1、BRITISH STANDARD BS ISO 16221:2001 BS 6068-5.29:2001 Water quality Guidance for determination of biodegradability in the marine environment ICS 13.060.70 NO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAWBS ISO 16221:2001 This British Standard, having been prepared under the dire
2、ction of the Health and Environment Sector Committee, was published under the authority of the Standards Committee and comes into effect on 15 April 2001 BSI 04-2001 ISBN 0 580 37200 6 National foreword This British Standard reproduces verbatim ISO 16221:2001 and implements it as the UK national sta
3、ndard. The UK participation in its preparation was entrusted by Technical Committee EH/3, Water quality, to Subcommittee EH/3/2, physical, chemical and biochemical methods, which has the responsibility to: A list of organizations represented on this subcommittee can be obtained on request to its sec
4、retary. This is one of a series of standards on water quality, others of which have been, or will be, published as sections of BS 6068. the various sections of BS 6068 are comprised within parts 1 to 7, which, together with Part 0, are listed below. Part 0 Introduction Part 1 Glossary Part 2 Physica
5、l, chemical and biochemical methods Part 3 Radiological methods Part 4 Microbiological methods Part 5 Biological methods Part 6 Sampling Part 7 Precision and accuracy NOTE The tests described in this British Standard should only be carried out by suitably qualified persons with an appropriate level
6、of biological expertise. Standard biological procedures should be followed throughout. Cross-references The British Standards which implement these international or European publications referred to in this document may be found in the BSI Standards Catalogue under the section entitled “Internationa
7、l Standards Correspondence Index”, or by using the “Find” facility of the BSI Standards Electronic Catalogue. A British Standard does not purport to include all the necessary provisions of a contract. Users of British Standards are responsible for their correct application. Compliance with a British
8、 Standard does not of itself confer immunity from legal obligations. aid enquirers to understand the text; present to the responsible international/European committee any enquiries on the interpretation, or proposals for change, and keep the UK interests informed; monitor related international and E
9、uropean developments and promulgate them in the UK. Summary of pages This document comprises a front cover, an inside front cover, the ISO title page, pages ii to v, a blank page, pages 1 to 12, an inside back cover and a back cover. The BSI copyright date displayed in this document indicates when t
10、he document was last issued. Amendments issued since publication Amd. No. Date CommentsReference number ISO 16221:2001(E) INTERNATIONAL STANDARD ISO 16221 First edition 2001-02-01 Water quality Guidance for determination of biodegradability in the marine environment Qualit de leau Lignes directrices
11、 pour la dtermination de la biodgradabilit en milieu marinISO 16221:2001(E) ii ISO 16221:2001(E)iii Contents Page Foreword.iv Introduction.v 1 Scope 1 2 Normative references 1 3 Terms and definitions .2 4 Principle4 5 Test environment.4 6 Reagents.4 7 Apparatus .8 8 Test procedure.8 9 Calculation an
12、d expression of results.9 10 Validity of results.10 11 Results of ring tests 10 12 Test report 10 Bibliography12ISO 16221:2001(E) iv Foreword ISO (the International Organization for Standardization) is a worldwide federation of national standards bodies (ISO member bodies). The work of preparing Int
13、ernational 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. International organizations, governmental and non-governmental, in liaison w
14、ith 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 given in the ISO/IEC Directives, Part 3. Draft International Sta
15、ndards adopted by the technical committees are circulated to the member bodies for voting. Publication as an International Standard requires approval by at least 75 % of the member bodies casting a vote. Attention is drawn to the possibility that some of the elements of this International Standard m
16、ay be the subject of patent rights. ISO shall not be held responsible for identifying any or all such patent rights. International Standard ISO 16221 was prepared by Technical Committee ISO/TC 147, Water quality, Subcommittee SC 5, Biological methods.ISO 16221:2001(E)v Introduction ISO/TC 147 has es
17、tablished International Standards for testing biodegradability of substances and waste water in the aquatic environment. All these methods, which are summarized in ISO 15462, can only be used for the determination and prediction of biodegradability in fresh water. There are, however, many cases, for
