1、Designation: F720 13F720 17Standard Practice forTesting Guinea Pigs for Contact Allergens: Guinea PigMaximization Test1This standard is issued under the fixed designation F720; the number immediately following the designation indicates the year of originaladoption or, in the case of revision, the ye
2、ar of last revision.Anumber in parentheses indicates the year of last reapproval.Asuperscriptepsilon () indicates an editorial change since the last revision or reapproval.1. Scope1.1 This practice is intended to determine the potential for a substance, or material extract, to elicit contact dermal
3、allergenicity.1.2 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.1.3 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
4、to establish appropriate safety safety, health and healthenvironmental practices and determine theapplicability of regulatory limitations prior to use.1.4 This international standard was developed in accordance with internationally recognized principles on standardizationestablished in the Decision
5、on Principles for the Development of International Standards, Guides and Recommendations issuedby the World Trade Organization Technical Barriers to Trade (TBT) Committee.2. Referenced Documents2.1 ASTM Standards:2F619 Practice for Extraction of Medical Plastics2.2 ISO Standards:3ISO 1099310 Biologi
6、cal evaluation of medical devicesPart 10: Tests for irritation and skin sensitization3. Summary of Practice3.1 After a two-stage induction employing Freunds complete adjuvant and sodium lauryl sulfate, the substance or extract isplaced on patches and then placed on the skin of guinea pigs.After 24 h
7、, the patches are removed and the skin examined for allergicreaction, and the intensity of the reaction scored at the time of removal and 24 and 48 h subsequent to removal.4. Significance and Use4.1 In selecting a new material for human contact in medical applications, it is important to ensure that
8、 the material will notstimulate the immune system to produce an allergic reaction. The reaction would be due to substances which could leach out ofa material. Therefore, this practice provides for using material extracts. The rationale for this practice is based on the fact that theguinea pig has be
9、en shown to be the best animal model for human allergic contact dermatitis. The use of Freunds completeadjuvant and sodium lauryl sulfate tends to enhance the potential of a material to cause an allergy. Therefore, this test, while notguaranteeing that a material is nonallergenic, is the most severe
10、 animal test in common use today.5. Materials and Manufacturer5.1 Hartley Strain Guinea Pigs, male, 300 to 500 g.5.1.1 Ten animals are used for each test material.5.2 Freunds Complete Adjuvant.5.3 Occlusive Surgical Tape, 3.75 cm in width.1 This practice is under the jurisdiction of ASTM Committee F
11、04 on Medical and Surgical Materials and Devices and is the direct responsibility of Subcommittee F04.16on Biocompatibility Test Methods.Current edition approved June 1, 2013Sept. 1, 2017. Published July 2013September 2017. Originally approved in 1981. Last previous edition approved in 20122013 asF7
12、20 81 (2012).F720 13. DOI: 10.1520/F0720-13.10.1520/F0720-17.2 For referencedASTM standards, visit theASTM website, www.astm.org, or contactASTM Customer Service at serviceastm.org. For Annual Book of ASTM Standardsvolume information, refer to the standards Document Summary page on the ASTM website.
13、3 Available from International Organization for Standardization (ISO), 1, ch. de la Voie-Creuse, CP 56, CH-1211 Geneva 20, Switzerland, http:/www.iso.org.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
14、 the previous version. Becauseit may not be technically 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.Copyrigh
15、t ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States15.4 Elastic Bandage.5.5 Sodium Lauryl Sulfate (10 weight %) in USP petroleum jelly.5.6 Positive Control Substance.5.6.1 5 % formaldehyde for water-soluble test substances.6. Preparation of Test
16、Samples6.1 Samples for Intradermal Injection:6.1.1 Water-Soluble Constituents or Water Extract Liquids:6.1.1.1 Dissolve the water-soluble constituent up to its maximum solubility, not to exceed a concentration of 10 weight %, orobtain a water extraction liquid as described in Practice F619.6.1.1.2 C
17、ombine equal volumes of the liquid described in 6.1.1.1 and Freunds complete adjuvant. Homogenize by continuousand vigorous vortex mixing for a minimum of 5 min. Emulsification is complete when a drop placed on the surface of a water-icebath remains intact.6.1.1.3 Also prepare the constituent or ext
18、ract to the same concentration in water without Freunds complete adjuvant.6.1.2 Oil Soluble Constituents:6.1.2.1 Dissolve oil-soluble constituents in Freunds complete adjuvant to a concentration of 10 weight %.6.1.2.2 Combine equal volumes of the 10 % Freunds adjuvant solution with an equal volume o
19、f water by slowly adding thewater to the adjuvant while homogenizing with a rotating stirrer. Homogenize by continuous and vigorous mixing for a minimumof 5 min. Emulsification is complete when a drop placed on the surface of the water in a water-ice bath remains intact.6.1.2.3 Also prepare the cons
20、tituent to an equal concentration without Freunds complete adjuvant.6.1.3 Vegetable Oil Extract Liquids:6.1.3.1 Mix equal volumes of oil extract liquid obtained in accordance with Practice F619 with an equal volume of Freundscomplete adjuvant.6.1.3.2 Also prepare extract to an equal concentration in
21、 water without Freunds complete adjuvant.6.1.4 Alternative Sample Preparation: As an alternative method for sample extraction, the method described in ISO 1099310,Annex E is recommended for consideration for the extraction of polymeric materials.6.1.5 Prepare control substances consistent with 6.1.1
22、 6.1.3.2.NOTE 1If the final concentration of the suspected allergen results in deleterious effects such as ulceration, necrosis, or systemic toxicity, use themaximum tolerable concentration.6.2 Samples for Topical Application:6.2.1 LiquidsFor all liquids, use the highest concentration not causing ex
23、cessive irritation or deleterious to the general healthof test animals. (See Note 1.)6.2.1.1 For oil-miscible liquids, dilute with petroleum jelly if necessary.6.2.1.2 For water-miscible liquids, dilute with water if necessary.7. Procedure for Test and Control Substances7.1 Induction:7.1.1 Intraderm
24、al Injection:7.1.1.1 Clip the shoulder region of each guinea pig free of hair, exposing a 4 by 6-cm area.7.1.1.2 Three injection sites on each side of the spine should be identified with at least 1.5 cm between sites. Intradermalinjections are then made as indicated below, with Site 1 being closest
25、to the animals head.(1) Site 10.1 mL of adjuvant without test sample(2) Site 20.1 mL of test sample without adjuvant(3) Site 30.1 mL of test sample emulsified in complete adjuvant7.1.2 Topical Application:7.1.2.1 One week after the intradermal injections, clip the test area of the guinea pigs.7.1.2.
