ASTM D4576-01(2006)e1
(Test Method)Standard Test Method for Mold Growth Resistance of Wet Blue
Standard Test Method for Mold Growth Resistance of Wet Blue
SIGNIFICANCE AND USE
This test method provides a technique for evaluating mold growth resistance characteristics of wet blue, and should assist in the prediction of storage time before molding occurs.
The degree of correlation between this test and commercial quantities of wet blue in storage or shipment situations, or both, has not been fully determined.
SCOPE
1.1 This test method covers the determination of mold growth resistance of wet blue subject to storage and shipping requirements and intended for use in leather manufacturing. This test method may not be suitable to evaluate fungicides that are inactivated by proteins. This includes alkyldimethylbenzyl ammonium chlorides.
1.2 Conclusions about mold growth resistance are drawn from the results by comparing the test with a simultaneously run control of known resistance. Success or failure is determined by the amount of mold growth relative to the control.
1.3 To allow use of this test method by any laboratory, flexibility has been permitted in times, temperature, and humidity of incubation, inoculum, hide sampling area, and choice of control. These may be adjusted to fit local conditions but must be standardized.
1.4 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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e1
Designation:D4576–01(Reapproved2006)
Standard Test Method for
Mold Growth Resistance of Wet Blue
This standard is issued under the fixed designation D 4576; 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.
e NOTE—Footnote 3 was revised in July 2008.
1. Scope 4. Significance and Use
1.1 This test method covers the determination of mold 4.1 This test method provides a technique for evaluating
growth resistance of wet blue subject to storage and shipping mold growth resistance characteristics of wet blue, and should
requirements and intended for use in leather manufacturing. assist in the prediction of storage time before molding occurs.
Thistestmethodmaynotbesuitabletoevaluatefungicidesthat 4.2 The degree of correlation between this test and commer-
are inactivated by proteins. This includes alkyldimethylbenzyl cial quantities of wet blue in storage or shipment situations, or
ammonium chlorides. both, has not been fully determined.
1.2 Conclusions about mold growth resistance are drawn
5. Interferences
from the results by comparing the test with a simultaneously
run control of known resistance. Success or failure is deter- 5.1 Acommon interference is contamination of plates, agar,
or samples by unwanted organisms that settle in from the
mined by the amount of mold growth relative to the control.
1.3 To allow use of this test method by any laboratory, environment.
5.2 Volatility and Leachability of Biocides—A “zone of
flexibility has been permitted in times, temperature, and
humidity of incubation, inoculum, hide sampling area, and inhibition” where no mold grows on the agar adjacent to the
specimen indicates that the fungicide may leach.
choice of control. These may be adjusted to fit local conditions
but must be standardized.
6. Apparatus
1.4 This standard does not purport to address all of the
6.1 Petri Dishes, 120 mm diameter. Sterile plastic dispos-
safety concerns, if any, associated with its use. It is the
able dishes are preferred.
responsibility of the user of this standard to establish appro-
6.2 Incubator, or location providing similar conditions be-
priate safety and health practices and determine the applica-
ing free of drafts, and capable of a constant (6 2°C) tempera-
bility of regulatory limitations prior to use.
ture within the 26 to 30°C range.
2. Terminology
6.3 Medicine droppers,disposableplastictypedelivering30
2.1 Definition of Term Specific to This Standard: to 35 drops per mL.
2.1.1 wet blue—hide or skin, or split of a hide or skin,
7. Reagents and Materials
tanned with basic chromium sulfate, containing approximately
7.1 Potato Dextrose Agar, a dehydrated plating medium
50 % moisture and having an acidic pH.
used in culturing yeasts and molds from dairy products.
3 6
3. Summary of Test Method
7.2 Inoculum, Aspergillus niger 1 3 10 spores per mL, or
other organism or a combination of organisms known to be
3.1 Wet blue test specimens are surrounded by but not
covered with agar, inoculated, and incubated. indigenous to the storage area of the wet blue.
3.2 After various incubation periods, mold growth is rated
8. Sampling, Test Specimen, and Test Units
as a percentage of the wet blue surface covered by mold.
8.1 Take test specimens from equivalent hide locations (for
3.3 Resistance to mold growth of the wet blue test specimen
example, butt area) for both test and control.
is determined by comparison with wet blue of known resis-
tance characteristics (the control), that is tested simultaneously.
A product that meets the requirements of this method is Potato Dextrose Agar
stock no. 0013-01-4, available from Difco Labs, P.O. Box 1058A, Detroit,
This test method is under the jurisdiction ofASTM Committee D31 on Leather MI 28232.
and is the direct responsibility of Subcommittee D31.02 on Wet Blue. An inoculum that meets the requirements of this method is available as ATCC
Current edition approved Oct. 1, 2006. Published November 2006. Originally (AmericanTypeCultureCollection)16404,andisavailablefromseveralsourcesfor
approved in 1986. Last previous edition approved in 2001 as D 4576 - 01. laboratory supplies.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959, United States.
e1
D4576–01 (2006)
8.2 If unable to test immediately, hold test specimens in
separate plastic bags and keep cool.
8.3 Test specimens should be a square, with a side of 25.4
mm (1 in.).
8.4 Use three test specimens for each test unit of wet blue
surface to be evaluated.
9. Procedure
9.1 Agar Preparation:
9.1.1 Agar Requirements—A split wet blue test specimen
requires about 25 mL solution and an unsplit wet blue test
specimenrequiresabout40mL.Calculatenumberofmillilitres
of agar required for tests to be performed, allowing 50 mL for
vitality check.
