ASTM E1259-94

Standard Test Method for Evaluation of Antimicrobials in Liquid Fuels Boiling Below 390°C

SCOPE
1.1 This test method is designed to evaluate antimicrobial agents for the prevention of microbially influenced deterioration of liquid fuels (as defined by Specification D 396, D 910, D 975, D 1655, D 2069, D 2880, D 3699, D 4818 and D 6227), system deterioration, or both.
1.2 Knowledge of microbiological techniques is required for these procedures.
1.3 It is the responsibility of the investigator to determine whether Good Laboratory Practice (GLP) is required and to follow them where appropriate (40 CFR, 160), or as revised.
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.

General Information

• Status: Historical
• Publication Date: 09-Apr-2001
• ICS: 75.160.20 - Liquid fuels
• Technical Committee: E35 - Pesticides, Antimicrobials, and Alternative Control Agents
• Drafting Committee: E35.15 - Antimicrobial Agents

Relations

• Replaced By: ASTM E1259-01 - Standard Test Method for Evaluation of Antimicrobials in Liquid Fuels Boiling Below 390°C
• Effective Date: 10-Apr-2001
• Referred By: ASTM D6469-20 - Standard Guide for Microbial Contamination in Fuels and Fuel Systems
• Effective Date: 10-Apr-2001
• Referred By: ASTM D7826-23a - Standard Guide for Evaluation of New Aviation Gasolines and New Aviation Gasoline Additives
• Effective Date: 10-Apr-2001
• Referred By: ASTM E1259-23 - Standard Practice for Evaluation of Antimicrobials in Liquid Fuels Boiling Below 390 °C
• Effective Date: 10-Apr-2001
• Referred By: ASTM D7847-22 - Standard Guide for Interlaboratory Studies for Microbiological Test Methods
• Effective Date: 10-Apr-2001

Standards Content (Sample)

NOTICE: This standard has either been superseded and replaced by a new version or discontinued.
Contact ASTM International (www.astm.org) for the latest information.
Designation: E 1259 – 94
Standard Test Method for
Evaluation of Antimicrobials in Distillate Fuels (Based on
Preliminary Screening and Compatibility)
This standard is issued under the fixed designation E 1259; 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 (e) indicates an editorial change since the last revision or reapproval.
1. Scope test method allows for impact of fuel/water partitioning and
time on the antimicrobial agent as well as the effect of
1.1 This test method is designed to evaluate antimicrobial
continual rechallenge. Every 2 weeks, water phase is increased
agents for the prevention of microbial-induced deterioration of
by 0.25 % while concomitantly a paired system is destructively
distillate fuels (as defined by Specification D 396 as fuel) or
tested. Thus, at 4 weeks, there is an equivalent 0.5 %, at 6
system deterioration, or both.
weeks 0.75 %, and at 8 weeks 1.0 %. At each sampling time
NOTE 1—A knowledge of microbiological techniques is required for
interval, treated and untreated aliquots are checked for the
these procedures.
three types of organisms in the initial inoculum. These counts
1.2 It is the responsibility of the investigator to determine
are coupled with gross observations of each system for biofilm
whether Good Laboratory Practice (GLP) is required and to
formation and interfacial growth.
follow them where appropriate (40 CFR, 160) or as revised.
4. Significance and Use
1.3 This standard does not purport to address all of the
safety concerns, if any, associated with its use. It is the
4.1 The procedure should be used to evaluate the relative
responsibility of the user of this standard to establish appro-
efficacy of microbicides in distillate fuels. The effect of
priate safety and health practices and determine the applica-
environmental conditions including a variety of fuel additives,
bility of regulatory limitations prior to use. See caution
metal surfaces, and climatology are variables that can be
statement, Note 2, in Section 8.
included in specific tests using this protocol.
2. Referenced Documents
5. Apparatus
2.1 ASTM Standards:
5.1 Colony Counter—Any of several types, for example, a
D 396 Specification for Fuel Oils
Quebec Colony Counter may be used.
D 4054 Practice for Evaluating the Compatibility of Addi-
5.2 Incubator—Any incubator capable of maintaining tem-
tives with Aviation-Turbine Fuels and Aircraft Fuel Sys-
perature of 30 to 35 6 2°C may be used.
tem Materials
5.3 Sterilizer—Any suitable steam sterilizer capable of
2.2 Federal Standard:
producing the conditions of sterility is acceptable.
40 CFR, Part 160, Good Laboratory Practice Standards
5.4 Separatory Funnels—Eight 1-L funnels.
5.5 Ring Stand, suitable for supporting separatory funnel.
3. Summary of Test Method
5.6 Vortex—Mixer.
3.1 This test method is conducted on a reference fuel for
6. Reagents and Materials
determining antimicrobial efficacy under well-defined condi-
tions that include specific inocula Pseudomonas aeruginosa,
6.1 Petri Dishes—100 by 15 mm required for performing
American Type Culture Collection, (ATCC) No. 33988, Hor-
standard plate count.
moconis resinae, ATCC No. 20495, and Yarrowia tropicalis
6.2 Bacteriological Pipets—10.0 mL and 1.1, or 2.2 mL
(formerly Candida tropicalis, ATCC No. 18138, water/fuel
capacity.
ratios, and time of containment. It is designed for destructive
6.3 Water Dilution Bottles—Any sterilizable glass container
sampling at regular intervals during bottom water buildup. This
having a 150–200 mL capacity and tight closure may be used.
6.4 Distillate Fuel.
6.5 Synthetic Bottom Water.
This test method is under the jurisdiction of ASTM Committee E-35 on
6.6 Soy Peptone Casein Digest Agar.
Pesticides and is the direct responsibility of Subcommittee E35.15 on Antimicrobial
6.7 Sabouraud Dextrose Agar.
Agents.
Current edition approved June 15, 1994. Published August 1994. Originally
published as E 1259 – 88. Last previous edition E 1259 – 88.
2 5
Annual Book of ASTM Standards, Vol 05.01. I-H CAT diesel fuel is available from Howell Hydrocarbons Inc., San Antonio,
Annual Book of ASTM Standards, Vol 05.02. TX 78223-3531.
4 6
Available from the Superintendent of Documents, U.S. Government Printing Items 6.5-6.12 are available from a variety of media manufacturers and
Office, Washington, DC 20402. chemical supply companies.
Copyright © ASTM, 100 Barr Harbor Drive, West Conshohocken, PA 19428-2959, United States.
E 1259
6.8 Agar, Bacteriological Grade. and 8 weeks, the following protocol is observed. The bottom
6.9 Potassium Tellurite Solution—sterile 1 %. water fraction including the fuel/water interface and a minimal
6.10 Gentamicin Sulfate—50 μg/mL. amount of fuel is bled from the separatory funnel and mixed
6.11 Plate Count Agar. vigorously in a vortex for 10 s. Before settling, aliquots are
6.12 Potato Dextrose Agar. removed for plate counts for the yeast, bacteria, and molds with
dilutions into sterile synthetic bottom water solution as fol-
7. Inoculum
lows: 1.0 mL into 99 mL for 1:100 and from this dilution
7.1 Maintenance and Preparation of Inocula—All three
subsequent dilutions for 1:1 000 and 1:10 000 are made.
cultures are transferred from specific agar slants (a) Pseudomo- These will be used for pour platings for bacteria and yeast,
nas aeruginosa, plate count agar; (b) Hormoconis resinae, and
respectively. In addition, three by two 0.1 mL portions will be
(c) Yarrowia tropicalis, potato dextrose agar to synthetic used for bacteria and yeast pour plates and for spread plates for
bottom water medium in a suitabl
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