18、 example, substances used off-shore, where an urgent need exists for testing biodegradability in the marine environment. This International Standard describes biodegradation testing in marine test systems, and is based on an established OECD Guideline and the experience gained by a working group of
19、the Oslo and Paris Commission (OSPARCOM) which has selected suitable standardized ISO methods, adopted for marine conditions and checked in a ring test.INTERNATIONAL STANDARD ISO 16221:2001(E)1 Water quality Guidance for determination of biodegradability in t h em a r i n ee nv i ro nm e nt 1 Scope
20、This International Standard specifies five methods for determining the ultimate aerobic biodegradability of organic compounds in the marine environment by aerobic microorganisms in static aqueous test systems. Standard degradation methods developed for testing in fresh water are modified and adapted
21、 to marine conditions. These methods are the DOC die-away test (ISO 7827), the closed bottle test (ISO 10707), the two-phase closed bottle test (ISO 10708), the CO 2 evolution test (ISO 9439) and the CO 2 headspace test (ISO 14593). The methods apply to organic compounds which a) are water-soluble u
22、nder the conditions of the test used; b) are poorly water-soluble under the conditions of the test used, in which case special measures may be necessary to achieve good dispersion of the compound (see for example, ISO 10634); c) are volatile, provided that an appropriate test with suitable condition
23、s is used; d) are not inhibitory to the test microorganisms at the concentration chosen for the tests. The presence of inhibitory effects can be determined as specified in this International Standard. NOTE The conditions described in this International Standard do not always correspond to the optima
24、l conditions for allowing the maximum degree of biodegradation to occur. For biodegradation methods in fresh water see ISO 14593 and ISO 15462, and for biodegradation at low concentrations see ISO 14592. 2 Normative references The following normative documents contain provisions which, through refer
25、ence in this text, constitute provisions of this International Standard. For dated references, subsequent amendments to, or revisions of, any of these publications do not apply. However, parties to agreements based on this International Standard are encouraged to investigate the possibility of apply
26、ing the most recent editions of the normative documents indicated below. For undated references, the latest edition of the normative document referred to applies. Members of ISO and IEC maintain registers of currently valid International Standards. ISO 7827, Water quality Evaluation in an aqueous me
27、dium of the “ultimate” aerobic biodegradability of organic compounds Method by analysis of dissolved organic carbon (DOC). ISO 9439, Water quality Evaluation of ultimate aerobic biodegradability of organic compounds in aqueous medium Carbon dioxide evolution test. ISO 10707, Water quality Evaluation
28、 in an aqueous medium of the “ultimate” aerobic biodegradability of organic compounds Method by analysis of biochemical oxygen demand (closed bottle test). ISO 10708, Water quality Evaluation in an aqueous medium of the “ultimate” aerobic biodegradability of organic compounds Determination of bioche
29、mical oxygen demand in a two-phase closed bottle test.ISO 16221:2001(E) 2 ISO 14592-1, Water quality Evaluation of the aerobic biodegradability of organic compounds at low concentrations Part 1: Shake-flask batch test with surface water or surface water/sediment suspensions. ISO 14592-2, Water quali
30、ty Evaluation of the aerobic biodegradability of organic compounds at low concentrations Part 2: Continuous flow river model with attached biomass. ISO 14593, Water quality Evaluation of ultimate aerobic biodegradability of organic compounds in aqueous medium Method by analysis of inorganic carbon i
31、n sealed vessels (CO 2 headspace test). 3 Terms and definitions For the purposes of this International Standard, the following terms and definitions apply. 3.1 ultimate aerobic biodegradation breakdown of a chemical compound or organic matter by microorganisms in the presence of oxygen to carbon dio
32、xide, water and mineral salts of any other elements present (mineralization) and normally the production of new biomass 3.2 primary biodegradation structural change (transformation) of a chemical compound by microorganisms resulting in the loss of a specific property 3.3 total organic carbon TOC all
33、 that carbon present in organic matter which is dissolved and suspended in the water sample 3.4 dissolved organic carbon DOC that part of the organic carbon in the water sample which cannot be removed by specified phase separation NOTE Examples of specified phase separation are centrifugation at 40
34、000 ms -2 for 15 min or membrane filtration using membranes with pores of 0,2 mt o0, 45 mdia met er. 3.5 total inorganic carbon TIC all that carbon in the water sample deriving from carbon dioxide and carbonate. 3.6 dissolved inorganic carbon DIC that part of the carbon in the water sample which can
