### 12

D R A FT PR O J EC T C H A R T ER : Propos ed 5

- y ea r m oni t or i ng pl an f or cy cl i c v ol at i l e

m et hy l si l oxane ( cV MS) m at er i al s i n sur f ac e s edi m ent and a quat i c bi ot a of t he I nner O sl of

j or d, N or w ay

.
Date: 23February 2011

PROJECT NUMBER : 02/444/S040/07/ xxx

STUDY AREA : I nner Oslofjord, Norway

TEST MATERIALS Octamethylcyclotetrasiloxane (D4): CAS 556

- 67

- 2 Decamethylcyclopentasiloxane (D5): CAS 541

- 02

- 6 Dodecamethylcyclohexasiloxane (D6): CAS 540

- 97

- 6

Consideration should be given to making samples available for analys is of other materials such as legacy contaminants and metals. Results from such analyses may be used to increase the power to detect significant temporal changes in cVMS concentrations.

PURPOSE Determine if concentrations of cyclic volatile methylsiloxan e (cVMS) materials in surface sediment and aquatic biota are stable or changing following a decreasing or increasing temporal trend over the duration of the project.

OBJECTIVE Conduct temporal trend analyses to determine if cVMS concentrations are stable or changing, based on detecting with 80% power (β=0.20) a statistically significant (α=0.05) annual rate of change of ±6% per year over a 5 year period (net change of

- 27% to 34%). This is equivalent to detecting with 65% power (β=0.35; α=0.05) an annual rate of change of ±5.0% per year over a 5 year period (net change of

- 23% to 28%). Detecting a change of ±6% per year with 80% power is less sensitive but statistically more robust than detecting a change of ±5% per year with 65% power. Established monit oring programs, for example the Northern Contaminants Program (NCP) of Canada and the Arctic Monitoring and Assessment Programme (AMAP) of Arctic Countries, are typically designed to detect with 80% power an annual rate of decrease of 5% per year over a 10 to 15 year period for a net decrease of 40 to 55% (UNEP 2004, 2007; EU 2010).

Any process or change in experimental design that decreases variability associated with cVMS concentrations in the samples will increase the power to detect a statistically sig nificant trend or change in concentration. Specific procedures that will be implemented to reduce within

- year and between

- year variability are discussed in the appropriate sections that follow.

Project Duration The expected minimum project duration is 5 years, based on a temporal trend power analysis across a range of values for coefficient of variation (CV=20% to 50%). The power analysis was optimized to estimate the number of samples that would be needed to detect with 90% power an annual change in

co ncentration of ±10% per year if samples were annually collected over a period of 3 to 10 years. Time

- series monitoring programs are unlikely to detect changes with any confidence within a sampling period of less than five years unless the changes are very large (UNEP 2004, 2007; EU 2010). It is expected that a period of 10

- 15 years would be required to detect changes in cVMS concentrations of 5% per year or less.

Project duration was estimated on the basis of within

- year variability of cVMS concentratio ns for a limited number of samples (n=3 to 6 samples per matrix) from previous monitoring programs in Oslofjord (Powell 2009, 2010). However, at least 4

- 5 years of monitoring are required to obtain reliable estimates of random within

- year and between

- year variation and other components of

### 13

variance (UNEP 2004, 2007; EU 2010). Therefore, project duration and experimental design will be

re

- evaluated on an annual basis and modified as needed to maintain or improve upon the defined project objectives.

Sample Matrices to be Collected Sediment (collected annually):

o 0

- 5 cm of surface sediment, sectioned into five 1

- cm thick strata. Each stratum will be analyzed in order to minimize between

- year variability ( EU 2010) and to evaluate potential degradation within t he sediment column. Power to detect a change in concentration in the surface stratum may be improved by treating concentrations in lower strata as covariates.

o Rates of sediment accumulation at the study area are on the order of 0.3 cm per year or less. A s a result a 1 cm stratum of surface sediment will represent about 3 years or more of sediment accumulation. Collection of individual strata greater than 1 cm in thickness will not provide sufficient resolution to detect a significant change over the 5 ye ar program.

o Sediment samples to be collected by sediment core and extruded to the desired thickness of 1 cm.

o Locations where sediment cores will be collected will be identified using a systematic random sampling design from along a transect (Gilbert 1987) that runs parallel to the predominant bottom current, extending from a known point source of wastewater release.

