Vermont Department of Environmental Conservation

Quality Assurance Guidelines for

Wastewater Treatment Facility Laboratories

1996

Prepared by:

Andrew Fish, C.E.T.

Wastewater Laboratory Specialist
for the VT Dept. of Environmental Conservation
 PREFACE

These guidelines have been developed to assist operators and technicians at Vermont
Wastewater Treatment Facilities to prepare acceptable Quality Control Manuals specific to their
facilities. It is further hoped that the guidelines will encourage the implementation of these
procedures. The quality control procedures outlined in this manual represent the minimum
requirements considered acceptable by the Vermont Department of Environmental Conservation
for facilities regulated under the NPDES program.

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 The laboratory quality control manual described in this guidance document is an integral
part of the appropriate quality assurance procedures required of all NPDES regulated facilities
pursuant to Title 40 of the Code of Federal Regulations, Part 122.41 (e) any commercial
laboratory performing analyses for an NPDES regulated facility must certainly meet these
minimum requirements.

It is absolutely essential that all laboratories analyzing wastewater compliance samples

follow defined quality assurance procedures. Those procedures must be documented in a
laboratory quality control manual.

To satisfy minimum NPDES requirements a quality control manual must address the
following items. (Each of which will be discussed in detail in the following pages.)

I. Quality Assurance Program Plan Elements

II. Staff Organization and Responsibilities
III. Quality Assurance Objectives and Activities
IV. Sample Collection Procedures
V. Sample Handling and Documentation Procedures
VI. Laboratory Water Quality
VII. Standard Operating Procedure for Each Analytical Method
VIII. Reagent/Standard Quality Preparation and Traceability
IX. Calibration Procedures and Frequencies
X. Data Handling - Reduction, Validation and Reporting
XI. Preventive Maintenance, Procedures and Schedules
XII. Corrective Action Contingencies
XIII. Quality Control Procedures (General)

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 In order to produce a truly useful QA/QC manual a table of contents should be
included. You may also wish to include things such as laboratory utilities information,
electrical supply, compressed air source, etc. or anything that might effect the function of
the laboratory.

I. Quality Assurance Project Plan Elements

This section of the manual simply lists the topics which will be included in your
quality assurance plan. It is basically a table of contents without the page numbers. It
should be the first step in preparing the manual as it will help you to organize your
thoughts and establish distinct sections.

II. Staff Organization and Responsibilities

Laboratory analytical centers can obviously vary considerably in size, number,

complexity of analyses performed, and in the number of people employed.

This section of the manual should identify all persons responsible for laboratory
activities from sampling to preparation and signing of the Discharge Monitoring Report.
It should describe the responsibilities of each of the individuals in certain terms;
identifying each individual using that persons name (not just the position - i.e. Andy
Fish, QA coordinator not QA coordinator).

A typical municipal wastewater facilitys staff organization and responsibilities

section might include:

1). The town manager as the person responsible for signing the WR-43.
2) The chief operator as the preparer of the WR-43.
3). Assistant operator as responsible for sample collection.
4) Lab tech - responsible for performing analyses maintaining bench sheets,

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 assisting in data review etc.

Each individuals responsibilities and limitations should be spelled out in this

section.

As an example of limitations, an analysts responsibilities might include informing

the chief operator if accuracy values are beyond established warning and control limits.

----This clearly shows that the analysts responsibility ends with informing his
supervisor of a problem. It is presumably the chief operators responsibility to determine
the action taken from that point.

An organization chart is very useful in identifying positions (where they fit into
the scheme of things).

III. Quality Assurance Objectives

This section, besides being useful to inspectors and certification officials can be
very helpful to the person preparing the manual to figure out just what am I trying to
demonstrate in this manual? What am I really after here?

Here is an example of typical Quality Assurance Objectives.

1. To ensure data produced by the laboratory is accurate and defensible.

2. To ensure samples collected are representative of preferred universe.
3. To ensure that all laboratory procedures are EPA approved.
4. To ensure that all equipment is properly calibrated and meets EPA specifications.
5. To ensure that proper corrective actions are initiated when necessary.

III. Quality Assurance Objectives (continued)

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 6. To ensure that each sample is tracked from collection time until the report is
finalized - with records maintained for the required intervals. Of course in
addition to simply listing the objectives you might summarize referencing specific
sections of the manual, how these objectives will be met.

IV. Sample Collection Procedures

Careful and precise documentation of sampling procedures is absolutely

imperative to ensure that good representative samples are consistently collected. Untold
time and dollars are wasted on analysis of samples that are collected at an improper
location, time or in an unacceptable container. An exact sampling procedure should be
written for each parameter required at your facility. It is smart to include detailed
photographs and or diagrams along with sampling location descriptions.

The sampling procedures section of your QA manual should include:

1) Description of container - glass, plastic, etc.

2) Container cleaning requirements -- acid washed, DI rinsed, etc.
3) Sample preservation techniques .
4) Sample holding times.
5) Exact sampling location description.
6) Exact sampling time for most representative sample.
7) Collection method/technique - grab, composite (time-flow, auto sampler)
etc.
8) Volume of sample required for analysis.

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V. Sample Handling and Documentation Procedures

This section must include:

A written description of exactly how the samples are handled after collection.
The methods for documentation must be clearly stated. The following items must be
included in this section.

1) Documentation of exact time, date and location sample was collected.

2) Labeling of sample bottles.
3) Chain of Custody Procedures (if analysis is performed by an independent
laboratory).

a) Sample custody forms, labels, seals.

b) Sample transportation and delivery procedure.

V. Laboratory Water Quality

As many of us are painfully aware, the quality of the water we use in rinsing
glassware, preparing reagents and standards, sample dilution, and blank preparations can
be the determining factory between accurate and inaccurate analytical results.

There are rather strict monitoring requirements for laboratories analyzing

drinking water. These requirements are listed in EPA/570/9-90/008 document Manual
for the Certification of Laboratories Analyzing Drinking Water, and include:

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V. Laboratory Water Quality (continued)

1) General Lab Water

Parameter Limit Monitoring Frequency

Conductivity <2 umhos/cm @ 25C Monthly

2) Water for Microbiological Analyses

Tests Limit Monitoring Frequency

Conductivity <2 umhos/cm @ 25C Monthly
Total Residual Chlorine <detection limit Monthly
Heavy Metals, Single4
(Cd, Cr, Cu, Ni, Pb, Zn) <0.05 mg/L Annually
Heterotrophic Plate Count <500 CFU/ml Monthly
Water Quality Test5
(Biosuitability) 0.8 - 3.0 ratio Annually

In wastewater analysis our main concerns regarding laboratory water quality are
conductivity, chlorine and metals that might interfere with certain analyses (i.e. high
copper level can affect results in the BOD analysis).

Generally accepted monitoring frequencies and limits for those parameters are
listed below.

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V. Laboratory Water Quality (continued)

2) Water for Microbiological Analyses (continued)

Parameter Limit Monitoring Frequency

Conductivity <2 umhos/cm @ 25C Monthly
Total Residual Chlorine 0.0 or <detectable limit Monthly
Copper < .05 mg/L Semi-Annually
Other Metals < .05 mg/L Annually
(Cd, Cr, Ni, Pb, Zn)

* Reality Check: The Vermont Department of Environmental Conservation does

not expect a small wastewater treatment facility required to perform pH analyses
only to have their own laboratory water analyzed for all the parameters listed
above. Such a facility should consider purchasing commercially available
distilled water, where concentrations of these parameters are often listed on the
label. Larger
facilities, performing BODs and more sophisticated analyses would be expected
to verify conductivity, chlorine and copper limits. Commercial laboratories
performing numerous parameters more frequently than the average treatment
facility would be expected to perform more frequent laboratory water analyses.

VII. Standard Operating Procedures - For Each Analytical Method

This section of the manual should include a simple step - by step procedure which
details the exact method in which the analysis is performed. It should NOT simply be a
copy of an analytical method from Standard Methods but instead a practical What you
really do to perform this analysis, type of description in an easy to read format.

