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1、 Reference number ISO 11348-1:2007(E) ISO 2007 INTERNATIONAL STANDARD ISO 11348-1 Second edition 2007-12-01 Water quality Determination of the inhibitory effect of water samples on the light emission of Vibrio fischeri (Luminescent bacteria test) Part 1: Method using freshly prepared bacteria Qualit
2、 de leau Dtermination de leffet inhibiteur dchantillons deau sur la luminescence de Vibrio fischeri (Essai de bactries luminescentes) Partie 1: Mthode utilisant des bactries frachement prpares Copyright International Organization for Standardization Provided by IHS under license with ISO Licensee=IH
3、S Employees/1111111001, User=Japan, IHS Not for Resale, 12/17/2007 19:55:29 MSTNo reproduction or networking permitted without license from IHS -,-,- ISO 11348-1:2007(E) PDF disclaimer This PDF file may contain embedded typefaces. In accordance with Adobes licensing policy, this file may be printed
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6、table for use by ISO member bodies. In the unlikely event that a problem relating to it is found, please inform the Central Secretariat at the address given below. COPYRIGHT PROTECTED DOCUMENT ISO 2007 All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or
7、utilized in any form or by any means, electronic or mechanical, including photocopying and microfilm, without permission in 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
8、Fax + 41 22 749 09 47 E-mail copyrightiso.org Web www.iso.org Published in Switzerland ii ISO 2007 All rights reserved Copyright International Organization for Standardization Provided by IHS under license with ISO Licensee=IHS Employees/1111111001, User=Japan, IHS Not for Resale, 12/17/2007 19:55:2
9、9 MSTNo reproduction or networking permitted without license from IHS -,-,- ISO 11348-1:2007(E) ISO 2007 All rights reserved iii Contents Page Foreword iv Introduction v 1 Scope . 1 2 Normative references. 1 3 Principle. 2 4 Interferences . 2 5 Reagents and materials . 2 6 Apparatus 4 7 Sampling and
10、 sample pretreatment 5 8 Cultivation of luminescent bacteria 5 9 Procedure 7 10 Evaluation 8 11 Expression of results . 10 12 Criteria of validity 11 13 Precision 12 14 Test report . 12 Annex A (informative) Colour-correction method. 13 Annex B (informative) Dilution level D Preparation of the dilut
11、ion series. 16 Annex C (informative) Precision data. 19 Annex D (informative) Testing salt water samples with the luminescent bacteria test with freshly cultured bacteria. 20 Bibliography. 23 Copyright International Organization for Standardization Provided by IHS under license with ISO Licensee=IHS
12、 Employees/1111111001, User=Japan, IHS Not for Resale, 12/17/2007 19:55:29 MSTNo reproduction or networking permitted without license from IHS -,-,- ISO 11348-1:2007(E) iv ISO 2007 All rights reserved Foreword ISO (the International Organization for Standardization) is a worldwide federation of nati
13、onal 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. Internat
14、ional 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 ru
15、les 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
16、 least 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 11348-1 was prepared by Technical Committee ISO/
17、TC 147, Water quality, Subcommittee SC 5, Biological methods. This second edition cancels and replaces the first edition (ISO 11348-1:1998), which has been technically revised. ISO 11348 consists of the following parts, under the general title Water quality Determination of the inhibitory effect of
18、water samples on the light emission of Vibrio fischeri (Luminescent bacteria test): Part 1: Method using freshly prepared bacteria Part 2: Method using liquid-dried bacteria Part 3: Method using freeze-dried bacteria Copyright International Organization for Standardization Provided by IHS under lice
19、nse with ISO Licensee=IHS Employees/1111111001, User=Japan, IHS Not for Resale, 12/17/2007 19:55:29 MSTNo reproduction or networking permitted without license from IHS -,-,- ISO 11348-1:2007(E) ISO 2007 All rights reserved v Introduction The measurements specified in ISO 11348 can be carried out usi
20、ng freshly prepared bacteria, as well as freeze-dried or liquid-dried bacterial preparations. Standardized work carried out by DIN Normenausschuss Wasserwesen and ISO/TC 147/SC 5/WG 1 has shown that, in special cases, these different techniques may deliver different results, especially in the presen
