Issuance Date: __June 5, 2002__________

Effective Date: __July 5, 2002__________

Expiration Date: _July 5, 2007 _________

FISHERY RESOURCE MANAGEMENT 

NATIONAL POLLUTANT DISCHARGE ELIMINATION SYSTEM 

WASTE DISCHARGE INDIVIDUAL PERMIT No. WA0041009 

WASHINGTON DEPARTMENT OF FISH AND WILDLIFE 

State of Washington 

DEPARTMENT OF ECOLOGY 

Olympia, Washington 98504-7600 

In compliance with the provisions of 

The State of Washington Water Pollution Control Law 

Chapter 90.48 Revised Code of Washington 

and 

The Federal Water Pollution Control Act 

(The Clean Water Act) 

Title 33 United States Code, Section 1251 et seq. 

Washington Department of Fish and Wildlife 

600 Capitol Way N. 

Olympia, WA 98501-1091 

is authorized to discharge in accordance with the special and general conditions which follow. 

Megan White, P.E., Manager 

Water Quality Program 

Department of Ecology 
 Page 2 of 29 

Permit No. WA0041009 

TABLE OF CONTENTS 

Page

SUMMARY OF PERMIT REPORT SUBMITTALS.....................................................................4 

SPECIAL CONDITIONS

S1. DISCHARGE LIMITATIONS............................................................................................5 

A. Water Quality Standards 

B. Temporary Water Quality Modification 

S2. MONITORING REQUIREMENTS....................................................................................6 

A. Monitoring 

B. Monitoring Schedule 

C. Sampling and Analytical Procedures 

D. Laboratory Accreditation 

S3. REPORTING AND RECORDKEEPING REQUIREMENTS ...........................................8 

A. Reporting 

B. Records Retention 

C. Recording of Results 

D. Additional Monitoring by the Permittee 

E. Noncompliance Notification 

S4. ANNUAL SEPA PROCESS .............................................................................................11 

S5. ANTIMYCIN SEPA REVIEW .........................................................................................11 

S6. BEST MANAGEMENT PRACTICES .............................................................................11 

S7. SPILL PREVENTION AND RESPONSE ........................................................................12 

PUBLIC NOTICE PROCEDURES

P1. RESIDENTIAL AND BUSINESS NOTICE PROCEDURES .........................................13 

P2. LEGAL NOTIFICATION PROCEDURES ......................................................................13 

P3. POSTING PROCEDURES................................................................................................14 

GENERAL CONDITIONS ...........................................................................................................15

G1. SIGNATORY REQUIREMENTS.....................................................................................15 

G2. RIGHT OF INSPECTION AND ENTRY.........................................................................16 

G3. PERMIT ACTIONS...........................................................................................................16

G4. REPORTING A CAUSE FOR MODIFICATION............................................................17 

G5. PLAN REVIEW REQUIRED ...........................................................................................18 

G6. COMPLIANCE WITH OTHER LAWS AND STATUTES.............................................18 
 Page 3 of 29 

Permit No. WA0041009 

G7. DUTY TO REAPPLY .......................................................................................................18 

G8. TRANSFER OF THIS PERMIT .......................................................................................18 

G9. REDUCED PRODUCTION FOR COMPLIANCE ..........................................................19 

G10. REMOVED SUBSTANCES .............................................................................................19 

G11. DUTY TO PROVIDE INFORMATION...........................................................................19 

G12. OTHER REQUIREMENTS OF 40 CFR...........................................................................19 

G13. ADDITIONAL MONITORING........................................................................................19 

G14. PAYMENT OF FEES........................................................................................................19 

G15. PENALTIES FOR VIOLATING PERMIT CONDITIONS .............................................19 

G16. UPSET ...............................................................................................................................20 

G17. PROPERTY RIGHTS........................................................................................................20 

G18. DUTY TO COMPLY ........................................................................................................20 

G19. TOXIC POLLUTANTS.....................................................................................................20 

G20. PENALTIES FOR TAMPERING .....................................................................................21 

G21. REPORTING PLANNED CHANGES..............................................................................21 

G22. REPORTING ANTICIPATED NON-COMPLIANCE.....................................................21 

G23. REPORTING OTHER INFORMATION..........................................................................21 

G24. REPORTING REQUIREMENTS APPLICABLE TO EXISTING MANUFACTURING, 

COMMERCIAL, MINING, AND SILVICULTURAL DISCHARGERS....................................21 

G25. COMPLIANCE SCHEDULES .........................................................................................22 

APPENDIX A RAPID METHOD FOR MEASURING ROTENONE IN WATER AT 

PISCICIDAL CONCENTRATIONS ............................................................................................23 

APPENDIX B SAMPLING PROTOCOLS FOR ZOOPLANKTON..........................................28 
 Page 4 of 29 

Permit No. WA0041009 

SUMMARY OF PERMIT REPORT SUBMITTALS 

Refer to the Special and General Conditions of this permit for additional submittal requirements. 

Permit 

Section 

Submittal Frequency First Submittal Date 

S3.A Post-Treatment Report / Discharge 

Monitoring Report 

Annually June 1, 2003 

S3.E Noncompliance Notification As necessary 

S5. Antimycin SEPA Review One time June 1, 2005 

S7. Spill Prevention and Response Plan One time Prior to first treatment 

G1. Notice of Change in Authorization As necessary 

G4. Permit Application for Substantive Changes 

to the Discharge 

As necessary 

G7. Application for Permit Renewal 1/permit cycle January 5, 2007 

G8 Notice of Permit Transfer As necessary 

G21 Notice of Planned Changes As necessary 

G22. Reporting Anticipated Non-compliance As necessary 
 Page 5 of 29 

Permit No. WA0041009 

SPECIAL CONDITIONS 

S1. DISCHARGE LIMITATIONS 

A. Water Quality Standards

All discharges and activities authorized by this permit shall be consistent with the terms 

and conditions of this permit. 

Use of any liquid or powder rotenone formulation shall not result in an exceedance of 

water quality standards as specified in WAC 173-201A. 

B. Temporary Water Quality Modification

The application of chemicals listed in this permit to perform fish pest control activities is allowed 

so long as the conditions of this permit are satisfied and the transitory water quality impact is 

limited to the minimum time necessary to accomplish the desired pest control objectives. 

This temporary water quality modification is allowed throughout the permit term, but its effect 

shall be temporary in a specific location, though locations where it is in effect may be widespread 

throughout the state, anywhere lakes or streams are subject to fish control activities by WDFW. 

C. Permitted Chemicals

Rotenone is the only chemical permitted for use as a fish toxicant under this individual 

permit. The rotenone product used must be licensed for use as a fish toxicant in the State 

of Washington at the time of treatment. 

The use of liquid rotenone is only authorized for spot applications in areas that are not 

practicably accessible by boat. Open water areas that are accessible by boat will be 

treated with powdered rotenone that is mixed with water and applied as a slurry, as 

described in S.6. Best Management Practices. 

Potassium permanganate is the only chemical permitted to neutralize rotenone treated 

waters when necessary to prevent damage to non-targeted organisms and maintain water 

quality outside of the area intended for rotenone treatment. 

Other pesticides may be applied on a limited basis in the context of a research and 

development effort under the jurisdiction of the Washington State Department of 

Agriculture (WSDA) through the issuance of a Washington State Experimental Use 

Permit. Limited amounts of an experimental use pesticide may only be distributed or 

used for testing purposes after a written permit has been obtained from WSDA for 

purposes which include gathering data in support of registration under the Federal 

Insecticide Fungicide and Rodenticide Act (FIFRA) Section (3) or Section 24(c). 