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VII. Standard Operating Procedures - For Each Analytical Method (continued)

Include specific items such as Turn the blender to setting 6 and blend for 5
minutes to get a good homogenous sample or Use only Class A volumetric glassware
from drawer #2.

Items that must be included in this section are:

1) Reference Method, number and Title

2) Description of instrumentation
3) Specific glassware and equipment cleaning procedures
4) Reagent and Standard Preparation
5) Step by Step Procedures (SOPs)
6) Calibration and Standardization procedures - specific to the analysis
7) Equations and Calculations

You may reference an SOP manual to satisfy #5 if you have a separate and
complete manual.

VIII. Reagent/Standard Quality, Preparation and Traceability

This section of the manual should describe the general procedures for preparation
of reagents and standards. The types of questions that should be addressed include:

1) Grade of materials used i.e. ASC or Analytical Reagent grade chemicals primary
standards, etc.
2) Measuring methods i.e. calibrated analytical balance, class A volumetric
glassware etc.
3) Reagent labeling: Must Include

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VIII. Reagent/Standard Quality, Preparation and Traceability (continued)

A. Reagent name
B. Concentration
C. Preparation and Expiration dates and/or received the dates a chemical is
received as well as the date it is opened.

IX. Calibrations Procedures and Frequencies

Documentation of all instrument or equipment calibration is essential to any good

quality assurance plan.

This section should detail the procedures, frequencies and type of calibration as
well as whether it was routine or professionally performed and finally who performed the
calibration.

Written documentation must include at a minimum:

1) Established frequency of routine calibration for each piece of equipment i.e. daily,
weekly, monthly.
2) Established frequency of professional calibrations for each piece of equipment.
3) Calibration procedures - Standards used, etc.
4) Dates and times calibrations are performed.
5) The name (* not just initials of the person who calibrated the instrument.)

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X Data Handling - Reduction Validation and Reporting

Reduction of Data

Data reduction is the process of transforming raw data into final results that are
reported in standard units to some authority. An efficient method of data reduction must
exist to reduce transcription and calculation errors. That methodology, whatever it may
entail, should be described in this section.

Validation of Data

Data validation starts with the analyst. It is usually the analysts responsibility to
ensure that instruments have been properly calibrated and are operating properly. He or
she then records the results on a bench sheet. Generally someone then transfers the data
to another form after, perhaps performing some calculations.

At this point there should be some mechanism in place to ensure that the data has
been accurately transferred and that any calculations were performed properly.

At least one other person beside the analyst should look over and double check
the bench sheet data vs. final results. One or both of these individuals must understand
the concept of significant figures and units of measures.

* More than one technical violation could have been avoided had this knowledge
been applied.

This section should describe the procedure by which data is checked. Specific
responsibilities should be spelled out here.

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X Data Handling - Reduction Validation and Reporting (continued)

Reduction of Data (continued)

Each WR-43 report form contains a certification signed by the permittees

authorized agent which states I certify under penalty of law that I have personally
examined and am familiar with the information submitted herein.

It would seem reasonable then to involve this person in the data validation
process.

It is important to include in this section.

Examples on description of

1) 8Bench sheets, lab books

2) A description of the check system used to avoid transcription and
calculation errors.
3) A procedure of how invalid data is handled on bench sheets and on the
WR-43 report form (i.e. BOD results not meeting the R1 D2) requirement
- How is this handled?
4) Rules for consistent rounding off of numerical results.

XI. Preventive Maintenance, Procedures and Schedules

Preventive maintenance is very important in order to minimize instrument Down

Time and ensure the continued accuracy of analytical results.

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XI. Preventive Maintenance, Procedures and Schedules (continued)

The old excuse I couldnt report pH results for the month of April because my
pH probe was broken, just isnt acceptable as a defense.

Proper preventative maintenance and contingency plans are essential for

uninterrupted analyses.

This section must describe procedures for routine and scheduled contract services.
Include:

1) Comprehensive routine maintenance schedule - list of instruments.

2) Professional service/maintenance schedule - list of instruments.
3) A list of employees responsible for performing maintenance.
4) A check list to ensure duties have been completed.

XI. Corrective Action

This section of the manual will outline the steps taken when any portion of the
quality assurance process become questionable or invalid.

These steps should include:

1) Identifying and defining the problem i.e. QC is outside control limits,

what caused unacceptable results.
2) A list defining need for corrective actions - i.e. (Poor blank or standards
results)
3) Actions to eliminate the problem.
4) Actions to prevent recurrence . You may include a copy of the
checklist(s)

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XI. Corrective Action (4) (continued)

in the manual and include a reference describing where the actual list(s) is/are
located if you desire.
5) Procedure for informing superiors of other authorities (if applicable).
6) Protocol for resampling and retesting.

XIII. Quality Control Procedures:

Quality control is defined in the 18th edition of Standard Methods for the
Examination of Water and Wastewater, 1992.

As as set of measures within a sample analysis methodology to assure that the

process is in control. This section of the manual is dedicated to describing those
measures taken to ensure process control. In it you should define terms and describe
general procedures which can be referenced in the QC portion of each analytical method.

Quality control consists of analyzing and reporting the results of quality control
standards, sample duplicates and replicates, spikes and blanks.

EPA recommends that a frequency of 10% to 20% be applied for quality control
measures. That is, for every 5 to 10 samples analyzed there should be one quality control
sample analyzed for each parameter being tested.

For commercial laboratories analyzing multiple samples, a quality control

standard or spike, a sample duplicate or replicate and a blank is expected to be run with
each batch of samples analyzed.

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XIII. Quality Control Procedures: (continued)

The minimum frequency for quality control analyses in laboratories performing

one sample at a time would be one-in every 10 samples. *Note: Certain analyses are
required by the Vermont Department of Environmental Conservation to be run in
duplicate 100%. (T.S.S. and E. Coli).

The Vermont Department of Environmental Conservation has the following

minimum requirements regarding QC for the common parameters listed in the Table
below:

Calibration/ QC Duplicates/
Parameter Standardization Standards Replicates Spikes Blanks

Ammonia As per method 1/10 Tests 1/10 Tests 1/Yr 1/each

BOD Meter before each use 1/10 Tests 1/10 Tests 1/Yr 1/each
IGGA/Each
if seeded

Chlorine Check standard 1/month 1/10 Tests 1/Yr 1/each

Residual Meter curve daily standard for each use

Titrimetric FAS stand 1/month 1/month 1/10 Tests 1/Yr 1/each

COD as per method 1/10 Tests 1/10 Tests 1/Yr 1/each

Cyanide as per method 1/each 1/each 1/each 1/each

E. Coli Additional QC required for E. Coli 2/Yr 1/each N/A 1/each

includes equipment sterility checks /POS
(indicator tape, Kilit ampules) control/each
each tests and pH check of test
dilution water (7>1 .2) each tests

Metals as per method 1/each 1/each 1/each 1/each

Nitrate Nitrogen as per method 1/10 Tests 1/10 Tests 1/Yr 1/each

Oil & Grease as per method 1/each 1/each 1/Yr 1/each

pH Minimum 2 point calibration each (3rd buffer)

use 1/each 1/10 Tests N/A N/A

Phosphorus 5 Standards/each 1/10 Tests 1/10 Tests 1/Yr 1/each

Settleable Solids N/A N/A 1/10 Tests N/A N/A

Total Suspended constant weights before and after 1/10 Tests 1/each N/A 1/each
Solids filtering sample

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XIII. Quality Control Procedures: (continued)

XIII. 1B. Spikes

Spikes are prepared by adding a predetermined quantity of a known standard to a

sample. By analyzing an unspiked sample and a spiked sample it is possible to determine
if there is a substance in the sample that interferes with test results.

From the results of analysis of a spike and unspiked sample, percent recovery can
be calculated using the equation:

S&U
% Recovery ' 100% x
Csa

Where S = Measured concentration in spiked aliquot

U = Measured concentration in unspiked aliquot
Csa = Actual concentration of spike added

If the Percent Recovery is outside the range of 80% to 100% the contents of the
sample may be interfering with the method of measurement for that analysis.

