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This project has received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement No 731065
Project Title: AQUACOSM: Network of Leading European AQUAtic
MesoCOSM Facilities Connecting Mountains to Oceans from
the Arctic to the Mediterranean
Project number: 731065
Project Acronym: AQUACOSM
Proposal full title: Network of Leading European AQUAtic MesoCOSM Facilities
Connecting Mountains to Oceans from the Arctic to the
Mediterranean
Type: Research and innovation actions
Work program topics
addressed:
H2020-INFRAIA-2016-2017: Integrating and opening research
infrastructures of European interest
Standard Operating Protocol (SOP) on Water Chemistry
Version: V1.0; 29 May 2020
Main Authors: Christian Preiler (WCL), Robert Ptacnik (WCL), Deniz Başoğlu (METU), Meryem
Beklioğlu (METU), Henrik Larson (UMF), Sébastien Mas (CNRS-MARBEC)
This project has received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement No 731065
Abstract 4 Water
Chemistry
This deliverable contains Standard Operating Procedures (SOP) that describes the methods for sampling and storing samples for the analysis of water chemistry from mesocosm experiments carried out in all aquatic environments (fresh and marine waters). It gathers best practice advice with a focus on sampling and pre-analytical processing and provides an overview of analytical methods used by project partners.
This SOP points out relevant considerations regarding planning and execution of sampling mesocosms. The listing of analytical methods used by partners allows to identify frequently used methods as well as differences in analytical procedures in the AQUACOSM community.
Keywords • Water Chemistry, Sample Storage
This project has received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement No 731065
Table of Contents
I. Cross References ....................................................................................................................................... 5
II. Dissemination activities related to the Deliverable .................................................................................. 5
1. Water Chemistry ........................................................................................................................................ 6
1.1 Definitions and Terms........................................................................................................................ 6
1.2 Health and Safety Indications ............................................................................................................ 7
1.2.1 General Information .................................................................................................................. 7
1.2.2 Safety Instructions ..................................................................................................................... 8
1.2.3 Working and Personal Protection (Safety) Equipment ............................................................. 8
1.2.4 Use, Storage and Disposal of Reagents and Chemicals ............................................................. 8
1.2.5 Use, Storage and Disposal of the Equipment ............................................................................ 9
1.3 Environment Indications ................................................................................................................... 9
1.4 Sampling ............................................................................................................................................ 9
1.4.1 Introduction ............................................................................................................................... 9
1.4.2 Sampling Strategy/ Sampling Plan ........................................................................................... 10
1.4.3 Equipment for Sampling .......................................................................................................... 11
1.4.4 Techniques for representative sampling ................................................................................. 12
1.4.5 Quality Assurance Considerations ........................................................................................... 13
1.5 Filtration .......................................................................................................................................... 14
1.5.1 Introduction ............................................................................................................................. 14
1.5.2 Vacuum Filtration .................................................................................................................... 14
1.5.3 Pressure Filtration ................................................................................................................... 15
1.5.4 Filter Types .............................................................................................................................. 15
1.5.5 Cleaning procedure for glass fibre filters ................................................................................ 16
1.5.6 Cleaning procedure for synthetic membrane filters ............................................................... 17
1.5.7 Volume for filtration ................................................................................................................ 17
1.6 Sample Storage ................................................................................................................................ 17
1.7 Auxiliary measurements .................................................................................................................. 19
1.7.1 Water transparency (aka Secchi depth) .................................................................................. 19
1.7.2 Light measurements ................................................................................................................ 19
1.7.3 Standard physical parameters (Temperature, Oxygen concentration, Conductivity) ............ 20
1.8 Methods applied by Project Partners .............................................................................................. 21
1.8.1 Aarhus University (AU) ............................................................................................................ 21
1.8.2 Centro de Biodiversidade e Recursos Genéticos – Universidade de Évora (CIBIO)................. 22
1.8.3 MARine Biodiversity, Conservation and Exploitation (CNRS-MARBEC) .................................. 24
This project has received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement No 731065
1.8.4 Ecole Normale Superieure (ENS) ............................................................................................. 26
1.8.5 GEOMAR Helmholtz Centre for Ocean Research Kiel (GEOMAR) ........................................... 28
Hellenic Center for Marine Research (HCMR) ......................................................................................... 32
1.8.6 Ludwig-Maximilians-Universität Munich (LMU) ...................................................................... 35
1.8.7 Middle East Technical University (METU) ............................................................................... 37
1.8.8 Netherlands Institute of Ecology (NIOO) ................................................................................. 39
1.8.9 Umweltbundesamt (UBA) ........................................................................................................ 40
1.8.10 University of Helsinki, Tvärminne Zoological Station (UH) ...................................................... 46
1.8.11 University of Bergen (UIB) ....................................................................................................... 48
1.8.12 Umea Marine Science Center (UMF) ....................................................................................... 51
1.8.13 WasserCluster Lunz (WCL) ....................................................................................................... 54
1.9 References 1 – Water Chemistry ..................................................................................................... 56
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I. Cross References
The SOPs that will be provided by AQUACOSM will be listed here in the following versions when the different
SOPs are completed.
The SOPs that will be provided by AQUACOSM will be for:
1. Phytoplankton (this SOP)
2. Zooplankton (Deliverable 4.1.2)
3. Microbial Plankton (Deliverable 4.1.3
4. Periphyton (Phytobenthos) (Deliverable 4.1.4)
5. Water Chemistry (Physical and Chemical Elements of Water) (Deliverable 4.1.5)
6. High-Frequency Data Collection (Deliverable 4.1.6)
7. QA/QC (Deliverable 4.1.7)
A general description for water sampling will be covered under the Water Chemistry SOP.
II. Dissemination activities related to the Deliverable
The SOPs will be made available to all users of TA in AQUACOSM, and will also be publicly available for any
user through the AQUACOSM webpage (https://www.aquacosm.eu/project-information/deliverables/)
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1. Water Chemistry
1.1 Definitions and Terms
Alkalinity capacity of water to resist changes in pH that would make the water more acidic
Analyte The constituent or characteristic of a sample to be measured
Aphotic Zone Zone within a water body where photosynthetic production is not possible (gross
primary production < respiration)
Blank A blank contains little to no analyte of interest. It is included in measurements to
trace contaminations or signal drift.
Conductivity Inverse of electrical resistivity, increases with ions in solution and is temperature
dependent
Detritus Dead particulate organic material
Euphotic Zone Zone within a water body where photosynthetic production is possible (gross
primary production > respiration), it roughly corresponds to 2-2.5-times of the
transparency (Secchi depth) [1]
Macrophytes Water plants
Matrix Components of a sample other than the analyte
Phytoplankton Community of free-floating, predominantly photosynthetic protists and
cyanobacteria in aquatic systems, (in limnological analysis commonly excluding
ciliates). [1]
Seston Organisms and non-living matter swimming or floating in water
Standard (analytical) Standardized reference material containing a known amount of the analyte, used to
calibrate measured signal against analyte concentration
Stratification Formation of a vertical temperature gradient within a water column that due to
differences in density avoids vertical mixing
Turbidity Cloudiness or haziness of a fluid
Zooplankton Community of free-floating, heterotrophic organisms
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1.2 Health and Safety Indications
1.2.1 General Information
In this section, general guidance on the protection of health and safety while sampling and analysing water
samples from mesocosm experiments will be provided to minimize the risk of health impacts, injuries and
maximize safety. The users of this SOP are expected to be familiar with the Good Laboratory Practice (GLP)
of World Health Organization (WHO) [4] and Principles on GLP of Organisation for Economic Co-operation
and Development (OECD) [5]. Health and Safety Instructions of the mesocosm facility, if there are any, shall
be followed properly to protect the people from hazardous substances and the harmful effects of them.
According to preventive employment protection measures to avoid accidents and occupational diseases (on-
site or in the laboratory), the work should be practiced consistent with national and EU regulations (see the
OSH Framework Directive 89/391/EEC, [6]). Other regulations and guidelines can be found on the EU – OSHA
website (European directives on safety and health at work [7]). All necessary safety and protective measures
shall be taken by the users of this SOP and the scientist-in-charge shall ensure that those measures comply
with the legal requirements.
The table below summarizes the hazards, risks and safety measures for laboratory studies on water
chemistry.
Table 1-1: Hazards and risks associated with laboratory work
Occupations at
risk
Hazards/Risks Preventive Measures
Laboratories o Exposure (skin, eye,
inhaling) to harmful
chemicals
o Ingestion of harmful
chemicals
o Appropriate personal protection equipment
(gloves, goggles, lab coat)
o Ventilated working area
o Being informed about risks of applied
chemicals and providing SDSs in the facility
o Clear labelling of all chemical containers
o No eating or drinking in the lab
o Washing hands after leaving the lab
o No storage of sample/ chemicals in empty
food containers
o Keeping solvents away from heat sources
(ovens, open flame, autoclave)
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o Flame/ Explosion
o Hot surfaces/ steam
o Vacuum/ Implosion
o Overpressure
o Accidental release of
substances harmful to
the environment
o Use of tightly closed boxes to store solvents in
refrigerators
o Using heat – insulating gloves
o Using autoclaves with safety interlock
o Using exclusively thick-walled containers
approved for vacuum applications
o Using autoclaves with safety interlock
o Collection of chemicals and samples and
disposal according to national and local
regulations
1.2.2 Safety Instructions
Personnel involved in practical work, i.e. installation, sampling, analysis at an AQUACOSM facility has to
receive a safety instruction of the respective institute. The safety instruction summarizes rules, information
and advices related to work safety based on legislation and experience. The safety instruction needs to be
completed prior to practical work.
1.2.3 Working and Personal Protection (Safety) Equipment
Personal protective equipment (gloves, safety glasses, lab coat) has to be provided in the labs and must be
used for handling chemicals.
Gloves should be chosen according to permeation time which depends on material and thickness of gloves
and type of chemical to be used. Check lists of permeation time provided by the distributor of gloves to
choose the type of glove offering best protection. Still, some chemicals easily penetrate any kind of available
glove material. In this case gloves need to be changed immediately after contact with the chemical. Consult
the SDS for recommendations on which material of glove to use.