21、ce of heavy metals. Such varying sensitivity is caused by differences in media composition used in the preparation of freeze-dried or liquid-dried bacteria. These protective media influence the bioavailability of toxicants and/or the light emission of luminescent bacteria. This means that the origin
22、 and type of preparation need to be taken into account when interpreting the results. This may be difficult sometimes, as freeze-dried and liquid-dried bacteria may be obtained from different suppliers. This, in turn, can mean that the composition is not known in detail and therefore cannot be inter
23、preted by the user. For this reason, in addition to toxicity measurements with liquid-dried bacteria (ISO 11348-2) and freeze-dried bacteria (ISO 11348-3), a procedure with freshly prepared bacteria is described in this part of ISO 11348, the performance of which can be interpreted by the user in ev
24、ery detail. The laboratories responsible for the results have the opportunity to select the most suitable technique based on expert judgement and information about the water sample to be tested. Copyright International Organization for Standardization Provided by IHS under license with ISO Licensee=
25、IHS Employees/1111111001, User=Japan, IHS Not for Resale, 12/17/2007 19:55:29 MSTNo reproduction or networking permitted without license from IHS -,-,- Copyright International Organization for Standardization Provided by IHS under license with ISO Licensee=IHS Employees/1111111001, User=Japan, IHS N
26、ot for Resale, 12/17/2007 19:55:29 MSTNo reproduction or networking permitted without license from IHS -,-,- INTERNATIONAL STANDARD ISO 11348-1:2007(E) ISO 2007 All rights reserved 1 Water quality Determination of the inhibitory effect of water samples on the light emission of Vibrio fischeri (Lumin
27、escent bacteria test) Part 1: Method using freshly prepared bacteria WARNING Persons using this part of ISO 11348 should be familiar with normal laboratory practice. This standard does not purport to address all of the safety problems, if any, associated with its use. It is the responsibility of the
28、 user to establish appropriate safety and health practices and to ensure compliance with any national regulatory conditions. IMPORTANT It is absolutely essential that tests conducted in accordance with this part of ISO 11348 be carried out by suitably trained staff. 1 Scope ISO 11348 describes three
29、 methods for determining the inhibition of the luminescence emitted by the marine bacterium Vibrio fischeri (NRRL B-11177). This part of ISO 11348 specifies a method using freshly prepared bacteria. This method is applicable to: waste water; aqueous extracts and leachates; fresh water (surface and g
30、round water); sea and brackish water; eluates of sediment (fresh water, brackish and sea water); pore water; single substances, diluted in water. 2 Normative references The following referenced documents are indispensable for the application of this document. For dated references, only the edition c
31、ited applies. For undated references, the latest edition of the referenced document (including any amendments) applies. ISO 5667-16, Water quality Sampling Part 16: Guidance on biotesting of samples ISO 5814, Water quality Determination of dissolved oxygen Electrochemical probe method ISO 7027, Wate
32、r quality Determination of turbidity Copyright International Organization for Standardization Provided by IHS under license with ISO Licensee=IHS Employees/1111111001, User=Japan, IHS Not for Resale, 12/17/2007 19:55:29 MSTNo reproduction or networking permitted without license from IHS -,-,- ISO 11
33、348-1:2007(E) 2 ISO 2007 All rights reserved 3 Principle The inhibition of light emission by cultures of Vibrio fischeri is determined by means of a batch test. This is accomplished by combining specified volumes of the test sample or the diluted sample with the luminescent bacteria suspension in a
34、test tube. The test criterion is the luminescence, measured after a contact time of 15 min or 30 min and optionally 5 min, taking into account a correction factor (fkt), which is a measure of intensity changes of control samples during the exposure time. The inhibitory effect of the water sample can
