1.1 本试验方法提供了一种协议，用于捕获、浓缩和测试燃料系统子样本（即试样）中存在的三磷酸腺苷（ATP），该子样本与以下相关: 1.1.1 液体燃料中发现的微生物和亲水颗粒，如 表X6.1 或 1.1.2 在燃料和相关底水或仅相关底水的混合物中发现的微生物和亲水颗粒。 1.1.3 通过这种生物发光测试检测到的ATP可以来自细胞ATP、细胞外ATP或两者的某种组合。 1.1.4 用于执行ATP生物发光的细胞和细胞外ATP被捕获，并从燃料系统样品中浓缩到含水试样（即子样品）中进行测试。例如，对于不包含任何可见燃油相关底水的燃油系统样品，含水试样是中所述的捕获溶液本身 8.2.1.1 . 对于燃料和相关底水（即自由水）的混合物的燃料系统样品，试样是捕获溶液和相关底水的等量。 1.2 使用专利生物发光酶测定法测量ATP，由此产生的光量与样品中ATP的浓度成比例。使用专用ATP测试笔产生并定量测量光 2. 和专用光度计 2. 并以（仪器特定的）相对光单位报告。 1.3 本试验方法同样适用于实验室或现场。 1.4 虽然生物发光是一种可靠且经验证的技术，但该方法无法将ATP与细菌或真菌区分开来。 1.5 对于水或捕获溶液样品，本试验方法可检测到的ATP浓度范围为1×10 –11 M至3×10 –8 M，相当于1×10 –14 摩尔/毫升至3×10 –11 水样或捕获溶液的摩尔/毫升。假设在500 燃料的毫升体积燃料的当量浓度为:6×10 –11 M至2×10 –14 M 1.6 以国际单位制表示的数值应视为标准值。本标准不包括其他计量单位。 1.6.1 相对光单元（RLU）有一个例外，如中所定义 3.1.19 . 1.7 本标准并非旨在解决与其使用相关的所有安全问题（如有）。 本标准的用户有责任在使用前制定适当的安全、健康和环境实践，并确定监管限制的适用性。 1.8 本国际标准是根据世界贸易组织技术性贸易壁垒（TBT）委员会发布的《关于制定国际标准、指南和建议的原则的决定》中确立的国际公认标准化原则制定的。 ====意义和用途====== 5.1 本试验方法测量样品中ATP的浓度。ATP是包括细菌和真菌在内的所有活细胞的组成部分。因此，ATP的存在是燃料系统中微生物污染的可靠指标。ATP与非生物来源的物质无关。 5.2 本试验方法不同于试验方法 D4012年 具体如下: 5.2.1 通过快速测定燃料（石油）样品、燃料和水混合物样品、与燃料相关的底水样品和在燃料或含水样品基质中自由获得的细胞外ATP； 5.2.2 通过提供一种使用自包含的测试设备和光度计捕获、提取和量化ATP的方法； 5.2.3 通过提供一种在通常不到10分钟的时间内量化燃料或水基质中存在的ATP的方法；和 5.2.4 通过提供ATP与化学干扰的快速分离，之前化学干扰阻止了在含有碳氢化合物和其他有机分子的复杂流体中使用ATP测定。 5.3 本试验方法不需要使用危险材料，也不会产生生物危害废物。 5.4 该测试方法可用于估计可行的微生物生物量，评估抗菌农药的功效，并监测燃料储存和分配系统中的微生物污染。

1.1 This test method provides a protocol for capturing, concentrating, and testing the adenosine triphosphate (ATP) present in a fuel system sub-sample (that is, test specimen) associated with: 1.1.1 Microorganisms and hydrophilic particles found in liquid fuels as described in Table X6.1 , or 1.1.2 Microorganisms and hydrophilic particles found in mixture of fuel and associated bottom water or just associated bottom water. 1.1.3 ATP detected by this bioluminescence test can be derived from cellular ATP, extra-cellular ATP, or some combination of both. 1.1.4 Cellular and extra-cellular ATP utilized to perform ATP bioluminescence are captured and concentrated from a fuel system sample into an aqueous test specimen (that is, sub-sample) for testing. For example, for a fuel system sample that does not contain any visible fuel associated bottom water, the aqueous test specimen is the capture solution itself described in 8.2.1.1 . For fuel system samples that are a mixture of fuel and associated bottom water (that is, free water), the test specimen is an aliquant of the capture solution and associated bottom water. 1.2 The ATP is measured using a patented bioluminescence enzyme assay, whereby light is generated in amounts proportional to the concentration of ATP in the sample. The light is produced and measured quantitatively using dedicated ATP test pens 2 and a dedicated luminometer 2 and reported in (instrument specific) Relative Light Units. 1.3 This test method is equally suitable for use in the laboratory or field. 1.4 Although bioluminescence is a reliable and proven technology, this method does not differentiate ATP from bacteria or fungi. 1.5 For water or capture solution samples, the concentration range of ATP detectable by this test method is 1 × 10 –11 M to 3 × 10 –8 M which is equivalent to 1 × 10 –14 moles/mL to 3 × 10 –11 moles/mL for water samples or capture solution. Assuming testing on fuel phase is performed on a 500 mL volume of fuel the equivalent concentrations is fuel would be: 6 × 10 –11 M to 2 × 10 –14 M. 1.6 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard. 1.6.1 There is one exception—Relative Light Unit (RLU) as defined in 3.1.19 . 1.7 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. 1.8 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. ====== Significance And Use ====== 5.1 This test method measures the concentration of ATP present in the sample. ATP is a constituent of all living cells including bacteria and fungi. Consequently, the presence of ATP is a reliable indicator of microbial contamination in fuel systems. ATP is not associated with matter of non-biological origin. 5.2 This test method differs from Test Method D4012 as follows: 5.2.1 By providing for the rapid determination of ATP present in a fuel (petroleum) sample, a fuel and water mixture sample, fuel-associated bottom water sample, and extracellular ATP freely available in the fuel or aqueous sample matrix; 5.2.2 By providing for a method to capture, extract, and quantify ATP using self-contained test device and luminometer; 5.2.3 By providing a method of quantifying ATP present in fuel or water matrices in generally less than 10 min; and 5.2.4 By providing for the rapid separation of the ATP from chemical interferences that have previously prevented the use of ATP determinations in complex fluids containing hydrocarbons and other organic molecules. 5.3 This test method does not require the use of hazardous materials and does not generate biohazard waste. 5.4 This test method can be used to estimate viable microbial biomass, to evaluate the efficacy of antimicrobial pesticides, and to monitor microbial contamination in fuel storage and distribution systems.