All other conditions of this permit apply as to appropriate monitoring and public 

notification procedures. 
 Page 6 of 29 

Permit No. WA0041009 

S2. MONITORING REQUIREMENTS 

A. Monitoring 

The WDFW shall conduct monitoring on each water body treated with aquatic pesticides 

to determine the extent and duration of the short-term water quality reduction resulting 

from rotenone applications. 

B. Monitoring Schedule

TABLE 1. MONITORING – ROTENONE TREATED WATERS

Parameters Units Minimum Sampling Frequency Sample Type 

Rotenone Toxicity - 

Trout Bioassay: 48-hr 

live box test (5 trout); 

100% survival of 

rainbow trout 

Number of 

days until 

100% survival 

Once post-treatment, approximately 

3-8 weeks after treatment 

Observation 

(no lab 

accreditation 

required) 

*If liquid rotenone is 

used: VOC, semi-VOC, 

plus any other inert 

ingredients listed on 

MSDS 1

μg/L 1. 24 hours after treatment, and 2. 

four weeks after treatment 

Grab 

pH Standard Once pretreatment Grab 

temperature  ̊F Once pretreatment Grab 

Alkalinity 2 mg/L CaCO3 Once pretreatment 2 Grab 

Organic demand 2, 3 Standard 3 Once pretreatment 2 Grab 

Zooplankton sampling See below 4 1. Pre-treatment, 2. Six months after 

treatment, and 3. One year after 

treatment 

Composite 4 

1

 If liquid rotenone formulation is utilized, test for the following parameters: VOC (EPA method 8310) and semiVOC (EPA method 502.2). Also test for any other inert ingredients listed on MSDS (i.e. the MSDS for Prentox ®

PrenfishTM Toxicant lists naphthalene; 1,2,4-trimethylbenzene; and acetone).

2 Only if neutralization of rotenone with potassium permanganate is required. 

3 Use the guidelines provided in Engstrom-Heg (1971) to determine organic demand for KMnO4.

4 Lakes only. Zooplankton sampling protocols set forth on Page 4-5 of “Water Quality Assessments of Selected 

Lakes within Washington State - 1998” Department of Ecology, December 2000, Publication No. 00-03-039 

(Appendix B).
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Permit No. WA0041009 

TABLE 2. MONITORING – RECEIVING WATERS DOWNSTREAM OF TREATED 

WATERS AND NEUTRALIZATION ZONE

Parameters Units Minimum Sampling Frequency Sample Type 

Rotenone Toxicity - 

Trout Bioassay: 48-hour 

live box test (5 trout) 

% survival 1. Begin test at time of treatment and 

2. Repeat at one week intervals until 

upstream treated water is detoxified 

per upstream bioassay. 

Observation 

(no lab 

accreditation 

required) 

Rotenone 1 mg/L Once 24 hours following treatment Grab 

*If liquid rotenone is 

used: VOC, semi-VOC, 

plus any other inerts 2

μg/L 1. 24 hours after treatment, and 2. 

four weeks after treatment 

Grab 

pH Standard Once pretreatment Grab 

temperature  ̊F Once pretreatment Grab 

Alkalinity 3 mg/L CaCO3 Once pretreatment 3 Grab 

Organic demand 3, 4 Standard 4 Once pretreatment 3, 4 Grab 

Zooplankton sampling See below 5 1. Pre-treatment, 2. Six months after 

treatment, and 3. One year after 

treatment 

Composite 5 

Macroinvertebrate 

monitoring (Only 

required for wadeable

streams) 

See below 6

 1. Pre-treatment, between August and 

September and 2. Post-treatment, 

approximately 1 year after treatment 

See below 6

1

 Analyze using methods set forth in Dawson et al. (1983); Appendix A

2

 If liquid rotenone formulation is utilized, test for the following parameters: VOC (EPA method 8310) and semiVOC (EPA method 502.2). Also test for any other inert ingredients listed on MSDS (i.e. the MSDS for Prentox ®

PrenfishTM Toxicant lists naphthalene; 1,2,4-trimethylbenzene; and acetone).

3 Only if neutralization of rotenone with potassium permanganate is required. 

4 Use the guidelines in Engstrom-Heg (1971) for measuring organic demand for KMnO4.

5 Lakes only. Zooplankton sampling protocols set forth on Page 4-5 of “Water Quality Assessments of Selected 

Lakes within Washington State - 1998” Department of Ecology, December 2000, Publication No. 00-03-039; 

Appendix B

6

 “Macroinvertebrate monitoring” includes gathering benthic invertebrate samples and summarizing the data using 

the benthic index of biotic integrity (B-IBI) and a ratio measure of the number of observed taxa divided by the 

number of expected taxa, the River Invertebrate Prediction and Classification System (RIVPACS). 

All bioassessment sampling and related habitat survey data, laboratory analysis, quality assurance, and data 

analysis shall follow the protocols in Benthic Macroinvertebrate Biological Monitoring Protocols for Rivers and 

Streams: 2001 Revision, Plotnikoff and Wiseman, August 2001 (http://www.ecy.wa.gov/biblio/0103028.html).
 Page 8 of 29 

Permit No. WA0041009 

C. Sampling and Analytical Procedures

Samples and measurements taken to meet the requirements of this permit shall be 

representative of the volume and nature of the monitored parameters, including 

representative sampling of any unusual discharge or discharge condition, including spills, 

upsets, and maintenance-related conditions affecting effluent quality. 

Sampling and analytical methods used to meet the monitoring requirements specified in 

this permit shall conform to the latest revision of the Guidelines Establishing Test 

Procedures for the Analysis of Pollutants contained in 40 CFR Part 136 or to the latest 

revision of Standard Methods for the Examination of Water and Wastewater (APHA), 

unless otherwise specified in this permit or approved in writing by the Department of 

Ecology (Department). 

D. Laboratory Accreditation

All monitoring data required by the Department shall be prepared by a laboratory 

registered or accredited under the provisions of, Accreditation of Environmental 

Laboratories, Chapter 173-50 WAC. Flow, temperature, settleable solids, conductivity, 

pH, and internal process control parameters are exempt from this requirement. 

Conductivity and pH shall be accredited if the laboratory must otherwise be registered or 

accredited. The Department exempts crops, soils, and hazardous waste data from this 

requirement pending accreditation of laboratories for analysis of these media. 

S3. REPORTING AND RECORDKEEPING REQUIREMENTS 

The Permittee shall monitor and report in accordance with the following conditions. The 

falsification of information submitted to the Department shall constitute a violation of the terms 

and conditions of this permit. 

A. Reporting

The first monitoring period begins on the effective date of the permit. Monitoring results 

shall be submitted annually. Monitoring data obtained during each monitoring period 

shall be summarized, reported, and submitted in an annual report form approved by the 

Department. 

Post Treatment Discharge Monitoring Report forms shall be received no later than June 

1, the year following each treatment. The report(s) shall be sent to Nancy C. Weller, 

Department of Ecology, Eastern Regional Office, N. 4601 Monroe, Spokane, WA 

99205-1295. 