1.2.4 Use, Storage and Disposal of Reagents and Chemicals
Before using a chemical the first time the Safety Data Sheet (SDS) needs to be consulted to be informed about
the hazard potential of the substance. For each chemical a SDS in its most recent version has to be provided
by the distributor. In addition, SDS for all chemicals used in a lab should be made available to all users, i.e.
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hard copies collected in a folder. The SDS provides information on safe handling, storing and disposing of a
chemical. Based on the hazard classifications the user has to apply appropriate personal protective
equipment and needs to work under the chemical fume hood if required to eliminate the risk of exposure.
Chemicals should be stored in original containers if possible. Other containers than original, including
samples containing chemicals need to be labelled clearly. The label should indicate name and concentration
of chemicals as well as date and person responsible. All containers need to be closed tightly and stored in a
ventilated area. Hazardous chemicals have to be stored in specific cabinets separated from other chemicals
and meeting the safety requirements of the respective hazard class. This applies for flammables, explosives,
oxidizing chemicals, acids, bases, and toxic chemicals.
Only the quantities of daily consumption should be stored directly at the working area. Corrosive chemicals
must never be stored above eye height.
Collection vessels for disposal must be clearly labelled with a systematic description of their contents. To
avoid dangerous chemical reactions, consult the SDS before mixing chemicals. Entrust waste chemicals to the
appropriate authorities for disposal.
1.2.5 Use, Storage and Disposal of the Equipment
Consult your local head of the lab about rules for disposal of equipment.
1.3 Environment Indications
A plan for the disposal of chemical waste needs to be prepared prior to the experiments. The plan must be
in competence with the EU Waste Legislation ([8]) and The List of Hazardous Wastes ([9]) provided by the
European Commission. The SDS needs to be revisited for the disposal of reagents and chemicals prior to
waste disposal.
1.4 Sampling
1.4.1 Introduction
Good quality of analytical data relies on (1) representative sampling, (2) suitable storage conditions, and (3)
accurate and precise measurements.
“Progress in analytical protocols results in the taking of samples increasingly becoming quality-determining
step in water quality assessment. Poor sampling design or mistakes in sampling technique or sample handling
during the sampling process inevitably lead to erroneous results, which cannot be corrected afterward.”
(Handbook of water analysis, Nollet, L., M., L., De Gelder, L., S., P., 2014)
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1.4.2 Sampling Strategy/ Sampling Plan
Prior to a sampling event a sampling strategy has to be defined in consideration of selected analytes and aims
of the study. This includes specification of analytes, spatial aspects (surface sample or depth-integrated
sample), sampling frequency, sampling equipment, volume and number of samples, type and size of sample
containers, processing and storage of sample, sample coding, and standardized documentation.
Figure 1-1: Elements of Sampling Strategy (from: Practical guidelines for the analysis of seawater [2])
A. Mixed mesocosms
In case mesocosms are mixed continuously, one water sample can be considered representative for the
entire mesocosm. Here, samples should be taken near the centre of the mesocosm. Care should be taken to
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avoid macrophytes, if present, while sampling. If it is unclear whether the whole water volume is efficiently
mixed, an initial series of samples can be taken in transects across the mesocosm and along the vertical axis.
B. Stratified mesocosms
If mesocosms waters are stratified or partially mixed, the sampling procedure must be determined after
careful considerations on the water layer(s) to be sampled (e.g., sampling discrete water layers; sampling
multiple depths with subsequent pooling to one combined representative sample; utilization of tube
sampler). The absence of vertical temperature profiles indicates vertical mixing, but NOT necessarily
homogenous distribution of motile organisms like (micro-) zooplankton. Hence, especially particulate matter
(chlorophyll-a, particulate nutrients) may be non-homogenously distributed even in the absence of a
thermocline, and require careful consideration of the appropriate sampling design.
1.4.3 Equipment for Sampling
The following specific pieces of equipment are suggested for collecting water samples from mesocosms
✓ Appropriate water sampler, depending on the type of sampling (stratified or integrated), and (e.g.
Schroder/Schindler/Ruttner sampler for single strata; tube sampler for depth integrated samples)
✓ As an alternative to a water sampler a sample can be retrieved by pumping, i.e. by a silicone tube
connected to a carboy which in turn is connected to a vacuum pump. Land based mesocosms can be
equipped with sampling ports, avoiding the risk of contamination from sampling equipment.
✓ Clean & rinsed sampling containers in the field
✓ Sampling containers needs to be labelled properly in the laboratory prior to sampling. The labels on
the sampling bottles need to be standardized and provide information on name of the experiment,
sampling date, mesocosm ID, and possibly depth with appropriate abbreviations of the treatments
of the experiments. Labels can be either printed or written using a permanent waterproof marker
Figure 1-2: Devices for water sampling - Ruttner Sampler, Schindler Patalas, and Tube Sampler
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1.4.4 Techniques for representative sampling
Representative sampling – samples need to be representative for the mesocosm unit of interest. In case
multiple parameters (e.g. dissolved & particulate nutrients, chlorophyll-a) are sampled on the same event, it
is mandatory that all parameters are analysed from the same water sample (multiple sub-samples from one
water sample). This implies that homogenous distribution of particles (phytoplankton, bacteria, detritus) is
ensured whenever a sub-sample is taken from the sampling container.
A. Mixed mesocosms
In case mesocosms are mixed continuously, one water sample can be considered representative for the
entire mesocosm. Here, samples should be taken near the centre of the mesocosm as stated in both [10] and
[12]. Care should be taken to avoid macrophytes, if present, while sampling. If it is unclear whether the whole
water volume is efficiently mixed an initial series of samples can be taken in transects across the mesocosm
and along the vertical axis (see point 6.4.B below).
B. Stratified mesocosms
If mesocosms waters are stratified or partially mixed, the sampling procedure must be determined after
careful considerations on the water layer(s) to be sampled (e.g. sampling discrete water layers; sampling
multiple depths with subsequent pooling to one combined representative sample; utilization of tube
sampler). Absence of vertical temperature gradients indicates vertical mixing, but NOT necessarily
homogenous distribution of motile organisms like (micro-) zooplankton. Heterogeneous distribution of
phytoplankton must also be assumed if mesocosms contain structuring elements (such as macrophytes), and
has not been proven to be (practically) homogenous (see 6.4.A). C. Composite and discrete sampling
Careful consideration of the sampling design is required in case the mesocosms are stratified and include an
aphotic zone (mesocosm depth > Zeu1). In this case the integrated water sample would typically be taken from
the euphotic zone (Zeu).
For shallow mesocosms containing sediment and possibly macrophytes, a detailed sampling design for
collecting samples at multiple horizontal and vertical positions might be needed [12]:
Best Practice Advice: “The entire water column, from the surface to approx. 5 cm above the sediment, is
sampled from three positions in each enclosure: 10, 30 and 60 cm from the enclosure wall. Two samples are
prepared: one to be used for chemical and phytoplankton analyses where water is sampled without touching
the plants – and one to be used for zooplankton analysis, where water is sampled also close to the plants.”
“The best way to sample from the entire water column is by using tube samplers which sample from top to
bottom. The diameter of the tube should not be too small to avoid zooplankton escaping during sampling (>
6 cm). If it is not possible to use a tube sampler, samples can be taken with a Ruttner water sampler from the
surface (20 cm below the water surface), middle and the bottom (20 cm above the sediment). The sample in
the middle should be adjusted according to the enclosure type and actual water depth [11].
1 Zeu corresponds to 2-2.5 x Secchi depth
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1.4.5 Quality Assurance Considerations
Any treatment of a sample, like transfer into another container, preservation, filtration, dilution may
introduce a contamination or alter the sample in another way. Consequently, the ideal procedure would be
to transfer the sample directly into the container for storage and analyse it immediately.
Sampling Technique
Already the technique of sampling can alter the quality of a sample. Pumping or temperature change may
already affect concentration of dissolved gases.
Sampling Sequence
Following sequence is recommended for subsampling water from a water sampler:
O2, pH/DIC/Alkalinity/ Nutrients
Contaminations
Potential sources for contamination of samples throughout the entire sampling procedure need to be
identified and avoided. Problematic contaminations can be either the chemical compound of interest or any
other substance interfering with the chemical analysis.
Sources for contaminations may be:
• Water Sampler/ Tube: Make sure it is clean and the materials suite your purpose.
• Sample Vials: Materials may release/ adsorb compounds into/ from your sample. Use clean
containers of appropriate material and consider rinsing them with sample first.
• Skin: Wear gloves to avoid contaminations with sweat, remains of soap, sun screen, etc.
• Boat Exhaust/ Cigarette Smoke: Rich in ammonia
Best Practice Advice: Use silicone tubing for O2, pH, DIC
Sample Matrix
The matrix of a sample (turbidity, salinity, colour, alkalinity, other chemical constituents) can affect the
chemical analysis in various ways. Optical characteristics of the sample can influence measurements based
on absorbance and fluorescence or may interfere with the signal-producing reaction. If sample and standard
do not share the same matrix, the calibration is incorrect.
Turbidity: If possible, turbidity should be removed by filtration. Alternatively, the chemically untreated
sample can be used as blank to correct for turbidity.
Salinity: Sea salt may suppress analyte absorbance in spectrophotometric measurements like phosphate,
silicate and ammonia. This problem can be addressed by preparing reference solutions of standards with
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nutrient-free or low-nutrient sea water, or artificial sea water. The salinity should be equal to that of the
samples. If salinity varies a lot, a mathematical correction of salt effects can be applied by establishing a
correction function for each analyte showing salt effects.
1.5 Filtration
1.5.1 Introduction
By means of filtration, the water sample is separated into a particulate and dissolved fraction for separate
analysis. The water sample can be forced through the filter material either with vacuum or over-pressure.