In this event the cause of the interference must be determined and corrective
actions must be initiated (i.e. change methods, etc.)

* One spike or Quality Control Standard should be run with one of every 10
samples run.

* For laboratories performing sample analysis in batches one spike or quality

control standard should be run with each batch.

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XIII. Quality Control Procedures: (continued)

XIII. 2B. Quality Control Standards

Quality control standards are substances of known concentration. They are used
to verify the accuracy of your analysis.

Quality control standards:

1) Must be prepared from a source different than what was used to calibrate
the instrument or prepare the standard curve.

2) The quality control standard must be specific for the parameter being
analyzed.

3) The quality control standard must approximate the concentration of the

sample and must have a value between that of the highest and lowest
calibration standards used.

For Example: If a sample is expected to have a pH of

approximately 6, calibration standards of pH 4 and 7 might be used along
with a quality control standard with a pH value of 6.

4) Remember: Quality control standards are not used to adjust the instrument
as are calibration standards. The resulted quality control standards are
simply recorded an compared to the actual known value.

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XIII. Quality Control Procedures: (continued)

XIII. 2B. Quality Control Standards (continued)

5) Unless control charts are used to determine the acceptable quality control
standard range. Results of the quality control standard must fall within a
range of 20% of the known value. That is the Percent Error must be
from 80% to 120% of the actual standard

Observed Value & Known Value

Percent Error ' x 100
Known Value

6) If results of a quality control standard do not fall within the acceptable

range the cause must be determined and recorded on bench sheets or other
appropriate record books. A not should be placed in the comments section
of the WR-43 report form. Corrective actions must be implemented.

III. Duplicates/Replicates
1) Duplicate samples are samples which are collected in two separate containers at
the same time and place.

2) Replicate samples are samples that are collected in a single container and are
poured off into a second container for separate analyses of the same parameter(s).

3) An absolute minimum of one duplicate or replicate per 10 samples is required.

4) The result of duplicate/replicate samples should not vary by more than 20% from
the original sample.

XIII. 2B. Quality Control Standards (continued)

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III. Duplicates/Replicates (continued)

5) Do not average original and duplicate/replicate results. Record the result of the
original sample. Then record on bench sheets or other appropriate record book
the Percent error for the duplicate/replicate.

Percent error is determined by applying the following equation.

Observed Value & Known Value

Percent Error ' x 100
Known Value

If the duplicate/replicate results fall outside the acceptable ( 20%) range of the
original sample a note should be made in the comments section of the WR-43
report form and on the bench sheet. Corrective action should be implemented.

IV. Blanks

1) A blank consists of either laboratory water or parameter specific dilution water.

Blanks provide a check of the laboratory water quality as well as a check of the
analysts analytical technique.

2) Blank must be analyzed with each analysis where applicable.

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XIII. 2B. Quality Control Standards (continued)

IV. Blanks (continued)

3) Acceptable blank values are included with each analytical method.

For example:

BOD = # 0.2 mg/L

E. Coli = 0 colonies/100 mls

If blank limit values are exceeded a note must be included in the comment section of the
WR-43 report form, and on the bench sheet. In some instances such as in the E. Coli
analysis, this invalidates the sample data. (Check the method info) In any blank
exceedance corrective action contingencies must be implemented.

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 The following pages contain an

Example QC Manual

Hopefully this example will help you in preparing

your own manual.

Remember: The manual must be specific to your own

facility and will remain a work in progress requiring
periodic updating and revisions.

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 Table of Contents
A. Title Page Page 17A
B. Introduction Page 18
1. Quality Assurance Project Plan Elements Page 19
2. Laboratory Staff Organization and Responsibilities Page 20
2.1. Town Manager Page 21
2.2 Chief Operator Page 21
2.3 Lab Supervisor Page 21
2.4 Operators Page 22
. 2.5 Skills and Training Page 22
3.0 Quality Assurance Objectives and Activities Page 23
4.0 Sample Collection Procedures Page 23
4.1 General Guidelines Page 23
4.2 Sample Type and Holding Times Page 24
4.3 Sample Preservation Page 25
5.0 Sample Handling and Documentation Procedures Pages 27 & 28
6.0 Laboratory Water Quality Page 28
6.1 Deionized Water Pages 28 & 29
7.0 Standard Operating Procedure for Each Analytical Method Pages 30
7.1 pH Pages 30 to 32
7.2 Total Residual Chlorine Pages 32 to 35
7.3 Settleable Solids Pages 35 to 36
7.4 Total Suspended Solids Pages 36 to 40
8.0 Reagent/Standard Quality, Preparation and Traceability Pages 40 to 41
9.0 Calibration Procedures and Frequencies Page 41
9.1 Instrument Calibration Procedures Pages 41 to 43
10.0 Data Handling-Reduction, Validation and Reporting Page 43
10.1 Data Reduction Pages 43 to 44
10.2 Data Validation Page 44
10.3 Reporting Pages 45 to 46
11.0 Preventive Maintenance, Procedures and Schedules Page 47
12.0 Corrective Action Contingencies Pages 48 to 49
13.0 Quality Control Procedures (General) Pages 50 to 55

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 INTRODUCTION

This Quality Control/Assurance Manual is written

specifically for the WWW Wastewater Facility in an attempt to
assure the continued high quality and accuracy of analytical
results. This will be accomplished by monitoring the accuracy
and precision of these results, by providing clear, concise
Standard Operating Procedures defining sampling and analytical
procedures specific to the WWW facility and by clearly defining
the responsibilities of all personnel regarding required
performance and documentation.

The WWW facility performs analyses required by the

NPDES permit for discharge to the WWW River, in-house
progress control analyses, and occasionally pH and chlorine
analyses for the Water Supply Department.

Analytical results generated for NPDES permit are

submitted to the State of Vermont on a monthly basis. Results
of process control analyses are recorded on bench sheets and
maintained in-house files for a period of three years.

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I. Quality Assurance Project Plan Elements

The fourteen Quality Assurance Project Plan Elements addressed in this manual
are:

Title Page
Introduction
Staff Organization and Responsibilities
Quality Assurance Objectives and Activities
Sample Collection Procedures
Sample Handling and Documentation Procedures
Laboratory Water Quality
Standard Operating Procedure for Each Analytical Method
Reagent/Standard Quality Preparation and Traceability
Calibration Procedures and Frequencies
Data Handling - Reduction, Validation and Reporting
Preventive Maintenance, Procedures and Schedules
Corrective Action Contingencies
Quality Control Procedures (General)

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2.0 Laboratory Staff Organization and Responsibilities

The Fuzzyville Wastewater Treatment facility has 5 permanent full-time

employees. At some time, all employees are responsible for collection of samples either
on a daily basis or on a rotating weekend schedule.

One employee is dedicated to laboratory work only, while all 5 might perform
daily analyses including pH, Cl2 and settleable solids.

The organizational structure is summarized in Table I below.

Table I

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2.1 Town Manager

The town manager is responsible for signing the WR-43 report form.

Before signing he/she questions the chief operator or to the reports accuracy and
any special comments.

Reviews all data.

Compares WR-43 results to bench sheet results.

Provides final data check.

Sends report to the proper state authority.

Makes final decision on equipment purchases after conferring with chief operator.

Reviews and is familiar with the laboratory QC manual.

2.2 Chief Operator

The chief operator is responsible for signing the WR-43 report form as the
preparer.

Transfer all results from bench sheets to the WR-43 report form.

Compares final WR-43 results to bench sheets to guard against transcription

errors etc.

Reviews an is familiar with the laboratory QC manual.

Schedule special sampling and analytical projects.

Reviews bench sheet data to ensure proper WC was practiced.

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 Is responsible for purchasing laboratory equipment.

Occasionally collect and analyzes effluent samples for pH.

On occasion when pH is analyzed he/she properly calibrates the pH meter.

2.3 Lab Supervisor

Is responsible for overall technical quality of the work performed in the

laboratory.

