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1、 Reference number ISO 14442:2006(E) ISO 2006 INTERNATIONAL STANDARD ISO 14442 Second edition 2006-04-01 Water quality Guidelines for algal growth inhibition tests with poorly soluble materials, volatile compounds, metals and waste water Qualit de leau Lignes directrices pour essais dinhibition de la
2、 croissance algale avec des matires peu solubles, des composs volatils, des mtaux et des eaux rsiduaires Copyright International Organization for Standardization Provided by IHS under license with ISO Licensee=NASA Technical Standards 1/9972545001 Not for Resale, 04/19/2007 06:30:16 MDTNo reproducti
3、on or networking permitted without license from IHS -,-,- ISO 14442:2006(E) PDF disclaimer This PDF file may contain embedded typefaces. In accordance with Adobes licensing policy, this file may be printed or viewed but shall not be edited unless the typefaces which are embedded are licensed to and
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6、nd, please inform the Central Secretariat at the address given below. ISO 2006 All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying and microfilm, without permission in
7、 writing from either ISO at the address below or ISOs 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 2006 All rights reserved Copy
8、right International Organization for Standardization Provided by IHS under license with ISO Licensee=NASA Technical Standards 1/9972545001 Not for Resale, 04/19/2007 06:30:16 MDTNo reproduction or networking permitted without license from IHS -,-,- ISO 14442:2006(E) ISO 2006 All rights reserved iii
9、Contents Page Foreword iv 1 Scope . 1 2 Normative references. 2 3 Analytical characterization of test materials and confirmation of concentrations and stability 2 4 Poorly soluble organic substances 3 4.1 General. 3 4.2 Preparation of saturated and supersaturated solutions. 3 4.3 Solvent addition 4
10、4.4 Dispersion using an emulsifying agent 5 4.5 Interference with algal growth and its measurement 5 5 Poorly soluble mixtures of organic substances 6 5.1 General. 6 5.2 Preparation of test media. 6 5.3 Test performance 7 6 Poorly soluble inorganic materials. 7 7 Volatile substances 8 7.1 General. 8
11、 7.2 Test system and growth medium 8 7.3 Test performance 9 7.4 Interference with algal growth. 9 8 Waste waters and environmental aqueous samples 9 9 Coloured and/or turbid samples . 10 10 Metals and metal compounds . 11 10.1 Introduction. 11 10.2 Modification of algal growth inhibition test proced
12、ures for testing materials containing heavy metals . 11 11 pH buffering. 12 12 Interpretation of the results. 13 Bibliography. 14 Copyright International Organization for Standardization Provided by IHS under license with ISO Licensee=NASA Technical Standards 1/9972545001 Not for Resale, 04/19/2007
13、06:30:16 MDTNo reproduction or networking permitted without license from IHS -,-,- ISO 14442:2006(E) iv ISO 2006 All rights reserved Foreword ISO (the International Organization for Standardization) is a worldwide federation of national standards bodies (ISO member bodies). The work of preparing Int
14、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
15、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 2. The main task of techni
16、cal 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 least 75 % of the member bodies casting a vote. Attention is drawn
17、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 14442 was prepared by Technical Committee ISO/TC 147, Water quality, Subcommittee SC 5, Biological methods. This sec
18、ond edition cancels and replaces the first edition (ISO 14442:1999), which has been technically revised. Copyright International Organization for Standardization Provided by IHS under license with ISO Licensee=NASA Technical Standards 1/9972545001 Not for Resale, 04/19/2007 06:30:16 MDTNo reproducti
19、on or networking permitted without license from IHS -,-,- INTERNATIONAL STANDARD ISO 14442:2006(E) ISO 2006 All rights reserved 1 Water quality Guidelines for algal growth inhibition tests with poorly soluble materials, volatile compounds, metals and waste water WARNING Persons using this Internatio
20、nal Standard should be familiar with normal laboratory practice. This International Standard does not purport to address all of the safety problems, if any, associated with its use. It is the responsibility of the user to establish appropriate safety and health practices and to ensure compliance wit
21、h any national regulatory conditions. IMPORTANT It is absolutely essential that tests conducted according to this standard be carried out by suitably trained staff. 1 Scope This International Standard provides procedures, not covered by the methods described in ISO 8692 and ISO 10253, for testing di
22、fficult substances for inhibition of algal growth. The main subjects covered by the guideline are the methods for preparing the test substance for testing and the procedures needed to carry out an appropriate test. The following test substances are covered by this guideline: a) poorly soluble pure o
23、rganic compounds; b) poorly soluble mixtures of organic substances; c) poorly soluble inorganic materials; d) volatile substances; e) waste waters and environmental samples containing water and sediments; f) coloured and/or turbid samples; g) compounds of heavy metals. The following methods of addit
24、ion are covered: direct; dispersion; water-soluble and water-accommodated fractions. Some guidelines related to the analytical procedures and to the interpretation of the results have been included. References to documents describing the background for the testing of difficult substances are given i
25、n the Bibliography. Copyright International Organization for Standardization Provided by IHS under license with ISO Licensee=NASA Technical Standards 1/9972545001 Not for Resale, 04/19/2007 06:30:16 MDTNo reproduction or networking permitted without license from IHS -,-,- ISO 14442:2006(E) 2 ISO 200
26、6 All rights reserved 2 Normative references The following referenced documents are indispensable for the application of this document. For dated references, only the edition cited applies. For undated references, the latest edition of the referenced document (including any amendments) applies. ISO
27、5667-16, Water quality Sampling Part 16: Guidance on biotesting of samples ISO 8692, Water quality Freshwater algal growth inhibition test with unicellular green algae ISO 10253, Water quality Marine algal growth inhibition test with Skeletonema costatum and Phaeodactylum tricornutum 3 Analytical ch