The filter type affects minimal particle size retained and filtering capacity. Filter material needs to be chosen
to minimize interaction with the analyte and to allow required cleaning procedures (see 7.5 and 7.6). Any
meaningful analysis of a set of filtered samples must include at least 4 blanks. Blank filters should be cleaned
according to established procedures and from the same batch as the filters used for the samples.
1.5.2 Vacuum Filtration
The circulate filter is placed between the filter support and the funnel held in place with a clamp. Filtrate is
collected in the receiving flask while particles are retained on the filter. Applied vacuum should not exceed
200 m bar to avoid rupture of cells and leaching of particulate material [13].
Figure 1-3: Vacuum filtration (from: Practical guidelines for the analysis of seawater [2])
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1.5.3 Pressure Filtration
As an alternative to vacuum filtration a sample can be pressurized and passed through a filter. Filtration with
syringe and filter holder may be an optimal procedure if samples need to be filtered immediately. Required
equipment is small and filtration does not depend on infrastructure, hence filtration with a syringe can be
easily performed in the field. Still it is only appropriate when rather small volumes of sample need to be
filtered.
To minimize sample carry-over syringe and filter need to be flushed with new sample, especially because
filter holders contain some dead volume.
Figure 1-4: Disposable filter discs and reusable filter holders used for filtration with the syringe
1.5.4 Filter Types
Glass fibre filters are best choice for organic carbon (DOC/POC), nitrogen (DON/PON), and phosphorus
(DOP/POP). Glass fibre filters have a poor uniform pore size, but they can be easily cleaned by baking at high
temperature (typically 450°C) for several hours to produce low blanks for these elements and at the same
time they provide good flow rates for high-volume samples. Glass fibre filters are the classical filter material
for the determination of chlorophyll pigments and are also suitable for the filtration of nutrient samples
except silica, in which polycarbonate filters are mostly used [2].
Besides glass fibre filters (GF/F ~ 0,7µm and GF/C ~ 1,2µm) membrane filters of 0,45 and 0,2µm pore size are
used to separate particulate and dissolved phases of water samples [3].
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Best practice advice:
● For efficient retention of seston including cyanobacteria GF/F filters should be used.
● Filters used for quantification of particulate carbon should be pre-combusted at 450°C
● Filters for quantification of particulate phosphorus should be acid washed.
Table 1-2: Filter Materials and their characteristics (from: Practical guidelines for the analysis of seawater [2])
1.5.5 Cleaning procedure for glass fibre filters
To assure glass fibre filters are free of organic traces they have to undergo a cleaning procedure.
Bake filters at 450°C for 4 hours. Place filters in a glass beaker and cover them with diluted acid (i.e. 10-15%
Hydrochloric Acid). Rinse filters repeatedly with analytical grade water and dry them at 60°C.
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✓ Best practice advice: Apply the above cleaning procedure (acid washing/ baking) to all glass fibre
filters even if not required for the subsequent analysis (i.e. chlorophyll). This will assure identical
filtration results/ comparability of chemical parameters since the high temperature and washing can
affect the effective pore size of the filters.
1.5.6 Cleaning procedure for synthetic membrane filters
Rinse with analytical grade water (i.e. MilliQ) and allow the filters to stand soaked in MilliQ for 30 minutes,
rinse again with MilliQ and press out the remaining water with air. A test tube rack can be used as support.
Always discard the first millilitres of sample to waste.
✓ Best practice advice: Use syringes with plastic plungers, avoid syringes with rubber plungers. The
rubber or the grease on the rubber is a potential source for contamination.
1.5.7 Volume for filtration
The water volume required in order to collect sufficient material on a filter depends both on the parameter,
the sensitivity of the methodology, and esp. on the density of particles in the water, which in turn depends
on the trophic state of the experimental system.
✓ As a rule of thumb, a clearly visible coloration on the filter (well visible against the white background
of a glass fibre filter) ensures reasonable quantity of material for most common analytical protocols
(PON/C/P, Chlorophyll-a).
✓ A careful documentation of filtered volume is mandatory in order to calculate concentrations (e.g.
µg Chl-a L-1).
1.6 Sample Storage
The biological activity in water does not stop with sample collection, since bacteria and micro- and nano-
plankton continue to digest and excrete material [13].
Nutrients are subject to rapid changes in their concentration within a few hours in unpreserved samples [2].
Sample preservation is needed whenever measurements cannot be performed immediately or when a
backup for potential reanalysis is required.
Each analyte has its own reaction chemistry and consequently different requirements for storage in solution.
Therefore, no general procedure can be recommended for the storage of water samples [3].
Optimum storage conditions differ largely among parameters (see Table 1-3 below).
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Table 1-3: Best Practice Advice for Storage of Water Samples
Analyte Container Preservation Notes
Soluble Reactive
Phosphorus
Glass, acid washed
LDPE, acid washed
Filter and measure immediately or store ≤4°C in the dark if
analysed within 24hrs.
For longer storage freeze filtered samples at -20°C.
Calcareous water (Ca2+ > 100mgL-1)
must be acidified with 1ml
concentrated hydrochloric acid L-1
before freezing to prevent co-
precipitation of phosphate
Particulate Phosphorus Plastic petri dish or
Eppendorf tube
Filter immediately, freeze filters at -20°C
Total Phosphorus Glass, acid washed
LDPE, acid washed
Store in containers and volumes desired for digestion ≤4°C in
the dark.
For long term storage freeze samples at -20°C.
Calcareous water (Ca2+ > 100mgL-1)
must be acidified with 1ml
concentrated hydrochloric acid L-1
before freezing to prevent co-
precipitation of phosphate
Total Dissolved
Phosphorus
Glass, acid washed
Store filtered samples in containers and volumes desired for
digestion ≤4°C in the dark.
For long term storage freeze filtered samples at -20°C.
Calcareous water (Ca2+ > 100mgL-1)
must be acidified with 1ml
concentrated hydrochloric acid L-1
before freezing to prevent co-
precipitation of phosphate
Total Reactive
Phosphorus
Glass, acid washed
Measure immediately, or store at 4°C in the dark if analysed
within 24hrs.
Ammonia Plastic or Glass Measure immediately, if necessary filter and store at 4°C for
up to 24hrs, or filter and freeze unacidified at -20°C for up to
28d
Note that samples that have been
measured immediately are not
necessarily comparable with
samples that have been filtered and
frozen. this is especially true for
ammonia.
DOC (NPOC) Polycarbonate or
cell culture flasks
Filter through combusted GF/F or 0,2µm membrane filter into
combusted glass vial and preserve immediately with H3PO4,
store in the dark at 4o C.
Nitrate Plastic or Glass Measure immediately, if necessary filter and store at 4°C up to
2d. For longer storage filter and store at -20°C.
In samples preserved with acid, NO3-
and NO2- cannot be determined as
individual species.
Nitrite Plastic or Glass Measure immediately, or filter and store at -20°C for longer
storage
Never acidify for storage.
Total Nitrogen Plastic or Glass Acidified to pH 1-2 with H2SO4 samples can be stored 1 month
Chlorophyll a Plastic petri dish or
Eppendorf tube
Filter immediately after sampling and freeze at -20C Protect from light, ideally store at -
80°C
Dissolved Inorganic
Silicate
Plastic Filter immediately. Store at 4°C for up to 1 month. Do not acidify, silicate precipitates
under acidic conditions. Avoid
freezing.
Total alkalinity Glass, acid washed Filter immediately after sampling and store at 4°C up to 24h or
up to a month if the samples is poisoned.
Samples can be poisoned with HgCl2
solution.
Oxygen, dissolved Plastic or Glass Cool, protect from air and light, store up to 6hrs On-site measurement preferable
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pH Plastic or Glass Cool, protect from air (fill bottle to cap) On-site measurement preferable
Alkalinity Plastic or Glass Cool, protect from air (fill bottle to cap), store up to 24h On-site measurement preferable,
especially for samples high in
dissolved gases
Best practice advice: Samples preserved by acidification should be neutralized prior to analysis. Acid and
base may introduce background to the sample, hence standards used in subsequent measurements must
undergo the same treatment.
1.7 Auxiliary measurements
Here we briefly outline measurements that are often conducted in combination with sampling for water
chemistry. Transparency and basic physical parameters are often taken alongside water sampling for water
chemistry and phytoplankton.
Best practice advice: The relevance of measuring irradiation and transparency depends on the depth of the
mesocosms. In shallow mesocosms (d < 2m), light very likely will not be limiting during the growing season.
Moreover, the influence of the suspended particles including phytoplankton on under water light climate is
very limited in shallow water columns.
1.7.1 Water transparency (aka Secchi depth)
Water transparency is a key parameter in limnology and oceanography, informing especially about the optical
depth of the water column, which affects e.g. the vertical structure of biota and biological processes, such as
primary production, but also vertical migration of the zooplankton.
Transparency is typically measured by lowering a white disk vertically into the water until it is not visible
anymore. Now the disk is slowly lifted, until it becomes visible. The depth where the disk just becomes visible
is defined Secchi depth. A detailed outline is given in
http://www.helcom.fi/Lists/Publications/Guidelines%20for%20measuring%20Secchi%20depth.pdf.
1.7.2 Light measurements
Light is a key resource for primary production. In aquatic ecology, it is commonly measured as photo-
synthetically active radiation (PAR; https://www.licor.com/documents/liuswfuvtqn7e9loxaut ). For proper
quantification of light, irradiation needs to be measured by an appropriate probe at various positions inside
the mesocosm, taking the optical structure (illuminated vs. shaded side etc.) and day-time into account.
Especially in narrow mesocosms with opaque walls, a proper quantification of irradiation may be very
difficult.
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If light intensity is a key parameter and e.g. manipulated by shading, comparing irradiance at a fixed position
(e.g. middle of water column) may be a good proxy. An example for light-manipulation inside mesocosms
can be found here https://www.nature.com/articles/srep29286
1.7.3 Standard physical parameters (Temperature, Oxygen concentration, Conductivity)
Standard physical parameters, especially water temperature, conductivity, pH and oxygen concentration are
often measured using handheld probes. For measuring all of these parameters, the same recommendations
regarding representativeness apply, as outlined above (→ 1.4.4 Techniques for representative sampling). If
mesocosms are stratified, layers must be sampled separately. Measurements of physical parameters using
submersible probes are described in section Fehler! Verweisquelle konnte nicht gefunden werden. Fehler!