Ensures the use of acceptable Standard analytical methods.

Provides training to all persons responsible for sampling.

Is responsible for preparation and revisions of laboratory QC manual.

Informs chief operator of any equipment needs.

Performs all instrument calibrations and reagent preparations.

Maintains proper bench sheets and equipment maintenance logs.

Responsible for maintaining current SOPs.

Collects and composites effluent samples.

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 2.4 Operators

Are responsible for sampling of specific parameters.

Are properly training and responsible for analysis of pH, Total Residual Chlorine
and Settleable solids when necessary.

Maintain proper bench sheets and calibration logs.

2.5 Skills and Training

EDUCATION
EXPERIENCE
Time & Type
RESPONSIBILITIES
REQUIREMENTS SPECIFIC TO A POSITION

Example:

SPECIAL
NAME POSITION EDUCATION EXPERIENCE SKILL RESPONSIBILITIES REQUIREMENTS

Kim Lab B.S. 15 yrs. as lab Phosphorus As described in None

Istry Supervisor Chemistry analyst determination organizational
using auto chart
analyzer

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 EXAMPLE

3.0 Quality Assurance Objectives

a. To ensure data produced by the laboratory is accurate and defensible.

b. To ensure samples collected are representative of preferred universe.

c. To ensure that all laboratory procedures are E.P.A. approved.

d. To ensure that all equipment is properly calibrated and meets E.P.A.

specifications.

e. To ensure that proper corrective actions are initiated when necessary.

f. To ensure that each sample is tracked from collection time until the report is
finalized - with records maintained for the required intervals.

4.0 Sample Collection Procedures

4.1 General Guidelines

a. Samples are collected in a well mixed area at the center of the channel avoiding
eddies, backwaters and area where settling might take place.

b. Influent samples are collected after the comminuter but above the RAS line. See
Diagram.

c. Effluent samples are collected after all treatment processes just before discharge
to the stream. See Diagram for specific parameters.

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4.1 General Guidelines (continued)

d. Grab samples

Grab samples are collected via a Nalgene container attached to an 8 foot

extendable aluminum pole. All grab samples are collected and immediately
returned to the lab for analysis.

*Note* E. Coli - collected directly into sterilized 250 ml. Plastic bottle containing
4 drops 10% Sodium Thiosulfate solution.

e. Composite samples

All composite samples consist of 24 disrete samples collected hourly via an Isco
automatic sampler. Each disrete sample (approximately 500 mls) is distributed
into properly cleaned, 1 liter Isco sample bottles. Composite samples are cooled
by ice packs which are placed in the sampler wells. Samples are composited at
the lab based on readings from flow charts.

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 SAMPLING PROCEDURES

4.2 Type of Sample and Holding Time

Type of Holding Sample

Parameter Sample Time Container Location
pH Grab 15 Minutes Plastic Bottle Refer to Diagram
Temperature Grab At Site Plastic Bottle Refer to Diagram
Dissolved Oxygen Grab 15 Minutes B.O.D. Bottle Refer to Diagram
B.O.D. Composite Flow 6 Hours Composite Refer to Diagram
Proportional Sampler Plastic
Bottles
Total Coliform Grab 1 Hour Sterile Sample Refer to Diagram
Fecal Coliform Plastic Bottle
E. Coli w/Sodium
Thiosulfate
Chlorine Residual Grab 15 Minutes Opaque B.O.D. Refer to Diagram
Glass Bottle
Total Suspended Composite Flow 6 Hours Composite Refer to Diagram
Solids Proportional Sampler Plastic
Bottles
Specific Grab 15 Minutes Glass Refer to Diagram
Conductance
Metals Grab/Comp 6 Months Amber Glass Refer to Diagram
TKN Grab 1 Hour Plastic Bottle Refer to Diagram
Settleable Solids Grab 30 Minutes Plastic Bottle Refer to Diagram

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 4.3 Sample Preservation

If samples are taken and not run within the time frame as in Chart 9.1 then the following
chart applies.

PRESERVATION CONDITIONS

Parameter Container Volume Preservation Holding Representative Sampling

Time Time
BOD, TSS P 1L 4 C 24 Hours 8 AM - 8 AM
TKN P .5L H2SO4 pH < 2.0 28 Days 8 AM - 8 AM
4C
Oils & Grease G 1L HCl pH < 2.0 24 Hours Between 8 AM and 12 PM
Metals GA .2L HN03 pH < 2.0 6 Months 8 AM - 8 AM
Phenols G .5L H3PO4 pH 24 Hours 8 AM - 8 AM
< 2.0 + 1t CuSO4 4C
Sulfides P .5L 2 ml Zinc 24 Hours 8 AM - 8 AM
Acetate 4C
Cyanides (T) P 1L NaOH pH > 12.0 4 C 24 Hours 2 PM
VOC V 40 ml 4C 24 Hours 2 PM

G = Glass bottle with Teflon lined lid

GA = Amber bottle with Teflon lined lid

P = Plastic Bottle

V = Approved glass vials with Teflon and pure rubber seals

Note: All samples are refrigerated at 4C after preservation.

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4.4 Sample Collection Location

BAD EXAMPLE : Effluent sample is collected at end of chlorine contact tank.

BETTER : The effluent grab samples for pH and Total Residual Chlorine are
collected at the outfall end of the chlorine contact tank at a point
approximately 1 foot upstream of the v-notch weir.

BEST : The effluent grab samples for pH and Total Residual Chlorine are
collected at the outfall end of the chlorine contact tank at a point
approximately 1 foot upstream of the v-notch weir, in the center of
the channel at a depth of approximately 1 foot. The open container
attached to a 6 foot aluminum pole is lowered into the waste
stream with the open end facing down stream as shown in pictures
#1 and #2.

Picture #1 Picture #2
Be specific and include
descriptions for each different type of sample (re: grab, composite)

33
-33-
5. Sample Handling Documentation Procedures

5.1 All grab samples collected for analysis of pH, Total Residual Chlorine and
Settleable Solids are immediately taken to the lab. The pH and Chlorine analyses
are begun immediately. The settleable solids sample is poured immediately into
the Imhoff cone.

5.2 The grab sample collected for analysis of E. Coli is immediately placed into a
cooler with four blue ice packs and sealed. It is immediately transported by WWW
to WWW laboratory. The approximate travel time is 35 minutes.

Each bottle is properly labeled and accompanied with a chain of custody form.
One copy of the COC form is maintained at the facility.

5.3 Composite samples

Immediately after the last 24 disrete sample is collected the entire sampler is
brought into the laboratory where the hourly sample volumes are calculated
measured and poured into a four liter jug. Aliquots are poured from the container
for specific parameters (BOD, TSS etc.) After thorough shaking. Sample
containers are then placed into the refrigerator at 4C until the analysis is
performed.

34
-34-
5. Sample Handling Documentation Procedures (continued)

5.4 Sample Identification/Labeling

a) All sample containers are labeled with the following information.

1) Type of sample (influent, effluent, grab, composite)

2) Parameter to be analyzed

3) Time and date collected

4) Initials of person who collected it

5) Preservation information

6) Any special instructions or remarks

b) All data from the label is transferred to bench sheets.

c) A chain of Custody form is completed. One copy accompanies the sample

to the lab. One copy is retained at the wastewater facility.

*Include a copy of the form in the manual*

d) The sample is delivered directly to the laboratory by name, assistant

operator

35
-35-
5. Sample Handling Documentation Procedures (continued)

5.4 Sample Identification/Labeling (continued)

e) When the sample is handed over to the commercial laboratory the

recipient signs the chain of custody form indicating the time the sample
was received.

f) A copy of the chain of custody form is returned to the wastewater facility

along with analytical results.

6.0 Laboratory Water Quality

This example was borrowed from a QC manual (prepared by personnel of the

Nashua NH Wastewater Treatment Facility (copied here with their kind permission).

6.1 Deionized Water

R/O Deionized water is used in the laboratory for reagent preparation, glassware
rinsing, and BOD dilution water.