28、aracterization of test materials and confirmation of concentrations and stability Analytical characterization of test substances and materials and the confirmation of their concentrations and stability in the testing environment is of major concern of regulatory authorities. Such activities are usua
29、lly not an integral part of this International Standard algal growth inhibition test methods. However, there may be situations where analysis may assist in defining the appropriate exposure conditions of test materials and chemicals and/or in the interpretation of the results. The relevant propertie
30、s of substances and materials can be assessed from basic properties such as solubility in water, partition coefficient (lg Pow), Henrys constant, photochemical and hydrolytic stability and biodegradability. Analytical confirmation is strongly recommended in order to confirm test substance concentrat
31、ions and is required for the calculation of effective concentration (EC) values of volatile substances (Clause 7). If losses due to adsorption on the test vessels or during transfer of test solutions and media occur, then analytical confirmation are of particular importance. This aspect is also spec
32、ified in ISO 5667-16. Due to the batch test system used for algal growth inhibition tests, loss of substances due to biodegradation (nearly all algal cultures contain bacteria), photodegradation, hydrolysis and/or adsorption cannot always be avoided. A decrease in measured concentrations is difficul
33、t to prevent by technical means, and is therefore considered acceptable for algal growth inhibition tests. The following precautions are suggested for maintaining test substance concentrations in algal growth inhibition tests: a) sterilization of media and equipment to reduce the effect of bacterial
34、 growth; b) change of the light quality to prevent photodegradation of test substances; c) avoidance of contact of test substance with water prior to testing to reduce hydrolytic decomposition; d) treatment of glassware (e.g. silanization); the effectiveness of such a treatment varies from one chemi
35、cal to the other; e) pre-conditioning of the glassware, before addition of the test media, with the test substance at concentrations to be used in the test. The effect of such technical measures is, if relevant and if possible, monitored by chemical analysis. Water, waste water and organic/inorganic
36、 solids/liquids may contain components that may modify the composition of the algal growth medium (by precipitation of a limiting nutrient, complexation of essential elements, addition of nutrients), and subsequently may cause effects on algal growth not related to toxic Copyright International Orga
37、nization for Standardization Provided by IHS under license with ISO Licensee=NASA Technical Standards 1/9972545001 Not for Resale, 04/19/2007 06:30:16 MDTNo reproduction or networking permitted without license from IHS -,-,- ISO 14442:2006(E) ISO 2006 All rights reserved 3 components. If such proble
38、ms occur, it may be advisable to determine the content of key components of the test material. Some relevant components are: calcium, magnesium, sodium, potassium, sulfate, chloride, ammonium, nitrate, phosphate, copper, cobalt, nickel, zinc, cadmium, organic matter i.e. measured as Chemical Oxygen
39、Demand (COD) and/or Total Organic Carbon (TOC). If the material contains a high concentration of readily degradable organic material, the subsequent bacterial growth may disturb the algal growth measurement. When untreated (not filtered or centrifuged) waste water is tested, contamination with other
40、 algal species may occur. 4 Poorly soluble organic substances 4.1 General A pure substance is a substance with one major component containing minor components as impurities. Poorly soluble substances are those with solubility limits below 100 mg/l in water. If, however, growth inhibition occurs at c
41、oncentrations much lower than the solubility limits in water or algal growth medium (the limit in the medium may be different), then the poorly soluble substance can be tested as a water soluble substance (added via a stock solution in test medium). This approach is usually not applicable to substan
42、ces with a water solubility below 1 mg/l to 10 mg/l (substances with a very low solubility). The methods described in this clause therefore refer to testing of substances causing effects on algal growth at concentrations at or around the solubility limit in water and to very low solubility substance
43、s. Testing of nominal concentrations markedly above the solubility limit is not recommended. It may, however, be unavoidable if the solubility limit in water or algal growth medium (which may be different) is not well established, or if a substance spontaneously forms dispersions in the test medium.
44、 NOTE Terminology according to Reference 3: water solubility below 100 mg/l: “sparingly soluble”; water solubility below 1 mg/l: “very low solubility”. A number of methods which are available for preparing test solutions of pure substances, are described in ISO 5667-16. Generally, it is preferred to
45、 use mechanical means to prepare stock solutions. 4.2 Preparation of saturated and supersaturated solutions If the solubility of a substance in water is between 1 mg/l and 100 mg/l, saturated solutions can be prepared by direct addition of the test substance. A saturated solution is usually prepared
46、 by stirring (e.g. magnetic stirrer or shaking, see also 5.1) an excess amount of the test substance in water for a period in test medium. A period of 20 h is practical for most substances, but a stirring period of up to three days may be considered to ensure saturation provided the substance is sta
47、ble. Lengthy stirring should be carried out in the dark and in the same temperature range as the growth inhibition test is carried out. Preferably, the equilibrium should be confirmed by chemical analysis. After a phase separation period of varying length, the clear phase is collected and tested as
48、the highest concentration. Filtration (through a 0,45 m membrane filter) or centrifugation may be useful for removing particulate matter. Certain membrane filters may interfere with the test substance. The type of filter should be chosen according to the physico-chemical properties of the test substance and the recommendations of the filter supplier. Further test concentrations can be prepared by dilution of the saturated solution with test medium. A small volume of a concentrated suspension of algal culture is then added to the test media to start the t