1、 Reference number ISO 27368:2008(E) ISO 2008INTERNATIONAL STANDARD ISO 27368 First edition 2008-08-15 Analysis of blood for asphyxiant toxicants Carbon monoxide and hydrogen cyanide Analyse du sang pour substances toxiques asphyxiantes Monoxyde de carbone et acide cyanhydrique ISO 27368:2008(E) PDF
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6、Os member body in the country of the requester. ISO copyright office Case postale 56 CH-1211 Geneva 20 Tel. + 41 22 749 01 11 Fax + 41 22 749 09 47 E-mail copyrightiso.org Web www.iso.org Published in Switzerland ii ISO 2008 All rights reservedISO 27368:2008(E) ISO 2008 All rights reserved iii Conte
7、nts Page Foreword iv Introduction v 1 Scope . 1 2 Normative references . 1 3 Terms and definitions. 2 4 Symbols and abbreviated terms . 5 5 Blood samples 6 5.1 General. 6 5.2 Sample condition 6 5.3 Sample collection . 6 5.4 Sample storage . 7 5.5 Sample analysis 7 6 Materials 7 7 Common quality anal
8、ytical elements 7 7.1 General. 7 7.2 Qualitative, quantitative and confirmatory analyses 7 7.3 Replicate analyses 7 7.4 Analytical batch 8 7.5 Open controls 8 7.6 Calibrators . 8 8 Measurement of CO in blood as COHb. 8 8.1 COHb by whole-blood oximeters 8 8.2 COHb by palladium chloride reduction 10 8
9、.3 COHb by visible spectrophotometry (using calibration curve) . 12 8.4 COHb by visible spectrophotometry (with CO saturation). 14 8.5 COHb by visible spectrophotometry (without CO saturation) . 16 8.6 COHb by headspace gas chromatography Nickel-hydrogen reduction and flame ionization detection 19 8
10、.7 COHb by headspace gas chromatography Thermal conductivity detection. 22 9 Measurement of HCN in blood as CN 23 9.1 CN by colourimetric method (p-nitrobenzaldehyde and o-dinitrobenzene) 23 9.2 CN by visible spectrophotometry 25 9.3 CN as HCN by headspace gas chromatography Nitrogen phosphorous det
11、ection 29 9.4 CN by headspace gas chromatography Electron capture detection 31 9.5 CN by spectrophotofluorimetry or high-performance liquid chromatography using a fluorescence detector. 33 9.6 CN by high-performance liquid chromatographymass spectrometry. 37 Annex A (normative) Analytical report pro
12、 forma. 41 Annex B (informative) Additional aspects of analytical methods . 43 Annex C (informative) Interpretation of results. 47 Bibliography . 52 ISO 27368:2008(E) iv ISO 2008 All rights reservedForeword ISO (the International Organization for Standardization) is a worldwide federation of nationa
13、l standards bodies (ISO member bodies). The work of preparing International Standards is normally carried out through ISO technical committees. Each member body interested in a subject for which a technical committee has been established has the right to be represented on that committee. Internation
14、al organizations, governmental and non-governmental, in liaison with ISO, also take part in the work. ISO collaborates closely with the International Electrotechnical Commission (IEC) on all matters of electrotechnical standardization. International Standards are drafted in accordance with the rules
15、 given in the ISO/IEC Directives, Part 2. The main task of technical committees is to prepare International Standards. Draft International Standards adopted by the technical committees are circulated to the member bodies for voting. Publication as an International Standard requires approval by at le
16、ast 75 % of the member bodies casting a vote. Attention is drawn to the possibility that some of the elements of this document may be the subject of patent rights. ISO shall not be held responsible for identifying any or all such patent rights. ISO 27368 was prepared by Technical Committee ISO/TC 92
17、, Fire safety, Subcommittee SC 3, Fire threat to people and environment. ISO 27368:2008(E) ISO 2008 All rights reserved v Introduction Carbon monoxide (CO) and hydrogen cyanide (HCN) are two of the primary toxic combustion gases present in fire atmospheres. Upon burning, carbon-containing substances
18、 generate CO, whereas nitrogen-containing substances also produce HCN. Since structures surrounding human beings are composed of polymeric materials containing carbon and nitrogen elements as their constituents, these materials generate CO and HCN upon burning and fire victims are exposed to these g
19、ases by inhaling smoke. Although ISO 19701 documents methods for the analysis of CO and HCN in fire effluents, the actual toxic insult to exposed persons can be assessed only by the analysis of the fire casualties blood for CO as carboxyhaemoglobin (COHb) and HCN as cyanide ion (CN ). These analytic