Biota (collected annually):

o Three target species are proposed for the monitoring program in the Inner Oslofjord: 1. Atlantic cod ( Gadus morhua ): age

- 2 fish (size

- class about 46

- 55 cm total length; weight class generally > 1000 g fresh weight); represents a high trophic level predator species. 2. Atlantic herring ( Clupea harengus ): age

- 0 fish (size

- class generally < 15 cm total length); represents a low trophic level forage species for cod. 3. Northern shrimp ( Pandalus borealis ): represents a low trophic level forage species for cod.

o The proposed target species were selected because: 1. Trend analysis monitoring of biota should be based on target species t hat include (UNEP 2004; EU 2010): A high trophic level predator species. A low trophic level forage species near the base of the food chain of the predator species. 2. These species are representative of the pelagic zone food chain (Atlantic cod and Atlantic herring) and the demersal zone food chain (Atlantic cod and Northern shrimp) in the Inner Oslofjord food web (Powell 2010). Northern shrimp are the preferred forage for Atlantic cod in the Inner Oslofjord (Heggelund 2001). The importance of Atlantic herr ing to the diet of Atlantic cod in the Inner Oslofjord is not known. However, age

- 0 Atlantic herring are the preferred forage for Atlantic cod in the Barents Sea (Johnsen et al. 2004). 3. These species are recommended for monitoring studies (UNEP 2004; EU 2010) and may be collected in sufficient numbers needed to meet the objectives of the experimental design. 4. Concentrations of cVMS materials in these species generally bracket the range in cVMS concentrations observed in the Inner Oslofjord (Powell 2010) an d are at least 5 to 10 times the analytical method detection limits of 0.5 ng/g ww. 5. These species have a high ecological significance in the Oslofjord and are commercially important as a food resource.

o Biota will be collected from the inner Oslofjord at th e CEMP stations used for the Norwegian Coordinated Environmental Monitoring Programme (Green et al. 2010).

### 14

The within

- year and between

- year variability resulting from confounding factors associated with sample collection will be reduced by:

o Collecting sam

ples from the same locations during the same timeframe (month or season) of every year;

o Collecting samples by the same field team (or under direction of the same team leader) using the same procedures every year;

o Collecting samples of the same size

- class o r age class every year;

o Selecting representative target species that can be collected every year in sufficient numbers required for the defined experimental design;

o Selecting representative target species that are expected to have concentrations at least 5 to 10 times the analytical method detection limit (MDL). For the cVMS materials MDL should be less than 0.5 ng/g ww and concentrations in the target species at least 2.5 to 5 ng/g ww.

Frequency of Sample Collection and Sample Collection Times Sediment ( individual samples)

o Sediment cores will be collected from five locations (to be determined) in the Inner Oslofjord. Samples will be collected from locations along a transect that runs parallel to the predominant bottom current, extending from a known poin t source of wastewater release;

o Samples will consist of 0

- 5 cm of surface sediment sectioned into five 1

- cm thick strata;

o Samples will be collected on an annual basis from the same locations (i.e., GPS coordinates) during the same month or season of each y ear (to be determined), including
year

- 0 and year

- 5.

Biota (individual or pooled samples)

o Top predator species (Atlantic cod) and lower trophic level forage species (Atlantic herring and Northern shrimp) will be collected by trawl;

o Samples will be collect ed on an annual basis from the same areas during the same month or season of each year (to be determined), including year

- 0 and year

- 5;

o Samples will be field processed for species identification, total length, and fresh weight. Condition factors will be d etermined from the length and weight of each individual specimen;

o Atlantic cod (age

- 2 fish; size

- class about 46

- 55 cm total length; weight class generally > 1000 g fresh weight) will be retained as individual samples. Atlantic herring (age

- 0 fish; size

- cl ass generally < 15 cm total length) and Northern shrimp will be retained as pooled samples (n=3 to 5 pooled samples per species with n=10 to 20 individual specimens per pooled sample).

Number of Samples A temporal trend power analysis was used to estimate the number of samples needed to detect with 80% power (β=0.20) a statistically significant (α=0.05) annual rate of change of ±6% per year over a 5 year period (net change of

- 27% to 34%). A constant coefficient of variation (CV) for concentrations of the cVMS materials was assumed for each collection interval (samples collected annually). Results from previous monitoring studies on the Oslofjord (Powell 2009, 2010) were used to estimate within

- year CV values for annual mean whole

- body lipid

- equivalent co ncentrations in biota and annual mean organic carbon

- equivalent concentrations in sediments. Concentrations of the cVMS materials were expressed on the basis of lipid content and organic carbon content to reduce the CV values used for each power analysis.