35、 be determined as LID (see Annex B) or as EC20- and/or EC50-values by means of a dilution series. (EC is the effective concentration). 4 Interferences Insoluble, slightly soluble or volatile substances or substances which react with the dilution water or the suspension, or alter their state during t
36、he test period, may affect the result or impair the reproducibility of the test results. Losses of luminescence caused by light absorption or light scattering may occur in the case of strongly coloured or turbid waters. This interference can be compensated by a sample treatment for turbidity (7.2) o
37、r, for example, by using a double-chambered absorption correction test tube (see Annex A). Since oxygen is required for the bioluminescence 6, samples with a high oxygen demand (and/or a low oxygen concentration) may cause a deficiency of oxygen and be inhibitory. Readily biodegradable nutrients in
38、the sample may cause a pollutant-independent reduction in bioluminescence 1. Samples with a pH outside the range of pH = 6,0 and pH = 8,5 affect the luminescence of the bacteria 6, 7. An adjustment of the sample is required when the toxic effect of pH is not wanted. As the test organism Vibrio fisch
39、eri is a marine bacterium, testing salt-water samples with the standard procedure often leads to stimulation effects of bioluminescence, which may mask inhibition effects (see Annex D). Salt concentrations in the initial sample exceeding 30 g/l NaCl, or contents of other compounds giving equal osmol
40、arity may lead, together with the salt spiking required by the test, to hyperosmotic effects. The resulting salt concentration in the test samples should not exceed the osmolarity of a 35 g/l NaCl solution in order to avoid these effects. 5 Reagents and materials Use chemicals of recognized analytic
41、al grade quality. Use distilled water or water of equivalent purity. 5.1 Test bacteria. Use a strain of luminescence bacteria belonging to the species Vibrio fischeri NRRL B-11177. The bacteria strain can be taken from commercially available freeze-dried or liquid-dried reagents or from culture coll
42、ections, e.g. Deutsche Sammlung fr Mikrorganismen und Zellkulturen GmbH, Mascheroder Weg 10, D-38124 Braunschweig, Germany, or NRRL, ARS Culture collection NCAUR, 1815 N, University Street, Peoria, Illinois 61604, USA. The bacterial suspensions used for toxicity measurements shall be freshly prepare
43、d from cultures. 5.2 Sodium chloride solution, as diluent. Dissolve 20 g of sodium chloride (NaCl) in water and make up to 1 l with water. Copyright International Organization for Standardization Provided by IHS under license with ISO Licensee=IHS Employees/1111111001, User=Japan, IHS Not for Resale
44、, 12/17/2007 19:55:29 MSTNo reproduction or networking permitted without license from IHS -,-,- ISO 11348-1:2007(E) ISO 2007 All rights reserved 3 5.3 Sodium hydroxide solution, e.g. c(NaOH) = 1 mol/l. 5.4 Hydrochloric acid, e.g. c(HCl) = 1 mol/l. For the adjustment of the pH, it may be necessary to
45、 use acids or bases of lower or higher concentration. 5.5 Solution for freshly prepared bacteria. 8,0 g D(+)-Glucose monohydrate (C6H12O6H2O) 20,0 g Sodium chloride (NaCl) 2,035 g Magnesium chloride hexahydrate (MgCl26 H2O) 0,30 g Potassium chloride (KCl) 11,9 g N-(2-Hydroxyethyl)piperazine-N-(2-eth
46、anesulfonic acid) (HEPES) Dissolve in water, stir for about 30 min and adjust the pH to 7,0 0,2 with sodium hydroxide solution (5.3) or hydrochloric acid (5.4). Make up to 1 l with water. This solution may be stored in portions at 18 C to 20 C. 5.6 Reference substances. Prepare the following referen
47、ce-substance stock solutions with sodium chloride solution (5.2) as diluent separately, without adjustment of the pH: 219,8 mg/l Zinc sulfate heptahydrate (ZnSO47 H2O) 9 mg/l 3,5-Dichlorophenol (C6H4OCl2) (purity W 99 %) 22,6 mg/l Potassium dichromate (K2Cr2O7) These concentrations are approximately
48、 twice the expected EC50-values for the respective reference substances in this part of ISO 11348. The volumes required depend on the test set-up. NOTE It is possible to use commercially available chemical preparations with defined concentrations of ZnSO4 and K2Cr2O7 (titrisol) for the preparation of the stock solutions of the reference substances. 5.7 Liquid broth for pre- and main cultures. 30 g Sodium chloride (NaCl) 6,10 g Sodiu