20、al findings are useful for estimating life-threatening characteristics of fire atmospheres, evaluating the degree of toxicity caused by smoke inhalation in fire victims, determining the cause and manner of death of fire victims, improving understanding of the direct causes of fire injury and death,
21、enhancing understanding of acute and delayed adverse effects of smoke on fire casualties, administering immediate treatment for smoke poisoning and monitoring delayed adverse effects of smoke, choosing appropriate emergency, long-term and/or follow-up treatments for surviving fire casualties, settin
22、g priorities for emergency treatment of multiple fire casualties, establishing relationships between the concentrations of CO and HCN in a fire atmosphere, blood COHb and CN levels, and the degree of toxicity and performance impairment, achieving correlations between concentrations of the two gases
23、in fire atmospheres and of COHb and CN in blood in order to improve tenability models, identifying deficiencies with materials, products, assemblies, structures and escape routes, and improving forensic toxicology analytical processes and procedures. Compliance with this International Standard can h
24、elp ensure a consistent data set for use in a variety of fields such as a) fire statistics, which themselves are frequently used to develop regulatory policy, b) international collaboration on improved design, materials and use of habitable structures, and, c) ultimately, improvement of internationa
25、l relations and trades. Such compliance can further assist in developing better and safer fire-safety instruments and structures (residential and commercial buildings; locomotive passenger vans, automobiles, aerospace vehicles and other vehicular structures). Various different methods are currently
26、used for obtaining blood analysis data for these two fire toxicants and the lack of standardized procedures can result in a wide variation of interpretation. It is, therefore, proposed to set out best-practice, standardized procedures for blood sample collection, sample storage, sample processing/pr
27、eparation, sample treatment and transfer to analytical instrumentation, analytical instrumentation ISO 27368:2008(E) vi ISO 2008 All rights reservedand techniques, data presentation and reporting, and guidance for data interpretation. The analytical methods included herein are based upon their suita
28、bility for performing an analysis on ante-mortem and post-mortem blood samples from fire victims and are commonly used in forensic toxicological analytical operations. This International Standard is structured as follows. Clause 1 describes the scope of this International Standard. Clause 2 cites th
29、e normative references. Clause 3 provides terms and their definitions. Clause 4 lists symbols and abbreviated terms. Clause 5 provides a general description of collecting, storing and analysing blood samples. Clause 6 covers the quality of materials used during an analysis. Clause 7 summarizes commo
30、n quality analytical elements. Clause 8 describes analytical methods for measuring CO as COHb. Clause 9 delineates analytical methods for measuring HCN as CN in blood. Annex A (normative) lists the information crucial for reporting blood analysis results. Annex B (informative) outlines additional as
31、pects of analytical methods. Annex C (informative) discusses the interpretation of results, including the interactive effects of CO and HCN. The bibliography includes references cited in this International Standard. INTERNATIONAL STANDARD ISO 27368:2008(E) ISO 2008 All rights reserved 1 Analysis of
32、blood for asphyxiant toxicants Carbon monoxide and hydrogen cyanide SAFETY PRECAUTIONS Due consideration shall be given to the fact that both the blood samples for the analyses of asphyxiant toxicants, carbon monoxide (CO) and hydrogen cyanide (HCN), and many of the reagents used for their analyses
33、can be biohazardous and/or toxic and can thereby pose serious health hazards. It is recommended that the collection of blood samples from fire victims be performed by medical practitioners and in accordance with best practices established by the medical authorities in the area. Additionally, it is a
34、ssumed that the procedures described herein are carried out by suitably qualified professional personnel, adequately trained in the hazards and risks associated with the handling of biological samples and such analyses and aware of any safety regulations that can be in effect. Consideration shall al