Post Treatment Discharge Monitoring Reports shall contain the following information: 

1. Name of lake or stream 

2. County 
 Page 9 of 29 

Permit No. WA0041009 

3. Section, Township and Range 

4. Date(s) of treatment 

5. Purpose of treatment 

6. Name of licensed applicator 

7. Lake description: Surface acreage, number of acre-feet, maximum depth and 

average depth 

8. Stream description: Width, length, flow rate of stream/outlet (cu. ft. per sec.); 

Volume and weight of water treated (gallons, pounds) 

9. Name of fish toxicant product used 

10. Description of treatment method(s) 

11. Quantity of fish toxicant used (pounds and/or gallons) 

12. Concentration of active rotenone in formulated rotenone product (%) 

13. Concentration of active rotenone in water (ppm) 

14. Water conditions/quality (temperature, pH, hardness, alkalinity – and any 

other additional data collected) 

15. Detoxification of rotenone treated water (if required): Description of 

detoxification methods/equipment; potassium permanganate application rate 

(pounds per hour); flow rate of stream/outlet (cu. ft. per sec.); estimate of 

average concentration (ppm) 

16. Description of lake inlet(s)/outlet(s) and any temporary water control 

measures (if required) 

17. Period of toxicity (duration of water quality reduction) 

18. Eradicated fish species 

19. Results of pre- and post-treatment monitoring 

20. Impact on non-targeted organisms 

21. Brief description of treatment/detoxification and other comments 

22. A copy of the amended FSEIS for the lakes/streams treated during the 

reporting period including all SEPA comments, results and decisions. 

23. A list of the lakes/streams proposed for treatment during the upcoming year 
 Page 10 of 29 

Permit No. WA0041009 

B. Records Retention

The Permittee shall retain records of all monitoring information for a minimum of three 

(3) years. Such information shall include all calibration and maintenance records and all 

original recordings for continuous monitoring instrumentation, copies of all reports 

required by this permit, and records of all data used to complete the application for this 

permit. This period of retention shall be extended during the course of any unresolved 

litigation regarding the discharge of pollutants by the Permittee or when requested by the 

Director. 

C. Recording of Results

For each measurement or sample taken, the Permittee shall record the following 

information: (1) the date, exact place, method, and time of sampling or measurement; (2) 

the individual who performed the sampling or measurement; (3) the dates the analyses 

were performed; (4) the individual who performed the analyses; (5) the analytical 

techniques or methods used; and (6) the results of all analyses. 

D. Additional Monitoring by the Permittee

If the Permittee monitors any pollutant more frequently than required by this permit using 

test procedures specified by Condition S2. of this permit, then the results of this 

monitoring shall be included in the calculation and reporting of the data submitted in the 

Permittee's Post Treatment Discharge Monitoring Report. 

E. Noncompliance Notification

In the event the Permittee is unable to comply with any of the terms and conditions of 

this permit due to any cause, the Permittee shall: 

1. Immediately take action to stop, contain, and clean up unauthorized discharges or 

otherwise stop the noncompliance, correct the problem and, if applicable, repeat 

sampling and analysis of any noncompliance immediately and submit the results 

to the Department within thirty (30) days after becoming aware of the violation. 

2. Immediately notify the Department of the failure to comply. 

3. Submit a detailed written report to the Department within thirty (30) days, unless 

requested earlier by the Department. The report shall contain a description of the 

noncompliance, including exact dates and times, and if the noncompliance has 

not been corrected, the anticipated time it is expected to continue; and the steps 

taken or planned to reduce, eliminate, and prevent reoccurrence of the 

noncompliance. 

Compliance with these requirements does not relieve the Permittee from responsibility to 

maintain continuous compliance with the terms and conditions of this permit or the 

resulting liability for failure to comply. 
 Page 11 of 29 

Permit No. WA0041009 

S4. ANNUAL SEPA PROCESS 

All lakes proposed for treatment are included in an addendum to the Final Supplemental 

Environmental Impact Statement (FSEIS). The FSEIS is subject to a 30 day public comment 

period. The annual SEPA process must be completed prior to conducting lake or stream 

rehabilitation activities. 

S5. ANTIMYCIN SEPA REVIEW 

On or before June 1, 2005, WDFW shall the complete the SEPA review process regarding the use 

of Antimycin as a fish toxicant for certain fish management projects. Based upon the outcome of 

the SEPA process, Ecology may modify the permit to include the use of Antimycin. 

S6. BEST MANAGEMENT PRACTICES 

A. In order to prevent unnecessary damage to the environment, the permittee shall follow the 

best management practices defined below on the day of application. 

B. The permittee shall comply with all product label instructions. When application 

requirements specified in this permit differ from those on the label, the more stringent of 

the two requirements must be complied with. However, no condition in this permit or 

any amended Order shall reduce or modify the label instruction. All applicable federal, 

state and local laws and ordinances shall be followed. 

C. Powdered rotenone formulations shall be applied in such a way that minimizes airborne 

dust, using the best available technology such as the method outlined in “Utah’s 

Procedure for Mixing Powdered Rotenone into a Slurry” (Thompson et al. 2001). 

D. In order to prevent an exceedance of water quality standards outside the area intended for 

rehabilitation, rotenone treatment should only take place in lakes that are not discharging 

to downstream waters. This is accomplished by limiting treatments to lakes with closed 

basins or conducting treatments only during periods of low water, usually September or 

October. 

In instances where treated waters may potentially discharge to downstream waters 

resulting in an exceedance of water quality standards outside the treatment area, such 

discharge shall be prevented by installing adequate temporary water control measures. 

When it is necessary and unavoidable to discharge rotenone treated waters to downstream 

waters, the permittee shall conduct pretreatment water quality and biological monitoring, 

as specified in the monitoring plan. 

Treated waters shall be effectively neutralized and detoxified using potassium 

permanganate so that water quality standards are not exceeded below the neutralization 

zone. For purposes of this section, neutralization zone is defined as the downstream 

waters where potassium permanganate has been applied but has not yet fully neutralized 

the rotenone, due to the lag time normally associated with detoxification. The 

neutralization zone is typically considered the distance that water can be expected to 

travel in 20 minutes. Since the neutralization zone may contain toxic levels of rotenone 

and potassium permanganate, some fish mortalities may occur in this zone. 
 Page 12 of 29 

Permit No. WA0041009 

Below the neutralization zone, rotenone must be totally neutralized and residual 

potassium permanganate levels maintained at a non-toxic level of 1 mg/L, not to exceed 2 

mg/L. Live trout cars will be set up below the neutralization zone to monitor the 

effectiveness of detoxification measures. 

Detoxification procedures must utilize calibrated equipment to achieve the minimum 

effective concentration of potassium permanganate to oxidize the rotenone within the 

neutralization zone. Potassium permanganate concentrations must be closely monitored 

using a field calibrated spectrophotometer to keep residual permanganate levels at a level 

that effectively neutralizes rotenone while preventing damage to aquatic life downstream 

of the treatment area and neutralization zone. 

F. In order to minimize the discharge of inert ingredients contained in liquid rotenone 

formulations, only powdered rotenone formulations shall be utilized, except in very 

limited cases when the WDFW finds it necessary to treat waters that are inaccessible by 

boat, such as weedy shorelines or marshy areas. 