Verweisquelle konnte nicht gefunden werden. (High Frequency Measurements).
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1.8 Methods applied by Project Partners
1.8.1 Aarhus University (AU)
Parameter
Detection Mode
Method Name Reference Comments Manual Automated
Alkalinity X Titration DS 253, 1977
Chlorophyll a X
Spectrophotometric determination in ethanol
extract
DS 2201, 1986
X Chlorophyll a sensors - Turner designs - Cyclops 7F
Conductivity X YSI 6600 Xylem Analytics
Nitrogen
NH4
X
Photometric method, ammonia-nitrogen
(indophenol blue)
DS 224, 1975
NO3
X
Automated Hydrazine Reduction method with FIA
Star 5000 Foss FIA Star 5000
TN
X
Automated Hydrazine Reduction method with FIA
Star 5000, digestion with peroxodisulfate/NaOH
DS 221/Foss FIA Star 5000
Oxygen X Oxygen probe Oxyguard
pH X pH probe Oxyguard
Phosphorus
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SRP
X
Orthophosphate photometric method (ascorbic
acid/molybdate) DS 291, 1985
TP
X
Total Phosphorus photometric method (ascorbic
acid/molybdate), digestion with peroxodisulfate DS 292, 1985
Silica X Photometric method, molybdosilicate DMU T.A. nr. 22
1.8.2 Centro de Biodiversidade e Recursos Genéticos – Universidade de Évora (CIBIO)
Parameter
Detection Mode Method Name
Reference
Comments
Manual Automated
Conductivity X Digital rugged conductivity probe CDC40105, Hach
Oxygen X
Digital, luminescent/optical dissolved
oxygen (LDO) probe LDO101, Hach
pH X
Digital combination pH electrode with
built-in temperature sensor PHC101, Hach
Chlorophyll a X Handheld Fluorometer /Chlorophyll in vivo Aquafluor®, Arar 1997 (EPA Method 445.0)
No cell disruption and
acidification is applied
Nitrogen
NH3 X Indophenol Blue Method Ivancic & Deggobis 1984
NO2 X Automated Hydrazine Reduction Method ISO 13395:1996
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NO3 X Automated Hydrazine Reduction Method ISO 13395:1996
TN X
Persulfate Digestion, Hydrazine Reduction
Method
Clesceri 1999 (4500-P, chapter J);
ISO 13395:1996
Phosphorus
TP/ TDP X
Persulfate Digestion and Ascorbic Acid
Method Grasshoff 1999 (chapter 10.2.13)
References
Arar, E. J., & Collins, G. B. (1997). Method 445.0: In vitro determination of chlorophyll a and pheophytin a in marine and freshwater algae by fluorescence.
Cincinnati: United States Environmental Protection Agency, Office of Research and Development, National Exposure Research Laboratory.
Clesceri, L. S., Greenberg, A. E., & Eaton, A.D. (1996). Standard methods for the examination of water and wastewater. APHA, AWWA and WPCF, Washington DC.
DIN EN ISO 15681-2, Water quality - Determination of orthophosphate and total phosphorus contents by flow analysis (FIA and CFA) - Part 2: Method by
continuous flow analysis (CFA), 06-2001
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1.8.3 MARine Biodiversity, Conservation and Exploitation (CNRS-MARBEC)
Parameter
Detection Mode
Method Name Reference Comments Manual Automated
Chlorophyll a X Fluorometric Detection Strickland and Parson 1972
Acetone extraction
Ultrasonic cell disruption
Pigments X High-Performance Liquid Chromatography Method Zapata et al. 2000
Suspended
particulate matter
(SPM)
X Gravimetric Strickland and Parson 1972
Nitrogen
NH4 X Ortho-phthaldialdehyde fluorometric Method (OPA) Holmes et al. 1999
NO2
X
CFA-based photometric detection, diazotization of
NO2 (Gries-Ilosvay reaction) with sulphanilamide
produce a reddish-purple colour, which is measured
at 540 nm.
ISO 13395:1996
NO3
X
CFA-based photometric detection, nitrate reduction
to nitrite by use of coppered Cd-granules and
detection as nitrite (see above)
ISO 13395:1996
Oxygen (dissolved) X Titration – Winkler method Strickland and Parsons 1972
- use of a silicone tube for sampling
to avoid air bubbles
- use of calibrated bottles/flasks.
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pHT (total scale) X
Spectrophotometric method (based on the
absorption ratio of the sulfonephthalein dye, m-
cresole purple
Byrne 1993
Liu et al 2011
- use of a silicone tube for sampling
to avoid air bubbles
Phosphorus
SRP
X
CFA-based photometric detection, reduction to
molybdenum blue complex by use of ascorbic acid.
The complex is measured at 660 nm.
ISO 15681-2
Salinity X
Silica X
CFA-based photometric detection, reduction to
molybdenum blue complex by use of ascorbic acid ISO-16264
Oxalic acid is added to avoid
phosphate interference
Temperature X
References
Byrne R.H. (1993). Spectrophotometric seawater pH measurements: total hydrogen ion concentration scale calibration of m-cresol purple and at-sea results.
Deep-Sea Research, Vol. 40, No 10, pp 2215-2129.
Holmes, R. M., Aminot, A., Kérouel, R., Hooker, B. A., & and Peterson, B. J. (1999). A simple and precise method for measuring ammonium in marine and freshwater
ecosystems. Can. J. Fish. Aquat. Sci., 56, 1801–1808
ISO 13395:1996, Water quality -- Determination of nitrite nitrogen and nitrate nitrogen and the sum of both by flow analysis (FIA and CFA) and spectrometric
detection.
ISO 15681-2, Determination of orthophosphate and total phosphorus contents by flow analysis (FIA and CFA), Part 2: Method by continuous flow analysis (CFA).
ISO-16264: Determination of soluble silicates by flow analysis (FIA and CFA) and photometric detection.
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Liu X., Patsavas M.C and Byrne R.H. (2011). Purification and characterization of meta-cresol purple for spectrophotometric seawater pH measurements. Environ.
Sci. Technol., 2011, 45 (11), pp 4862–4868.
Strickland, J.D.H., & Parsons, T.R. (1972). A Practical Handbook of Seawater Analysis. 2nd Edition, Fisheries Research Board of Canada Bulletin, 167, 310 p.
Zapata, M., Rodriguez, F., & Garrido, J. (2000). Separation of chlorophylls and carotenoids from marine phytoplankton: A new HPLC method using a reversed
phase C-8 column and pyridine-containing mobile phases. Mar. Ecol. Prog. Ser., 195, 29–45, doi:10.3354/meps195029
1.8.4 Ecole Normale Superieure (ENS)
Parameter
Detection Mode
Method Name Reference Comments Manual Automated
Alkalinity,
total
X Potentiometric automated titration with open cell
method
Dickson et al. 2007
Chlorophyll a X Fluorometric Detection after Acetone Extraction Jespersen and Christoffersen 1987
Dissolved oxygen X Optical method (optode sensor)
Nitrogen
NO2
X Colorimetric with Sulfanilamide and NEDD Grasshoff et al. 1983 N° G-173-96 Rev. 10 Seal analytical AA3
autoanalyzer method
X Colorimetric Strickland and Parsons, 1972 UV VIS
spectrophotometer
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NO3 X
Colorimetric with Sodium salicylate Strickland and Parsons, 1972 UV VIS
spectrophotometer
X Cd reduction and colorimetric method with
Sulfanilamide and NEDD
Grasshoff et al. 1983 N° G-392-08 Rev. 5 Seal analytical AA3
autoanalyzer method
NH4+ X Ortho-phthaldialdehyde (OPA) fluorometric Method Kerouel and Amniot 1997 NH4+
TN
X Persulfate digestion, Cd reduction and colorimetric
method
Grasshoff et al. 1983
N° G-392-08 Rev. 5
Seal analytical AA3
autoanalyzer method
pH X
Potentiometric with glass/reference electrode cell
(total scale)
Dickson et al. 2007
Phosphorus
SRP X Colorimetric with molybdate and ascorbic acid Strickland and Parsons, 1972 UV VIS
spectrophotometer
X Colorimetric with molybdate and ascorbic acid Murphy and Riley 1962
Drummon and Maher 1995
N° G-175-96 Rev. 15 (Multitest MT 18)
TP/TDP
X Persulfate and sulfuric acid digestion then molybdate
and ascorbic acid method
Grasshoff et al. 1983 N° G-393-08 Rev. 4 Seal analytical AA3
autoanalyzer method
Silicate X Ascorbic Acid - Molybdenum – Oxalic - Blue Complex Grasshoff et al. 1983 N°. G-177-96 Rev. 11
(Multitest MT19)
Seal analytical AA3
autoanalyzer method
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References
Dickson A.G., Sabine, C.L. and Christian, J.R. (Eds.) 2007. Guide to best practices for ocean CO2 measurements. PICES Special Publication 3, 191 pp.
Jespersen, A-M. & K. Christophersen, 1987. Measurements of chlorophyll- a from phytoplankton using ethyl alcohol as extraction solvent. Arch.Hydrobiol. 109:
445-454.
Kerouel, R.and Amniot, A. Marine Chemistry Vol. 57, no 3-4, pp.265-275, Jul 1997.
Strickland, J. D. H., and Parsons, T. R. (1972). A practical handbook of seawater analysis. B. Fish. Res. Board Can. 167, 311.
K. Grasshoff et al., Methods of Seawater Analysis, 2nd edition, Verlag Chemie, 1983.
Murphy, J. and Riley J.P., 1962. A modified single solution method for the determination of phosphate in natural waters. Analytica Chimica Acta 27:31-36.
Drummon, L. and Maher, W., 1995. Re-examination of the optimum conditions for the analysis of phosphate. Analytica Chimica Acta 302: 69-74.