35、so be given to the safe and ecologically acceptable disposal of all biological samples and chemicals used for analyses. This can require extensive and specific treatment prior to release of the waste into the environment. Again, it is assumed in this International Standard that the personnel respons
36、ible for the safe disposal of such bio-samples and reagents are suitably qualified and trained in these procedures and techniques and are aware of the regulations that can be in force. 1 Scope This International Standard details analytical methods suitable for analysing the two primary toxic combust
37、ion gases, carbon monoxide (CO) and hydrogen cyanide (HCN), in blood samples collected from fire casualties. In blood, CO is measured as carboxyhaemoglobin (COHb) and HCN as cyanide ion (CN ). Although numerous methods are reported in the literature for performing blood COHb and CN analyses, the ana
38、lytical methods included herein are based upon their suitability for performing the analysis on ante-mortem and post- mortem blood samples from fire casualties. The analytical principle, analysis time, repeatability, reproducibility, robustness, effectiveness and instruments used are considered for
39、those methods. Some of the methods described herein might not be suitable for analysing putrid or clotted blood. Burned (solid) blood can be analysed after homogenization. 2 Normative references The following referenced documents are indispensable for the application of this document. For dated refe
40、rences, only the edition cited applies. For undated references, the latest edition of the referenced document (including any amendments) applies. ISO 3696:1987, Water for analytical laboratory use Specification and test methods ISO 13344, Estimation of the lethal toxic potency of fire effluents ISO/
41、TS 13571, Life-threatening components of fire Guidelines for the estimation of time available for escape using fire data ISO 13943, Fire safety Vocabulary ISO 19701, Methods for sampling and analysis of fire effluents ISO 27368:2008(E) 2 ISO 2008 All rights reserved3 Terms and definitions For the pu
42、rposes of this document, the terms and definitions given in ISO 19701, ISO 13344, ISO/TS 13571, ISO 13943, ISO 3696, and the following apply. 3.1 analyte substance that is being identified or determined in a specimen during an analysis EXAMPLES COHb and CN . 3.2 analytical batch set of aliquots take
43、n out from the specimens associated with various cases (fire casualties) and from negative and positive blind controls for performing a particular type of analysis 3.3 asphyxiant toxicant causing loss of consciousness and ultimately death resulting from hypoxic (deficiency-of-oxygen) effects, partic
44、ularly on the central nervous and/or cardiovascular systems 3.4 blind controls open controls but their identity is unknown to the analysts See open controls (3.20). 3.5 calibrator material that is based on, or traceable to, a reference preparation or material and whose values are determined by accep
45、table reference methods 3.6 carboxyhaemoglobin compound formed when CO combines with haemoglobin NOTE Haemoglobin has an affinity for binding to CO that is approximately 245 times higher than that for binding to oxygen; thereby the ability of haemoglobin to carry oxygen is seriously compromised duri
46、ng CO poisonings (see C.3.3 and Reference 73). 3.7 Cheyne-Stokes respiration breathing pattern characterized by rhythmic waxing and waning of the depth of respiration, with regularly recurring periods of breathing cessation 3.8 cutaneous blood vessels blood vessels relating to, or affecting, the ski
47、n 3.9 cyanogenic glycosides group of molecules containing a sugar moiety and a cyanide (CN) group NOTE Cyanogenic glycoside can release the poisonous HCN gas if acted upon by some enzyme. EXAMPLE Amygadlin from almond. ISO 27368:2008(E) ISO 2008 All rights reserved 3 3.10 cyanomethaemoglobin compoun
48、d formed when CN combines with methaemoglobin NOTE During the treatment of CN poisonings, haemoglobin is chemically converted to methaemoglobin, which easily binds with CN , producing cyanomethaemoglobin. The formation of cyanomethaemoglobin is an essential and critical step in the CN detoxification
49、 process (see Reference 71). 3.11 cyanosis bluish discoloration of the skin caused by the lack of oxygen in the blood 3.12 deoxyhaemoglobin form of haemoglobin without oxygen, the predominant protein in the red blood cells NOTE Haemoglobin forms an unstable, reversible bond with oxygen. The oxygen-bonded haemoglobin is known as oxyhaemoglobin. In the oxygen-unloaded form, it is called deoxyhaemoglobin and is purple-blue. 3.13 fire effluent totality of gas