System includes: Millipore R/O system followed by a Milli-Q deionization

system and a 0.02 micron final filter.

TYPE: I : R/O, Deionization and 0.02 um. filter.

TYPE: II: R/O only

36
-36-
6.1 Deionized Water (continued)

Quality Control Tests:

Parameter Requirements Frequency

(Required) Conductivity > 0.5 megohms resistance or Once Per Month
< 2 microhms/cm at 25C
(Suggested) Total Chlorine Non-detectable Once Per Month
Residual
(Suggested) Biosuitability of Ratio of 0.8 - 3.0 Semi-annually
Laboratory Pure (April & Oct.)
Water
(Suggested) Metals Lead, Nickel, Zinc, Copper, Semi-Annually
Cadmium, Chromium, Iron, (April & Oct.)
Silver
(Suggested) Total Plate Count < 200 colonies/100 ml. Once Per Month

* All QC data generated is recorded in the bound QC Manual.

Other Quality Control Measures

R/O system is back flushed daily for one half of an hour. Water for BOD dilution
water and reagent dilution is made fresh after morning flush cycle.
Tubing and associated hardware (if in contact with the water) are replaced as needed.

6.2 Distilled Water

Occasionally bottled distilled water is purchased (Poland Springs) for

preparation of reagents, BOD dilution water and glassware rinsing.

37
-37-
6.1 Deionized Water (continued)

Other Quality Control Tests: (continued)

The quality specifications for this bottled water are included on the product label
and meets all requirements for analytical use.

7. Standard Operating Procedures for Each Analytical Method

7.1 pH

7.1 A. Method Reference

Electrometric Method #4500 - HB. 18th Edition Standard Methods pages

for the Examination of Water and Wastewater 4 - 65 ... 4 - 69.

7.1 B. Apparatus
pH meter (make and model)
pH temperature probe (type)
Buffer solutions Fisher, NIST certified, 4.0, 7.0, 10.0
Glass beakers (50 ml)
Magnetic stirrer and stir bars
Safety Glasses, gloves
Squeeze bottle for RO/DI water

7.1 C. Standardization

- The meter is standardized before each use with fresh pH 4.0 and 10.0
buffer solutions.

38
-38-
7.1 C. Standardization (continued)

- The pH of a fresh pH 7.0 buffer is checked and the result is recorded in the
QC log.

- If the result of the checked buffer varies by more than .1 standard unit.
The meter is restandardized and the problem is corrected.

- The Fill hole and electrode cap are removed during use and replaced
after each use.

7.1 D. Interferences

- It is important, even with temperature compensation capabilities to

approximate as closely as possible the sample temperature when
standardizing the meter. To accomplish this , buffers are stored in the
refrigerator during winter months and at room temperature during summer
months.

- Sodium error is not a problem at this facility.

- Oil material, especially from influent testing can cause development of a

film on the electrode causing poor response. This can be avoided by
periodically washing the probe with a mild detergent followed through
rinses with RO/DI water.

- pH Electrodes are replaced if proper standardization cannot be

accomplished.

39
-39-
7. Standard Operating Procedures for Each Analytical Method (continued)

7.1 E. Sample Preparation and Preservation

A minimum 25 ml sample is collected in a glass container. The

sample is taken immediately to the lab for analysis.

7.1 F. Procedure

Place pH and temperature probes into the sample. Allow meter to

equilibrate. Then record the reading in pH units at temp C.

7.1 G. Calculation

The pH meter reads directly in a pH standard units pH is reported

to the nearest 0.1 unit at a given temperature.

7.1 H. Quality Control

1. All required calibrations and other preventative maintenance on

instrumentation and equipment is performed regularly and
recorded on bench sheets and calibration logs.

2. Buffers are NIST Traceable, are changed daily and recorded.

3. 10% Duplication and Replication.

40
-40-
7. Standard Operating Procedures for Each Analytical Method (continued)

7.1 H. Quality Control (continued)

4. The pH meter will be calibrated before each use, and separate

standard will be processed as a control buffer to validate the
calibration, pH 10 buffer will be used as a control.

5. Grab samples will be delivered immediately to the lab for analysis.

6. Glassware will be thoroughly rinsed with purified water.

7. Temperature probe will be calibrated against a NIST traceable

thermometer.

8. Calibration sheet will include: date and time of calibration, buffers

used and order of use, exact meter results, analyst performing
calibration, buffer and sample temperature.

9. Meter range 0-14, accuracy 0.1 pH, repeatability 0.1 pH,

temperature compensation and capable of 2 point calibration.

7.2. Total Residual Chlorine

7.2 A. Method Reference

Hach DR 700 method which is based on the DPD colorimetric

method #4500-CL G 18th Edition Standard Methods pages 4-45 and 4-46.

41
-41-
7.2. Total Residual Chlorine (continued)

7.2 B. Apparatus

Spectro Photometer Model DR 700

DPD Total Residual Chlorine powder pillow/packets
Vials 10 ml - marked - dedicated currettes
Kimwipe Tissues
Safety glasses, gloves
Scissors or clippers

7.2 C. Standardization

The meter has an internal calibration which is automatically

enabled for each use.

In addition a Hach brand Voluette ampule certified standard is

used daily. Results are recorded and standard curve is plotted.

A blank consisting of the raw sample is used to zero the instrument

each use.

7.2 D. Interference

Color and turbidity interferences are reduced by the use of an

untreated sample as a blank to zero the device.

Organic contaminants may interfere but can usually be spotted by

the strange coloration - yellowish or greenish.

42
-42-
7.2 D. Interference (continued)

Oxidized manganese because of its orange color can create a

problem but is at least partially compensated for by use of a blank.

7.2 E. Sample Preparation and Preservation

Because of the unstable characteristics of chlorine in an aqueous

solution it is important to take great care in the sample collection not to
alter the sample chlorine concentration.

The sample is collected into an airtight BOD bottle which has been
covered with black electrical tape. The bottle is filled well into the neck
so as to overflow when the ground glass stopper is inserted. Make sure
there are no bubbles in the bottle. It is then taken to the dab for immediate
analysis (within 5 minutes from collection to analysis).

7.2 F. Procedure

Rinse the sample cells and caps thoroughly with RO/DI.

Pour sample into 2 sample cells to the 10 ml mark.

Empty the contents of one Total Cl2 powder pillow or packet into
one of the sample cells. Cap and invert 10 - 15 minutes to mix.

Wait 3 minutes.

43
-43-
7.2 F. Procedure (continued)

Using the up and down arrows set the device to mode 5207 (515
nanometers).

Wipe both sample cells carefully with a Kimwipe being careful to

remove any fingerprints or other marks.

Place the cell containing untreated sample (Blank) into the cell
compartment. Close the light shield and press the zero button.
The meter should read 0.00 mg/L.

Place the cell containing treated sample into the cell compartment
and push the read button.

Record the result in mg/L.

7.2 G. Calculations

The DR700 reads concentration of Total Residual Chlorine in

mg/L. Results are recorded directly to bench sheets.

Standard curves are plotted in mg/L.

7.2 H. Quality Control

1. All reagents are NIST traceable and preparation and expiration dates are
recorded.

44
-44-
7.2 H. Quality Control (continued)

2. The Hach DR700 colorimeter is maintained by QC Services with regularly

scheduled service every 6 months.

3. Glassware is thoroughly washed with a 2% solution of Micro brand

detergent, and rinsed with RO deionized water. Separate cells for
duplicate total chlorine determinations.

4. Hach Volette Ampule Certified Standards are used daily, and are recorded
as a standard curve for a control chart.

5. Duplicate samples are analyzed once per week.

6. Grab samples are recorded at the exact time of sampling and are analyzed
immediately after entering the laboratory. The time of analysis and date
are recorded.
NO HOLD TIME

Calculation for Voluette Ampule Standards:

ml of standard addition
x Chlorine conc. of Vol. Ampule
ml of standard additional % ml of sample

= Concentration of chlorine added to the sample.

