5.1 This test method is intended to provide a tool for assessing whether fuel storage and distribution facilities, or end user fuel tanks, are subject to microbial growth, and to alert fuel suppliers or users to the potential for fuel quality or operational problems or the requirement for preventative or remedial measures, or both.5.2 This test method allows assessment of whether antigens generated by microbial activity in the specimens are present within specific defined ranges.5.3 This test method measures the presence of microbial and metabolite antigens in a specimen. The antigens are generated from the living cells and metabolites created by fungi and bacteria during growth on fuel. Consequently, the presence of antigens is an indicator of microbial contamination in fuel systems. Antigens are not associated with matter of nonbiological origin.5.3.1 Some of the antigens detected by this test method can persist after treatment with a biocide. See 11.4.5.4 This test method is semi-quantitative and can be used to determine whether contamination in samples drawn from fuel tanks and systems is negligible or present at moderate or heavy levels.5.4.1 Further information on using the test to assess biodeterioration risk is provided in Appendix X1.5.5 The significance of these levels to the operator will depend on the fuel type, the sampling location, the equipment or facility sampled, and the specific operating circumstances.5.6 Further guidance on interpretation of test results can be found in Guide D6469, in Energy Institute guidelines for the investigation of the microbial content of petroleum fuels, and in the IATA Guidance Material on Microbial Contamination in Aircraft Fuel Tanks.5.7 Further guidance on sampling can be found in Practice D7464.5.8 Testing can be conducted on a routine basis or to investigate incidents.5.9 Microbiological tests are not intended to be used to determine compliance with fuel specifications or limits. The implementation of specification limits for microbiological contamination in fuels is generally not appropriate, and microbial contamination levels cannot be used alone or directly to make inferences about fuel quality or fitness for use.5.10 When interpreting results, it must be appreciated that the test result applies only to the specific sample and specimen tested and not necessarily to bulk fuel. Microbiological contamination usually shows a highly heterogeneous distribution in fuel systems, and therefore, analysis of a single sample will rarely provide a complete assessment of the overall levels of contamination present.5.11 Water phase will usually contain substantially higher amounts of microbial contamination than fuel phase and, consequently, a different interpretation of results is required. This is why this test method reports antigen concentration per mL for water and per L for fuel.5.12 This test method differs from some other methods (for example Test Methods D7687 and D7978) and practices (for example Practice D6974) in that it detects microbial activity in fuels or associated aqueous specimens in the field and does not need to be performed in a laboratory or in an aseptic environment. It may be used in a laboratory.5.13 This test method does not require specialist microbiological experience or knowledge.5.14 This test method provides rapid results that reflect the total active microbial contamination in the specimen, and enables result to be obtained within 15 min.5.15 This test method differentiates among three ranges of contamination for H. resinae, other fungi, and aerobic bacteria (see 1.3).1.1 This test method describes a procedure that can be used in the field or in a laboratory to detect antigens indicative of microbial contamination in liquid fuels, including those blended with synthesized hydrocarbons or biofuels, with kinematic viscosities (at 40 °C) of ≤24 mm2s–1 (for example, Specifications D396, D975, and D1655) and in fuel-associated water.1.1.1 This test method has been validated by an ILS for a range of middle distillate fuels meeting Specification D1655, EN590, Specification D975, and ISO 8217:2012.1.2 This test method semi-quantitatively assesses the concentration of specific antigens generated by commonly recovered, fuel-associated, aerobic microorganisms during active growth in fuel systems.1.2.1 A proprietary formulation of antibodies and antibody mixtures is used to detect three types of microbial antigen contamination: antigens generally found in aerobic bacteria, antigens generally present in common fungi (yeast and molds), and an antigen that is characteristic of Hormoconis resinae (the fungus most commonly associated with fuel biodeterioration).1.2.2 Although the antibodies and antibody mixtures are characteristic of diverse types of bacteria and fungi, it is unlikely that they are universal. Recognizing that for every microbe that has been isolated and characterized, it is likely that there are a billion that have not. Consequently, as is the case with all microbiological test methods, this test method does not purport to detect 100 % of the microbes present in a fuel or fuel-associated water sample.1.3 For each of the three sets of antigen detected (H. resinae, common fungi, and aerobic bacteria), the test detects whether the antigen concentration present is within set ranges representing negligible, moderate, or heavy microbial contamination.1.3.1 For fuel specimens, the antigen concentration ranges detected are <150 µg/L (negligible), 150 µg/L to 750 µg/L (moderate), and >750 µg/L (heavy).1.3.2 For specimens of water associated with fuel, the antigen concentration ranges detected are <33 µg/mL (negligible), 33 µg/mL to 166 µg/mL (moderate), and >166 µg/mL (heavy).1.4 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.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, health, and environmental practices and determine the applicability of regulatory limitations prior to use. For a specific hazard statement, see Section 8.1.6 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

