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1、BRITISH STANDARD BS 6068-5.0: 1998 ISO/TR 15462:1997 Water quality Part 5: Biological methods Section 5.0: Selection of tests for biodegradability ICS 13.060.01 Licensed Copy: sheffieldun sheffieldun, na, Tue Dec 05 01:30:48 GMT+00:00 2006, Uncontrolled Copy, (c) BSI BS 6068-5.0:1998 This British St
2、andard, having been prepared under the direction of the Health and Environment Sector Board, was published under the authority of the Standards Board and comes into effect on 15 April 1998 BSI 04-1999 ISBN 0 580 29786 1 Amendments issued since publication Amd. No.DateComments Licensed Copy: sheffiel
3、dun sheffieldun, na, Tue Dec 05 01:30:48 GMT+00:00 2006, Uncontrolled Copy, (c) BSI BS 6068-5.0:1998 BSI 04-1999i Contents Page National forewordii Forewordii 1Scope1 2References1 3Conclusions and recommendations2 Annex A Overview on ISO methods4 Annex B Comparison of International Standards with OE
4、CD guidelines9 Licensed Copy: sheffieldun sheffieldun, na, Tue Dec 05 01:30:48 GMT+00:00 2006, Uncontrolled Copy, (c) BSI BS 6068-5.0:1998 ii BSI 04-1999 National foreword This British Standard reproduces verbatim ISO/TR 15462:1997 and implements it as the UK national standard. The UK participation
5、in its preparation was entrusted by Technical Committee EH/3, Water quality, to Subcommittee EH/3/5, Biological methods, which has the responsibility to: aid enquirers to understand the text; present to the responsible international/European committee any enquiries on the interpretation, or proposal
6、s for change, and keep the UK interests informed; monitor related international and European developments and promulgate them in the UK. A list of organizations represented on this subcommittee can be obtained on request to its secretary. This is one of a series of standards on water quality, others
7、 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: Physical, chemical and biochemical methods; Part 3: Radiological method
8、s; Part 4: Microbiological methods; Part 5: Biological methods; Part 6: Sampling; Part 7: Precision and accuracy; Cross-references The British Standards which implement international or European publications referred to in this document may be found in the BSI Standards Catalogue under the section e
9、ntitled “International 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. Comp
10、liance with a British Standard does not of itself confer immunity from legal obligations. Summary of pages This document comprises a front cover, an inside front cover, pages i and ii, the ISO/TR title page, page ii, pages 1 to 9 and a back cover. This standard has been updated (see copyright date)
11、and may have had amendments incorporated. This will be indicated in the amendment table on the inside front cover. Licensed Copy: sheffieldun sheffieldun, na, Tue Dec 05 01:30:48 GMT+00:00 2006, Uncontrolled Copy, (c) BSI Licensed Copy: sheffieldun sheffieldun, na, Tue Dec 05 01:30:48 GMT+00:00 2006
12、, Uncontrolled Copy, (c) BSI BS 6068-5.0:1998 ii BSI 04-1999 Foreword ISO (the International Organization for Standardization) is a worldwide federation of national standards bodies (ISO member bodies). The work of preparing International Standards is normally carried out through ISO technical commi
13、ttees. 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 with ISO, also take part in the work. ISO collaborates closely with the I
14、nternational Electrotechnical Commission (IEC) on all matters of electrotechnical standardization. The main task of technical committees is to prepare International Standards, but in exceptional circumstances a technical committee may propose the publication of a Technical Report of one of the follo
15、wing types: type 1, when the required support cannot be obtained for the publication of an International Standard, despite repeated efforts; type 2, when the subject is still under technical development or where for any other reason there is the future but not immediate possibility of an agreement o
16、n an International Standard; type 3, when a technical committee has collected data of a different kind from that which is normally published as an International Standard (“state of the art”, for example). Technical Reports of types 1 and 2 are subject to review within three years of publication, to
17、decide whether they can be transformed into International Standards. Technical Reports of type 3 do not necessarily have to be reviewed until the data they provide are considered to be no longer valid or useful. ISO/TR 15462, which is a Technical Report of type 3, was prepared by Technical Committee
18、 ISO/TC 147, Water quality, Subcommittee SC 5, Biological methods. Descriptors: Water, quality, water pollution, tests, water tests, determination, biodegradability, selection, surveys. Licensed Copy: sheffieldun sheffieldun, na, Tue Dec 05 01:30:48 GMT+00:00 2006, Uncontrolled Copy, (c) BSI BS 6068
19、-5.0:1998 BSI 04-19991 1 Scope The biodegradability of substances and wastewater ingredients depends not only on the molecular structures of the test material, but also on important additional factors such as the aquatic or terrestrial test environment; aerobic or anaerobic test conditions; source a
