Testing Standard for the Collection, Sample Handling, Transfer and Analysis of Influent and Effluent

Samples for D4

August 16, 2013

Silicones Environmental, Health and Safety Center 1

Synopsis: 

This testing standard provides guidelines for the collection and analysis of environmental samples for D4

in support of a Testing Program detailed in this Enforceable Consent Agreement (ECA). The purpose of 

the Testing Program is to “...develop environmental testing data for D4 in the vicinity of manufacturing 

and processing (including product formulating) sites and POTWs in the U.S. so that exposures and risks 

to sediment and aquatic organisms can be assessed.” This standard cites the requirements for the 

sampling of influent and effluent as described in Section VII of the ECA and Appendices 1, 2, and 3, and 

identifies specific guidelines and references that will be used in the preparation of site-specific Sampling 

Plans for the collection of influent and effluent samples, which will be included in a Study Plan for the 

Testing Program. 

Requirements for sampling influents and effluents: 

Per Appendix 1, Environmental Testing at D4 Direct Discharge Sites:

“With respect to the wastewater treatment effluent (outfall), sampling is to occur at the compliance 

point in accordance with the facility’s discharge permit (specific IDs will be provided in the Study 

Plan). In addition to the concentration of D4 in the effluent, total effluent flow rate shall be 

recorded and reported.”

Per Appendix 2, Environmental Testing for Indirect Discharge Sites and Appendix 3, POTWs Not 

Receiving D4 from Industrial Facilities: 

“The concentration of D4 in the influent at the headworks of the POTW (i.e., regulated compliance 

point per the POTW’s discharge permit) shall be measured along with the total influent flow rate. “

“The concentration of D4 in the effluent at the regulated compliance point per the POTW’s 

discharge permit shall be measured along with the total effluent flow rate. ”

Establishment of the location for influent sample collection: 

At each POTW, influent samples should be collected at the headworks of the POTW. The exact location 

for collection of an influent sample will be facility-specific and specified in the POTW’s discharge permit. 

Each site-specific Sampling Plan shall include a description of the portions of the POTW where influent is 

contained, including a basis for determining the appropriate sampling location. 
Testing Standard for the Collection, Sample Handling, Transfer and Analysis of Influent and Effluent

Samples for D4

August 16, 2013

Silicones Environmental, Health and Safety Center 2

Establishment of the location for effluent sample collection: 

At each site (i.e., POTW or direct discharge facility), effluent samples shall be collected at the regulated 

compliance point per the POTW’s discharge permit. Each site-specific Sampling Plan shall include a 

complete description of the effluent discharge portions of the POTW, including the established sampling 

location. 

Special considerations for the sampling and handling of influent and effluent samples for D4: 

Cyclic volatile methyl siloxane (cVMS) materials, such as D4, are widely used in consumer products and 

may be present in personal care products, commercial products, and lubricants, and may be 

components of antifoams widely used in waste water treatment plants. To avoid the significant 

potential for sample contamination during collection, processing, storage, and analysis for D4 in 

environmental matrices, appropriate quality control measures should be implemented to the extent 

possible in the field. For example, field crew shall refrain from using any personal care products (e.g. , 

sun-block, sun-screen, hand lotion, antiperspirants, etc.) while preparing for or conducting any field 

activities or during collection of samples to minimize the possibility for contamination with cVMS 

materials. Appropriate requirements for collecting and handling influent and effluent samples that 

comply with the special considerations related to D4 shall be specified in the site-specific Sampling Plan. 

Collection of influent samples for analysis of D4: 

At each site, the time that influent samples shall be collected will be chosen in consultation with the 

POTW operator and should reflect normal operations and conditions. The time of collection of the 

influent should correspond with the time of collection of effluent, surface water, and sediment samples 

at each facility. 

During each sampling event at a given site, four grab samples of influent (two for analysis of D4 and two 

for use in preparing field spikes) shall be prepared once during the day of field sampling. Influent flow 

rate shall be measured at the time of sample collection. Specific requirements for collection of grab

samples, including sample bottle preparation, sample preservation, shipping and storage of the influent 

samples shall be specified in the Study Plan and will be based on USEPA 2013 (Wastewater Sampling, 

Section 4, Sample Types and Section 6, Manual Sampling), USEPA 2004 (NPDES Compliance Inspection 

Manual, Section 5B, Sampling Procedures and Techniques), Knoerr 2013 (Sample Collection section, p. 