S7. SPILL PREVENTION AND RESPONSE 

A. Prior to the first rotenone treatment, WDFW shall submit a Spill Prevention and Response 

Plan to the Department of Ecology, Eastern Regional Office, N. 4601 Monroe, Spokane, 

WA 99205-1295. The spill plan should cover a plan for the prevention, containment, and 

control of spills or unplanned discharges from the application, storage and transportation 

of the pesticides. It should also include spills of oil and gasoline from application 

equipment including boats. According to the severity of the spill, it should tell when to 

report certain magnitudes of spills along with a list of names and telephone numbers of 

spill respondent teams at both WDFW and Ecology. 

B. Spills into state waters, spills onto land with a potential for entry into state waters, or 

other significant spills that may effect health, the environment, or property must 

immediately be reported to the following state and federal contacts: 

National Response Center (Federal): 1-800-424-8802, and 

Emergency Management Division (State): 1-800-258-5990, and 

The appropriate Dept. of Ecology regional office: 

Northwest Office, Bellevue: 1-425-649-7000 

Southwest Office, Olympia: 1-360-407-6300 

Central Office, Yakima: 1-509-575-2490 

Eastern Office, Spokane: 1-509-456-2926 

Within 5 days the event must be also be reported to Nancy C. Weller, Permit Manager, 

Department of Ecology, Eastern Regional Office, N. 4601 Monroe, Spokane, WA 99205-

1295. It should be a written report that includes a description of the event, including 

exact date and time, and the actions taken to correct the problem 

C. In the event of a spill, containment and clean-up efforts shall begin immediately and be 

completed as soon as possible, taking precedence over normal work. Clean-up shall 

include proper disposal of any spilled material and used clean-up material. 
 Page 13 of 29 

Permit No. WA0041009 

PUBLIC NOTICE PROCEDURES 

P1. RESIDENTIAL AND BUSINESS NOTICE PROCEDURES 

A. Prior to the initiation of rotenone treatment, the WDFW shall notify all property owners 

located within one-quarter (1/4) mile of the shoreline or stream bank radius and fivehundred (500) feet upland of the waters affected by rotenone treatment, including waters 

treated with potassium permanganate to detoxify or neutralize rotenone treated waters. 

B. This notification may be done by mail, e-mail, or by handbills given directly to the 

residences or businesses. If hand bills are used, the applicator shall secure the 

notices to the residences or businesses doorknob in a fashion that will hold them 

in place but will not damage property. If the residence or business is gated or guarded by 

watch dogs, the applicator may secure the notice in clear view on the outside of the 

gateway or may attach the notice to the outside of the residence in a fashion that will hold 

it in place but will not damage property. 

C. A copy of the notice and a list of names and addresses where they were sent shall be kept 

by the applicator for seven (7) years and be hand delivered or mailed to Ecology 

immediately upon request. Where notices were delivered by hand (hand bills), names are 

not required to be recorded; only the address where notification was made. 

D. Notification must take place at least 10 days, but not more than 21 days prior to initial 

treatment. 

E. Notification information must include: 

1. The lake or stream to be treated. 

2. The name of the pesticide (and oxidizer, when applicable) to be used. 

3. The internet address (URL) of the Material Safety Data Sheet (MSDS) for the 

chemical products used. For example, the URL for the MSDS for PrenfishTM 

Fish Toxicant Powder is http://www.prentiss.com/msds/pdf/655_691.pdf . 

4. The purpose of the treatment. 

5. Any public use or water use restrictions. 

6. The date(s) of treatment / restricted use. 

7. The names and phone numbers of designated contact people at WDFW and 

Department of Ecology so people can obtain additional information. 

P2. LEGAL NOTIFICATION PROCEDURES 

A. The Department of Fish and Wildlife shall publish announcements in the legal section of 

the local newspaper of general circulation (or nearest regional paper if local paper does 

not exist) 10 to 21 days prior to initial treatment. The legal notice shall include: 

1. The lake or stream to be treated. 

2. The name of the pesticide (and oxidizer, when applicable) to be used. 

3. The purpose of the treatment. 

4. Any public use or water use restrictions. 

5. The posting procedures. 
 Page 14 of 29 

Permit No. WA0041009 

6. The date(s) of treatment / restricted use. 

7. The names and phone numbers of designated contact people at WDFW and 

Department of Ecology so people can obtain additional information. 

B. An original affidavit from the legal department of the newspaper shall be kept by the 

Department of Fish and Wildlife for seven years and be mailed to the Department of 

Ecology upon request. 

P3. POSTING PROCEDURES 

A. The WDFW shall post signs prior to the application of any pesticide(s) no more than 

forty-eight (48) hours prior to application. The Department of Fish and Wildlife shall 

use good faith and reasonable effort to ensure that posted signs remain in place until the 

end of the period of water use restrictions, or 30 days, whichever is longer. The 

Department of Fish and Wildlife shall be responsible for removal of all signs before the 

following treatment of the waterbody. 

B. When the EPA label restricts human consumption of fish, swimming, irrigation, 

livestock watering, or any other precaution(s) relevant to public or private water use, all 

posted signs shall explicitly state the restriction(s) or precaution(s). 

C. The WDFW shall construct and post signs as follows: 

1. Public Property 

Signs shall be a minimum of eight and one-half (8 1⁄2) by eleven (11) inches in size and be 

made of durable weather-resistant material. Lettering shall be bold black type with the 

word “CAUTION” at least one (1) inch high and all other words at least one-quarter (1/4) 

inch high. The sign board shall be white, yellow, or orange. Signs shall be placed facing 

all points of normal public access to the shoreline or stream bank; or one every onehundred (100) feet of public shoreline within ten (10) feet of the mean high water mark. 

Signs shall be posted so that they are secure from the normal effects of weather and water 

currents but cause no damage to private or public property. The applicator shall post all 

signs within 24 hours of initial treatment. 

2. Boat Access Areas 

Boat launches are defined as publicly designated and privately owned community access 

boat launches. Signs shall be posted at all boat launches on the waterbody to be treated. 

Signs shall be a minimum of two (2) feet by three (3) feet in size and be made of durable 

weather-resistant material. Lettering shall be bold black type with the word “CAUTION” 

at least two (2) inches high and all other words at least one-half (1/2) inch high. The 

colors used for the sign board shall be white, yellow, or orange. 

Signs must be placed within twenty-five (25) feet of the shoreline, facing the entrance to 

the boat launch. Where the public access has a shoreline length greater than one hundredfifty (150) feet, the applicator shall place signs so that they are clearly readable by all 

people using the access areas. Signs shall be posted so that they are secure from the 

normal effects of weather and water currents but cause no damage to private or public 

property. 
 Page 15 of 29 

Permit No. WA0041009 

GENERAL CONDITIONS 

G1. SIGNATORY REQUIREMENTS 

All applications, reports, or information submitted to the Department shall be signed and 

certified. 

A. All permit applications shall be signed by either a responsible corporate officer of at 

least the level of vice president of a corporation, a general partner of a partnership, or 

the proprietor of a sole proprietorship. 

B. All reports required by this permit and other information requested by the Department 

shall be signed by a person described above or by a duly authorized representative of 

that person. A person is a duly authorized representative only if: 

1. The authorization is made in writing by a person described above and submitted to 

the Department. 

2. The authorization specifies either an individual or a position having responsibility 

for the overall operation of the regulated facility, such as the position of plant 

manager, superintendent, position of equivalent responsibility, or an individual or 

position having overall responsibility for environmental matters. (A duly 

authorized representative may thus be either a named individual or any individual 

occupying a named position.) 