1.8.5 GEOMAR Helmholtz Centre for Ocean Research Kiel (GEOMAR)
Parameter
Detection Mode Method Name
Reference
Comments
Manual Automated
Alkalinity,
total X Potentiometric titration, open-cell method Dickson et. al., 2003
Carbon
DIC X Acidification, gas stripping, Infrared absorption e.g. Goyet & Snover, 1993
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POC X
Treated with fuming HCL in a desiccator for 2h,
elemental analysis accomplished by combustion
analysis
Sharp 1974, Hansen and Koroleff 1999;
Grasshoff “Methods of seawater
analysis”, 1999
TPC X
Elemental analysis accomplished by combustion
analysis
Sharp 1974, Hansen and Koroleff 1999;
Grasshoff “Methods of seawater
analysis”, 2001
Chlorophyll a X Fluorometric Detection after acetone extraction Welschmeyer 1994
Pigments X
Reverse-phase high-performance liquid
chromatography Barlow et al., 1994
Nitrogen
DON X Determination by alkaline persulphate oxidation Hansen and Koroleff, 1999
NH4 X Determined fluorometrically Holmes et. al. 1999
NO2 X
Automated Camium Reduction Method,
photometrically
Murphey and Riley et.al., 1962; Hansen
and Koroleff, 1999; Grasshoff “Methods of
seawater analysis”, 1999; NIOZ –
Nederlands Instituut for Onderzoek der
Zee (Royal Netherlands Institute ), Den
Hoorn (Texel), The Netherlands
NO3 X
Automated Cadmium Reduction Method,
photometrically
Murphey and Riley et.al., 1962; Hansen
and Koroleff, 1999, modified by Keroul
and Aminot 1997, Grasshoff “Methods of
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seawater analysis”, 1999; NIOZ –
Nederlands Instituut for Onderzoek der
Zee (Royal Netherlands Institute for Sea
Reserach), Den Hoorn (Texel), The
Netherlands
PON
Elemental analysis accomplished by combustion
analysis
Sharp 1974, Hansen and Koroleff 1999;
Grasshoff “Methods of seawater
analysis”, 2000
TPN
X
Elemental analysis accomplished by combustion
analysis
Sharp 1974, Hansen and Koroleff 1999;
Grasshoff “Methods of seawater
analysis”, 2002
pH T (total
scale) X
Spectrophotometric method (based on the
absorption ratio of the sulfonephthalein dey, m-
cresole purple Clayton and Byrne, 1993
Phosphorus
DOP X Determination by alkaline persulphate oxidation Hansen and Koroleff, 1999
SRP X Automated Cadmium Reduction Method
Murphey and Riley et.al., 1962; Hansen
and Koroleff, 1999, modified by Keroul
and Aminot 1997, Grasshoff “Methods of
seawater analysis”, 1999; NIOZ –
Nederlands Instituut for Onderzoek der
Zee (Royal Netherlands Institute for Sea
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Reserach), Den Hoorn (Texel), The
Netherlands
TPP X Spectrophotometrically
Hansen and Koroleff, 1999; Holmes et al.,
1999
Silica
Biogenic
silica X
Spectrophotometrically, leaching method (135
minutes, 85°C with 0.1M NaOH) Hansen and Koroleff, 1999
Silic acid X Automated Cadmium Reduction Method
Murphey and Riley et.al., 1962; Hansen
and Koroleff, 1999, modified by Keroul
and Aminot 1997, Grasshoff “Methods of
seawater analysis”, 1999; NIOZ –
Nederlands Instituut for Onderzoek der
Zee (Royal Netherlands Institute for Sea
Reserach), Den Hoorn (Texel), The
Netherlands
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Hellenic Center for Marine Research (HCMR)
Parameter
Detection Mode Method Name
Reference
Comments
Manual Automated
Carbon X
POC X
CHN analyzer Hedges and Stern (1984)
TOC
X
high-temperature catalytic oxidation
method Sempéré et al. (2002)
Chlorophyll a X
Fluorometric Detection after acetone
extraction Holm-Hansen et al. (1965)
Nitrogen
NH3
X
Vis/UV spectrophotometric
determination Ivancic & Deggobis 1984
NO2
X
Vis/UV spectrophotometric
determination Strickland and Parsons (1972)
NO3
X
Vis/UV spectrophotometric
determination Strickland and Parsons (1972)
PON X
CHN analyzer Hedges and Stern (1984)
TN
X
Wet-oxidation
Pujo-Pay & Raimbault (1994) and
Raimbault et al. (1999)
Oxygen,
dissolved X Winkler Carpenter (1965a, b)
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Phosphorus
SRP
X
Vis/UV spectrophotometric
determination Strickland and Parsons (1972) micromolar level
SRP X
MAGIC method Rimmelin and Moutin, 2005 nanomolar level
TP
X
Wet-oxidation
Pujo-Pay & Raimbault (1994) and
Raimbault et al. (1999)
Silica X
Vis/UV spectrophotometric
determination Strickland and Parsons (1972)
References
Carpenter, J. H., 1965(a). The accuracy of the Winkler method for the dissolved oxygen analysis. Limnology and Oceanography, 10, 135-140.
Carpenter, J. H., 1965(b). The Chesapeake Bay Institute technique for dissolved oxygen method. Limnology and Oceanography, 10, 141-143.
Hedges, J. I., and Stern, J. H. (1984). Carbon and Nitrogen determination of carbonate-containing solids. Limnol. Oceanogr. 29, 657–663.
Holm-Hansen, O., Lorenzen, C. J., Holmes, R. W., and Strickland, J. D. H. (1965). Fluorometric determination of chlorophyll. J. Cons. Perm. Int. Explor. Mer. 30, 3–
15.
Invancic, I., and Degobbis, D. (1984). An optimal manual procedure for ammonia analysis in natural waters by the indophenol blue method. Water Res. 18, 1143–
1147.
Kirchmann, D. L., Newell, S. Y., and Hodson, R.,E. (1986). Incorporation versus biosynthesis of leucine: implications for measuring rates of protein syntheis and
biomass production by bacteria in marine systems. Mar. Ecol. Prog. Ser. 32, 47–59.
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Lin, P., Chen, M., and Guo, L. (2012). Speciation and transformation of phosphorus and its mixing behavior in the Bay of St. Louis estuary in the northern Gulf of
Mexico. Geochim. Cosmochim. Acta 87, 283–298.
Miyazaki, Y., Kawamura, K., Jung, J., Furutani, H., and Uematsu, M. (2011). Latitudinal distributions of organic nitrogen and organic carbon in marine aerosols
over the western North Pacific. Atmos. Chem. Phys. 11, 3037–3049. doi:10.5194/acp-11-3037-2011.
Pujo-Pay, M., Raimbault, P., 1994. Improvement of the wet-oxidation procedure for simultaneous determination of particulate organic nitrogen and phosphorus
collected on filters. Mar. Ecol. Prog. Ser. 105, 203-207.
Raimbault, P., Pouvesle, W., Diaz, F., Garcia, N., Sempere R., 1999. Wet oxidation and automated colorimetry for simultaneous determination of organic carbon,
nitrogen and phosphorus dissolved in seawater. Marine Chemistry, 66, 161-169.
Sempéré, R., Panagiotopoulos, C., Lafont, R., Marroni, B., and Van Wambeke, F. (2002). Total organic carbon dynamics in the Aegean Sea. J. Mar. Syst. 33–34,
355–364.
Smith, D. C., and Azam, F. (1992). A simple, economical method for measuring bacterial protein synthesis rates in seawater using 3H-leucine. Mar. Microb. Food
Webs 6, 107–114.
Steeman-Nielsen, E. (1952). The use of radio-active carbon (C14) for measuring organic production in the sea. Journal du Cons 18, 117–140.
Strickland, J. D. H., and Parsons, T. R. (1972). A practical handbook of seawater analysis. B. Fish. Res. Board Can. 167, 311.
Rimmelin, P., and Moutin, T. (2005). Re-examination of the MAGIC method to determine low orthophosphate concentration in seawater. Anal. Chim. Acta 548,
174–182.
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1.8.6 Ludwig-Maximilians-Universität Munich (LMU)
Parameter
Detection Mode
Method Name Reference Comments Manual Automated
Alkalinity X Acidic titration DIN ISO 9963-1/2
Carbon (POC) X Elemental analyser DIN 38409-46; Hedges & Stern 1984
Chlorophyll a X
In vitro: fluorometric detection after acetone
extraction DIN 38412-16
X
In vitro: fluorometric detection after ethanol
extraction DIN 38412-16
X
In vivo: fluorometric detection (Algal lab
analyser, Turner, AquaPen)
Chlorophyll content is excited
by coloured LEDs and allocated
to the different algal classes
Cl- X Ion chromatography DIN ISO 10304-1; Smith & Chang 1983
Nitrogen
NO2 X Ion chromatography DIN ISO 10304-1; Smith & Chang 1983
NO3 X Ion chromatography DIN ISO 10304-1; Smith & Chang 1983
NH4 X Fluorometrical Holmes et al. 1999
PON X Elemental analyser Hedges & Stern 1984
Oxygen, dissolved X Winkler Carpenter (1965a, b)
Phosphorus
PP X Photometric with ammoniummolybdate DIN ISO 6878:2004; Grasshoff et al. 1999
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SRP X Photometric with ammoniummolybdate DIN ISO 6878:2004; Grasshoff et al. 1999
TP X Photometric with ammoniummolybdate DIN ISO 6878:2004; Grasshoff et al. 1999
Salinity X 2 graphite electrodes WTW multi probe
SiO2 X Photometric detection DIN ISO 15923-1
SO4 Ion chromatography DIN ISO 10304-1; Smith & Chang 1983
References
Carpenter, J. H., 1965(a). The accuracy of the Winkler method for the dissolved oxygen analysis. Limnology and Oceanography, 10, 135-140.
Carpenter, J. H., 1965(b). The Chesapeake Bay Institute technique for dissolved oxygen method. Limnology and Oceanography, 10, 141-143.