45
-45-
7.3. Settleable Solids

7.3 A. Method Reference

Volumetric Method #2540 F 18th Edition Standard Methods for the

Examination of Water and Wastewater pages 2-57.

7.3 B. Apparatus

1 Liter Imhoff Cones

7.3 C. Standardization N/A

7.3 D. Interferences N/A

7.3 E. Sample Preparation and Preservation

Mix the sample thoroughly before pouring off.

7.3 F. Procedure

Place one liter of well mixed sample into a clean Imhoff cone.

Allow the sample to settle for 45 minutes.

Rotate the cone to dislodge solids on the side of the cone.

Allow to settle for an additional 15 minutes.

46
-46-
7.3 F. Procedure (continued)

After a total of elapsed time of 60 minutes from the time the sample was
poured. The settled solids are read in ml/liter and recorded.

7.3 G. Calculations

Results are recorded as ml/L.

7.3 H. Quality Control

Duplicate samples are collected and analyzed weekly.

7.4 Total Suspended Solids

7.4 A. Method Reference

Total Suspended Solids Dried at 103 - 105C Method #2540 D.

18th Edition Standard Methods for the Examination of Waste and

Wastewater pages 2-56.

7.4 B. Apparatus

Drying Oven 104 C ( 1C)

Filter Paper, glass fiber 934AH

Buchner Funnel

47
-47-
 7.4 B. Apparatus (continued)

Rubber stopper (to fit Buchner funnel to 1 liter side arm flask)

(2) 1 foot sections of rubber hose (one to connect side arm to

filtration flask to safety trap) One to connect trap to vacuum pump.

(2) 1 liter side arm flasks - 1 filtration 1 safety vacuum pump.

Graduated Cylinders

Desiccator - fresh desiccant

Forceps

Safety glasses and gloves

Aluminum weighing tray

(4) Place analytical balance squeeze bottle for RO/DI water.

7.4. C. Standardization (or more accurately pre-test preparation)

The balance must be checked to be sure it is level and then

properly zeroed after brushing any debris off the pan.

48
-48-
7.4. C. Standardization (or more accurately pre-test preparation)
(continued)

Filters must be rinsed with RO/DI water dried at 104C for

approximately 15 minutes, weighed and then redried, redesiccated and
reweighed to establish constant weight ( 0.5 mg) before the analysis is
performed.

- Make sure the drying oven is maintained at a constant temperature

of 104C 1.

7.4. D. Interferences

Water with every high mineral content might need to be dried

longer, desiccated and weighed quickly as moisture will be quickly
absorbed from the atmosphere. This affect can be reduced by placing
desiccant in a container inside the balance chamber.

Dont include large floating particles in the sample unless they are
truly representative of the actual sample conditions.

7.4 E. Sample Preparation and Preservation

Collect a sample volume of at least 2 liters.

Keep samples refrigerated at 4C up until the time of analysis to

reduce the microbiological decomposition of solids.

49
-49-
7.4 E. Sample Preparation and Preservation (continued)

Try to perform the analysis within 24 hours (7 days absolute

maximum)

Bring the sample to room temperature before beginning analysis.

7.4 F. Procedure

1. Prepare the funnel and vacuum apparatus. Make sure the vacuum
is not excessive so as to rip the filter.

2. Put the pre-weighed filters into the funnel wrinkled side

3. Seat the filter by rinsing the funnel and filter with about 25 mls of
RO/DI water. Turn on the vacuum pump until DI water is drawn
through the filter.

4. Shake sample thoroughly, then pour off 1 liter of effluent (100 ml

influent) into the graduated cylinder.

5. Pour the sample slowly into the center of the filter with the
vacuum on.

- Rinse the graduated cylinder with at least 3 successive 20

ml DI water rinses turn the graduated cylinder while
pouring rinsewater into funnel to insure that all solids are
rinsed out.

50
-50-
7.4 F. Procedure (continued)

- Wash down the sides of the funnel into the filter.

- Remove the filter from the funnel with the forceps.

- Place the filter into an aluminum weighing pan and place onto the
center rack of the drying oven at 104C for 2 - 3 hours (preferably
overnight).

- Cool to room temperature by putting the filter into the desiccator

for 15 to 30 minutes.

- Weigh (if the aluminum pan was included in the initial weighing it
must certainly be included in all weighings.)

- Redry at 104C for at least 1 hour.

Redesiccate for 15 to 30 minutes.

- Reweigh

- Repeat drying, desiccating and weighing if necessary until

constant weight is achieved (<.5 mg difference)

7.4 G. Calculations

A & B x 100,000
TSS in mg/L =
C

51
-51-
7.4 G. Calculations (continued)

Where A = weight of filter and residual (in grams)

B = weight of filter (in grams)

C = volume of sample filtered (in milliliters)

7.4 H. Quality Control

1. The oven temperatures are closely monitored.

Temperatures are recorded every four hours when in use.

2. The analytical balance is checked for level, auto calibrated

before each use and professionally calibrated every 6
months.

3. The temperature of the sample storage refrigerator is

checked and recorded every 4 hours when in use.

4. A replicate sample is analyzed each time the analysis is

performed. If results are not within a specified range
(determined by control chart) corrective actions are
initiated.

5. One duplicate sample is analyzed for every ten TSS

analyses performed. Results are recorded in QC log and
are plotted on control charts.

52
-52-
 7.4 H. Quality Control (continued)

6. A RO/DI water blank is run each time the analysis is

performed. Results are plotted on a blank control chart.

7. Sufficient sample, up to one liter, is filtered to produce at

least 2.5 mg residual.

8. Results are recorded in QC log and bench sheets.

7.5 5 Day BOD

7.5 A. Method Reference: 5 Day BOD Method 521 OB 18th Edition

Standard Methods for Examination of Water
and Wastewater pages 5-2 through 5-6.
7.5 B. Apparatus

Dissolved Oxygen Bottles

Overcaps
One Liter Graduated Cylinders
Assorted Sizes of Graduated Cylinders
Beakers, Assorted Sizes
Pipettes
Pipette Bulb
Carboy
Siphon
Propeller Mixer
DO Meter
Thermometer

53
-53-
7.5 B. Apparatus (continued)

Air Pump, Tubing, Filter

Stand
pH Meter
Incubator
Refrigerator
DO Bottles with Auto Pipettes
500 ml Erlenmeyer Flasks
25 ml Burette and Burette Stand
Dropping Bottles

54
-54-
7.5 C. Reagents:

Reagent Recipe Hold Time

Phosphate Buffer - 8.5 KH2PO4 - Indefinite @ 4 C
(Dilution water - 21.75g K2HPO4 - Throw out if biological growth
component.) - 33.4g Na2HPO4 .7H2O appears.
- 1.7g NH4Cl
- Dilute to 1R DI water.
- Autoclave 45 minutes.
Magnesium Sulfate - 22.5g Mg SO4.7H2O in 1R DI water - Indefinite @ 4 C
- Autoclave 45 minutes. - Throw out if biological growth
appears.
Calcium Chloride - 27.5g CaCl2 in 1R DI water. - Indefinite @ 4 C
- Autoclave 45 minutes. - Throwout if biological growth
appears.
Ferric Chloride - 0.25g FeCl3 .6H2O in 1R DI water. - Indefinite @ 4 C
(Dilution water - Autoclave 45 minutes. - Throw out if biological growth
component) appears.
1N Sulfuric Acid - 28 ml conc. H2SO4 to 1R DI water.
(To adjust pH sample)
1N Sodium Hydroxide - 40g NaOH to 1R DI water.
(To adjust pH sample)
GGA ampules Order from: Hach Company
- 300 mg/l glucose PO Box 907
- 300 mg/l glutanic acid Ames, Iowa
Polyseed Order from: Polybac Corp. 1 year at 20 C
3894 Courtney Street
Bethlehem, PA
Sodium Sulfite - Prepare small vials each containing 1 - 2 hours
(Dechlorination reagent) 0.79g of Na2SO3
- Dissolve 1 vial in 500 ml DI water
prior to use.