Sediment: A minimum sample replication of n=5 samples/stratum/year would be required to detect with 80% power (β=0.20; α=0.05) an annual rate of change of ±6% per year in carbon

equivalent concentration (CV=20%) over a 5 year period (net change of

- 27% to 34%).

### 15

Biota: A min imum sample replication of n=15 samples/species/year would be required to detect with 80% power (β=0.20; α=0.05) an annual rate of change of ±6% per year in whole

body lipid

- equivalent concentration (CV=35%) over a 5 year period (net change of

- 27% to 34%) . For temporal trend studies of contaminants in fish, monitoring guidelines (UNEP 2004) recommend that a minimum of 12 individual samples be collected each year. Atlantic cod (age

- 2 fish; size

- class about 46

- 55 cm total length; weight class generally >10 00 g fresh weight) will be retained as individual samples (n=15 to 20 individual specimens). Atlantic herring (age

- 0 fish; size

- class < 15 cm total length) and Northern shrimp will be retained as pooled samples (n=3 to 5 pooled samples per species with n= 10 to 20 individual specimens per pooled sample).

The numbers of samples to be collected were estimated on the basis of within

- year variability of cVMS concentrations for a limited number of individual samples (n=3 to 6 samples per matrix) from previous m onitoring programs in Oslofjord (Powell 2009, 2010). However, at least 4

- 5 years of monitoring are required to obtain reliable estimates of random within

- year and between

- year variation and other components of variance (UNEP 2004, 2007; EU 2010). Therefo re, the numbers of samples to be collected will be re

- evaluated and modified as needed on an annual basis to maintain or improve upon the project objectives.

Analysis of Samples Sediments and biota will be processed and analyzed following previously vali dated methods and QA procedures (Powell et. al. 2010).

Atlantic cod will be processed and analyzed as individual samples. Atlantic herring and Northern shrimp will be processed and analyzed as pooled samples (n=3 to 5 pooled samples per species with n=10 to 20 individual specimens per pooled sample). Except for Atlantic cod, all samples will be processed as whole

- body homogenates. Atlantic cod will be processed without the head, which is expected to result in a much more homogeneous matrix that will red uce within

- year variability for both cVMS concentration and lipid content.

In addition to analysis of cVMS, samples will be analyzed for lipid content (biota) or organic carbon content (sediments), water content, and the isotopic signatures of stable isot opes of carbon ( 13 C) and nitrogen ( 15 N).

If samples will be analyzed by more than one facility then one laboratory must be responsible for processing and distributing the processed samples to the participating laboratories for analysis.

A standardized QA program, including the analysis of standard reference materials (SRMs) should be followed by all laboratories involved with the study.

Certified SRMs for cVMS materials do not presently exist and would need to be prepared for each matrix in sufficient mass for use over the duration of the monitoring program.

Measurement Basis:

o In order to meet the defined objective of the study, it is important that the within

- year variability by species across samples be minimized. Therefore, temporal trend analyses of the biological data will be based on whole

- body lipid equivalent concentrations of the cVMS materials. Temporal trend analyses of the sediment data will be based on organic carbon equivalent concentrations of the cVMS materials.

o All biota samples will be processed as whole

- body homogenates except for Atlantic cod, which will be processed without the head. Processing the cod without the head is expected to result in a much more homogeneous matrix that will reduce within

- year variability for both cVMS co ncentration and lipid content;

o Within species variation across samples is larger for cVMS concentrations in other matrices or for concentrations not expressed on the basis of lipid content or organic

### 16 carbon content. For example, a mean coefficient of vari ation (CV) of 35% was obtained for whole

- body lipid

- equivalent concentrations across individual samples by species (n=3 to 6 samples for each species) in the Oslofjord food web that was sampled in 2008 (Powell 2010). In contrast, the mean CV value based o n whole

- body wet weight concentrations was about 65% (Powell 2010). Similarly, CV values based on lipid

equivalent concentrations in cod liver were about 50% (Powell 2009).

The within

- year and between

- year variability resulting from confounding factors a ssociated with sample analysis will be reduced by:

o Processing and analyzing samples by the same laboratory or laboratories using the same procedures every year;

o Analyzing sample matrices (e.g., whole

- body) that minimize between

- sample variability

o Analyzing sample matrices with concentrations that are at least 5 to 10 times the analytical method detection limit (MDL). For the cVMS materials MDL should be less than 0.5 ng/g ww and concentrations in the target species at least 2.5 to 5 ng/g ww.

o Analyzing pool ed samples in order to increase sample replication without increasing the number of analyses.

o Consistent use of concentrations that are reported on a normalized basis (e.g., lipid weight) that minimizes the between

- sample variability.