C. Changes to authorization. If an authorization under paragraph B.2 above is no longer 

accurate because a different individual or position has responsibility for the overall 

operation of the facility, a new authorization satisfying the requirements of paragraph

B.2 above must be submitted to the Department prior to or together with any reports, 

information, or applications to be signed by an authorized representative. 

C. Certification. Any person signing a document under this section shall make the 

following certification: 

 I certify under penalty of law, that this document and all 

attachments were prepared under my direction or supervision in 

accordance with a system designed to assure that qualified 

personnel properly gathered and evaluated the information 

submitted. Based on my inquiry of the person or persons who 

manage the system, or those persons directly responsible for 

gathering information, the information submitted is, to the best of 

my knowledge and belief, true, accurate, and complete. I am 

aware that there are significant penalties for submitting false 

information, including the possibility of fine and imprisonment for 

knowing violations. 
 Page 16 of 29 

Permit No. WA0041009 

G2. RIGHT OF INSPECTION AND ENTRY 

The Permittee shall allow an authorized representative of the Department, upon the 

presentation of credentials and such other documents as may be required by law: 

A. To enter upon the premises where a discharge is located or where any records must be 

kept under the terms and conditions of this permit. 

B. To have access to and copy - at reasonable times and at reasonable cost - any records 

required to be kept under the terms and conditions of this permit. 

C. To inspect - at reasonable times - any facilities, equipment (including monitoring and 

control equipment), practices, methods, or operations regulated or required under this 

permit. 

D. To sample or monitor - at reasonable times - any substances or parameters at any 

location for purposes of assuring permit compliance or as otherwise authorized by the 

Clean Water Act. 

G3. PERMIT ACTIONS 

This permit may be modified, revoked and reissued, or terminated either at the request of 

any interested person (including the permittee) or upon the Department’s initiative. 

However, the permit may only be modified, revoked and reissued, or terminated for the 

reasons specified in 40 CFR 122.62, 122.64 or WAC 173-220-150 according to the 

procedures of 40 CFR 124.5. 

A. The following are causes for terminating this permit during its term, or for denying a 

permit renewal application: 

1. Violation of any permit term or condition. 

2. Obtaining a permit by misrepresentation or failure to disclose all relevant facts. 

3. A material change in quantity or type of waste disposal. 

4. A determination that the permitted activity endangers human health or the 

environment or contributes to water quality standards violations and can only be 

regulated to acceptable levels by permit modification or termination [40 CFR part 

122.64(3)]. 

5. A change in any condition that requires either a temporary or permanent reduction or 

elimination of any discharge or sludge use or disposal practice controlled by the 

permit [40 CFR part 122.64(4)]. 

6. Nonpayment of fees assessed pursuant to RCW 90.48.465. 

7. Failure or refusal of the permittee to allow entry as required in RCW 90.48.090. 
 Page 17 of 29 

Permit No. WA0041009 

B. The following are causes for modification but not revocation and reissuance except 

when the permittee requests or agrees: 

1. A material change in the condition of the waters of the state. 

2. New information not available at the time of permit issuance that would have 

justified the application of different permit conditions. 

3. Material and substantial alterations or additions to the permitted facility or activities 

which occurred after this permit issuance. 

4. Promulgation of new or amended standards or regulations having a direct bearing 

upon permit conditions, or requiring permit revision. 

5. The Permittee has requested a modification based on other rationale meeting the 

criteria of 40 CFR part 122.62. 

6. The Department has determined that good cause exists for modification of a 

compliance schedule, and the modification will not violate statutory deadlines. 

7. Incorporation of an approved local pretreatment program into a municipality’s 

permit. 

C. The following are causes for modification or alternatively revocation and reissuance: 

1. Cause exists for termination for reasons listed in A1 through A7, of this section, and 

the Department determines that modification or revocation and reissuance is 

appropriate. 

2. The Department has received notification of a proposed transfer of the permit. A 

permit may also be modified to reflect a transfer after the effective date of an 

automatic transfer (General Condition G8) but will not be revoked and reissued after 

the effective date of the transfer except upon the request of the new permittee. 

G4. REPORTING A CAUSE FOR MODIFICATION 

The Permittee shall submit a new application, or a supplement to the previous application, 

along with required engineering plans and reports whenever a material change to the facility 

or in the quantity or type of discharge is anticipated which is not specifically authorized by 

this permit. This application shall be submitted at least sixty (60) days prior to any proposed 

changes. The filing of a request by the Permittee for a permit modification, revocation and 

reissuance, or termination, or a notification of planned changes or anticipated 

noncompliance does not relieve the Permittee of the duty to comply with the existing permit 

until it is modified or reissued. 
 Page 18 of 29 

Permit No. WA0041009 

G5. PLAN REVIEW REQUIRED 

Prior to constructing or modifying any wastewater control facilities, an engineering report 

and detailed plans and specifications shall be submitted to the Department for approval in 

accordance with Chapter 173-240 WAC. Engineering reports, plans, and specifications 

shall be submitted at least one hundred eighty (180) days prior to the planned start of 

construction unless a shorter time is approved by Ecology. Facilities shall be constructed 

and operated in accordance with the approved plans. 

G6. COMPLIANCE WITH OTHER LAWS AND STATUTES 

Nothing in this permit shall be construed as excusing the Permittee from compliance with 

any applicable federal, state, or local statutes, ordinances, or regulations. 

G7. DUTY TO REAPPLY 

The Permittee shall apply for permit renewal at least 180 days prior to the specified 

expiration date of this permit. 

G8. TRANSFER OF THIS PERMIT 

In the event of any change in control or ownership of facilities from which the authorized 

discharge emanate, the Permittee shall notify the succeeding owner or controller of the 

existence of this permit by letter, a copy of which shall be forwarded to the Department. 

A. Transfers by Modification

Except as provided in paragraph B below, this permit may be transferred by the 

Permittee to a new owner or operator only if this permit has been modified or revoked 

and reissued under 40 CFR 122.62(b)(2), or a minor modification made under 40 CFR 

122.63(d), to identify the new Permittee and incorporate such other requirements as 

may be necessary under the Clean Water Act. 

B. Automatic Transfers

This permit may be automatically transferred to a new Permittee if: 

1. The Permittee notifies the Department at least 30 days in advance of the proposed 

transfer date. 

2. The notice includes a written agreement between the existing and new Permittee’s 

containing a specific date transfer of permit responsibility, coverage, and liability 

between them. 

3. The Department does not notify the existing Permittee and the proposed new 

Permittee of its intent to modify or revoke and reissue this permit. A modification 

under the subparagraph may also be minor modification under 40 CFR 122.63. If 

this notice is not received, the transfer is effective on the date specified in the 

written agreement. 
 Page 19 of 29 

Permit No. WA0041009 

G9. REDUCED PRODUCTION FOR COMPLIANCE 

The Permittee, in order to maintain compliance with its permit, shall control production 

and/or all discharges upon reduction, loss, failure, or bypass of the treatment facility until 

the facility is restored or an alternative method of treatment is provided. This requirement 

applies in the situation where, among other things, the primary source of power of the 

treatment facility is reduced, lost, or fails. 

G10. REMOVED SUBSTANCES 

Collected screenings, grit, solids, sludges, filter backwash, or other pollutants removed in 

the course of treatment or control of wastewaters shall not be resuspended or reintroduced to 

the final effluent stream for discharge to state waters. 