9.1.2 Weighout3.9gpotatodextroseagarforevery100mL
of agar required.
FIG. 1 Specimen with inoculum locations shown (X)
9.1.3 Pour a volume of water equivalent to total millilitre
...
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. Because
it 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 version
of the standard as published by ASTM is to be considered the official document.
e1
Designation:D4576–86(Reapproved1996) Designation:D4576–01(Reapproved2006)
Standard Test Method for
Mold Growth Resistance of Blue Stock (Leather)Wet Blue
This standard is issued under the fixed designation D 4576; 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.
e NOTE—Footnote 3 was revised in July 2008.
1. Scope
1.1 This test method covers the determination of mold growth resistance of blue stockwet blue subject to storage and shipping
requirements and intended for use in leather manufacturing. This test method may not be suitable to evaluate fungicides that are
inactivated by proteins. This includes alkyldimethylbenzyl ammonium chlorides.
1.2 Conclusions about mold growth resistance are drawn from the results by comparing the test with a simultaneously run
control of known resistance. Success or failure is determined by the amount of mold growth relative to the control.
1.3 To allow use of this test method by any laboratory, flexibility has been permitted in times, temperature, and humidity of
incubation, inoculum, hide sampling area, and choice of control. These may be adjusted to fit local conditions but must be
standardized.
1.4 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.
2. Terminology
2.1 Definition of Term Specific to This Standard:
2.1.1 blue stockwet blue—hide or skin, or split of a hide or skin, tanned with basic chromium sulfate, containing approximately
50 % moisture and having an acidic pH.
3. Summary of Test Method
3.1Blue stock3.1 Wet blue test specimens are surrounded by but not covered with agar, inoculated, and incubated.
3.2 After various incubation periods, mold growth is rated as a percentage of the wet blue stock covered. surface covered by
mold.
3.3 Resistance to mold growth of the subjectwet blue stock test specimen is determined by comparison with blue stockwet blue
of known resistance characteristics (the control), that is tested simultaneously.
4. Significance and Use
4.1 This test method provides a technique for evaluating mold growth resistance characteristics of blue stockwet blue, and
should assist in the prediction of storage time before molding occurs.
4.2 The degree of correlation between this test and blue stock in commercial quantitiescommercial quantities of wet blue in
storage or shipment situations, or both, has not been fully determined.
5. Interferences
5.1 A common interference is contamination of plates, agar, or samples by unwanted organisms that settle in from the
environment.
5.2 Volatility and Leachability of Biocides—A“zone of inhibition” where no mold grows on the agar adjacent to the specimen
indicates that the fungicide may leach.
6. Apparatus
6.1 Petri Dishes—120, 120 mm diameter. Sterile plastic disposable dishes are preferred.
6.2 Incubator, or other location, location providing similar conditions being free of drafts, and capable of a constant (6 2°C)
This test method is under the jurisdiction of ASTM Committee D31 on Leather and is the direct responsibility of Subcommittee D31.02 on Blue Stock.
Current edition approved June 9, 1986. Published July 1986.
This test method is under the jurisdiction of ASTM Committee D31 on Leather and is the direct responsibility of Subcommittee D31.02 on Wet Blue.
Current edition approved Oct. 1, 2006. Published November 2006. Originally approved in 1986. Last previous edition approved in 2001 as D 4576 - 01.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959, United States.
e1
D4576–01 (2006)
temperature within the 26 to 30°C range, range.
6.3 Medicine droppers, disposable plastic type delivering 30 to 35 drops per mL.
7. Reagents and Materials
7.1 Potato Dextrose Agar, a dehydrated plating medium used in culturing yeasts and molds from dairy products.
3 6
7.2 Inoculum, Aspergillus niger 1 3 10 spores per mL, or other organism or a combination of organisms known to be
indigenous to the storage area orof the blue stock. wet blue.
8. Sampling, Test Specimen, and Test Units
8.1 Take test specimens from equivalent hide locations (for example, butt area) for both test and control.
8.2 If unable to test immediately, hold test specimens in separate plastic bags and keep cool.
8.3Cut test specimens one in. square.
8.4Use three test specimens for each test unit or blue stock surface to be evaluated.
8.3 Test specimens should be a square, with a side of 25.4 mm (1 in.).
8.4 Use three test specimens for each test unit of wet blue surface to be evaluated.
9. Procedure
9.1 Agar Preparation:
9.1.1 Agar Requirements—A split wet blue stock sampletest specimen requires about 25 mL solution and an unsplit wet blue
stock test specimen requires about 40 mL. Calculate number of mLmillilitres of agar required for tests to be performed, allowing
50 mL for vitality check.
9.1.2 Weigh out 3.9 g potato dextrose agar for every 100 mL of agar required.
9.1.3Addmillilitre9.1.3 Pouravolumeofwaterequivalenttototalmillilitresofagarrequirementsolutiontobeaker.Bringwater
to boiling on hot plate equipped with magnetic stirrer mechanism. While stirring, slowly add dry agar.
9.1.4 Boil agar for 20 min.
NOTE 1—Pressure cooking for 20 min. is preferable to open boiling.
9.1.5 Cover with aluminum foil to prevent contamination and cool to 50°C before pouring.
NOTE 2—This temperature is critical, as 50°C allows some water of condensation to develop on petri dish cover providing humidity for growth of
fung
...
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