1、Designation: F2064 14F2064 17Standard Guide forCharacterization and Testing of Alginates as StartingMaterials Intended for Use in Biomedical and TissueEngineered Medical Product Applications1This standard is issued under the fixed designation F2064; the number immediately following the designation i
2、ndicates the year oforiginal adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. Asuperscript epsilon () indicates an editorial change since the last revision or reapproval.INTRODUCTIONAlginate has found uses in a variety of
3、 products ranging from simple technical applications such asviscosifiers to advanced biomedical matrices providing controlled drug delivery from immobilizedliving cells. As for most hydrocolloids, the functionality of alginate is related to its chemical andstructural composition. The aim of this gui
4、de is to identify key parameters relevant for thefunctionality and characterization of alginates for the development of new commercial applications ofalginates for the biomedical and pharmaceutical industries.1. Scope1.1 This guide covers the evaluation of alginates suitable for use in biomedical or
5、 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 alginates.1.3 As with any material, some characteristics of alginates may be altered
6、 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 offabricated forms of this polymer should be evaluated using test methods that are appropriate to ensure safety and
7、efficacy and arenot addressed in this guide.1.4 WarningMercury has been designated by EPA and many state agencies as a hazardous material that can cause centralnervous system, kidney, and liver damage. Mercury, or its vapor, may be hazardous to health and corrosive to materials. Cautionshould be tak
8、en when handling mercury and mercury-containing products. See the applicable product Material Safety Data Sheet(MSDS) for details and EPAs website (http:/www.epa.gov/mercury/faq.htm) for additional information. Users should be awarethat selling mercury or mercury-containing products, or both, in you
9、r 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 responsibilityof the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatorylim
10、itations prior to use.1.6 This international standard was developed in accordance with internationally recognized principles on standardizationestablished in the Decision on Principles for the Development of International Standards, Guides and Recommendations issuedby the World Trade Organization Te
11、chnical Barriers to Trade (TBT) Committee.2. Referenced Documents2.1 ASTM Standards:2D2196E2975 Test Methods for Rheological Properties of Non-Newtonian Materials by Rotational ViscometerMethod forCalibration or Calibration Verification of Concentric Cylinder Rotational ViscometersF619 Practice for
12、Extraction of Medical Plastics1 This guide is under the jurisdiction of ASTM Committee F04 on Medical and Surgical Materials and Devices and is the direct responsibility of Subcommittee F04.42on Biomaterials and Biomolecules for TEMPs.Current edition approved Oct. 1, 2014March 1, 2017. Published Feb
13、ruary 2015April 2017. Originally approved in 2000. Last previous edition approved in 20002014 asF2064 00 (2006)F2064 14.1. DOI: 10.1520/F2064-14.10.1520/F2064-17.2 For referencedASTM standards, visit theASTM website, www.astm.org, or contactASTM Customer Service at serviceastm.org. For Annual Book o
14、f ASTM Standardsvolume information, refer to the standards Document Summary page on the ASTM website.This document is not an ASTM standard and is intended only to provide the user of an ASTM standard an indication of what changes have been made to the previous version. Becauseit may not be technical
15、ly possible to adequately depict all changes accurately, ASTM recommends that users consult prior editions as appropriate. In all cases only the current versionof the standard as published by ASTM is to be considered the official document.Copyright ASTM International, 100 Barr Harbor Drive, PO Box C
16、700, West Conshohocken, PA 19428-2959. United States1F748 Practice for Selecting Generic Biological Test Methods for Materials and DevicesF749 Practice for Evaluating Material Extracts by Intracutaneous Injection in the RabbitF756 Practice for Assessment of Hemolytic Properties of MaterialsF763 Prac
17、tice for Short-Term Screening of Implant MaterialsF813 Practice for Direct Contact Cell Culture Evaluation of Materials for Medical DevicesF895 Test Method for Agar Diffusion Cell Culture Screening for CytotoxicityF981 Practice for Assessment of Compatibility of Biomaterials for Surgical Implants wi
18、th Respect to Effect of Materials onMuscle and Insertion into BoneF1251 Terminology Relating to Polymeric Biomaterials in Medical and Surgical Devices (Withdrawn 2012)3F1439 Guide for Performance of Lifetime Bioassay for the Tumorigenic Potential of Implant MaterialsF1903 Practice for Testing For Bi
