1、December 2015 English price group 16No 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).ICS 13.060.70!%Jw“2398458www.din.de
2、DIN EN 16698Water quality Guidance on quantitative and qualitative sampling of phytoplankton from inland waters;English version EN 16698:2015,English translation of DIN EN 16698:2015-12Wasserbeschaffenheit Anleitung fr die quantitative und qualitative Probenahme von Phytoplankton aus Binnengewssern;
3、Englische Fassung EN 16698:2015,Englische bersetzung von DIN EN 16698:2015-12Qualit de leau Lignes directrices sur lchantillonnage quantitatif et qualitatif du phytoplancton dans les eaux intrieures;Version anglaise EN 16698:2015,Traduction anglaise de DIN EN 16698:2015-12www.beuth.deDocument compri
4、ses 39 pagesDTranslation by DIN-Sprachendienst.In case of doubt, the German-language original shall be considered authoritative.12.15 DIN EN 16698:2015-12 2 A comma is used as the decimal marker. National foreword This document (EN 16698:2015) has been prepared by Technical Committee CEN/TC 230 “Wat
5、er analysis” (Secretariat: DIN, Germany). The responsible German body involved in its preparation was DIN-Normenausschuss Wasserwesen (DIN Standards Committee Water Practice), Working Committee NA 119-01-03-05-06 AK Biologisch-kologische Gewsseruntersuchung of NA 119-01-03 AA Wasseruntersuchung. Des
6、ignation of the method: Guidance on quantitative and qualitative sampling of phytoplankton from inland waters: Method DIN EN 16698 M 38 This standard has been prepared under mandate M/424, which was given to CEN by the European Commission for the development and enhancement of standard methods in su
7、pport of the Water Framework Directive (WFD) (Directive 2000/60/EC). For the application of this standard, the following information should be taken into account: in deviation from the English reference document EN 16698:2015-10, 6.3.4, 1). the statement “with euphotic depth epilimnetic depth (zeu z
8、epi)” has been technically corrected and brought in line with Figure 1. The DIN Standards corresponding to the International Standards referred to in this document are as follows: ISO 17289 DIN ISO 17289 DIN EN 16698:2015-12 3 Expert assistance and specialized laboratories will be required to perfor
9、m the analyses 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 specified. This standard has been prepared by D
10、IN-Normenausschuss Wasserwesen (DIN Standards Committee Water Practice) in collaboration with the Wasserchemische Gesellschaft Fachgruppe in der Gesellschaft Deutscher Chemiker (Water Chemistry Society Division of the German Chemical Society). It is part of the series Deutsche Einheitsverfahren zur
11、Wasser-, Abwasser- und Schlammuntersuchung (German standard methods for the examination of water, waste water and sludge): Guidance on quantitative and qualitative sampling of phytoplankton from inland waters (M 38). Standard methods published as DIN Standards are obtainable from Beuth Verlag GmbH,
12、either individually or grouped in volumes. The standard methods included in the loose-leaf publication entitled Deutsche Einheitsverfahren zur Wasser-, Abwasser- und Schlammuntersuchung will continue to be published by Beuth Verlag GmbH and Wiley-VCH Verlag GmbH zseccis the Secchi depth in metres (m
13、); t is an empiric constant ranging from 0,8 to 2,5, depending on the turbidity of the lake 12: under usual conditions in clear and turbid lakes: t = 2,5; in humic lakes usually: t = 1,0 (see Annex A, Figure A1). NOTE Humic lakes have a colouration value 30 mg Pt/l. Figure 1 provides a decision sche
14、me for sampling in lakes and reservoirs. N1)National footnote: Take note of information given in National foreword. N1)DIN EN 16698:2015-12 EN 16698:2015 (E) 14 Figure 1 Decision scheme for sampling in lakes and reservoirs 6.4 Preparation of mixed samples of a water column 6.4.1 Preparation of mixed
15、 samples using an integral water sampler With an integral water sampler a sample of the complete water column can be taken continuously and in a time-saving manner. Prior to bottling the samples, the whole content of the integral water sampler should be filled into a mixing container (5.1.9) to elim
16、inate any layering in the sampler. For sampling very small depth ranges integral water samplers are not recommended due to the very low lowering velocity required or the small volumes obtained, respectively. 6.4.2 Preparation of mixed samples using other water samplers To collect a complete represen
17、tative sample of the whole water column, universal water samplers or tube samplers with a length of 0,5 m to 2 m (5.1.1) should be used (see Annex B for examples). Samplers with a closing lid which does not uncover the complete cross-section area of the sampling tube while lowered in the water (e.g.