20、nd concentration of the microorganisms of the inoculum; acclimatisation and adaptation of the inoculum; concentration of the test material; availability of inorganic nutrients and additional organic material for co-metabolic processes; possible toxic effects of the test material under the test condi
21、tions; physical and chemical properties and bioavailability of the test material (e.g. volatility, water solubility, adsorption); conditions and physical and chemical properties of the test system (volume of test vessels, static, dynamic, closed, CO2 removal, temperature, mixing, shaking, oxygen sup
22、ply); test duration; analytical parameters used (summary parameters such as DOC, BOD, CO2 or substance specific analysis). As so many factors can influence the test results it is not possible to define a “true” or “reference” method. The reproducibility of the test results using different methods or
23、 conditions or even using identical test methods can be low and differing test results can be obtained. Normally a test material which is either easily biodegradable or poorly biodegradable will produce similar test results. So-called “grey” substances, which are moderately biodegradable and need, f
24、or example, special bacteria or long adaptation periods, will often produce disparate results. Biodegradability tests are used to establish the biodegradability of chemical structures and to predict the biodegradation behaviour of a test material in a natural or technical environment. The aim of eac
25、h test method should be to simulate, to a certain degree, such an environment. Therefore, as so many factors can influence or even exclude certain methods, it is necessary to have a sufficient number of different standardized test methods to allow the choice of the best one for the specific purpose.
26、 It is also to be remembered that it was ISOs intention to standardize the biodegradation tests of the OECD Guidelines for Testing of Chemicals (see Annex B). 2 References ISO 7827:1994, Water quality Evaluation in an aqueous medium of the “ultimate” aerobic biodegradability of organic compounds Met
27、hod by analysis of dissolved organic carbon (DOC). ISO 8192:1986, Water quality Test for inhibition of oxygen consumption by activated sludge. ISO 9408:1991, Water quality Evaluation in an aqueous medium of the “ultimate” aerobic biodegradability of organic compounds Method by determining the oxygen
28、 demand in a closed respirometer. ISO 9439:1990, Water quality Evaluation in an aqueous medium of the “ultimate” aerobic biodegradability of organic compounds Method by analysis of released carbon dioxide. ISO 9887:1992, Water quality Evaluation of the aerobic biodegradability of organic compounds i
29、n an aqueous medium Semi-continuous activated sludge method (SCAS). ISO 9888:1991, Water quality Evaluation of the aerobic biodegradability of organic compounds in an aqueous medium Static test (Zahn-Wellens method). ISO 10634:1995, Water quality Guidance for the preparation and treatment of poorly
30、water-soluble organic compounds for the subsequent evaluation of their biodegradability in an aqueous medium. ISO 10707:1994, Water quality Evaluation in an aqueous medium of the ultimate aerobic biodegradability of organic compounds Method by analysis of biochemical oxygen demand (closed bottle tes
31、t). ISO 10708:1997, Water quality Evaluation in an aqueous medium of the ultimate aerobic biodegradability of organic compounds Method by determining the biochemical oxygen demand in a two-phase closed bottle test. ISO 10712:1995, Water quality Pseudomonas putida growth inhibition test (Pseudomonas
32、cell multiplication inhibition test). ISO 11733:1995, Water quality Evaluation of the elimination and the biodegradability of organic compounds in an aqueous medium Activated sludge simulation test. ISO 11734:1995, Water quality Evaluation of the ultimate anaerobic biodegradability of organic compou
33、nds in digested sludge Method by measurement of the biogas production. ISO 14592-1 and ISO 14592-2, Water quality Evaluation of aerobic biodegradability of organic compounds at low concentrations in water Part 1: Shake flask method; Part 2: River simulation test (in preparation). Licensed Copy: shef
34、fieldun sheffieldun, na, Tue Dec 05 01:30:48 GMT+00:00 2006, Uncontrolled Copy, (c) BSI BS 6068-5-0:1998 2 BSI 04-1999 ISO 14593, Water quality Evaluation in aqueous medium of ultimate aerobic biodegradability of organic compounds Determination of released inorganic carbon in sealed vessels (in prep
35、aration). ISO 15522, Water quality Determination of inhibitory effect of water constituents on growth of sludge microorganisms (in preparation). 3 Conclusions and recommendations 3.1 Differences between test methods The test methods for aerobic biodegradability are not of equal potential, largely be
36、cause of the different microbial densities and test durations used. ISO 7827, ISO 9439, ISO 9408, ISO 10708 and ISO 14593 are of roughly equal potential. ISO 10707 has a lower potential and is especially applicable to volatile and inhibitory test compounds. These methods correspond in principle to t
37、he tests on ready biodegradability of OECD (see 3.4). ISO 9887 and ISO 9888 are tests with a high degradation potential and may be used to determine the intrinsic or inherent biodegradability of a chemical. ISO 11733 is a simulation test for wastewater treatment plants and ISO 14592 for river water.