19、ological Responses to Particles In VitroF1904 Practice for Testing the Biological Responses to Particles in vivoF1905 Practice For Selecting Tests for Determining the Propensity of Materials to Cause Immunotoxicity (Withdrawn 2011)3F1906 Practice for Evaluation of Immune Responses In Biocompatibilit
20、y Testing Using ELISATests, Lymphocyte Proliferation,and Cell Migration (Withdrawn 2011)3F2259 Test Method for Determining the Chemical Composition and Sequence in Alginate by Proton Nuclear MagneticResonance (1H NMR) SpectroscopyF2315 Guide for Immobilization or Encapsulation of Living Cells or Tis
21、sue in Alginate GelsF2605 Test Method for Determining the Molar Mass of Sodium Alginate by Size Exclusion Chromatography with Multi-angleLight Scattering Detection (SEC-MALS)2.2 USP Document:4USP Monograph USP 35/NF 30 Sodium Alginate2.3 ISO Documents:5ISO 31-8 Quantities and units Part 8: Physical
22、chemistry and molecular physicsISO 10993 Biological Evaluation of Medical Devices:ISO 10993-1 Biological Evaluation of Medical DevicesPart 1: Evaluation and TestingISO 10993-3 Part 3: Tests for Genotoxicity, Carcinogenicity and Reproductive ToxicityISO 10993-9Part 9: Framework for Identification and
23、 Quantification of Potential Degradation ProductsISO 10993-17Part 17: Methods for Establishment of Allowable Limits for Leachable Substances Using Health-Based RiskAssessmentISO 13408-1: 1998: Aseptic Processing of Health Care ProductsPart 1: General Requirements.2.4 ICH Documents:6International Con
24、ference on Harmonization (ICH) S2 Guidance on Genotoxicity Testing and Data Interpretation forPharmaceuticals Intended for Human UseInternational Conference on Harmonization (ICH) Q1AICH Harmonized Tripartite Guidance for Stability Testing of New DrugSubstances and Products (2003)2.5 FDA Documents:7
25、FDA Interim Guidance for Human and Veterinary Drug Products and Biologicals. Kinetic LAL techniques. DHHS, July 15,19912.6 ANSI Documents:5ANSI/AAMI/ISO 11737-1: 2006 Sterilization of Medical DevicesMicrobiological MethodsPart 1: Estimation of Bioburdenon Product.ANSI/AAMI/ISO 11737-2: 1998 Steriliz
26、ation of Medical DevicesMicrobiological MethodsPart 2: Tests of SterilityPerformed in the Validation of a Sterilization Process2.7 AAMI Documents:8AAMI/ISO 141601998 Sterilization of Single-Use Medical Devices Incorporating Materials of Animal OriginValidationand Routine Control of Sterilization by
27、Liquid Chemical SterilantsAAMI ST67: 2011 Sterilization of Health Care ProductsRequirements and Guidance for Selecting a SterilityAssurance Level(SAL) for Products Labeled “Sterile”AAMI TIR No. 191998 Guidance for ANSI/AAMI/ISO 10993-7: 1995, Biological Evaluation of Medical DevicesPart 7:Ethylene O
28、xide Sterilization Residuals3 The last approved version of this historical standard is referenced on www.astm.org.4 Available from U.S. Pharmacopeia (USP), 12601 Twinbrook Pkwy., Rockville, MD 20852.5 Available from American National Standards Institute (ANSI), 25 W. 43rd St., 4th Floor, New York, N
29、Y 10036.6 Available from ICH Secretariat, c/o IFPMA, 30 rue de St-Jean, P.O. Box 758, 1211 Geneva 13, Switzerland.7 Available from U. S. Food and Drug Administration, 5600 Fishers Lane, Rockville MD 20857-0001.8 Association for the Advancement of Medical Instrumentation 1110 North Glebe Rd., Suite 2
30、20, Arlington, VA 222014795.F2064 1722.8 National Institute of Standards and Technology:9NIST SP811 Special Publication: Guide for the Use of the International System of Units2.9 Other Documents:21CFR184.1724 Listing of Specific Substances Affirmed as GRASSodium Alginate103. Terminology3.1 Definitio
31、ns of Terms Specific to This Standard: (see also Terminology F1251):3.1.1 alginate, na polysaccharide substance containing calcium, magnesium, sodium, and potassium salts obtained from someof the more common species of marine algae. Alginate exists in brown algae as the most abundant polysaccharide,
32、 mainlyoccurring in the cell walls and intercellular spaces of brown seaweed and kelp. Its main function is to contribute to the strengthand flexibility of the seaweed plant.Alginate is classified as a hydrocolloid. The most commonly used alginate is sodium alginate.3.1.2 decomposition, nstructural
33、changes of alginates due to exposure to environmental, chemical or thermal factors, such astemperatures greater than 180C. Decomposition can result in deleterious changes to the alginate.3.1.3 degradation, nchange in the chemical structure, physical properties, or appearance of a material. Degradati
34、on ofpolysaccharides occurs by means of cleavage of the glycosidic bonds, usually by acid catalyzed hydrolysis. Degradation can alsooccur thermally. It is important to note that degradation is not synonymous with decomposition. Degradation is often used as asynonym for depolymerization when referrin