8), and APHA 1999 (Part 1060 Collection and Preservation of Samples) (see Annexes 1 to 4). The sample 

containers will contain LDPE that will adsorb any dissolved D4 and prevent volatilization of any D4 

present. In the laboratory, the entire contents of the sample container will be extracted including LDPE 

sorbent and any solids present.
Testing Standard for the Collection, Sample Handling, Transfer and Analysis of Influent and Effluent

Samples for D4

August 16, 2013

Silicones Environmental, Health and Safety Center 3

Also during each sampling event at a given site, two field blanks and two field spiked-samples will be 

prepared as described below. Serving as field blanks, two samples of laboratory-grade water having very 

low or non-detectable concentrations of cVMS materials shall be taken into the field and processed like 

an influent sample. The laboratory-grade water samples will be used to evaluate contamination bias 

resulting from collection, processing, transport, storage, and analysis of samples. Two of the influent 

grab samples shall be spiked in the field with a known amount of D4. The field-spiked influent samples

will be used to evaluate loss of cVMS during collection, processing, transport, storage, and analysis of 

samples, and to evaluate analytical bias. Specific requirements for preparation, processing, and analysis 

of the field blanks and field spiked samples shall be detailed in the Study Plan and the Quality Assurance 

Project Plan (QAPP).

During the time of collection of the influent samples, two separate sample bottles shall be prepared for 

analysis of total organic carbon (TOC). The TOC analyses shall follow procedures equivalent to AHPA 

1999 Part 5310, Total Organic Carbon (TOC) (see Annex 5).

Collection of effluent samples for analysis of D4: 

At each site, the time that effluent samples shall be collected will be chosen in consultation with the 

POTW operator and should reflect normal operations and conditions. The time of collection of the 

effluent should correspond with the time of collection of influent, surface water, and sediment samples 

at each facility. If any facility has more than one outfall, then each outfall shall be sampled. 

During each sampling event at a given site, four grab samples of effluent (two for analysis of D4 and two 

for use in preparing field spikes) shall be prepared once during the day of field sampling. Effluent flow 

rate shall be measured at the time of sample collection. Specific requirements for collection of grab

samples, sample bottle preparation, sample preservation, shipping and storage of the effluent samples 

shall be based on USEPA 2013 (Wastewater Sampling, Section 4 Sample Types and Section 6 Manual 

Sampling), USEPA 2004 (NPDES Compliance Inspection Manual, Section 5B, Sampling Procedures and 

Techniques), Knoerr 2013 (Sample Collection section, p. 8), and APHA 1999 (Part 1060 Collection and 

Preservation of Samples) (see Annexes 1 to 4). The composite sample containers will contain LDPE that 

will adsorb any dissolved D4 and prevent volatilization of any D4 present. In the laboratory, the entire 

contents of the composited sample container will be extracted including LDPE sorbent and any solids 

present.

Also during each sampling event at a given site, two field blanks and two field spiked-samples will be 

prepared as described below. Serving as field blanks, two samples of laboratory-grade water having very 

low or non-detectable concentrations of cVMS materials shall be taken into the field and processed like 

an effluent sample. The laboratory-grade water samples will be used to evaluate contamination bias 

resulting from collection, processing, transport, storage, and analysis of samples. Two of the effluent 

composite samples shall be spiked in the field with a known amount of D4. The field-spiked effluent
Testing Standard for the Collection, Sample Handling, Transfer and Analysis of Influent and Effluent

Samples for D4

August 16, 2013

Silicones Environmental, Health and Safety Center 4

samples will be used to evaluate loss of cVMS during collection, processing, transport, storage, and 

analysis of samples, and to evaluate analytical bias. Specific requirements for preparation, processing, 

and analysis of the field blanks and field spiked samples shall be detailed in the Study Plan and the 

Quality Assurance Project Plan (QAPP).