Statistical Analysis Data will be specifically analyzed to detect changes or trends in cVMS concentrations that may occur over the duration of the program (i.e., temporal trend analysis). Temporal trends will be evaluated using procedures described in various guidance docume nts (Gilbert 1987; UNEP 2004, 2007; OSPAR 2008; EU 2010).

Contract To be developed on a year by year basis.

Project Team Team Leader: To be identified

Sample Collection: To be identified. It is proposed that samples be collected in cooperation with t he Norwegian Institute for Water Research (NIVA). An industry representative (e.g., the Team Leader) should participate on every sample collection.

Sample Analysis: To be identified. It is proposed that cVMS analyses be conducted by one of the followin g laboratories:

o Dow Corning Corporation, Auburn, Michigan, USA

o Evonik Goldschmidt GmbH, Essen, Germany

Statistical Analysis and Database Management: To be identified.

References

EU. 2010. Guidance Document No: 25 Guidance on chemical monitoring of s ediment and biota. Technical Report 2010.3991

Gilbert, R.O. 1987. Statistical Methods for Environmental Pollution Monitoring. Van Nostrand Reinhold, New York, NY. ISBN 0

- 442

- 23050

- 8.

Green, N.W., Schøyen, M., Øxnevad, S., Ruus, A., Berge, J.A., Høgå sen, T., Håvardstun, J., Rogne, Å.G., and Tveiten, L. 2010. Coordinated Environmental Monitoring Programme (CEMP). Levels, trends and effects of hazardous substances in fjords and coastal waters

– 2008.

### 17

(Norwegian State Pollution Monitoring Programme. Re port no. 5867/2010. TA

- no 2566/2010, SPFO

- report 1062/2010). NIVA O

- 29106, NIVA rapport, Lnr. 5867

- 2010, NIVA. Available at (as of 24 Jan 2011): http://www.klif.no/no/Tema/Miljoovervakning/Statlig

miljoovervakning/Miljogifter

- langs

- norskekysten/Rapporter /Hazardous

- substances

- in

- fjords

and

- coastal

- waters

- 2008

-- Levels

- trends

- and

- effects

-- Long

- term

- monitoring

- of

- environmental

quality

- in

- Norwegian

- coastal

- waters/?attachment=true.

Heggelund, H.I. 2001. Diett og matinntak til torsk, Gadus morhua L., i Oslofjor den. M.sc. thesis. (Diet and feeding of cod, Gadus morhua L. in the Oslofjord). University of Oslo. In Norwegian.

Johnsen, G.O., Bogstad, B., Mehl, S. & Ulltang, Ø. 2004. Consumption of juvenile herring ( Clupea harengus ) by cod ( Gadus morhua ) in the Baren ts Sea: a new apprach to estimating consumption in piscivorous fish. Can. J. fish. Aquat. Sci. 61: 343

- 359.

OSPAR. 2008. CEMP Assessment Manual. Co

- ordinated Environmental Monitoring Programme Assessment Manual for contaminants in sediment and biota. OSPAR Commission Publication Number No. 379/2008.

Powell, D.E. 2009. Cyclic volatile methylsiloxane materials (D3, D4, D5, and D6) in livers of Atlantic cod ( Gadus morhua ) from Oslofjord, Norway. Comparison and assessment of analytical methods utilized by Dow Corning Corporation, Evonik Goldschmidt, and the Norwegian Institute for Air Research. Final Report submitted to the Centre Européen des Silicones (CES). Dow Corning Corporation, technical report number 2009

- I0000

- 60983.

Powell, D.E. 2010. Bio accumulation and trophic transfer of cyclic volatile methylsiloxane (cVMS) materials in the aquatic marine food webs of the Inner and Outer Oslofjord, Norway. Final Report submitted to the Centre Européen des Silicones (CES). Dow Corning Corporation,
tec hnical report number 2010

- I0000

- 62594.

UNEP. 2004. Guidance for a Global Monitoring Programme for Persistent Organic Pollutants, 1 st Edition. United Nations Environment Programme. UNEP Chemicals, Geneva, Switzerland.

UNEP. 2007. Draft Guidance on t he Global Monitoring Plan for Persistent Organic Pollutants. United Nations Environment Programme. UNEP/POPS/COP.3/INF/14.