Grasshoff, K., Kremling, K., & Ehrhardt, M. (Eds.). (1999). Methods of seawater analysis. John Wiley & Sons
Hedges, J. I., & Stern, J. H. (1984). Carbon and nitrogen determinations of carbonate‐containing solids. Limnology and oceanography, 29(3), 657-663.
Holmes, R.M., Aminot, A., Kérouel, R., Hooker, B.A & Peterson, B.J. (1999) A simple and precise method for measuring ammonium in marine and freshwater
ecosystems. Can.J.Fish.Aquat.Sci. 56: 1801-1808.
Smith, F. C., & Chang, R. C. C. (1983). The practice of ion chromatography. Wiley.
DIN EN ISO 9963-1:1996-02: Wasserbeschaffenheit - Bestimmung der Alkalinität - Teil 1: Bestimmung der gesamten und der zusammengesetzten Alkalinität
(ISO 9963-1:1994); Deutsche Fassung EN ISO 9963-1:1995
DIN EN ISO 9963-2:1996-02: Wasserbeschaffenheit - Bestimmung der Alkalinität - Teil 2: Bestimmung der Carbonatalkalinität (ISO 9963-2:1994); Deutsche
Fassung EN ISO 9963-2:1995
DIN 38409-46:2012-12: Deutsche Einheitsverfahren zur Wasser-, Abwasser- und Schlammuntersuchung - Summarische Wirkungs- und Stoffkenngrößen
(Gruppe H) - Teil 46: Bestimmung des ausblasbaren organischen Kohlenstoffs (POC) (H 46)
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DIN 38412-16:1985-12: Deutsche Einheitsverfahren zur Wasser-, Abwasser- und Schlammuntersuchung; Testverfahren mit Wasserorganismen (Gruppe L);
Bestimmung des Chlorophyll-a-Gehaltes von Oberflächenwasser (L 16)
DIN EN ISO 10304-1:2009-07: Wasserbeschaffenheit - Bestimmung von gelösten Anionen mittels Flüssigkeits-Ionenchromatographie - Teil 1: Bestimmung von
Bromid, Chlorid, Fluorid, Nitrat, Nitrit, Phosphat und Sulfat (ISO 10304-1:2007); Deutsche Fassung EN ISO 10304-1:2009
DIN EN ISO 6878:2004-09: Wasserbeschaffenheit - Bestimmung von Phosphor - Photometrisches Verfahren mittels Ammoniummolybdat (ISO 6878:2004);
Deutsche Fassung EN ISO 6878:2004
DIN ISO 15923-1:2014-07: Wasserbeschaffenheit - Bestimmung von ausgewählten Parametern mittels Einzelanalysensystemen - Teil 1: Ammonium, Nitrat, Nitrit,
Chlorid, Orthophosphat, Sulfat und Silikat durch photometrische Detektion (ISO 15923-1:2013)
1.8.7 Middle East Technical University (METU)
Parameter
Detection Mode Method Name
Reference
Comments
Manual Automated
Alkalinity X Titration
Standard Methods, 22. Edition. American
Health Association, 1996.
Chlorophyll a X
Spectrophotometric Detection after dissolving in
ethanol
Jespersen, A-M. & K. Christophersen,
1987.
Nitrogen
NH3 X The Skalar Autoanalyzer method
San++ Automated Wet Chemistry
Analyzer, Skalar Analytical,
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B.V., Breda, The Netherlands
X Indophenol Blue Method Chaney, A. L. and Morbach, E. P., 1982.
NO2 X The Skalar Autoanalyzer method
San++ Automated Wet Chemistry
Analyzer, Skalar Analytical,
B.V., Breda, The Netherlands
X Spectrophotometric method (pink dye)
Mackereth, F.J., H. J. Heron & J. F. Talling,
1978.
NO3 X The Skalar Autoanalyzer method
San++ Automated Wet Chemistry
Analyzer, Skalar Analytical,
B.V., Breda, The Netherlands
X Spectrophotometric method (pink dye)
Mackereth, F.J., H. J. Heron & J. F. Talling,
1978.
TN
X The Skalar Autoanalyzer method
San++ Automated Wet Chemistry
Analyzer, Skalar Analytical,
B.V., Breda, The Netherlands
Phosphorus
SRP X Ascorbic Acid Method
Mackereth, F.J., H. J. Heron & J. F. Talling,
1978.
TP/ TDP X Persulfate Digestion, Ascorbic Acid Method
Mackereth, F.J., H. J. Heron & J. F. Talling,
1978.
Silica X
Molybdosilicic Acid Method/ Heteropoly Yellow
Method
Golterman, H., L. Clymo & M. A. M.
Ohnstad, 1978.)
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References
Chaney, A. L. and Morbach, E. P., 1982. Modified reagents for the determination of urea and ammonia. Clin. Chem. 8, 130-132.
Golterman, H., L. Clymo & M. A. M. Ohnstad, 1978. Methods for chemical and physical analyses of freshwaters. 2nd edition. Blackwell Scientific Publishers, Oxford.
Jespersen, A-M. & K. Christophersen, 1987. Measurements of chlorophyll a from phytoplankton using ethyl alcohol as extraction solvent. Arch.Hydrobiol. 109:
445-454.
Mackereth, F.J., H. J. Heron & J. F. Talling, 1978. Water analyses: some methods for limnologists. Freshwater Biological Assoc. Scientific Publication No: 36.
Standard Methods, 22. Edition. American Health Association, 1996.
San++ Automated Wet Chemistry Analyzer, Skalar Analytical,B.V., Breda, The Netherlands.
1.8.8 Netherlands Institute of Ecology (NIOO)
Parameter
Detection Mode Method Name
Reference
Comments
Manual Automated
Alkalinity X Si Analytics Titroline-7000, Titrasoft software 3.1 Si Analytics,
Carbon
Shimadzu TOC-L
Shimandzu Benelux, Den Bosch, The
Netherlands
DOC/ DIC X
TOC/ TIC
Chlorophyll a X HPLC ultimate 3000
Nitrogen
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NH3 X Quaatro method 541502714000
Quaatro Applications, Beun de Ronde,
Abcoude, The Netherlands
NO2 X Quaatro method 5415028714100
Quaatro Applications, Beun de Ronde,
Abcoude, The Netherlands
NO3 X Quaatro method 5415028714100
Quaatro Applications, Beun de Ronde,
Abcoude, The Netherlands
TN X Flash CN analyser Interscience, Breda, The Netherlands.
Phosphorus
TP/ TDP X Quaatro method Q-031-04 Rev. 1
Quaatro Applications, Beun de Ronde,
Abcoude, The Netherlands
Silica X Quaatro method Q-038-04 Rev 1
Quaatro Applications, Beun de Ronde,
Abcoude, The Netherlands
1.8.9 Umweltbundesamt (UBA)
Parameter
Detection Mode Method Name
Reference
Comments
Manual Automated
Alkalinity X Acidic titration by robotic titrosampler
Endpoints: DIN EN ISO 9963-1, optional:
Gran-Plot-titration acc. Sigg & Stumm
1989
Coupled with major
anion and cation
analyses (TitrIC-System)
(incl. ion mass balance)
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Carbon
DOC/ DIC
TOC/ TIC X
Combustion and IR-detection by through-flow or
automatic sampler DIN EN 1484
Chlorophyll,
total (chl-a +
pheophytin) X
Photometric detection after cell disruption and
subsequent hot ethanol extraction
Endpoints: Total chlorophyll acc. Parsons
& Strickland 1963, chl a + pheophytin acc.
DIN 38412-16
Ultrasonic cell
disruption
Filterable dry
matter X Gravimetric DIN 38409-2
Major
anionic
components:
F, Cl, Br, SO4,
PO4, NO2,
NO3 X
IC-automated detection by electric conductivity,
incl. inline dialysis, chemical and CO2-surpression DIN EN ISO 10304-1
Coupled with titration
and major anion
analyses (TitrIC-System)
(incl. ion mass balance)
Major
cationic
components:
Li, Na, K, Mg,
Ca, NH4 X
IC-automated detection by electric conductivity
incl. inline dialysis DIN EN ISO 14911
Coupled with titration
and major anion
analyses (TitrIC-System)
(incl. ion mass balance)
Nitrogen
NH3 X
CFA-based photometric detection, indophenol blue
method (Berthelot’s reagent)
Chaney & Marbach 1962, Bucur et al.
2006, DIN EN ISO 11732, Skalar Kat Nr.
155-002w/r
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NO2 X
CFA-based photometric detection, diazotization of
NO2 (Gries-Ilosvay reaction) with sulphanilamide
Bendschneider & Robinson 1952, acc. to
DIN EN ISO 13395, device specific: Skalar:
Katnr. 461-031
NO3 X
CFA-based photometric detection, reduction to
nitrite by use of coppered Cd-granules and
detection as nitrite (see above)
Wood et al. 1967, Nydahl 1976
TN/TDN
X
Pressure digestion by use of persulfate oxidation to
nitrate and subsequent detection as nitrite (see
above)
Koroleff 1983b
Joint digestion of
nitrogen + phosphor
feasible
Phosphorus
SRP X
CFA-based photometric detection, reduction to
molybdenum blue complex by use of ascorbic acid
and antimonyl tartrate
Murphy & Riley 1962, Walinga et al. 1995,
acc. to DIN EN ISO 15681-2: Skalar: Kat Nr.
503-010w/r, a + b
Optional: Low-level-
phosphate module used
below 2 µg/L PO4-P
TP/ TDP X
Pressure digestion by use of persulfate oxidation
and subsequent photometric detection as
phosphate (SRP) (see above)
Koroleff 1983a
Joint digestion of
nitrogen + phosphor
feasible
Silica X
CFA-based photometric detection, reduction to
molybdenum blue complex by use of ascorbic acid
Mullin & Riley 1955, DIN EN ISO 16264,
Skalar: Katnr. 563-052
CFA-based photometric
detection, reduction to
molybdenum blue
complex by use of
ascorbic acid
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References
Bendschneider, K., Robinson, R.J. (1952): A new spectrophotometric method for the determination of nitrite in sea water. J. Mar. Res. 11, 97-96.
