1、September 2012 Translation by DIN-Sprachendienst.English price group 14No part of this translation may be reproduced without prior permission ofDIN Deutsches Institut fr Normung e. V., Berlin. Beuth Verlag GmbH, 10772 Berlin, Germany,has the exclusive right of sale for German Standards (DIN-Normen).
2、ICS 13.060.70!$Vc“1915164www.din.deDDIN EN 16161Water quality Guidance on the use of in vivo absorption techniques for the estimationof chlorophyll-a concentration in marine and fresh water samples;English version EN 16161:2012,English translation of DIN EN 16161:2012-09Wasserbeschaffenheit Anleitun
3、g fr die Anwendung der in-vivo-Absorption zur Abschtzung der Chlorophylla-Konzentration in Meer- und Swasser;Englische Fassung EN 16161:2012,Englische bersetzung von DIN EN 16161:2012-09Qualit de leau Lignes directrices sur lutilisation des techniques dabsorption in vivo pour lestimation dela concen
4、tration de chlorophylle-a dans les eaux marines et eaux douces;Version anglaise EN 16161:2012,Traduction anglaise de DIN EN 16161:2012-09www.beuth.deIn case of doubt, the German-language original shall be considered authoritative.Document comprises 29 pages 08.12 DIN EN 16161:2012-09 2 A comma is us
5、ed as the decimal marker. National foreword This standard has been prepared by Technical Committee CEN/TC 230 “Water analysis” (Secretariat: DIN, Germany). The responsible German body involved in its preparation was the Normenausschuss Wasserwesen (Water Practice Standards Committee), Working Commit
6、tee NA 119-01-03-05-06 AK Biologisch-kologische Gewsseruntersuchung of NA 119-01-03 AA Wasseruntersuchung. This standard is part of the series Deutsche Einheitsverfahren zur Wasser-, Abwasser- und Schlamm-untersuchung Biologisch-kologische Gewsseruntersuchung (Gruppe M) (German standard methods for
7、the examination of water, waste water and sludge Biological-ecological methods of analysis (group M) and describes method M 39. Designation of method M 39 “Guidance on the use of in vivo absorption techniques for the estimation of chlorophyll-a concentration in marine and fresh water samples”: Metho
8、d DIN EN 16161 M 39 This standard has been published to implement the Water Framework Directive (WFD), Directive 2000/60/EC of the European Parliament and of the Council of 23 October 2000 establishing a framework for Community action in the field of water policy1). Expert assistance and specialized
9、 laboratories will be required to perform the analysis described in this standard. Existing safety requirements are to be observed. Depending on the objective of the analysis, a check shall be made on a case-by-case basis as to whether and to what extent additional conditions will have to be specifi
10、ed. German standard methods for the examination of water, waste water and sludge Standard methods published as DIN Standards are obtainable from Beuth Verlag GmbH, either individually or grouped in volumes. The standard methods included in the loose-leaf publication entitled Deutsche Einheitsverfahr
11、en zur Wasser-, Abwasser- und Schlammuntersuchung will continue to be published by Wiley- VCH Verlag and Beuth Verlag GmbH. All standard methods relevant to the Abwasserverordnung (Waste Water Regulation) (AbwV)2) included in the new Regulation on Section 57, paragraph 1, number 1, of the Gesetz zur
12、 Ordnung des Wasserhaushalts (German Water Management Act)2)concerning Anforderungen an das Einleiten von Abwasser in Gewsser - together with the Abwasserverordnung and the Gesetz zur Ordnung des Wasserhaushalts have been published by Beuth Verlag GmbH as a loose-leaf collection Analysenverfahren in
13、 der Abwasserverordnung Rechtsvorschriften und Normen (Supplements). Standards or draft standards bearing the group title “German standard methods for the examination of water, waste water and sludge” are classified under the following categories (main titles): 1)Registered in the DITR database of D
14、IN Software GmbH, obtainable from: Beuth Verlag GmbH, 10772 Berlin. 2) Registered in the DITR database of DIN Software GmbH, obtainable from: Beuth Verlag GmbH, 10772 Berlin. DIN EN 16161:2012-09 3 General information (group A) (DIN 38402) Sensory analysis (group B) (DIN 38403) Physical and physicoc
15、hemical parameters (group C) (DIN 38404) Anions (group D) (DIN 38405) Cations (group E) (DIN 38406) Substance group analysis (group F) (DIN 38407) Gaseous constituents (group G) (DIN 38408) Parameters characterizing effects and substances (group H) (DIN 38409) Biological-ecological methods of analys