18、 Ruttnersampler) are not appropriate. A depth profile should be produced from sub-samples covering equidistantly the water column. The distance between the sampling depths should not exceed 1 m in shallow (i.e. polymictic) lakes and 2 m in deep lakes. DIN EN 16698:2015-12 EN 16698:2015 (E) 15 In cas
19、es where a deep chlorophyll maximum (DCM) occurs (recognized by detection of a distinctive maximum of oxygen, chlorophyll-a or pH in the depth profile), it is essential to take a sample from the corresponding depth too. With regard to chlorophyll-a, the conditions for a DCM are met when the maximum
20、value is at least 1,5 times the average measured in the epilimnion. For the parameters oxygen and pH, no quantitative criterion is available. Sub-samples collected from the different depths should be combined in a mixing container (see 5.1.10). The mixed sample should be protected from direct sunlig
21、ht until it is filled into the sample bottle(s). The total volume of the mixed sample should be sufficient for the survey objectives. This should be observed especially when using an integrating sampler, where the sample volume can be very limited. The mixed sample should be homogenized by stirring
22、carefully, but avoiding vortex formation. Do not shake the container to avoid destruction of sensitive phytoplankton organisms. When samples are collected from the outflow or edge of lakes using a bottle on a long pole or weighted bottle on a long rope (see 4.2.3), it is assumed that the collected s
23、ample is fully mixed and sufficiently representative of the lake for the purposes of the sampling program. 6.5 Bottling and fixation of samples The bottling of the sub-sample should be carried out using a clean bailer. The first part of the content of the cup is used for rinsing the sample bottle an
24、d is discarded afterwards. The bottles should be filled up to not more than 90 % of their volume or 2 cm to 3 cm below the opening. This is necessary to allow shaking and homogenization of the samples for withdrawal of sub-samples (see EN 15204). For eutrophic to hypertrophic lakes 100 ml bottles an
25、d for oligotrophic to eutrophic lakes 250 ml bottles should be used. For lakes with a Secchi depth of more than 10 m a 500 ml sample should be bottled. 250 ml bottles are required when 50 ml sedimentation tubes are used so that more than one replicate filling of a counting chamber is possible. 500 m
26、l bottles allow pre-sedimentation procedure (see EN 15204), but this step should be avoided if possible because this may introduce another source of error into the process. The fixation using Lugols solution (5.2.1, 5.2.2) should lead to a cognac coloured or light brown colour of the sample. For ori
27、entation: a 225 ml sample (using a 250 ml bottle) should be fixed with 8 to 10 drops of Lugols solution (about 0,5 ml). After adding the solution, mix the sample by slowly tilting the closed bottle. The fixed sample should not be heated or exposed to direct sunlight during transport. It is recommend
28、ed to use backup bottles which should be fixed in the same way. 6.6 Storage and transport of the samples Preserved samples should be stored in clear narrow-necked glass bottles (not brown glass) in the dark at 4C to 10 C for not longer than 6 months prior to analysis (for details see EN 15204). The
29、narrow neck minimizes the loss of iodine (and thus loss of preservative effect) by diffusing through the screw cap of the bottle. Transparent, clear glass bottles should be used to allow a check for colouration (cognac colour) of the samples at regular intervals. Plastic bottles are not suitable, be
30、cause they do not allow for a visual control of the colouration as the plastic itself becomes tainted by the preservative. Another disadvantage is the loss of iodine diffusing through the plastic material. Beware of over-fixation (dark brown colour) and freezing of samples. The addition of formaldeh
31、yde as described by EN 15204 is only necessary for long term fixation and should be avoided. The samples should be stored preferably vibration free and necessarily standing upright (to avoid losses by leakage) in a dark place. If the samples are to be stored before transport, they should be stored i
32、n a dark and cool place. The samples should be packed and sent to the laboratory conducting the analyses within 6 months. A sampling form listing sampling position, date and type of sample and additional sampling information (see 6.1) should be given together with the samples. DIN EN 16698:2015-12 E
33、N 16698:2015 (E) 16 6.7 Additional samples for analysis of diatoms If pelagic diatoms are to be identified to species level, an additional sample is necessary: in rivers at least 0,5 l and in lakes at least 1,0 l additional sample volume. The preservation should be carried out with ethanol (5.2.3) a