55
-55-
 7.5 C. Reagents (continued)

Reagent Recipe Hold Time

Phosphate Buffer - 24g Na2HPO4 - Indefinite at 4 C
(For dechlorination) - 46g KH2PO4 - Throw out if biological growth
- 800mg disodium EDTA dihydrate appears.
into 100ml DI water.
- All of above into 1R DI water.
- 20mg H2Cl2 preservative.
- Autoclave 45 minutes.
DPD Indicator - 1g N,N-Diethyl-p-phenylenediamine - Discard if pink color appears.
oxalate - Store in dark bottle @ 4 C.
- 8ml 1 +3 H2SO4
- 200mg disodium EDTA dihydrate
- Dissolve all of the above in 1R DI
water
0.25N FAS titrant - 49g Fe (NH4)2(SO4)2.6H2O - 3 months @ 4 C
- 10ml H2SO4 - Must be standardized.
- Dissolve above in 1R boiled and
cooled DI water.
1 + 3 sulfuric acid - one part H2SO4 to three parts distilled
water.
0.25N potassium - 12.26g, K2Cr2O7 to 1R DI water Indefinite at 4 C
dichromate
Potassium iodide crystals - Prepare small vials each containing Indefinite
1g KI

7.5 D. DO Meter Standardization

1. Turn DO meter onto RED LINE, allow to warm up for 15 minutes.

2. Fill 1R graduated Cylinder with DI water and siphon into 3 BOD bottles,
without aerating water.

56
-56-
7.5 D. DO Meter Standardization (continued)

3. To two of the bottles add 2 mls of DO reagent #1 and 2 mls of DO

reagent #2 below surface.

4. Invert bottles until uniform consistency. Let stand until floc settles to
50% bottle volume. Invert a second time and let stand again.

5. Add 2 mls of DO #3, cap and invert until floc is complete gone.

6. Pour contents into 50 ml wide-mouth flask and titrate using .0375N

Na2S2O3 until pale yellow. Add 1 ml of starch indicator (1/2 dropper
full and swirl. Titrate slowly until blue becomes clear.

7. Add as many drops of black titrant (potassium bi-iodate) as are necessary

to bring blue color back. If one drop does this, record the reading on
the buret. If it takes more than 1 drop, multiply the number of drops
by 0.05 mls and subtract from buret reading.

Reading on buret is in milliliters and is equivalent to oxygen in

milligrams per liter. Report readings to nearest 0.05 ml.

8. Average two titrations and record results in record book.

9. Place DO probe into third bottle making sure there are no air bubbles
trapped. Turn knob to zero and adjust until zeroed.

57
-57-
7.5 D. DO Meter Standardization (continued)

10. Turn knob to calibrate 1-10, turn on stirrer.

11. Let sit 2 minutes and adjust until meter reads average of two Winkler
titrations. If the average DO of the Winkler Titrations and the DO from
the meter differ by more than .5, rerun Winkler. Run samples within 4
hours of calibration.

E. Preparation of seed - do earlier on day samples arrive.

1. Empty one capsule of Polyseed BOD seed inoculum into 500 ml of

dilution water.

7.5 E. Interferences

Presence of copper and other metals in the sample can adversely affect results.

See Sample Preparation for Treatment of samples for other interferences.

7.5 F. Sample Preparation and Preservation

24 Hour Composite Samples -

Sample is refrigerated at 4 C from the time the first sample is collected

until one hour before analysis is begun.

It is placed in a warm water bath before analysis until the sample temp
reaches 20 C. The sample is thoroughly mixed just before being poured

58
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7.5 F. Sample Preparation and Preservation (continued)

24 Hour Composite Samples - (continued)

off. Dechlorination: Samples are dechlorinated using sodium sulfite.

Dechlorination is described in the Procedure section (7.5 G).

Seeding - All samples are seeded. The process is described in the Procedure
section 7.5 G.

pH: The sample pH is adjusted to between 6.5 to 7.5 if necessary using sulfuric
acid or sodium hydroxide (usually not necessary).

7.5 G. Procedure

A. Dilution Water Prep - 24 hours before use.

1. Wash 20 R carboy and siphon tubing with micro and rinse very
well with DI water.

2. Fill carboy with DI water filtered through 0.2 Fm filter.

-10R for first sample

-3R for each sample thereafter.

ex: 7 samples would require 28R (10 for the first then 3 X 6 = 18
for rest.)

59
-59-
7.5 G. Procedure (continued)

3. Add 1 ml per R for each:

MgSO4

CaCl2

FeCl3

4. Aerate for about 15 minutes.

5. Store in 20 C incubator for at least 24 hours and no longer than

5 days.

6. Just prior to use add 1 ml phosphate buffer per liter; aerate 15

minutes; let sit for 1 hour, then use.

7. Be sure temp. Is 20 to 23 C.

8. Be sure DO is between 7.5 and 8.5 mg/1.

a) If >8.5 mg/l, shake closed container vigorously, open and

let sit.

b) If <7.5 mg/l, aerate some more.

60
-60-
7.5 G. Procedure (continued)

B. Glassware prep. - Label all ahead of time.

Bottles 1000 ml Graduates

Blank 3 1
GGA 3 1
Seed 9 (3/dilution) 3 (1/dilution)
Sample 1 9 (3/dilution) 3 (1/dilution)
Sample 2 9 (3/dilution) 3 (1/dilution)

C. Dechlorination of samples.

1. Just prior to use, prepare 0.78g Na2SO3 to 500 mls DI water.

2. To a 500 ml wide-mouth flask, add:

- 100 ml sample

- 5 mls DPD indicator

- 5 mls PO4 buffer

- 1 g KI crystals (may be pre-weighed in small vials)

3. A faint pink-red color after 2 minutes indicates chlorine is

present. Stop here if no color.

61
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7.5 G. Procedure (continued)
4. Using standardized 0.00282N FAS, titrate to clear endpoint. Ml
FAS used equals mg/l Cl2.

5. Dechlorinate with Na2SO3 using in ml 5 times the amount of

chlorine per liter.

Example: Cl2mg/l = FAS used

2.2 mls FAS used = 2.2 mg/l Cl2

2R sample: 2R x 2.2 x 5 = 22 ml Na2SO3

Add 22 mls Na2SO3 to 2R sample to dechlorinate,

shake sample. _________________________

6. After 15 minutes repeat steps 2 and 3. If no pink color, then

sample is

2. Aerate and stir for at least 60 minutes.

3. When pipetting be sure flakes of bran do not get into pipette.

62
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7.5 G. Procedure (continued)

F. Seeding Chart

Expected Seed BOD mg/l Seed Dilution (%)* Seed (ml) to dilutions +
50 (Polyseed range) 2, 5, 10 20
75 2, 5, 10 15
100 1, 2, 5 8
125 1, 2, 5 8
150 1, 2, 5 6
175 1, 2, 5 6
200 0.51, 2, 5
225 0.5, 1, 2 5
250-400 0.5, 1,2 3

* Seed dilutions calculated for an approximate DO depletion of 3 mg/l

+ calculated for a DO depletion of approximately 1 mg/l

2. Seed Calculation:
0.8 mg/l ml seed used in samples % GGA
x 1000 '
Expected seed BOD mg/l

63
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7.5 G. Procedure (continued)

G. Dilution Technique.

1. BOD rules:

a) Blank depletion must be <0.2 mg/l.

b) Want a dilution such that residual DO is at least 1.0 mg/l.

c) Want a dilution such that depletion is at least 2.0 mg/l..

2. Calculation for dilution:

2 mg/l x 1000 ml
' mls sample to dilute 1R
Expected sample BOD

Example: Expected BOD is about 15 mg/l, you want 2 mg/l depletion.

2 mg/l x 1000 ml
' 133 ml to 1R or 13.3%
15 mg/l

3. General ranges for dilutions:

0.0 - 1.0 % for strong industrial wastes

1 - 5% for raw and settled wastewater
5 - 25% for biologically treated effluent
25 - 50% for polluted river waters

H. Actual procedure on day samples started.

7.5 G. Procedure (continued)

64
-64-
1. Add 10 ml GGA ampule to 1R graduated cylinder (gives 2%
GGA).