G11. DUTY TO PROVIDE INFORMATION 

The Permittee shall submit to the Department, within a reasonable time, all information 

which the Department may request to determine whether cause exists for modifying, 

revoking and reissuing, or terminating this permit or to determine compliance with this 

permit. The Permittee shall also submit to the Department upon request, copies of records 

required to be kept by this permit [40 CFR 122.41(h)]. 

G12. OTHER REQUIREMENTS OF 40 CFR 

All other requirements of 40 CFR 122.41 and 122.42 are incorporated in this permit by 

reference. 

G13. ADDITIONAL MONITORING 

The Department may establish specific monitoring requirements in addition to those 

contained in this permit by administrative order or permit modification. 

G14. PAYMENT OF FEES 

The Permittee shall submit payment of fees associated with this permit as assessed by the 

Department. 

G15. PENALTIES FOR VIOLATING PERMIT CONDITIONS 

Any person who is found guilty of willfully violating the terms and conditions of this permit 

shall be deemed guilty of a crime, and upon conviction thereof shall be punished by a fine of 

up to ten thousand dollars ($10,000) and costs of prosecution, or by imprisonment in the 

discretion of the court. Each day upon which a willful violation occurs may be deemed a 

separate and additional violation. 

Any person who violates the terms and conditions of a waste discharge permit shall incur, in 

addition to any other penalty as provided by law, a civil penalty in the amount of up to ten 
 Page 20 of 29 

Permit No. WA0041009 

thousand dollars ($10,000) for every such violation. Each and every such violation shall be 

a separate and distinct offense, and in case of a continuing violation, every day's continuance 

shall be deemed to be a separate and distinct violation. 

G16. UPSET 

Definition – “Upset” means an exceptional incident in which there is unintentional and 

temporary noncompliance with technology-based permit effluent limitations because of 

factors beyond the reasonable control of the Permittee. An upset does not include 

noncompliance to the extent caused by operational error, improperly designed treatment 

facilities, inadequate treatment facilities, lack of preventive maintenance, or careless or 

improper operation. 

An upset constitutes an affirmative defense to an action brought for noncompliance with 

such technology-based permit effluent limitations if the requirements of the following 

paragraph are met. 

A Permittee who wishes to establish the affirmative defense of upset shall demonstrate, 

through properly signed, contemporaneous operating logs or other relevant evidence that: 1) 

an upset occurred and that the Permittee can identify the cause(s) of the upset; 2) the 

permitted facility was being properly operated at the time of the upset; 3) the Permittee 

submitted notice of the upset as required in condition S3.E; and 4) the Permittee complied 

with any remedial measures required under S5 of this permit. 

In any enforcement proceeding the Permittee seeking to establish the occurrence of an upset 

has the burden of proof. 

G17. PROPERTY RIGHTS 

This permit does not convey any property rights of any sort, or any exclusive privilege. 

G18. DUTY TO COMPLY 

The Permittee shall comply with all conditions of this permit. Any permit noncompliance 

constitutes a violation of the Clean Water Act and is grounds for enforcement action; for 

permit termination, revocation and reissuance, or modification; or denial of a permit renewal 

application. 

G19. TOXIC POLLUTANTS 

The Permittee shall comply with effluent standards or prohibitions established under Section 

307(a) of the Clean Water Act for toxic pollutants within the time provided in the 

regulations that establish those standards or prohibitions, even if this permit has not yet been 

modified to incorporate the requirement. 
 Page 21 of 29 

Permit No. WA0041009 

G20. PENALTIES FOR TAMPERING 

The Clean Water Act provides that any person who falsifies, tampers with, or knowingly 

renders inaccurate any monitoring device or method required to be maintained under this 

permit shall, upon conviction, be punished by a fine of not more than $10,000 per violation, 

or by imprisonment for not more than two years per violation, or by both. If a conviction of 

a person is for a violation committed after a first conviction of such person under this 

Condition, punishment shall be a fine of not more than $20,000 per day of violation, or by 

imprisonment of not more than four (4) years, or by both. 

G21. REPORTING PLANNED CHANGES 

The Permittee shall, as soon as possible, give notice to the Department of planned physical 

alterations or additions to the permitted facility, production increases, or process 

modification which will result in: 1) the permitted facility being determined to be a new 

source pursuant to 40 CFR 122.29(b); 2) a significant change in the nature or an increase in 

quantity of pollutants discharged; or 3) a significant change in the Permittee’s sludge use or 

disposal practices. Following such notice, this permit may be modified, or revoked and 

reissued pursuant to 40 CFR 122.62(a) to specify and limit any pollutants not previously 

limited. Until such modification is effective, any new or increased discharge in excess of 

permit limits or not specifically authorized by this permit constitutes a violation. 

G22. REPORTING ANTICIPATED NON-COMPLIANCE 

The Permittee shall give advance notice to the Department by submission of a new 

application or supplement thereto at least one hundred and eighty (180) days prior to 

commencement of such discharges, of any facility expansions, production increases, or other 

planned changes, such as process modifications, in the permitted facility or activity which 

may result in noncompliance with permit limits or conditions. Any maintenance of 

facilities, which might necessitate unavoidable interruption of operation and degradation of 

effluent quality, shall be scheduled during non-critical water quality periods and carried out 

in a manner approved by the Department. 

G23. REPORTING OTHER INFORMATION 

Where the Permittee becomes aware that it failed to submit any relevant facts in a permit 

application, or submitted incorrect information in a permit application or in any report to the 

Department, it shall promptly submit such facts or information. 

G24. REPORTING REQUIREMENTS APPLICABLE TO EXISTING 

MANUFACTURING, COMMERCIAL, MINING, AND SILVICULTURAL 

DISCHARGERS 

The Permittee belonging to the categories of existing manufacturing, commercial, mining, or 

silviculture must notify the Department as soon as they know or have reason to believe: 
 Page 22 of 29 

Permit No. WA0041009 

A. That any activity has occurred or will occur which would result in the discharge, on a 

routine or frequent basis, of any toxic pollutant which is not limited in this permit, if 

that discharge will exceed the highest of the following “notification levels:” 

1. One hundred micrograms per liter (100 μg/l). 

2. Two hundred micrograms per liter (200 μg/l) for acrolein and acrylonitrile; five 

hundred micrograms per liter (500 μg/l) for 2,4-dinitrophenol and for 2-methyl-4,6-

dinitrophenol; and one milligram per liter (1 mg/l) for antimony. 

3. Five (5) times the maximum concentration value reported for that pollutant in the 

permit application in accordance with 40 CFR 122.21(g)(7). 

4. The level established by the Director in accordance with 40 CFR 122.44(f). 

B. That any activity has occurred or will occur which would result in any discharge, on a 

non-routine or infrequent basis, of a toxic pollutant which is not limited in this permit, if 

that discharge will exceed the highest of the following “notification levels:” 

1. Five hundred micrograms per liter (500μg/L). 

2. One milligram per liter (1 mg/L) for antimony. 

3. Ten (10 ) times the maximum concentration value reported for that pollutant in the 

permit application in accordance with 40 CFR 122.21(g)(7). 