16、is (group M) (DIN 38410) Microbiological methods (group K) (DIN 38411) Test methods using water organisms (group L) (DIN 38412) Individual constituents (group P) (DIN 38413) Sludge and sediments (group S) (DIN 38414) Bio-assays with microorganisms (group T) (DIN 38415) In addition to the methods des
17、cribed in the DIN 38402 to DIN 38415 series of standards, there are a number of European and International Standards available as DIN EN, DIN EN ISO and DIN ISO Standards which also form part of the collection of German standard methods. Information on Parts of these series of standards that have al
18、ready been published can be obtained from the offices of the Normenausschuss Wasserwesen, telephone +49 (30) 2601-2448, or from Beuth Verlag GmbH, Burggrafenstrae 6, 10787 Berlin, Germany. DIN EN 16161:2012-09 4 This page is intentionally blank EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN 16
19、161 July 2012 ICS 13.060.70 English Version Water quality - Guidance on the use of in vivo absorption techniques for the estimation of chlorophyll-a concentration in marine and fresh water samples Qualit de leau - Lignes directrices sur lutilisation des techniques dabsorption in vivo pour lestimatio
20、n de la concentration de chlorophylle-a dans les eaux douces et eaux marines Wasserbeschaffenheit - Anleitung fr die Anwendung der in-vivo-Absorption zur Abschtzung der Chlorophyll a-Konzentration in Meer- und Swasser This European Standard was approved by CEN on 17 May 2012. CEN members are bound t
21、o comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this European Standard the status of a national standard without any alteration. Up-to-date lists and bibliographical references concerning such national standards may be obtained on application to the CEN-C
22、ENELEC Management Centre or to any CEN member. This European Standard exists in three official versions (English, French, German). A version in any other language made by translation under the responsibility of a CEN member into its own language and notified to the CEN-CENELEC Management Centre has
23、the same status as the official versions. CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxe
24、mbourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and United Kingdom. EUROPEAN COMMITTEE FOR STANDARDIZATION COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG Management Centre: Avenue Marnix 17, B-1000 Brussels 2012 CEN
25、 All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN 16161:2012: EEN 16161:2012 (E) 2 Contents Page Foreword 3Introduction .41 Scope 52 Normative references 53 Terms and definitions .54 Principle 65 Apparatus .66 Procedure .76.1 Calibratio
26、n 76.2 Blank Measurement .76.3 Sample measurement 87 Calculation and Expression of Results .87.1 General 87.2 Datum 87.3 Mathematical routine .97.4 Other factors influencing the chlorophyll-a estimation .98 Quality Assurance .98.1 Repeatability .98.2 Uncertainty .99 Test report . 10Annex A (normativ
27、e) Published in vivo specific chlorophyll a absorption spectrum 11Annex B (informative) Determination of the appropriate chlorophyll-a specific spectral absorption coefficient for the IVP system . 13Annex C (informative) Factors influencing the chlorophyll-a estimation . 16C.1 Breakdown pigments . 1
28、6C.2 Package effect . 17C.3 Chlorophyll-b . 18C.4 Phycocyanin 18Annex D (informative) Examples of paired sample method validation Comparison of extraction and in vivo methods under operational conditions 20D.1 General . 20D.2 The IRH Laboratory Data Set . 20D.3 The Adasa Sistemas Laboratory Data Set
29、 . 21Annex E (informative) Validation of the spectrometric technique by determining the chlorophyll-a specific absorption of a set of algal samples 22E.1 Laboratory sample data for validation a*chl_systhe appropriate system unit chlorophyll-a response peak height . 22E.2 In vivo photometric chloroph
30、yll-a and the package effect . 24Bibliography . 25DIN EN 16161:2012-09 EN 16161:2012 (E) 3 Foreword This document (EN 16161:2012) has been prepared by Technical Committee CEN/TC 230 “Water analysis”, the secretariat of which is held by DIN. This European Standard shall be given the status of a natio
31、nal standard, either by publication of an identical text or by endorsement, at the latest by January 2013, and conflicting national standards shall be withdrawn at the latest by January 2013. Attention is drawn to the possibility that some of the elements of this document may be the subject of paten
32、t rights. CEN and/or CENELEC shall not be held responsible for identifying any or all such patent rights. According to the CEN/CENELEC Internal Regulations, the national standards organisations of the following countries are bound to implement this European Standard: Austria, Belgium, Bulgaria, Croa