26、2 If the test sample is non-irritating, treat each test area with 10 % sodium lauryl sulfate (SLS) in petroleum jelly, 24 hbefore applying the test patches. Massage the SLS into the skin with a glass rod.7.1.2.3 Apply the test sample to a 2 by 4-cm patch of qualitative filter paper. Apply the test a
27、gent in petroleum jelly Foroil-miscible liquids, if they have been diluted in accordance with section 6.2.1.1, apply the diluted liquid to the filter in a thick,even layer. Apply Otherwise, apply the test liquid or the test extract neat to the test paper until saturated.7.1.2.4 Apply filter paper to
28、 the injection site of the guinea pigs. Cover with 3.75-cm occlusive surgical tape and wrap an elasticbandage around the torso to secure the tape.7.1.2.5 Leave in place for 48 h.7.2 Challenge:7.2.1 Two weeks following the induction described in 7.1, perform the following:7.2.2 Shave a 5 by 5-cm area
29、 on theanimals flank.F720 1727.2.3 Delivery of Test Sample:7.2.3.1 Test solids as a 25 % concentration (providing this concentration is not irritating) in petroleum jelly by weight. Applyon a 2 by 2-cm piece of filter paper as outlined in 7.1.2.3.7.2.3.2 Test liquids in undiluted form consistent wit
30、h 6.1.5. Apply on a 2 by 2-cm piece of filter paper as outlined in 7.1.2.3.7.2.4 Cover the filter paper and sample with occlusive surgical tape and an elastic bandage as described in 7.1.2.4.7.2.5 Leave the samples in place for 24 h.8. Interpretation of Results8.1 Read the challenge sites within 1 h
31、 after removing the patches and at 24 and 48 h after removing the patches. Grade eachsite for erythema and edema in accordance with Table 1.8.2 Prepare a tabular listing of the reactions at 1, 24, and 48 h. Any animal showing a reaction at 24 or 48 h of two or greaterfor erythema and edema shall be
32、considered sensitized.8.3 Rate the allergenicity of the test material in accordance with Table 2.8.3.1 If a significant number (more than 50 %) of animals show a reaction score of one, repeat the test with an additional tenanimals.8.3.2 If 60 % of the animals in the positive control group do not sho
33、w a reaction of two or greater, repeat the test.9. Report9.1 The report shall include the following:9.1.1 Test material description, generic name, product name, and lot number.9.1.2 Method of preparation, that is, extraction liquid used, and form of exposure.9.1.3 Any necessary dilutions due to seve
34、re reactions.9.1.4 General conditions of animal health.9.1.5 Scoring of erythema and edema for each animal.9.1.6 Rating of sensitization response.10. Keywords10.1 acute toxicity tests; allergenicity; biocompatibility; guinea pigs; sensitization; test animalsTABLE 1 Scoring Criteria for Test Reaction
35、sReaction Description ScoreErythema erythema and escharno erythema 0very slight erythema (barely perceptible) 1well-defined erythema (pale red in color) 2moderate to severe erythema (red and area well de-fined)3severe erythema (beet redness to slight eschar for-mation)4Edema edema formationno edema
36、0very slight edema (barely perceptible) 1slight edema (edges of area well-defined by definiteraising)2moderate edema (edges raised approximately 1 mm) 3severe edema (raised more than 1 mm and extendingbeyond the area of exposure)4F720 173ASTM International takes no position respecting the validity o
37、f any patent rights asserted in connection with any item mentionedin this standard. Users of this standard are expressly advised that determination of the validity of any such patent rights, and the riskof infringement of such rights, are entirely their own responsibility.This standard is subject to
38、 revision at any time by the responsible technical committee and must be reviewed every five years andif not revised, either reapproved or withdrawn.Your comments are invited either for revision of this standard or for additional standardsand should be addressed to ASTM International Headquarters. Y
39、our comments will receive careful consideration at a meeting of theresponsible technical committee, which you may attend. If you feel that your comments have not received a fair hearing you shouldmake your views known to the ASTM Committee on Standards, at the address shown below.This standard is co
40、pyrighted by ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959,United States. Individual reprints (single or multiple copies) of this standard may be obtained by contacting ASTM at the aboveaddress or at 610-832-9585 (phone), 610-832-9555 (fax), or serviceastm.
41、org (e-mail); or through the ASTM website(www.astm.org). Permission rights to photocopy the standard may also be secured from the Copyright Clearance Center, 222Rosewood Drive, Danvers, MA 01923, Tel: (978) 646-2600; http:/ 2 Rating of Sensitization Response% Sensitized Grades Classification0 to 8 I no different than control9 to 28 II mild29 to 64 III moderate65 to 80 IV strong81 to 100 V extremeF720 174