### 18

D R A FT PR O J EC T C H A R T ER : Propos ed m oni t or i ng pl an f or cy cl i c v ol at i l e

m et hy l si l oxane ( cV MS) m at er i al s i n t he i nf l uent , e f f l uent , and s l udg e of

a m uni ci pal w as t ewat er t r ea t m ent pl ant

.
Date: 25 February 2011

PROJECT NUMBER : 02/444/S040/07/ xxx

PROPOSED STUDY AREAS : Oslo, Norway Tokyo, Japan Toronto, Ontario, Canada St. Paul, Minnesota, USA

TEST MATERIALS Octamethylcyc lotetrasiloxane (D4): CAS 556

- 67

- 2 Decamethylcyclopentasiloxane (D5): CAS 541

- 02

- 6 Dodecamethylcyclohexasiloxane (D6): CAS 540

- 97

- 6

PURPOSE Determine representative concentrations of cVMS materials in the wastewater stream (influent, effluent, and slud ge) of a municipal wastewater treatment plant. The data will be used to estimate mass loadings of the cVMS materials to the local environment and to detect any temporal trends associated with the mass loading. It is anticipated that results from this pro ject will be used for environmental fate modelling and to provide a baseline for long

- term environmental monitoring
programs.

Guidelines and literature focusing on sampling of wastewater for pharmaceuticals and personal care products (PPCPs) are sparse an d rarely cited. The objectives and experimental designs discussed in the remainder of this document were based on information obtained from available guidance documents for monitoring of wastewater (U.S. EPA 1982; Clesceri et al 1998; Ort et al 2010).

OB JECTIVES The four basic factors which affect the quality of environmental data are sample collection, sample preservation, analyses, and recording. Improper actions in any one area may result in poor data from which poor judgments are certain. The object ives for the project were specifically defined to address these factors and were structured to transition three defined categories of wastewater studies

-- (i) method development, (ii) demonstrating presence or absence, and (iii) quantification of fate. Spe cific objectives for the project are to:

i. Phase 1: Method Development

Develop loss

- free methods for collection, storage, and analysis of cVMS materials in wastewater samples (influent, effluent, and sludge). Develop robust experimental designs that are ap propriate for collection of samples from municipal wastewater treatment plants that may exhibit unique transport characteristics, highly dynamic flows, and variable loads of the cVMS materials. Define and validate holding times that identify the maximum ti me allowed from the moment a sample is collected to when analysis of that sample must begin.

ii. Phase 2: Demonstrating Presence or Absence:

Implement results from Phase 1 and modify project duration and experimental designs as needed to maintain or improve upon the defined project objectives, or to account for unforeseen sample variability in terms of time and concentration. Determine representative concentrations of cVMS materials in the wastewater stream (influent, effluent, and sludge) of a municipal wast ewater treatment plant.

### 19 Determine the overall removal efficiency of cVMS materials in wastewater during the treatment process. Determine diurnal cycle (i.e., short

- term variability) of cVMS concentrations in the wastewater stream.

iii. Phase 3: Quantification of Fate:

Implement results from Phase 2 and modify project duration and experimental designs as needed to maintain or improve upon the defined project objectives, or to account for unforeseen sample variability in terms of time and concentration. Determin e seasonal and annual patterns or temporal trends associated with concentrations of cVMS materials in the wastewater stream (influent, effluent, and sludge). Estimate mass loadings of cVMS materials to the local environment. Determine any temporal trends a ssociated with mass loadings of cVMS materials to the local environment.

Project Duration The expected minimum project duration is 1.5 years. Project duration was estimated based on the assumption that 0.5 years would be required for method development (i.e., Phase 1 of the project) and that a minimum of 1 year of sample collection would be required to detect seasonal trends (i.e., Phase 3 of the project). Project duration and experimental design will be re

- evaluated after each phase of the project and modified as needed to maintain or improve upon the defined project objectives, or to account for unforeseen sample variability in terms of time and concentration. Modifications may include revised estimates of the number of samples to be collected, the fr equency of collection, and the desired power to detected changes over the duration of the project.

Experimental Design

– Phase 1 (Method Development) The four basic factors that affect the quality of environmental data are sample collection, sample
preser vation, analyses, and recording. For this project, it is expected that most of the effort associated with method development will be allocated to development of procedures for sample collection and preservation. It is anticipated that analytical methods and quality assurance and control (QA/QC) procedures developed for other matrices (e.g., water, sediment, soil, and biological tissues) may be directly applied and validated for analysis of cVMS materials in wastewater matrices (i.e., influent, effluent, a nd sludge).