33、tia, Cyprus, Czech Republic, Denmark, Estonia, Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and t
34、he United Kingdom. DIN EN 16161:2012-09 EN 16161:2012 (E) 4 Introduction Surveys of chlorophyll and nutrient concentration are fundamental descriptors of primary productivity and eutrophic threat in coastal and inland waters. Chlorophyll-a concentration can be determined by sampling and laboratory a
35、nalysis using the techniques described in ISO 10260. Achieving consistent results with this technique requires careful attention during the various steps of the process commonly used, such as during sampling, transport, filtering, freezing, storage and extraction and subsequent pigment estimation. T
36、he in vivo technique described here can be applied to surveys where a rapid non-destructive and repeatable measurement capability is required. It can be used either in the field or laboratory. No chemicals are required. Utilised in association with other methods of chlorophyll-a determination such a
37、s ISO 10260, HPLC pigment analysis and chlorophyll fluorescence measurements techniques, it can help identify sources of inconsistency or be used as an alternative technique in its own right. As chlorophyll-a estimates can be achieved in times as short as one minute, the technique can enhance survey
38、ing capability considerably. This standard describes procedures to implement and verify performance. DIN EN 16161:2012-09 EN 16161:2012 (E) 5 1 Scope This European Standard provides guidance in the use of in vivo absorption techniques to quantify chlorophyll-a concentration in marine and fresh water
39、s. This European Standard is comprised of the following: definition of the equipment requirement; a priori data and mathematical tools; recommendations for verification of measurement system performance and consideration of factors that can influence measurements; listing of the procedures to be imp
40、lemented. 2 Normative references The following documents, in whole or in part, are normatively referenced in this document and are indispensable for its application. For dated references, only the edition cited applies. For undated references, the latest edition of the referenced document (including
41、 any amendments) applies. ENV 13005, Guide to the expression of uncertainty in measurement 3 Terms and definitions For the purposes of this document, the following terms and definitions apply. 3.1 absorption coefficient a natural logarithm of the ratio between the light intensity entering and corres
42、ponding intensity emerging directly through a sample of water divided by the sample path length (in metres) in cases where the scattering of light is negligible Note 1 to entry: The unit is m-1. Note 2 to entry: A spectrophotometer often gives the Log10of the ratio in place of the natural logarithm.
43、 3.2 extinction c sum of losses of directly transmitted light by absorption and scattering Note 1 to entry: The unit is m-1. Note 2 to entry: The extinction c is related to absorption a and scattering b, by bac += . 3.3 extractive photometric EP method of chlorophyll concentration estimation involvi
44、ng extraction and absorption measurement DIN EN 16161:2012-09 EN 16161:2012 (E) 6 3.4 in vivo photometric IVP method of assessing chlorophyll-a concentration through the use of in vivo spectral photometry 3.5 package effect flattening of a spectral absorption feature arising from excessive absorbing
45、 molecule concentration within cells 3.6 resolution width at half height of the instrument response function 3.7 scattering coefficient b natural logarithm of the ratio between the light intensity entering and corresponding intensity emerging directly through a sample of water divided by the sample
46、path length (in metres) in cases where the absorption of light is negligible Note 1 to entry: The unit is m-1. 3.8 spectrum set of data of a sample taken over a defined wavelength range and by a defined resolution 3.9 wavelength range range from minimum to maximum wavelength over which a spectrum is
47、 described 4 Principle The in vivo photometric absorption technique (IVP) is based on: a) the additive nature of absorption of individual constituents within a suspension; b) the use of a priori knowledge about the absorption features of chlorophyll-a in the wavelength area of approximately 675 nm;
48、c) the absence of other components interfering with spectral features of chlorophyll-a in this region; d) the use of a measurement cell (a cuvette or other sample receptacle) of sufficient length and spectrophotometer of sufficient performance to enable the absorption feature of chlorophyll-a to be identified at