35、g to polymers.3.1.4 depolymerization, nreduction in length of a polymer chain to form shorter polymeric units. Depolymerization mayreduce the polymer chain to oligomeric or monomeric units, or both. In alginates, hydrolysis of the glycosidic bonds is the primarymechanism.3.1.5 Endotoxin, na high-mol
36、ecular weight lipopolysaccharide (LPS) complex associated with the cell wall of gram-negativebacteria that is pyrogenic in humans. Though endotoxins are pyrogens, not all pyrogens are endotoxins.3.1.6 Gabbreviation for -L-guluronic acid, one of the two monomers making up the alginate polysaccharide
37、molecule. G-richalginate has a greater than 50 % content of guluronate residues in the polymer chain. G-block refers to a homopolymeric blockof G residues.3.1.7 hydrocolloid, na water-soluble polymer of colloidal nature when hydrated.3.1.8 Mabbreviation for -D-mannuronic acid, one of the two monomer
38、s making up the alginate polysaccharide chain. M-richalginate has a greater than 50% content of mannuronate residues in the polymer chain.3.1.9 molar mass average, nthe given mass-average molar mass (Mw) of an alginate will always represent an average of allof the molecules in the population. The mo
39、st common ways to express the Mw are as the number average M n! and the weightaverage M w!. The two averages are defined by the following equations:Mn 5(iNiMi(iNiand Mw 5(iwiMi(iwi5(iNiMi2(iNiMi(1)where:Ni = number of molecules having a specific molar mass, Mi, andwi = mass of molecules having a spe
40、cific molar mass, Mi.In a polydisperse molecular population the relation Mw Mn is always valid. The coefficient Mw/Mn is referred to as thepolydispersity index, and will typically be in the range from 1.5 to 3.0 for commercial alginates.3.1.9.1 DiscussionThe term molecular weight (abbreviated MS) is
41、 obsolete and should be replaced by the SI (Systme Internationale) equivalent ofeither relative molecular mass (Mr), which reflects the dimensionless ratio of the mass of a single molecule to an atomic mass unit(see ISO 31-8), or molar mass (M), which refers to the mass of a mole of a substance and
42、is typically expressed as grams/mole.For polymers and other macromolecules, use of the symbols Mw, Mn, and Mz continue, referring to mass-average molar mass,number-average molar mass, and z-average molar mass, respectively. For more information regarding proper utilization of SI units,see NIST SP811
43、.3.1.10 pyrogen, nany substance that produces fever when administered parenterally.9 Available from National Institute of Standards and Technology (NIST), 100 Bureau Dr., Stop 1070, Gaithersburg, MD 20899-1070, http:/physics.nist.gov/cuu/Units/bibliography.html.10 Available from Superintendent of Do
44、cuments, U.S. Government Printing Office, Washington, DC 20402.F2064 1734. Significance and Use4.1 This guide contains a listing of those characterization parameters that are directly related to the functionality of alginate.This guide can be used as an aid in the selection and characterization of t
45、he appropriate alginate for a particular application. Thisguide is intended to give guidance in the methods and types of testing necessary to properly characterize, assess, and ensureconsistency in the performance of a particular alginate. It may have use in the regulation of these devices by approp
46、riate authorities.4.2 The alginate covered by this guide may be gelled, extruded, or otherwise formulated into biomedical devices for use intissue-engineered medical products or drug delivery devices for implantation as determined to be appropriate, based on supportingbiocompatibility and physical t
47、est data. Recommendations in this guide should not be interpreted as a guarantee of clinical successin any tissue engineered medical product or drug delivery application. Further guidance for immobilizing or encapsulating livingcells or tissue in alginate gels can be found in Guide F2315.4.3 To ensu
48、re that the material supplied satisfies requirements for use in TEMPS, several general areas of characterizationshould be considered. These are: identity of alginate, physical and chemical characterization and testing, impurities profile, andperformance-related tests.5. Chemical and Physical Test Me
49、thods5.1 Identity of AlginateThe identity of alginates can be established by several methods including, but not limited to thefollowing:5.1.1 Sodium alginate monograph USP 35/NF30.5.1.2 Fourier Transform Infrared Spectroscopy (FT-IR)Almost all organic chemical compounds absorb infrared radiation atfrequencies characteristic for the functional groups in the compound. A FT-IR spectrum will show absorption bands relating tobond stretching and bending and can therefore serve as a unique fingerprint of a specific compound. Identity o