The analytical results for a sample are only as accurate as the quality of the sample itself. If the sample was improperly collected, the uncertainty associated with the analytical results will be high regardless of how accurate the analytica l procedures are. Several studies (reviewed by Ort et al, 2010) have demonstrated that the uncertainty associated with sampling wastewater can be very significant, and that insufficient emphasis is placed on development of methods that are appropriate for sample collection. As a result, the importance of short

- term variations are ignored and inappropriate sampling procedures are employed that lead to over interpretation of the measured data and ultimately, wrong conclusions. Inappropriate sampling repres ents the major source of inaccuracy for monitoring studies on personal care products in wastewaters. However, sampling errors and sampling biases can be greatly reduced or eliminated by choosing an appropriate sampling mode and frequency. In addition, pr oper sampling maximizes the value of measured data for experimental assessment of fate in wastewaters and is required for formulation and validation of fate models.

It is anticipated that a combination of various sample types will be collected over the du ration of the proposed project. As such, sample collection methods appropriate for cVMS materials should be developed for each sample type. Wastewater sampling is generally performed by one of two methods; grab sampling or composite sampling. Grab sampl ing consists of collecting a discrete sample at a specific point of time. As such, a grab sample reflects conditions only at the point in time that the sample was collected, and then only if the sample was properly collected. Composite sampling consists of a collection of numerous individual discrete samples taken at regular intervals over a

### 20

period of time that are then pooled into a single composite sample. Sample to be collected may include any of the following sample types:

Grab Samples : A grab samp le can only represent the composition of its source at the time and place of collection, and should not exceed a sampling time of 15 minutes. Because of widely varying flow and intermittent pulses of materials to the wastewater stream, a grab sample taken at one specific time will not be representative of conditions throughout the daily flow pattern. By definition, a single grab sample cannot be used to evaluate the long

- term conditions of a wastewater stream. Grab samples must be collected at locations where the wastewater stream is well mixed and has sufficient velocity to keep the solids in suspension, while avoiding locations where the settling of solids occurs or floating debris is present.

The collection of a grab sample is appropriate when it is d esired to:

o Characterize water quality at a particular time.

o Provide information about minimum and maximum concentrations.

o Allow collection of variable sample volumes.

o Corroborate composite samples.

Grab samples should be used when:

o The wastewater stream d oes not flow continuously.

o The characteristics of the wastewater stream are relatively constant.

o The parameters to be analyzed are likely to deteriorate rapidly or change with storage (for example, volatile organic compounds (e.g., VOCs).

o Information on ma ximum, minimum, or variability is desired.

o The history of water quality is to be established based on relatively short time intervals.

o The spatial variability for a parameter is to be determined.

Composite Samples : Individual samples that are taken and d eposited in the same collection bottle. There are two methods that are most common to collecting composite samples. Time

proportional samples are taken at set increments of time. Flow

- proportional samples are taken when a measured volume of water flows over the sensor of a flow meter. The preferred method is flow

- proportional sampling, which gives the most representative sample. Without appropriate modifications, composite sampling is not valid for collection of samples when the parameter to be measu red deteriorates rapidly (e.g., VOCs).

Composite samples should be used when:

o Determining average concentrations.

o Calculating mass/unit time loadings.

Volatile Organic Compound (VOCs) Grab Samples : These grab samples are collected in clean glass vials, usually containing a preservative, with Teflon

- lined caps having a flexible septum. There must not be any air bubbles in the vial, so the cap must not have an air space under it. The vial must be filled until the liquid crowns and the cap screwed until t he septum bulges.

Grab Composite Samples : This procedure is not commonly used but is useful for such parameters as total oil and grease released to the sewer system in a 24

- hour period. Individual samples are grabbed in the field and then composited in the lab or in the field for analysis. A variation of this sample type is the flow

- proportional grab composite sample. Samples are grabbed in the usual manner but the flow level is recorded at the time of sampling. After all the predetermined times of s ampling are done the composite is made with portions of each grab sample according to the amount of flow at the time of sampling.

### 21

Discrete Samples : Grab samples collected and used to monitor characteristics of the wastewater flow at certain times of the day, for certain parameters, such as during high or low pH, or during high or low flow events. Samples are taken in individual bottles at the time of the event and each sample is analyzed. This method can also be used for flow

- proportional composite samp les if the flow rate is recorded at the time of sampling.