Bucur, B., Catala Icardo, M., Martinez Calatyud, J. (2006): Spectrometric determination of ammonia by an rFIA assembly. Revue Roumaine de Chimie 51, 101-108.
Chaney, A.L., Marbach, E.P. (1962): Modified reagents for determination of urea and ammonia. Clin. Chem. 8, 130-132.
DIN 38409-2: Summarische Wirkungs- und Stoffkenngrößen (Gruppe H), Bestimmung der abfiltrierbaren Stoffe und des Glührückstandes (H 2).
German standard methods for the examination of water, waste water and sludge; parameters characterizing effects and substances (group H); determination of
filterable matter and the residue on ignition (H 2).
DIN 38412-16: Testverfahren mit Wasserorganismen (Gruppe L), Bestimmung des Chlorophyll-a- Gehaltes von Oberflächenwasser (L16)
German standard methods for the examination of water, waste water and sludge; test methods using water organisms (group L); determination of chlorophyll a
in surface water (L 16)
DIN EN 1484: Wasseranalytik - Anleitungen zur Bestimmung des gesamten organischen Kohlenstoffs (TOC) und des gelösten organischen Kohlenstoffs (DOC);
Deutsche Fassung EN 1484-1997
Water analysis - Guidelines for the determination of total organic carbon (TOC) and dissolved organic carbon (DOC)
DIN EN ISO 11732: Wasserbeschaffenheit - Bestimmung von Ammoniumstickstoff - Verfahren mittels Fließanalytik (CFA und FIA) und spektrometrischer Detektion
(ISO 11732:2005); Deutsche Fassung EN ISO 11732: 2005.
Water quality - Determination of ammonium nitrogen - Method by flow analysis (CFA and FIA) and spectrometric detection (ISO 11732:2005)
DIN EN ISO 13395: Wasserbeschaffenheit. Bestimmung von Nitritstickstoff, Nitratstickstoff und der Summe von beiden mit der Fließanalytik (CFA und FIA) und
spektrometrischer Detektion (ISO 13395: 1996). Deutsche Fassung EN ISO 13395: 1996.
Water quality - Determination of nitrite nitrogen and nitrate nitrogen and the sum of both by flow analysis (CFA and FIA) and spectrometric detection (ISO 13395:
1996)
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DIN EN ISO 15681-2: Wasserbeschaffenheit. Bestimmung von Orthophosphat und Gesamtphosphor mittels Fließanalytik (FIA und CFA). Teil 2: Verfahren mittels
kontinuierlicher Durchflussanalyse (CFA) (ISO 15681-2: 2003). Deutsche Fassung EN ISO 15681-2: 2004.
Water quality - Determination of orthophosphate and total phosphorus contents by flow analysis (FIA and CFA) - Part 2: Method by continuous flow analysis
(CFA) (ISO 15681-2: 2003)
DIN EN ISO 16264: Wasserbeschaffenheit. Bestimmung löslicher Silicate mittels Fließanalytik (FIA und CFA) und photometrischer Detektion (ISO 16264:2002).
Deutsche Fassung EN ISO 16264: 2004.
Water quality - Determination of soluble silicates by flow analysis (FIA and CFA) and photometric detection (ISO 16264: 2002)
DIN EN ISO 9963-1: Wasserbeschaffenheit-Bestimmung der Alkalinität, Teil 1: Bestimmung der gesamten und der zusammengesetzten Alkalinität (ISO 9963-1):
1994, Deutsche Fassung EN ISO 9963-1: 1995
Water quality - Determination of alkalinity - Part 1: Determination of total and composite alkalinity (ISO 9963-1: 1994)
DIN EN ISO 10304-1: Wasserbeschaffenheit- Bestimmung von gelösten Anionen mittels Flüssigkeits- Ionenchromatographie, Teil 1: Bestimmung von Bromid,
Chlorid, Fluorid, Nitrat, Nitrit, Phosphat und Sulfat (ISO 10304-1:2007), Deutsche Fassung EN ISO 10304-1:2009
Water quality - Determination of dissolved anions by liquid chromatography of ions - Part 1: Determination of bromide, chloride, fluoride, nitrate, nitrite,
phosphate and sulfate (ISO 10304-1:2007)
DIN EN ISO 14911: Wasserbeschaffenheit- Bestimmung der gelösten Kationen Li+, Na+, NH4+ , K+, Mn2+, Ca2+, Mg2+, Sr2+ und Ba2+ mittels
Ionenchromatographie, Verfahren für Wasser und Abwasser (ISO 14911:1998), Deutsche Fassung EN ISO 14911: 1999
Water quality - Determination of dissolved Li⁺, Na⁺, NH₄⁺, K⁺, Mn²⁺, Ca²⁺, Mg²⁺, Sr²⁺ and Ba²⁺ using ion chromatography - Method for water and waste water (ISO
14911: 1998)
Koroleff, F. (1983a): Determination of phosphorus by acid persulphate oxidation. - In Grasshoff, K. (ed.): Methods of sea water analysis (2nd ed.), p. 134-136.
Weinheim: Verlag Chemie.
Koroleff, F. (1983b): Determination of total and organic nitrogen after persulphate oxidation. - In Grasshoff, K. (ed.): Methods of sea water analysis (2nd ed.), p.
164-168. Weinheim: Verlag Chemie.
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Mullin, J.B., Riley J.P. (1955): The colorimetric determination of silicate with special reference to sea and natural waters. - Anal. Chim. Acta 12, 162-176.
Murphy, J., Riley J.P. (1962): A modified method for the determination of phosphate in natural waters. - Anal. Chim. Acta 27, 31-36.
Nydahl, F. (1976): On the optimum conditions for the reduction of nitrate to nitrite by cadmium. Talanta 23: 349-357.
Parsons T.R. & Strickland J.D.H. (1963): Discussion of spectrophotometric determination of marine-plant pigments, with revised equations for ascertaining
chlorophylls and carotenoids. J. Mar. Res. 21: 155-63.
Skalar Kat Nr. 155-002w/r, Analyse: Ammonium, Bereich: 2 - 100 ppb P, Matrix: Abwasser. - Issue 102197/MH/97202624 97-0411.
Skalar Kat Nr. 461-031, Analyse: Nitrat + Nitrit, Meßbereich: 2 - 100 ppb P, Matrix: Seewasser. - Issue 0690899/MH/99207147. 990728 Institut
für Meereskunde
Skalar Kat Nr. 503-010w/r, a: Analyse: Phosphat, Meßbereich: 2 - 100 ppb P, Matrix: Seewasser. - Issue 060899/MH/99207147.
Skalar Kat Nr. 503-010w/r, b: Analysis: Phosphate, Range: 5 - 100 ppb P, Matrix: Surface- & drinking water. - Issue 0690803/MH/99226559.
Skalar Kat Nr. 563-052: Analyse: Silikat, Meßbereich: 0.02 - 1 ppm Si, Matrix: Wasser. - Issue 111397/MH/97202944 97-0541.
Sigg, L. & Stumm, W. (1989): Aquatische Chemie. - 396 S., 128 Abb., 37 Tab. Zürich: Verlag der Fachvereine 1989.
Walinga, I., Van Der Lee, J.J., Houba, V.J.G., Van Vark. W & Novozamsky, I. (1995): 1.7.2 Determination of phosphorus by colorimetry (automated, by flow
analyzer). - In: Walinga, I., Van Der Lee, J.J., Houba, V.J.G., Van Vark. W & Novozamsky, I. (eds.) (1995): Plant analysis manual: PANA-A1/34 - 39. Dordrecht:
Kluwer.
Wood, E.D., Armstrong, F.A.J. & Richards, F.A. (1967): Determination of nitrate in sea water by cadmium-copper reduction to nitrite. J. mar. biol. Ass. U.K. 47, 23-
31.
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1.8.10 University of Helsinki, Tvärminne Zoological Station (UH)
Parameter
Detection Mode Method Name
Reference
Comments
Manual Automated
Carbon
DOC X
High-temperature catalytic oxidation and
infrared detection
Grasshoff et al. (1999) (chapter
15)
POC X
High-temperature catalytic oxidation and
mass spectrometric detection
Grasshoff et al. (1999) (chapter
17)
Chlorophyll a X
Fluorometric detection after ethanol
extraction
Baltic marine environment
protection commission (1988)
Nitrogen
NH3 X Indophenol Blue Method
Modification of Grasshoff et al.
(1999) (chapter 10.2.10);
Grasshoff (1976) (chapter 9.2)
Dichloroisocyanuric acid
as hypochlorite donor
NO2 X Automated Colorimetric Method
Modification of Grasshoff et al.
(1999) (chapter 10.2.8);
Grasshoff (1976) (chapter 9.3)
NO3 X
Automated Colorimetric Method, Vanadine
Chloride Reduction Method
Modification of Grasshoff et al.
(1999) (chapter 10.2.9);
Grasshoff (1976) (chapter 9.4)
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PON X
High-temperature catalytic oxidation and
chemiluminescence detection
Grasshoff et al. (1999) (chapter
17)
TDN
X
High-temperature catalytic oxidation and
chemiluminescence detection
Grasshoff et al. (1999) (chapter
15)
TN X
Persulfate Digestion, Vanadine Chloride
Reduction Method
Modification of Grasshoff et al.
(1999) (chapter 10.2.16);
Grasshoff (1976) (chapter 9.8.3)
Phosphorus
SRP X Automated Ascorbic Acid Method
Modification of Grasshoff et al.
(1999) (chapter 10.2.5);
Grasshoff (1976) (chapter 9.1.2)
PP
X
Dry Ashing, Ascorbic Acid Method
Solorzano 1980a; Modification
of Grasshoff et al. (1999)
(chapter 10.2.12); Grasshoff
(1976) (chapter 9.1.2)
TP X
Persulfate Digestion, Automated Ascorbic
Acid Method
Modification of Grasshoff et al.