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4.1 The purpose of this code is to simplify the reporting of solid particle counts and water droplet concentrations present in petroleum products when measured by imaging instruments. Industry is accustomed to using a three number code to report contamination levels and so this reporting method for imaging instruments is presented to organize results in a similar format.4.2 Particle count results as described by this classification are reported per the preferred number ranges in Table 1. Preferred numbers were originally developed by Charles Renard and codified in ISO 3. This format is the preferred reporting format since ISO 4406 has no ability to report water.4.3 Imaging instruments are capable of identifying insoluble water droplets separate from solids, and therefore a suffix code is added after the three (or four) solid particle codes to report water content in parts per million (ppm (v)). To report water content, detected droplets will be converted to ppm (v). The distribution of water droplet size may be reported if it is useful, but it is not required.1.1 This classification identifies a concise code which can be used by imaging instruments for reporting of solid particles and insoluble water content in hydrocarbon-based petroleum products. The coding system includes the reporting of water content and particle counts in the ≥1 µm range.NOTE 1: Calibration is not in accordance with ISO 11171. Comparability to ISO 4406 and its requirements is not inferred.1.2 It is valid for imaging instruments measuring particle size by projected equivalent particle diameter (see 3.1.1), and which are calibrated and verified using particle size and count standards traceable to NIST.21.3 Due to the variation of method and instrument types and resolutions, the reporting of results will include the ASTM standard used to conduct the analysis.1.4 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.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.1.6 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

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5.1 The two procedures in the test method provide rapid methods for field detection of free water and solid contaminants, or any other visually apparent contamination. Uncertain or marginal results by either of these methods would normally result in the performance of methods such as D2276, D5452, or D3240 for quantitative determination of contaminants.5.1.1 Particulate determination in appearance tests is sensitive to sampling procedures. The presence of a small number of particles may indicate, for example, that the sample line was not flushed to provide a representative sample. The persistent presence of even a small number of particles, however, may be cause for further investigation depending on the situation.5.2 Experience has shown that an experienced tester using a clear bottle can detect as little as 40 ppm of free, suspended water in the fuel. Thus, a fuel rated as clear and bright can still fail lower limits set by quantitative methods. A rater will also have difficulty resolving particles smaller than 40 μm. Smaller particles must be determined by other than visual methods such as D2276, D5452 or chemical field tests listed in Manual 5.55.3 Experience has shown the visual appearance of fuel in a white porcelain bucket to be the most suitable method for the detection of dye contamination or other unusual discoloration. In the U.S., the white porcelain bucket is used to detect the dye.1.1 This test method covers two procedures for establishing the presence of suspended free water, solid particulate, and other contaminants in aviation gasoline and aviation turbine fuels.1.1.1 Both procedures are intended primarily for use as field tests with the fuel at handling temperature.1.1.2 Procedure A uses transparent sample containers; Procedure B uses opaque containers.1.2 Both procedures are rapid methods for contamination detection and include ratings of haze appearance and particulate presence.1.3 The values stated in SI units are to be regarded as the standard. The values given in parentheses are for information only.1.4 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

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4.1 This practice describes and defines factors to be taken into consideration when designing and fabricating a cleanroom or controlled area that is used for aerospace operations and fabrication. Following the suggestions herein should provide a facility that is more capable of meeting performance requirements and that will offer protection against contamination for objects fabricated and processed in such a facility.1.1 The purpose of this practice is to provide design and construction guidelines for contamination controlled facilities used in the assembly and integration of aerospace hardware. The guidelines herein are intended to ensure that the facilities, when used properly, will meet the cleanliness requirements of aerospace hardware and processes. The objective is to limit contamination due to the deposition of particulate and molecular contaminants on flight hardware surfaces.1.2 One cleanliness classification of a facility is the airborne particle concentrations in accordance with ISO 14644-1 and 14644-2. Airborne particle concentrations in accordance with FED-STD-209E are included for reference. This simple classification is inadequate to describe a facility that will support the assembly and integration of spacecraft. The extended duration of hardware exposure during fabrication and testing, the sensitivity of the hardware to hydrocarbons and other molecular contaminants, and the changing requirements during assembly and integration must be considered in addition to the airborne particle concentrations.1.3 The guidelines specified herein are intended to provide facilities that will effectively restrict contaminants from entering the facility, limit contamination generated by and within the facility, and continuously remove airborne contaminants generated during normal operations. Some items of support hardware, such as lifting equipment, stands, and shoe cleaners, are addressed since these items are often purchased and installed with the facility and may require accommodation in the design of the facility.1.4 Active filtration of molecular contaminants (such as hydrocarbons, silicones, and other chemicals) is discussed. Such active filtration of molecular contaminants may be required for the processing of highly sensitive optical devices, especially infrared and cryogenic sensors. Control of microbiological contamination is not included although HEPA (High Efficiency Particulate Air) filtration will provide some control of airborne bacteria, spores, and other viable contaminants that are typically carried on particles of sizes 0.3 μm and larger. Control of radioactive contamination and accommodation of very hazardous materials such as propellants, strong acids or caustics, or carcinogens are not addressed.1.5 No facility will compensate for excessive contamination generated inside the facility. In addition to an effective facility design, the user must also institute a routine maintenance program (see Practice E2042) for the facility, and personnel and operational disciplines that limit the transfer of contaminants through entry doors and contaminant generation inside the facility.1.6 This practice only addresses guidelines for contamination control in facility design. It must be implemented in compliance with all mandatory government and regulatory building and safety codes. References to related cleanroom standards and U.S. building codes and standards may be found in IEST-RP-CC012.1.7 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.1.7.1 The values given in parentheses are provided for information only and are not considered standard.1.8 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, health, and environmental practices and determine the applicability of regulatory limitations prior to use.1.9 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