38、 ISO 11734 is a test for anaerobic biodegradability. As yet only one method for assessing anaerobic biodegradability has been agreed, and is applied independently of tests for aerobic biodegradability. Priority for applying the test could be given to those chemicals which preferentially adsorb onto
39、solids such as activated sludge and which are not sufficiently degraded aerobically. Chemicals which degrade readily or easily in the aquatic environment will normally degrade equally well in all of the aerobic ISO tests, while less easily degradable chemicals degrade only in those with a higher pot
40、ential, but not in any of the tests using lower cell densities. Similarly, some chemicals which degrade in the more powerful batch tests (ISO 9887 and ISO 9888) do not necessarily degrade in the continuously performed activated sludge simulation test, while other chemicals do and there may be of cou
41、rse differences between aerobic and anaerobic biodegradability. 3.2 Analytical parameters Normally, summary parameters such as dissolved organic carbon, biochemical oxygen demand or biologically released carbon dioxide are used as analytical tools to obtain information on ultimate biodegradability (
42、mineralization). However, the ISO methods also allow the use of substance-specific analytical techniques to investigate the biological transformation of a chemical, the so-called primary biodegradability. A list of test methods is given in Annex A. 3.3 Bacterial toxicity It has also been observed th
43、at chemicals which are inhibitory towards bacteria and do not degrade at the normal test concentrations may degrade in tests using lower concentrations of the test compound. If a chemical is suspected of not degrading because of toxicity to the test bacteria, additional vessels may be set up within
44、the test to assess any inhibition. Otherwise specific tests may be used, such as the test for inhibition of oxygen consumption by activated sludge (ISO 8192) or a test on the growth of sewage bacteria (ISO 15522) or Pseudomonas putida (ISO 10712). 3.4 Test strategies As there are differences between
45、 the complexity, duration, quality of results and costs of the various tests, these should be considered when choosing which test or test strategy to use when commencing a study. The choice of a test depends normally on the purpose of testing or on legal requirements which have to be fulfilled. One
46、strategy is to apply a simple low-cell density method. If a chemical degrades adequately, further testing is normally unnecessary. A low or zero value for biodegradation may be sufficient for the purpose, otherwise the test could be repeated with an inoculum pre-exposed to the chemical, or a test wi
47、th a higher cell density and a longer test period could be applied. Another strategy is to start with a high-density method to determine whether biodegradation potential is available at all and to answer the question, “Is the test compound easily biodegradable in environmental compartments?” by usin
48、g a low-cell density method in the next step. Because of its complexity, the activated sludge simulation test (ISO 11733) is not usually applied until after other tests have first been carried out. Failure to degrade in one of the more powerful tests, ISO 9887 and ISO 9888, makes it often unnecessar
49、y to apply a simulation test. The choice of a method also depends on the physical and chemical properties of the test chemical. Volatile test compounds can only be tested in closed systems such as ISO 10707 or ISO 10708. Insoluble and poorly soluble chemicals cannot be assessed by all the available methods and guidance in preparing such chemicals for testing is given in ISO 10634. In some cases it may be helpful to use an adapted inoculum. Pre-adaptation could be achieved usin