Samples cannot be analyzed for cVMS concentrations on

- site immediately after collection. Depending on location of the study area, the samples will be shipped to Evonik Goldschmidt GmbH (Essen, Ger many) or to Dow Corning Corporation (Auburn, Michigan, USA) for analysis. Therefore, in conjunction with the development of methods for sample collection, appropriate methods for sample preservation must also be developed and validated for each sample typ e. For the cVMS materials, the purpose of sample preservation should be to prevent volatile loss and to minimize degradation until the sample can be analyzed.

Holding times for each sample type will also need to be identified and validated, based on the method of preservation. The holding time is the maximum time allowed from the moment a sample is collected to when analysis on that sample must begin. All samples must be analyzed within the validated holding time for the analytical results to be conside red acceptable. A validated holding time in conjunction with proper sample preservation is required to retain the integrity of the sample and ensure that accurate results are obtained. It is anticipated that a minimum holding time of 72 hours will be req uired for this project.

Experimental Design

– Phase 2 (Demonstrating Presence or Absence) The experimental design for Phase 2 of the project will be developed based on the results obtained from Phase 1 of the project. The focus of the Phase 2 experimenta l design will be on identification of sample collection intervals and sample collection frequency. Inappropriate sampling (in terms of collection procedures, intervals, and frequency) represents the major source of inaccuracy for monitoring studies on pe rsonal care products in wastewaters (Ort et al, 2010).

Short

- term variations in the composition of the wastewater stream resulting from widely varying flow and intermittent pulses of materials may confound interpretation of the measured data and lead to i ncorrect and misleading conclusions. Consequently, sampling intervals and frequency are crucial parameters for monitoring compounds in the wastewater stream that are released through the use and disposal of personal care products. Sampling intervals shou ld be selected on the basis of the expected frequency of changes, which may vary from as little as 5 min to as long as 1 h or more. Because of analytical costs, high sampling frequencies may become prohibitive. However, low sampling frequencies increase random uncertainty, which leads to systematic overestimations or underestimations of concentrations and loads in the wastewater stream. Sampling intervals and frequencies for monitoring a wastewater stream can only be determined after detailed preliminary work that involves a high sampling frequency is completed.

It is anticipated that large diurnal variations will be observed for concentrations of cVMS materials in the wastewater stream that are associated with intermittent “wastewater pulses” or dischar ges to the sewer. At the source, these pulses may last a few seconds (toilet flushes) to minutes (emptying a bathtub or pumping water out of washing machines, etc.). Depending on hydraulic conditions and flow distances, the number of wastewater pulses th at contain the cVMS materials will depend on the number of people in the catchment and the frequency that personal care products containing the cVMS materials are used. If only a few such wastewater pulses are expected over the course of a day a high samp ling frequency is necessary to capture these pulses.

It is assumed that major pulses of cVMS materials in the wastewater stream will occur twice a day, with much smaller pulses intermittently occurring in between the major pulses. Major pulses of cVMS ma terials are anticipated to occur during weekday mornings in association with the morning activities of individuals preparing for the workday. Major pulses of cVMS materials, but of smaller magnitude, are also anticipated to occur during the early evening in association with recreational

### 22

activities after the workday. It is also anticipated that the major pulses of cVMS materials may be associated with seasonal cycles that are primarily related to temperature and daylight hours.

Based on these assumption s it is anticipated that discrete grab samples collected at 1

- hour intervals over a 24

- hour period will initially be used to identify presence of cVMS materials in the wastewater stream. Ideally, this sampling routine should be conducted over a number of consecutive days rather than non

- consecutive days (Ort et al 2010). Results from the analysis of these samples may also be used to identify any diurnal cycles that may influence cVMS concentrations in the waste stream. Repeated sampling over a longer per iod would be required to determine weekly and seasonal variations. Results may also be used to estimate the number of samples and frequency of collection for Phase 3 of the project.

Experimental Design

– Phase 3 (Quantification of Fate) The experimental design for Phase 3 of the project will be developed based on the results obtained from Phase 2 of the project. The focus of the Phase 3 experimental design will be identification of seasonal and annual patterns or temporal trends associated with concentra tions of cVMS materials in the wastewater stream. Results from Phase 3 of the project will be used to estimate mass loadings of cVMS materials to the local environment.

