de St-Jean, PO Box 758, Association for the Advancement of Medical Instrumentation, 111 N. Glebe
1211 Geneva 13, Switzerland. Rd., Suite 220, Arlington, VA 22201–4795.
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F2103–01 (2007)
2.8 EN Documents:
where:
N = number of molecules having a specific molecular
EN 12442-1 Animal Tissues and Their Derivative Utilized
i
in the Manufacture of Medical Devices—Part 1:Analysis weight M and
i
w = weight of molecules having a specific molecular
and Management of Risk
i
weight M. In a polydisperse molecular population the
i
EN 12442-Part 3: Validation of the Elimination and/or
– – –
Inactivation of Virus and Transmissible Agents relation M > M is always valid. The coefficient M /
w n w
–
M is referred to as the polydispersity index, and will
n
3. Terminology
typically be in the range 1.5 to 3.0 for commercial
3.1 Definitions: chitosans.
3.1.1 chitosan, n—a linear polysaccharide consisting of 3.1.8 pyrogen, n—any substance that produces fever when
b(1→4) linked 2-acetamido-2-deoxy-D-glucopyranose administered parenterally.
(GlcNAc) and 2-amino-2-deoxy-D-glucopyranose (GlcN).
4. Significance and Use
3.1.1.1 Discussion—Chitosan is a polysaccharide derived
by N-deacetylation of chitin.
4.1 This guide contains a listing of those characterization
3.1.2 decomposition,n—structuralchangesofchitosansasa
parameters that are directly related to the functionality of
result of exposure to environmental, chemical, or thermal
chitosan. This guide can be used as an aid in the selection and
factors, such as temperatures greater than 200°C.
characterization of the appropriate chitosan or chitosan salt for
3.1.2.1 Discussion—Decomposition can result in deleteri-
a particular application. This standard is intended to give
ous changes to the chitosan.
guidance in the methods and types of testing necessary to
3.1.3 degradation, n—change in the chemical structure,
properly characterize, assess, and ensure consistency in the
physical properties, or appearance of a material. performance of a particular chitosan. It may have use in the
3.1.3.1 Discussion—Degradation of polysaccharides occurs
regulation of devices containing chitosan by appropriate au-
by means of cleavage of the glycosidic bonds, usually by acid thorities.
—catalyzed hydrolysis. Degradation can also occur thermally.
4.2 The chitosan salts covered by this guide may be gelled,
Note that degradation is not synonymous with decomposition. extruded, or otherwise formulated into biomedical devices for
Degradation is often used as a synonym for depolymerization
use as tissue-engineered medical products or drug delivery
when referring to polymers. devicesforimplantationasdeterminedtobeappropriate,based
3.1.4 degree of deacetylation, n—the fraction or percentage on supporting biocompatibility and physical test data. Recom-
of glucosamine units (deacetylated monomers) in a chitosan mendations in this guide should not be interpreted as a
polymer molecule. guarantee of clinical success in any tissue-engineered medical
3.1.5 depolymerization, n—reductioninlengthofapolymer product or drug delivery application.
4.3 To ensure that the material supplied satisfies require-
chain to form shorter polymeric units.
3.1.5.1 Discussion—Depolymerization may reduce the ments for use in TEMPs, several general areas of characteriza-
tion should be considered. These include identity of chitosan,
polymer chain to oligomeric or monomeric units, or both. In
chitosan, hydrolysis of the glycosidic bonds is the primary physical and chemical characterization and testing, impurities
profile, and performance-related tests.
mechanism.
3.1.6 endotoxin, n—a high-molecular-weight lipopolysac-
5. Chemical and Physical Test Methods
charide (LPS) complex associated with the cell wall of
gram-negative bacteria that is pyrogenic in humans. 5.1 Identity of Chitosan—The identity of chitosan and
3.1.6.1 Discussion—Though endotoxins are pyrogens, not
chitosan salts can be established by several methods including,
all pyrogens are endotoxins. but not limited to the following:
3.1.7 molecular mass average (molecular weight average), 5.1.1 Chitosan chloride monograph Ph. Eur.
n—the given molecular weight (Mw) of a chitosan will always 5.1.2 Fourier Transform Infrared Spectroscopy (FT-IR)—
represent an average of all of the molecules in the population. Almost all organic chemical compounds absorb infrared radia-
The most common ways to express the Mw are as the number tion at frequencies characteristic for the functional groups in
– –
the compound. A FT-IR spectrum will show absorption bands
average (M ) and the weight average (M ). The two averages
n w
relating to bond stretching and bending and can therefore serve
are defined by the following equations:
as a unique fingerprint of a specific compound. Cast a chitosan
NM
(i i i
–
film from a 0.25 % (w/v) solution of chitosan (in 1 % acetic
M 5
n
N
(i i
acid) or chitosan salt (dissolved in water) by drying approxi-
mately 500 µL of the sample onto a disposable IR card for 3
and
to4hat 60°C. Record a background spectrum between 4000
WM NM
( (
– i i i i i i -1
and400cm-1using128scansataresolutionof4cm .Record
M 5 5
w
W NM
(i i (i i i
the IR spectrum of a dried blank IR card, then record the IR
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Available from European Committee for Standardization, CEN Management NosuitablecommerciallyavailableIRcar
...
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