35、not be removed by specified phase separation NOTE Examples of specified phase separation are centrifugation at 40 000 ms -2 for 15 min or membrane filtration using membranes with pores of 0,2 mt o0, 45 mdia met er. 3.7 chemical oxygen demand COD mass concentration of oxygen equivalent to the amount
36、of a specified oxidant consumed by a chemical compound or organic matter when a water sample is treated with that oxidant under defined conditions NOTE It is expressed in this case as milligrams oxygen uptake per milligram (or gram) test compound.ISO 16221:2001(E)3 3.8 biochemical oxygen demand BOD
37、mass concentration of dissolved oxygen consumed under specified conditions by the aerobic biological oxidation of a chemical compound or organic matter in the water sample NOTE It is expressed in this case as milligrams oxygen uptake per milligram (or gram) test compound. 3.9 theoretical oxygen dema
38、nd ThOD theoretical amount of oxygen required to oxidize a chemical compound completely, calculated from the molecular formula NOTE It is expressed in this case as milligrams oxygen uptake per milligram (or gram) test compound. 3.10 theoretical amount of formed carbon dioxide ThCO 2 theoretical amou
39、nt of carbon dioxide formed after oxidizing a chemical compound completely, calculated from the molecular formula NOTE It is expressed in this case as milligrams oxygen uptake per milligram (or gram) test compound. 3.11 theoretical amount of inorganic carbon ThIC theoretical amount of inorganic carb
40、on formed after oxidizing a chemical compound completely, calculated from the molecular formula NOTE It is expressed in this case as milligrams oxygen uptake per milligram (or gram) test compound. 3.12 lag phase time from the start of a test until adaptation and/or selection of the degrading microor
41、ganisms are achieved and the biodegradation degree of a chemical compound or organic matter has increased to about 10 % biodegradation NOTE It is expressed in days. 3.13 maximum level of biodegradation degree of biodegradation of a chemical compound or organic matter in a test, above which no furthe
42、r biodegradation takes place during the test NOTE It is expressed as a percent. 3.14 biodegradation phase time from the end of the lag phase of a test until about 90 % of the maximum level of biodegradation has been reached NOTE It is expressed in days. 3.15 plateau phase time from the end of the bi
43、odegradation phase until the end of the test NOTE It is expressed in days.ISO 16221:2001(E) 4 3.16 pre-exposure pre-incubation of an inoculum in the presence of the test chemical compound or organic matter, with the aim of enhancing the ability of this inoculum to biodegrade the test material by ada
44、ptation and/or selection of the microorganisms 3.17 preconditioning pre-incubation of an inoculum under the conditions of the subsequent test in the absence of the test chemical compound or organic matter, with the aim of improving the performance of the test by acclimatization of the microorganisms
45、 to the test conditions 4P r i n c i p l e This International Standard describes five methods for determining the biodegradability of organic compounds in the marine environment by aerobic microorganisms using static aqueous test systems. Standard degradation methods developed for testing in fresh w
46、ater are modified, adapted and used for this purpose. Test mixtures are prepared containing natural or artificial seawater, marine bacteria and the organic compound, at a suitable concentration, as the sole source of carbon and energy. The test mixtures and controls are incubated at the desired temp
47、erature. Ultimate biodegradation is followed over a specified period by measuring summary parameters as described in the basic test methods. Biodegradation based on DOC (dissolved organic carbon) removal is determined by comparing the measured concentrations at the start and the end of the test as s
48、pecified in the DOC die-away test (ISO 7827). BOD (biochemical oxygen demand) is measured and compared with the theoretical oxygen demand (ThOD) or the measured chemical oxygen demand (COD) as specified in the closed bottle test (ISO 10707) and the two-phase closed bottle test (ISO 10708). The evolu
49、tion of carbon dioxide (CO 2 )i s determined and compared with the theoretical carbon dioxide evolution (ThCO 2 )u s i n gt h eC O 2 evolution test (ISO 9439), and TIC (total inorganic carbon) is determined and compared with the theoretical inorganic carbon (ThIC) in accordance with the CO 2 headspace test (ISO 14593). If required and if a substance-specific analytical method is available, information on primary degradability may be obtained by measuring the loss of the test compound during the test, or biodegradation may be determined at