It is anticipated that the flow rate of the wastewater stream will vary significantly over time during the course of a day when sampling takes place. Therefore, flow

- proportional composite sampling would be used if appropriate methods can be developed (i.e., Phase 1). Flow

- proportional composite sampling is the only sampling mode that co rrectly weights the individual subsamples, which are combined to form a composite sample, and analyzed to obtain an average concentration. If appropriate methods cannot be developed for composite sampling, then flow

- proportional grab sampling may be used. If concentrations of the cVMS materials in the wastewater stream were relatively constant over the course of a day (as demonstrated from the Phase 2 results), a grab sample would be equal to the average concentration in a flow

- proportional composite samp le. However, to demonstrate that concentrations of the cVMS materials were relatively constant would require a large number of grab samples to be collected at very short intervals (1

- 5 min over several hours) during the Phase 2 program.

Sample Matrices t o be Collected Wastewater influent, effluent, and sludge (collectively referred to as the wastewater stream) of a municipal wastewater treatment plant will be collected and analyzed for this project. Sludge is the mud

- like residual material generated duri ng the wastewater treatment process that consists of organic and inorganic solids present in raw sewage. Sludge should not be confused with bio

- solids, which are processed and treated sludges. Samples will be collected from locations specified by the fac ility operator or by individuals having experience with the facility being monitored.

Frequency of Sample Collection and Sample Collection Times The frequency of sample collection will be based on results from Phase 1 and Phase 2 of the project.

Number o f Samples The number of sample to be collected will be determined by the type of samples collected (grab vs composite), by the sampling mode used to collect the samples (continuous vs discrete), and on results from Phase 1 and Phase 2 of the project.

Anal ysis of Samples Analytical methods and quality assurance and control (QA/QC) procedures will developed as part of Phase 1 of the project. It is anticipated that analytical methods and QA/QC procedures developed for other matrices (e.g., water, sediment, soil, and biological tissues) may be directly applied and validated for analysis of cVMS materials in the wastewater matrices (i.e., influent, effluent, and sludge).

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Statistical Analysis Data will be specifically analyzed to detect changes or trends in cV MS concentrations that may occur over the short

- term and long

- term for the duration of the program. Temporal trends will be evaluated using procedures described in various guidance documents (Gilbert 1987; UNEP 2004, 2007; OSPAR 2008; EU 2010).

Contract To be developed on a semi

- annual basis, and re

- evaluated after completion of Phase 1 and Phase 2 of the project.

Project Team Team Leader: To be identified. The Team Leader should work closely with the facility operator where the monitoring study is con ducted.

Sample Collection: To be identified. It is proposed that samples be collected by individuals having substantial experience with monitoring wastewater treatment facilities. The individuals should also be experienced with development of experimen tal designs for monitoring wastewater treatment facilities. An industry representative (e.g., the Team Leader) should participate on initial sample collections.

Sample Analysis: To be identified. It is proposed that cVMS analyses be conducted by one of the following laboratories:

o Dow Corning Corporation, Auburn, Michigan, USA

o Evonik Goldschmidt GmbH, Essen, Germany

Statistical Analysis and Database Management: To be identified.

References

Clesceri, L.S.; Greenberg, A.E.; Eaton, A D. 1998. Standar d Methods for the Examination of Water and Wastewater ; American Public Health Association: Washington, DC; Vol. 20, ISBN 0875532357.

EU. 2010. Guidance Document No: 25 Guidance on chemical monitoring of sediment and biota. Technical Report 2010.3991

G ilbert, R.O. 1987. Statistical Methods for Environmental Pollution Monitoring. Van Nostrand Reinhold, New York, NY. ISBN 0

- 442

- 23050

- 8.

Ort , C.; Lawrence, M.G.; Rieckermann, J.; Joss, A. 2010. Sampling for pharmaceuticals and personal ccare product s (PPCPs) and illicit drugs in wastyewater systems: are your conclusions valid? A critical review. Environmental Science and Technology 44: 6024

– 6035.

OSPAR. 2008. CEMP Assessment Manual. Co

- ordinated Environmental Monitoring Programme Assessment Ma nual for contaminants in sediment and biota. OSPAR Commission Publication Number No. 379/2008.

UNEP. 2004. Guidance for a Global Monitoring Programme for Persistent Organic Pollutants, 1 st Edition. United Nations Environment Programme. UNEP Chemicals , Geneva, Switzerland.

UNEP. 2007. Draft Guidance on the Global Monitoring Plan for Persistent Organic Pollutants. United Nations Environment Programme. UNEP/POPS/COP.3/INF/14.

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U.S. EPA 1982. Handbook for Sampling and Sample Preservation of Water a nd Wastewater ; 600/4

- 82

- 029; Environmental Monitoring and Support Laboratory.