4. The level established by the Director in accordance with 40 CFR 122.44(f). 

G25. COMPLIANCE SCHEDULES 

Reports of compliance or noncompliance with, or any progress reports on, interim and final 

requirements contained in any compliance schedule of this permit shall be submitted no later 

than fourteen (14) days following each schedule date. 
 Page 23 of 29 

Permit No. WA0041009 

APPENDIX A 

Rapid Method for Measuring 

Rotenone in Water at Piscicidal Concentrations 

V. K. DAWSON, P. D. HARMAN, D. P. SCHULTZ, AND J. L. ALLEN 

United States Fish and Wildlife Service 

National Fishery Research Laboratory 

Post Office Box 818, La Crosse, Wisconsin 54601 

Abstract 

A high-performance liquid chromatography (HPLC) procedure that is rapid, specific, and sensitive 

(limit of detection <0.005 mg/liter) was developed for monitoring application and degradation rates of 

rotenone. For analysis, a water sample is buffered to pH 5 and injected through a Sep Pak ® C18

disposable cartridge. The cartridge adsorbs and retains the rotenone which then can be eluted 

quantitatively from the cartridge with a small volume of methanol. This step effectively concentrates the 

sample and provides sample cleanup. The methanol extract is analyzed directly by HPLC on an MCH 10 

reverse-phase column; methanol: water (75:25, volume: volume) is the mobile phase and flow rate is 1.5 

ml/minute. The rotenone is detected by ultraviolet spectrophotometry at a wavelength of 295 nm. 

Received November 29, 1982 Accepted May 24, 1983 

Rotenone, the active constituent of derris root, has been used widely as an insecticide and piscicide. 

Its use for removing undesired fish populations in the United States began in the 1930s (Schnick 1974). 

Several analytical procedures have been reported for the analysis of rotenone, including colorimetry 

(Gross and Smith 1934; Goodhue 1936), infrared spectrometry (Delfel 1976), thin-layer chromatography 

(Delfel and Tallent 1969), gas chroma- tography (Delfel 1973), and high-performance liquid 

chromatography (HPLC) (Bushway et al. 1975; Freudenthal and Emmerling 1977; Bowman et al. 1978; 

Kobayashi et al. 1980). These procedures are either costly, time-consuming, insensitive, or lack 

specificity for monitoring concentrations of rotenone in water during fish- eradication projects. 

We describe a simple, rapid HPLC procedure with a sample-concentrating step that can be used to 

determine residues of rotenone in water at piscicidal concentrations. 

Methods 

Apparatus 

1. HPLC–Varian 5000 equipped with varichrom ultraviolet-light detector and optional Model CDS111L data system.1

 Operating conditions: 

stationary phase––30 cm X 4 mm Varian micropak MCH- 10 reverse phase; 

mobile phase––methanol: water (75:25, volume: volume); 

flow rate––1.5 ml/minute; chart 

speed––1 cm/minute; 

wavelength––295 nm; 

attenuation––0.04 absorbance full scale. 

 

1

 Mention of commercial products does not imply endorsement by the United States Government. 
 Page 24 of 29 

Permit No. WA0041009 

2. Sep Pak® C18 disposable cartridges from Waters Associates, Incorporated. 

3. Vortex stirrer. 

4. Disposable syringes (50 ml). 

5. Test tubes (15 ml) with Teflon-lined screw caps. 

Reagents 

1. Methanol––HPLC grade. 

2. Water––HPLC grade. 

3. Rotenone––purified grade from Aldrich Chemical Company; 0.0 1 g/ 100 ml methanol (made fresh 

daily). 

4. Acetic acid (glacial)––American Chemical Society (ACS) reagent grade, 0.2 M; 11.6 ml diluted to 1 

liter with water. 

5. Sodium acetate––ACS reagent grade, 0.2 M; 2.72 g of C2H3O2Na 3H2O diluted to 100 ml with water. 

6. Buffer reagent––0.1 M; 14.8 ml of 0.2 M acetic acid + 35.2 ml of 0.2 M sodium acetate diluted to 100 

ml with water. 

Procedure 

1. Precondition Sep Paks with 2 ml methanol and 5 ml water according to instruction sheet supplied by 

manufacturer. 

2. Add 1 ml buffer reagent for each 50 ml of water sample (if expected concentration is less than 0.02 

mg/liter, more than 50 ml of sample may have to be extracted). 

3. Attach preconditioned Sep Pak to 50-ml syringe with plunger removed. 

4. Transfer sample to syringe, insert plunger, and force sample through Sep Pak at a rate of not more 

than 40 ml/minute. Discard water. 

5. Remove Sep Pak, remove plunger, and replace Sep Pak on syringe. 

6. Add 2 ml methanol, insert plunger, and slowly force methanol through Sep Pak into test tube. 

7. Cap tube and mix on vortex stirrer. 

8. Analyze by HPLC against 50 ml of a standard containing a known concentration of rotenone in water 

solution processed as above. 

Results and Discussion 

The ultraviolet spectrum of rotenone has an absorption maximum at 295 nm (Fig. 1). A monochromatic 

detector (254 nm) can be used, but a considerable loss of sensitivity and potential loss of specificity will 

occur. Performance of monochromatic detectors can be enhanced by the use of 313-nm filters, but a 

grating monochrometer set at 295 nm provides optimal results. 

As indicated in step 1 of the procedure, the Sep Paks must be prerinsed with methanol followed by 

water before they are used in the analyses. Previous tests (Dawson 1982) indicated that, for best results, 

this step should not be completed more than 2 hours before an analysis. 

Rotenone recovery is influenced by the rate water samples flow through the Sep Paks during extraction 

and by the elution rate of methanol. The recovery was less than 70% at a flow of 100 ml/minute but 

exceeded 90% at flows of 40 ml/minute or less. Several volumes of methanol were evaluated for most 

efficient elution of adsorbed rotenone from Sep Paks. Small volumes of methanol provided more 
 Page 25 of 29 

Permit No. WA0041009 

concentrated samples, but recoveries were consistently better when 2 ml or more of methanol were used 

for elution. 

Recoveries of rotenone were evaluated at various pH values to determine whether or not acidity of 

water samples affected the utility of the method. Water samples were fortified with 0.08 mg/liter of 

rotenone and buffered to pH 5, 7, and 9 before the Sep Pak extraction. Recoveries of rotenone from the 

buffered samples were 98, 94, and 73%, respectively, indicating that acidification is essential for optimal 

performance of the Sep Paks. 

FIGURE 1.––Chemical structure and ultraviolet spectrum of rotenone (10 mg/liter) in methanol : water (75:25, 

volume : volume). 

A water sample volume of 50 ml is sufficient for the analysis of rotenone concentrations of 0.02 

mg/liter or greater. However, as much as 200 ml of sample may have to be extracted to achieve a 

sensitivity of 0.005 mg/liter. The limiting factors for greater sample concentration are the tedium of 

extracting large volumes of sample and possible interferences that may develop in samples as a result of 

the extraction and concentration of contaminants. Recently, J. T. Baker Chemical Company developed the 

Baker-10® extraction system, in which similar adsorption chromatography is used and the tedium of 

analysis is reduced by a vacuum manifold that extracts up to 10 samples simultaneous. 

Samples extracted on Sep Paks are stable for only a few hours. However, if the sample is eluted from 

the Sep Pak and stored in the methanol eluate, the samples are stable for up to 2 days. For best results, 

samples should be kept cool and in the dark. 

Retention time for rotenone from a sample of spiked pond water injected on the reverse-phase column 

was 5.7 minutes (Fig. 2). Unfortified pond water had no interfering peaks. The pen deflection at 2 minutes 

in Fig. 2 is the solvent injection peak. 