During the time of collection of the effluent samples, two separate sample bottles shall be prepared for 

analysis of total organic carbon (TOC). The TOC analyses shall follow procedures equivalent to AHPA 

1999 Part 5310, Total Organic Carbon (TOC) (see Annex 5).

Analysis of influent and effluent samples for D4: 

All influent and effluent samples shall be analyzed for D4 at a commercial testing laboratory. The 

commercial laboratory will validate a method for the analysis of D4 in the influent and effluent samples 

that shall be adapted from the analytical method specified in Knoerr (2013) (see Annex 3). The specific 

level of detection shall be based on the Practical Quantitation Limit (PQL) (USEPA 1985 and APHA 1999, 

see Annexes 6 and 7). The PQL is in turn based on the matrix-specific Method Detection Limit (MDL). 

These terms are defined as follows:

Practical Quantitation Limit (PQL): Minimum concentration of a constituent in a sample that can be 

measured among laboratories within specified limits during routine laboratory operations. The PQL 

is proposed because different laboratories may produce different MDLs even though they may be 

using the same analytical procedures, instruments and sample matrices. The PQL is typically defined 

as 5 to 10 times the MDL.

Method Detection Limit (MDL): Concentration of a constituent in a sample, that when processed 

through the complete analytical method, can be measured and reported with 99% probability that it 

is greater than 0 or statistically different from a blank. The MDL is a measure of a methods ability to 

quantify a constituent in a sample matrix.

During method development, the Knoerr (2013) analytical method for D4 achieved an average matrixspecific Method Detection Limit (MDL) of 0.037 μg/L. For use at the commercial laboratory, the targeted 

PQL for D4 in aqueous matrices shall be calculated as 5 times the Knoerr (2013) MDL. Therefore, when 

rounded to one significant figure, the PQL for D4 in aqueous matrices shall be 0.20 μg/L.

Compliance with Good Laboratory Practices:

Sample collection, handling, and transfer activities should be conducted in accordance with GLP 

requirements, as applicable. The analytical laboratory conducting the analyses of D4 shall be a GLP 

certified facility and all analytical work shall be conducted in accordance with GLP requirements. The 
Testing Standard for the Collection, Sample Handling, Transfer and Analysis of Influent and Effluent

Samples for D4

August 16, 2013

Silicones Environmental, Health and Safety Center 5

QAPP shall be prepared according to USEPA guidance and shall include information on all field and 

laboratory quality control measures to be followed. 

Annex 1 – USEPA 2013. Wastewater Sampling, Section 4 Sample Types, Section 6 Manual Sampling. 

Operating Procedure No. SESDPROC-306-R3. US Environmental Protection Agency, Region 4. February, 

2013.

Annex 2 – USEPA 2004. NPDES Compliance Inspection Manual, Section 5B, Sampling Procedures and 

Techniques . EPA 305-X-04-001, US Environmental Protection Agency, July, 2004.

Annex 3 – Knoerr SM. 2013. NON REGULATED STUDY: Development of Collection, Storage and Analysis 

Procedures for the Quantification of D4, D5, and D6 in Waste Water Treatment Plant Effluent and 

Surface Water Samples. Dow Corning Corporation, HES Study No. 11936-108, April 18, 2013.

Annex 4 – APHA 1999. Standard Methods for the Examination of Water and Wastewater, Part 1060, 

Collection and Preservation of Samples. American Public Health Association, American Water Works 

Association, Water Environment Federation.

Annex 5 – AHPA 1999. Standard Methods for the Examination of Water and Wastewater, Part 5310 C. 

Total Organic Carbon (TOC). American Public Health Association, American Water Works Association, 

Water Environment Federation.

Annex 6 – USEPA 1985. National primary water drinking standards: synthetic organics, inorganics, and 

bacteriologicals. 40 CFR Part 141; Federal Register 50: No. 219, November 13. 1985.

Annex 7 – APHA 1999. Standard Methods for the Examination of Water and Wastewater, Part 1030 C. 

Method Detection Level, Section 2 Determining Detection Levels. American Public Health Association, 

American Water Works Association, Water Environment Federation.