34、nd should yield an ethanol concentration of 10 % or higher. Attention should be paid that the storage time after fixation with ethanol does not exceed 3 months. Preservation with formaldehyde may keep the samples longer but may also cause loss of fine structure elements of the valvae. Another method
35、 for diatom preservation is the storage of a filtered unfixed sample. An unfixed sample should be filtered using a filter with smooth surface and a maximum pore diameter of 2 m. In case of filter clogging, more than one filter for one sample should be used. After having filtered the sample, the filt
36、er should be stored air-dried and at room temperature 5, 6, 7. 6.8 Qualitative sampling In some cases it is necessary to determine phytoplankton organisms alive (e.g. see EN 15204, 8). In these cases a separate sample, taken according to 6.2 and 6.3, with a minimum volume of 500 ml and without any f
37、ixation, should be stored dark and cool for subsequent sedimentation in an Utermhl chamber. Alternatively a sample can be taken using a plankton net (5.1.12) to determine living phytoplankton. Samples with living plankton organisms should be analysed within 24 h. In case of studying ontogenetic stag
38、es and to enrich rare forms, net samples should be fixed using Lugols solution (5.2.1, 5.2.2). 7 Measurements of accompanying parameters 7.1 General Prior to sampling of phytoplankton in lakes, the sampling depth (epilimnion/euphotic zone) should be determined by measuring the following parameters:
39、Secchi depth; water temperature; dissolved oxygen; chlorophyll-a, using a fluorescence probe (optional); pH (optional). The results (especially for temperature and oxygen) should be recorded in a depth profile table (see Annex D). The relevant depth range for sampling should be measured using depth
40、intervals of 1 m. 7.2 Secchi depth Use a white disk attached to a measuring tape with a length of 20 m; the accuracy of the Secchi depth information should be 2 % of the mean reading value. Lower the disk, on its measuring tape, into the water until the disk is barely visible. Read the length of imm
41、ersed measuring tape. Then lift the disk until it becomes visible again. Repeat the test several times. Calculate the average of the two readings. If no viewscope is used, choose the shaded side of the boat. DIN EN 16698:2015-12 EN 16698:2015 (E) 17 7.3 Water temperature Use a device equipped with d
42、epth probe and a measurement range of 0 C to 35 C; accuracy 0,1 C (see EN ISO 5814). 7.4 Dissolved oxygen Use a device equipped with depth probe and a detection limit of 0,1 mg/l; accuracy 0,1 mg/l (see EN ISO 5814 or ISO 17289). 7.5 pH Use a device equipped with depth probe with pH range from 3 to
43、12 and accuracy 0,2 pH units (see EN ISO 10523). 7.6 Chlorophyll-a Use a device equipped with depth probe and a fluorescent sensor featuring a detection limit of 0,5 g/l; accuracy 0,1 g/l. 8 Quality Assurance The quality assurance should be in accordance with EN 14996. An example for a sampling prot
44、ocol is given in Annex D. DIN EN 16698:2015-12 EN 16698:2015 (E) 18 Annex A (informative) Description of methodology A.1 Water colour, Secchi-depth and euphotic depth Figure A.1 displays the dependency of Secchi-depth and the euphotic depth (depth featuring 1 % of subsurface illumination) from water
45、 colour in humic lakes 13. Key Y depth in meters (m) X water colour (g Pt/m3) 1 Secchi depth visibility: sdv = 17,54 colour -0,572 1 % of subsurface illumination: z1 %= 41,85 colour-0,77Figure A.1 Relations between water colour and Secchi disk visibility and water colour and the depth with 1 % of su
46、bsurface illumination 13 DIN EN 16698:2015-12 EN 16698:2015 (E) 19 Table A.1 Relations between Colour, absorption coefficient at 430 nm (g430), Secchi depth (sdv) and the depth with 1 % of subsurface illumination (z1 %) Colour Absorption coefficient at 430 nm sdv z1 %Ratio z1 %/sdv g Pt/m3m1m m 2 0,
47、150 11,8 24,3 2,1 5 0,376 7,0 12,1 1,7 20 1,502 3,2 4,2 1,3 40 3,004 2,1 2,4 1,1 100 7,511 1,3 1,2 0,9 200 15,022 0,9 0,7 0,8 colour (g Pt/m3) = 5,19 105 g430 e(0,415 + 0,028 430)16 A.2 Secchi depth practical hints The minimum diameter of the disc is 20 cm for Secchi depths 2 m) shall be conical, en
48、larged at the end with transparent acrylic glass. On ships with high vessel sides a viewscope is strongly recommended. If no viewscope is available the shaded side of the boat should be used. From bridges, the distance to the water surface usually will be too far for exact measurements. In this case
49、, the use of a viewscope of appropriate length is required (practicable for distances up to approximately 3 m). A.3 Sampling frequency examples The frequency and time of sampling for phytoplankton will be determined by the purpose of the study. For a general investigation of phytoplankton in lakes, taking a sample in the spring, summer and late summer will give an impression of the phytoplankton community with minimal resourc