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5.1 This test method indicates and measures the amount of insoluble contamination of hydraulic fluids. Minimizing the levels of insoluble contamination of hydraulic fluids is essential for the satisfactory performance and long life of the equipment. Insoluble contamination can not only plug filters but can damage functional system components resulting in wear and eventual system failure.1.1 This test method covers the determination of insoluble contamination in hydraulic fluids by gravimetric analysis. The contamination determined includes both particulate and gel-like matter, organic and inorganic, which is retained on a membrane filter disk of pore diameter as required by applicable specifications (usually 0.45 μm or 0.80 μm).1.2 To indicate the nature and distribution of the particulate contamination, the gravimetric method should be supplemented by occasional particle counts of typical samples in accordance with Test Method F312.1.3 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.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, health, and environmental practices and determine the applicability of regulatory limitations prior to use. For a specific warning statement, see 7.1.1.5 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

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This test method covers the apparatuses required, sampling methods, standard procedures and calculations, and test reports for counting and sizing airborne microparticulate matter, the sampling areas for which are specifically those with contamination levels typical of cleanrooms and dust-controlled areas. The test method is based on the microscopical examination of particles impinged upon a membrane filter with the aid of a vacuum. Sampling may be done in a cleanroom, clean zone, or other controlle areas, or in a duct or pipe, wherein the number of sampling points is proportional to the floor area of the enclosure to be checked. The apparatus and facilities required are typical of a laboratory for the study of macroparticle contamination. The operator must have adequate basic training in microscopy and the techniques of particle sizing and counting.1.1 This test method covers counting and sizing airborne particulate matter 5 µm and larger (macroparticles). The sampling areas are specifically those with contamination levels typical of cleanrooms and dust-controlled areas.1.2 Units—The values stated in either SI units or inch-pound units are to be regarded separately as standard. The values stated in each system are not necessarily exact equivalents; therefore, to ensure conformance with the standard, each system shall be used independently of the other, and values from the two systems shall not be combined.1.3 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, health, and environmental practices and determine the applicability of regulatory limitations prior to use.1.4 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

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5.1 This guide provides information addressing the conditions that lead to fuel microbial contamination and biodegradation and the general characteristics of and strategies for controlling microbial contamination. It compliments and amplifies information provided in Practice D4418 on handling gas-turbine fuels. More detailed information may be found in Guidelines for the Investigation of Microbial Content of Liquid Fuels and for the Implementation of Avoidance and Remedial Strategies, 3rd Ed.,10 ASTM Manual 47, and Passman, 2019.115.2 This guide focuses on microbial contamination in refined petroleum products and product handling systems. Uncontrolled microbial contamination in fuels and fuel systems remains a largely unrecognized but costly problem at all stages of the petroleum industry from crude oil production through fleet operations and consumer use. This guide introduces the fundamental concepts of fuel microbiology and biodeterioration control.5.3 This guide provides personnel who are responsible for fuel and fuel system stewardship with the background necessary to make informed decisions regarding the possible economic or safety, or both, impact of microbial contamination in their products or systems.1.1 This guide provides personnel who have a limited microbiological background with an understanding of the symptoms, occurrence, and consequences of chronic microbial contamination. The guide also suggests means for detection and control of microbial contamination in fuels and fuel systems. This guide applies primarily to gasoline, aviation, boiler, industrial gas turbine, diesel, marine, furnace fuels and blend stocks (see Specifications D396, D910, D975, D1655, D2069, D2880, D3699, D4814, D6227, and D6751), and fuel systems. However, the principles discussed herein also apply generally to crude oil and all liquid petroleum fuels. ASTM Manual 472 provides a more detailed treatment of the concepts introduced in this guide; it also provides a compilation of all of the standards referenced herein that are not found in the Annual Book of ASTM Standards, Section Five on Petroleum Products and Lubricants.1.2 This guide is not a compilation of all of the concepts and terminology used by microbiologists, but it does provide a general understanding of microbial fuel contamination.1.3 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.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, health, and environmental practices and determine the applicability of regulatory limitations prior to use.1.5 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

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