(1999) (chapter 10.2.13);
Grasshoff (1976) (chapter 9.1.4)
Silica X Automated Molybdosilicate Method
Modification of Grasshoff et al.
(1999) (chapter 10.2.11);
Grasshoff (1976) (chapter 9.6.2)
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References
Guidelines for the Baltic monitoring programme for the third stage (1988). Part D. Biological determinands. Baltic Sea Environment Proceedings No. 27 D. Baltic
Marine Environment Protection Commission – Helsinki Commission.
Grasshoff, K. (Ed.) (1976). Methods of seawater analysis. Verlag Chemie.
Grasshoff, K., Kremling, K., & Ehrhardt, M. (Eds.). (1999). Methods of seawater analysis. John Wiley & Sons.
SOLORZANO, L., SHARP, J. H. (1980a). Determination of total dissolved phosphorus and particulate phosphorus in natural waters. Limnol. Oceanogr., 25(4), 754-
758.
1.8.11 University of Bergen (UIB)
Parameter
Detection Mode Method Name
Reference
Comments
Manual Automated
Carbon
POC X Flash elemental analyses Pella & Colombo 1973
TOC1 X
High temperature catalytic
oxidation Børsheim 2000
Chlorophyll a X
Fluorometric Detection after
Acetone Extraction Parsons et al. (1984)
X
Fluorometric Detection after
Methanol Extraction Holm-Hansen and Riemann (1978)
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Nitrogen
NH4+ X
Ortho-phthaldialdehyde
fluorometric Method (OPA)
Holmes et al. 1999
Adapted for microwell plate according
to Poulin & Pelletier 2007
NO3 X2
Cadmium reducing column
(nitrate to nitrite)
Parsons et al. (1992) adapted to an
autoanalyzer (San1 Segmented Flow
Analyser, Skalar Analytical B.V., The
Netherlands) as described in Rey et al.
(2000).
PON X Flash elemental analyses Pella & Colombo 1973
Phosphorus
SRP X
Ascorbic Acid - molybdenum
- tartrate blue complex Koroleff 1983
Salinity X
Direct measurement with
SAIV SD204 CTD -
Silicate
Ascorbic Acid - Molybdenum
– Oxalic - Blue Complex Valderrama 1995
1 No routine analysis
2 Performed by Institute of Marine Research (IMR) in Bergen
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References
Børsheim, K. Y. 2000. Bacterial production rates and concentrations of organic carbon at the end of the growing season in the Greenland Sea. Aquat. Microb.
Ecol. 21: 115– 123. doi:10.3354/ame021115
Holm-Hansen O, Riemann B (1978). Chlorophyll a determination: improvements in methodology. Oikos 30: 438-447.
Holmes RM, Aminot A, Keroul R, Hooker AH, Peterson BJ (1999). A simple and precise method for measuring ammonium in marine and freshwater ecosystems.
Aquat Sci 56: 1801-1808.
Koroleff F (1983). Determination of nutrients. In: Grasshoff K, Ehrhardt M, Kremling K (eds). Methods in seawater analyses. Verlag Chemie: Weinheim/Deerfield
Beach, Florida. pp 125-131.
Parsons, T. R., Y. Maita, and C. M. Lalli. 1984. A manual of chemical and biological methods for seawater analysis, p.
Pergamon Press.
Pella E, Colombo B (1973). Study of carbon, hydrogen and nitrogen determination by combustion-gas chromatography. Microchimica Acta 61: 697-719.
Rey, F., T. T. Noji, and L. A. Miller. 2000. Seasonal phytoplankton development and new production in the central Greenland Sea. Sarsia 85: 329–344. doi:10.1080/
00364827.2000.10414584
Valderrama JC (1995). Methods of nutrient analysis. In: Hallograeff GM, Anderson DM, Cembella AD (eds). Manual of harmful marine microalgae. IOC manuals
and guides. UNESCO: Paris. pp 262-265.
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1.8.12 Umea Marine Science Center (UMF)
Parameter
Detection Mode Method Name
Reference
Comments
Manual Automated
Alkalinity X Potentiometric Titration
SS-EN ISO 9963-
1:1994 modified
HC-B-B151
Carbon
DOC X
High temperature combustion with NDIR
detection
HC-C-C21 / SS-EN
1484 ed. 1
modified
Chlorophyll a X Spectrofluorometry, ex 433nm/em 673nm ICES / HC-C-C21 Ethanol extraction
C and N X
Elemental analysis: High temperature
combustion with IR-detection
LECO
Corporation5
Nitrogen
NH4 X CFA (QuAAtro, Autoanalyzer ) ”Grasshoff”2 Photometric Phenol method
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NO2 X CFA (QuAAtro, Autoanalyzer ) ”Grasshoff”2
Photometric
sulfanilamide/Ethylenediamine
NO3 X CFA (QuAAtro, Autoanalyzer ) ”Grasshoff”2
Photometric CD-reductor,
sulfanilamide/Ethylenediamine
TN X
Simultaneous N and P Oxidative digestion
with peroxodisulfate using the borate buffer
system followed by CFA (QuAAtro,
Autoanalyzer ). ”Grasshoff”2
Photometric CD-reductor,
sulfanilamide/Ethylenediamine
TDN X
Simultaneous N and P Oxidative digestion
with peroxodisulfate using the borate buffer
system followed by CFA (QuAAtro,
Autoanalyzer ). ”Grasshoff”2
Photometric CD-reductor,
sulfanilamide/Ethylenediamine
Oxygen X Winkler Titration SS-EN 25813:1992
pH
HC-B-B141 / SS-EN
ISO 10523:2012 pH 7 – 10
Phosphorus
PP X Photometric Dry combustion
Solarzano3 S-EN
ISO 6878:20054
Photometric Molybdate-
ascorbic acid
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TP X
Simultaneous N and P Oxidative digestion
with peroxodisulfate using the borate buffer
system followed by CFA (QuAAtro,
Autoanalyzer ). ”Grasshoff”2
Photometric Molybdate-
ascorbic acid
TDP X
Simultaneous N and P Oxidative digestion
with peroxodisulfate using the borate buffer
system followed by CFA (QuAAtro,
Autoanalyzer ). ”Grasshoff”2
Photometric Molybdate-
ascorbic acid
Silica X CFA (QuAAtro, Autoanalyzer) ”Grasshoff”2
Photometric Molybdate-Oxalic
acid
References
1HELCOM Combined Manual for Marine Monitoring (2015) Letters B or C refers to actual part and annex
2K. Grasshoff et al, Methods of Seawater Analysis, 2nd edition, Verlag Chemie, 1983, page 125-187; 347-376
3L.Solarzano, J. H. Sharp, Limnol. Oceanogr.., 25(4) 1980 754-758,
4SS-EN ISO 6878:2005, Water quality—Determination of phosphorus - Ammonium molybdate spectrometric method
5LECO Corporation, Thru Spec CHN/CHNS Micro Carbon/Hydrogen/Nitrogen/Sulfur Determinators. Instruction Manual Version 2.7X. Part Number 200-716, July
2015
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1.8.13 WasserCluster Lunz (WCL)
Parameter
Detection Mode Method Name
Reference
Comments
Manual Automated
Alkalinity X Titration Schwoerbel (chapter 1.2.5)
Carbon
DOC/ DIC X Oxidation and Conductometric Detection U.S. Patent No. 5,132,094
TOC/ TIC X Oxidation and Conductometric Detection U.S. Patent No. 5,132,094
Chlorophyll a X Fluorometric Detection after Acetone Extraction Arar 1997 (EPA Method 445.0)
No cell disruption and
acidification is applied
Nitrogen
NH3 X Indophenol Blue Method Ivancic & Deggobis 1984
X Indophenol Blue Method ISO 7150
NO2 X Automated Hydrazine Reduction Method ISO 13395:1996
X Colorimetric Method ISO 13395:1996
NO3 X Automated Hydrazine Reduction Method ISO 13395:1996
X Sodiumsalicylate Method Schwoerbel (chapter 1.2.14)
TN
X Persulfate Digestion, Hydrazine Reduction Method
Clesceri 1999 (4500-P, chapter J);
ISO 13395:1996
Phosphorus
SRP X Ascorbic Acid Method Grasshoff 1999 (chapter 10.2.5)
X Automated Ascorbic Acid Method ISO 15681-2
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PP
X
Dry Ashing and Ascorbic Acid Method
Solorzano 1980a; Grasshoff 1999 (chapter
10.2.5)
Digestion modified from
Solorzano
TP/ TDP X Persulfate Digestion and Ascorbic Acid Method Grasshoff 1999 (chapter 10.2.13)
Silica X Molybdosilicate Method or Heteropoly Blue Method Clesceri 1999 (4500-SiO2, chapter C or D)
References
Arar, E. J., & Collins, G. B. (1997). Method 445.0: In vitro determination of chlorophyll a and pheophytin a in marine and freshwater algae by fluorescence.
Cincinnati: United States Environmental Protection Agency, Office of Research and Development, National Exposure Research Laboratory.
Clesceri, L. S., Greenberg, A. E., & Eaton, A.D. (1996). Standard methods for the examination of water and wastewater. APHA, AWWA and WPCF, Washington DC.
DIN EN ISO 15681-2, Water quality - Determination of orthophosphate and total phosphorus contents by flow analysis (FIA and CFA) - Part 2: Method by
continuous flow analysis (CFA), 06-2001
Grasshoff, K., Kremling, K., & Ehrhardt, M. (Eds.). (1999). Methods of seawater analysis. John Wiley & Sons
ISO 7150-1: 1984, Water quality—Determination of ammonium, manual spectrometric method
ISO 13395:1996, Water quality—Determination of nitrite nitrogen and nitrate nitrogen and the sum of both by flow analysis (CFA and FIA) and spectrometric
detection.
Ivančič, I., & Degobbis, D. (1984). An optimal manual procedure for ammonia analysis in natural waters by the indophenol blue method. Water Research, 18(9),
1143-1147.
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