Water samples from ponds treated with rotenone in summer and late fall were analyzed for residues of 

rotenone by this HPLC method (unpublished data). No interference problems were encountered and the 

measured concentration agreed closely with that calculated on the basis of the application rate. 
 Page 26 of 29 

Permit No. WA0041009 

FIGURE 2.––Liquid chromatograms of (A) pond water sample fortified with rotenone (0.1 mg/liter) and 

(B) unfortified pond water; mobile phase––methanol:water (75:25, volume:volume); flow rate––1.5 

ml/minute; wavelength––295 nm; attenuation––0.04 absorbance full scale. The water sample (50 ml) 

was concentrated 25 times on Sep Pak by elution with 2 ml methanol 

Five replicate sample of pond water fortified with rotenone (0.1 mg/liter) were analyzed to evaluate the 

consistence of the method. The mean percentage of recovery and standard error were 97.6 + 1.6. 

The use of a micro-processor data system, such as a Varian CDS-111L, greatly facilitates the analysis 

by integrating peak areas and converting values directly into concentration units. 
 Page 27 of 29 

Permit No. WA0041009 

References 

BOWMAN, M. C., C. L. HOLDER, AND L. 1. BONE. 1978. High pressure liquid chromatographic 

determination of rotenone and degradation products in animal chow and tissues. journal of the 

Association of Official Analytical Chemists 61:1445- 1455. 

BUSHWAY, R. J., B. S. ENGDAHL, B. M. COLVIN, AND A. R. HANKS. 1975. Separation of 

rotenoids and the determination of rotenone in pesticide formulations by high-performance liquid 

chromatography. Journal of the Association of Official Analytical Chemists 58:965-970. 

DAWSON, V. K. 1982. A rapid high-performance liquid-chromatographic method for simul-taneouslv 

determining the concentrations of TFM and Bayer 73 in water during lampricide treatments. Canadian 

Journal of Fisheries and Aquatic Sciences 39:778-782. 

DELFEL, N. E. 1973. Gas-liquid chromatographic determination of rotenone and cleguelin in plant 

extracts and commercial insecticides. Journal of the Association of Official Analytical Chemists 

56:1343-1349. 

DELFEL, N. E. 1976. Ultraviolet and infrared analysis of rotenone: effect of other rotenoids. Journal of 

the Association of Official Analytical Chemists 59:703-707. 

DELFEL, N. E., AND W. H. TALLENT. 1969. Thin layer densitometric determination of rotenone and 

deguelin. Journal of the Association of Official Analytical Chemists 52:182-187. 

FREUDENTHAL, R. I., AND D. C. EMMERLING. 1977. Separation of rotenoids by high-pressure 

liquid chromatography. Journal of Chromato-graphy 134:207-209. 

GOODHUE, L. D. 1936. An improvement on the Gross and Smith colorimetric method for the 

determination of rotenone and deguelin. Journal of the Association of Official Agricultural Chemists 

19:703-707. 

GROSS, C. R., AND C. M. Smith. 1934. Colorimetric method for determination of rotenone. Journal of 

the Association of Official Agricultural Chemists 17:336-339. 

KOBAYASHI, H., O. MATANO, AND S. GOTO. 1980. Determination of rotenoids in soil and crops by 

high-performance liquid chromato-graphy. Journal of Pesticide Science 5:89-92. 

SCHNICK, R. A. 1974. A review of the literature on use of rotenone in fisheries. United States Fish and 

Wildlife Service Literature Review 74-15. NTIS (National Technical Information Service) PB-235 

454/AS, Springfield, Virginia, USA. 
 Page 28 of 29 

Permit No. WA0041009 

APPENDIX B 

Sampling Protocols for Zooplankton 

The new approach to measure uses on a given lake includes a need to evaluate the health of a fishery. A 

widely utilized tool on the east coast of the United States is the measuring of zooplankton as a cost 

effective surrogate to collecting and measuring fish. An index has been developed in order to determine 

the predator/prey balance in the fish communities within a given lake (Mills and Schavone, 1982). In a 

study of 18 natural lakes in upstate New York, Mills and Schavone (1982) demonstrated a strong 

correlation between mean length of cladocerans and planktivore weight (r2

 = .70; P<0.05). In other 

words, the presence of large zooplankton indicate predator fish are keeping prey species in balance. 

Dominance of smaller zooplankton suggests an ineffective amount of predators to suppress planktivore 

density. 

A standard approach to sampling zooplankton was followed in the field. Methods for collecting, storage, 

and enumeration are patterned after the “Zooplankton Workshop Reference Guide” prepared by BSA 

Environmental Services, Inc. (Beaver, 1997). 

Sampling Period 

Five vertical tows were pulled in June and five were pulled in August from the deep site of each selected 

lake. Tows were composited into one 125ml sample bottle. Five tows were not necessary if there were 

an abundance of zooplankton in the first few tows. Duplicate samples (again, five tows if necessary) 

were taken at duplicate TP sample sites. 

Field Procedure 

The Wisconsin net was pulled from a depth of one meter off the bottom or 20 meters, whichever was less. 

The depth was rounded to the most conservative meter to ensure bottom sediments weren’t disturbed. 

Nets were retrieved at a rate of one meter per three seconds. 

Upon retrieval of the sample, a squirt bottle filled with tap or distilled water was used to dislodge any 

zooplankton that may have been clinging to the mesh. Samples were discharged into a 125 mL amber, 

Lugol-treated sample container and preserved with approximately 15 mls of Lugol’s solution. 

Sample Analysis 

Samples were analyzed for relative abundance of cladocerans and copepods and their mean length. 

Relative abundance was determined using a Sedgewick-Rafter counting chamber and a compound 

microscope. Subsamples were analyzed to estimate mean length of the zooplankton using a compound 

microscope and an ocular equipped with a reticle. Measurements were recorded to the nearest 0.2mm. 

Results were tabulated as the ratio of total cladocerans:total copepeods. 
 Page 29 of 29 

Permit No. WA0041009 

Data Interpretation 

Due to time constraints in 1998, it was not possible to fully analyze the collected samples of zooplankton 

but the following provides possibilities for future data interpretation. A subset of lakes sampled by 

Ecology in 1998 were also surveyed by the Washington State Department of Fish and Wildlife. 

Zooplankton mean length data should be compared to fish length-frequency distributions to evaluate 

whether there is a particular mean zooplankton length that could be used as a pivot indicator of a balanced 

predator/prey fish population in a given lake (e.g. 1.0 mm is used in some states). An index with a range 

of mean lengths within given categories may be the most effective use of the zooplankton data. For 

example, mean zooplankton lengths between 0.9 mm and 1.0 mm may be rated as “fair” for predator/prey 

populations and mean lengths between 1.0 mm and 1.1 mm may rate “good” and above 1.1 mm rated as 

“fair,” etc. A fair rating or worse could then be used to demonstrate impairment of a beneficial use. 

Relative zooplankton abundance data may be correlated with nutrient and Secchi data. It is uncertain at 

this time whether or not zooplankton abundance is a good indicator of predator/prey balance. However, 

correlations with traditional water chemistry data and additional fish population data may demonstrate 

whether or not trophic cascade effects are present in Washington lakes (Brett and Goldman, 1996). 

Zooplankton data may also explain differences between total phosphorus concentrations and expected 

correlated values for chlorophyll a/Secchi measurements. 

*Appendix B Sampling Protocols for Zooplankton was taken from pages 4-5 of Water Quality 

Assessments of Selected Lakes Within Washington State 1998, Washington State Department of 

Ecology. December 2000. Publication No. 00-03-039.