2. Add seed (usually 20 ml for polyseed) to GGA and sample

cylinders.

3. Prepare dilution of samples in graduated cylinders (be sure

samples at 20 C).

4. Set up sample sheet to receive data (see sample data sheet).

5. Transfer sample dilutions, GGA, blank and seed dilutions to BOD

bottles (3 bottles per dilution).

6. Put plastic caps on 2 bottles for each dilution and place in 20

incubator for 5 days 3 hours.

7. Read DO of third bottle for each group and record as Initial DO

(see sample data sheet).

8. Read DO final after 5 days 3 hours and record both values for
each dilution.

65
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7.5 G. Procedure (continued)

Calculations:

BOD of seed x ml seed used

A. Seed factor:
1000 ml

40 mg/l x 15 ml
Example: ' 0.6 mg/l
1000 ml

B. DO depletion:

DOI & DOF ' DO depletion

DO1' Initial DO

DOF ' average final DO

Example: 8.20 mg/l & 5.4 mg/l ' 2.8 mg/l (depletion)

DO depletion & Seed Factor

C. BOD mg/l:
% Dilution

2.8 mg/l & 0.6 mg/l

Example: ' 4.4 mg/l
.5 (50% dilution)

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7.5 G. Procedure (continued)

Quality Control:

1. Blanks are run as a bottle and dilution water check. Values #0.2 mg/l are
acceptable.

2. GGA should run 198 30.5 mg/l.

3. A duplicate should be run in each batch.

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 5 Day BOD Data Sheet
Sample Locations:Sample Data Sheet
Lab ID Numbers: 96274 99385
Date & Time Sampled: 10-21-92 10:00 a.m. Date & Time Sample arrived at Lab: 10-
12-92 1:00 pm
Date & Time BOD Setup: 10-21-92 3:00 p.m. Date & Time Final DO Read: 10-26-92

1:00 p.m.
Analyst:

Sample % Dil ml Seed DOI DOF DOF Avg. DO Seed DO BOD

DOF Depl. Factor Depl. mg/l

Blank --- --- 8.80 8.75 8.65 8.70 0.10 --- --- ---
GGA 2 20 8.75 4.30 4.25 4.28 4.47 0.46 4.01 200.5
S1 2 20 8.75 8.20 8.20 8.20 0.55 --- --- <2
Depl.
S2 5 50 8.75 7.70 7.65 7.68 1.07 --- --- <2
Depl.
S3 10 100 8.80 6.60 6.40 6.50 2.30 --- --- 23
96274 A 5 20 8.75 6.00 6.05 6.07 2.68 0.46 2.22 44.4
B 10 20 8.75 3.65 3.70 3.67 5.08 0.46 4.62 46.2
C 30 80 <1 <1 --- --- --- --- --- ---

99385 D 5 20 8.75 6.55 6.65 6.60 2.15 0.46 1.69 <2

Depl.
E 10 20 8.75 5.10 4.95 5.03 3.72 0.46 3.26 32.6
F 30 20 8.80 <1 <1 --- --- --- ---

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 Sample % Dil ml Seed DOI DOF DOF Avg. DO Seed DO BOD
DOF Depl. Factor Depl. mg/l

Notes and Calculations:

23 mg/l x 20 mg/l
Seed Factor: ' 0.46 mg/l
1000 ml

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 5 Day BOD Data Sheet - Continued

Sample % Dil ml Seed DOI DOF DOF Avg. DO Seed DO BOD

DOF Depl. Factor Depl. mg/l

Notes and Calculations:

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8.0 Reagent/Standard Quality, Preparation and Traceability

8.1 All standards and reagents are prepared from reagent grade materials, primary standards or they
are purchased from reputable vendors. Reagents are stored according to manufacturers
instructions and discarded upon expiration. Standards and reagents are prepared using bala
nces that are calibrated daily, Class A volumetric glassware and ASTM Type II reagent water.

Once a solution is prepared it is labeled with the solution name or description, storage
requirements, concentration or normality, preparation and expiration dates and initials of
preparer. Expiration dates for standards and reagents are specified in methods that are adhered
to unless degradation prior to this date is observed. Log books are utilized to record the
preparation of standards.

Calibration standards (working standards) are dilutions or mixtures of stock standards

used to calibrate an instrument. These standards are prepared or restandardized frequently as
specified in Laboratory Standard Operating Procedures (SOPs). New standards are checked
again old standards to insure there has not been an error in preparation.

Quality control reference samples are analyzed along with most analytes, depending upon
availability to validate standards, technique and methodology. Quality control reference samples
are prepared from a different source than that used in the preparation of standards for use in the
standard curve and are US EPA certified, if possible.

9.0 Calibration Procedures and Frequencies

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9.1 Instrument Calibration Procedures

All instruments and equipment used are routinely calibrated by laboratory personnel or by
external calibration agencies or equipment manufacturers. Maintenance schedules can be found in the
Preventative Maintenance Section of this manual (Section 1.0). Instrument calibration procedures,
frequencies, standards and traceability are summarized in Table 9.2 To insure that instruments have
performed adequately throughout the analysis, it is laboratory practice to run a standard or quality
control reference sample at the end of an extended run.

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9.1 Instrument Calibration Procedures (continued)

Table 9.2

Instrument/ Calibration Procedure Calibration Quality Control

Analytes Frequency Standard Standard
(Traceability) (Traceability)
pH/Millivolt Meter Daily 2 point Certified buffers EPA Reference
- pH calibration (NIST traceable) Sample (US EPA)
bracketing
sample
Conductance meter Daily .01M Potassium Primary Grade Conductivity
- conductivity Chloride Reagents Standards (NIST)
Analytical Balances Daily Calibrated Calibration Weights NBS
according to verified using Traceable
manufacturers Class -S weights
directions.
Spectrophotometer Daily 3 - 5 Point Primary Grade EPA Reference
- COD Calibration Reagents Sample (US EPA)
Turbidity Meter Daily Calibrated AMCO Sealed AMCO Sealed
- turbidity According to Primary Secondary Standards
AMCO Calibration
instructions Standards
Dissolved Oxygen 4 Hours Winkler Primary Grade (NIST traceable)
Meter Titration Reagents
- BOD
- DO
Cloroimeter (Fixed Each use Internal NIST SMR 930 EPA Reference
Photometer) instrument S/M99 Sample (US EPA)
- Chlorine calibration and
Hach Spec T
Color Standard
COD Reactor Daily 5 Point Primary Grade EPA Reference
- COD Calibration Reagents Sample (US EPA)
Millivolt Meter Daily 4 Point Primary Grade EPA Reference
- TKN Calibration Reagents Sample (US EPA)

2 Point
Calibration
Probe

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 9.1 Instrument Calibration Procedures (continued)

Table 9.2

Instrument/ Calibration Procedure Calibration Quality Control

Analytes Frequency Standard Standard
(Traceability) (Traceability)
Thermometers Annually/ Accuracy Thermometer
- incubators Semi- checked over NIST
- pH Annually range to be used.
- conductivity (bacteriology) Deviations
recorded on
thermometer.
Incubators Twice Daily Temperature Thermometer
- bacteriology Check NIST
Autoclave Weekly Sterility Check Spore Strips
- bacteriology Temperature Maximum
Check Thermometer
NIST
Refrigeration Units Daily Temperature Thermometer
- reagent sample storage Check (NBS)

10.0 Data Handling - Reduction, Validation and Reporting

10.1 Data Reduction

Raw data is recorded directly onto bench sheets by the technician who actually
performed the analysis.

If more than one analyst is involved in setting up an analysis or reading results (Ex. BOD
initials DCs determined by one person and final DOs by another) both analysts initial the bench
sheet. All calculations are included on bench sheets.

Only the chief operator can transfer information from the bench sheets to WR-43 report
forms.

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Data Handling - Reduction, Validation and Reporting (continued)

10.2 Data Validation

The analyst who generates the data has the prime responsibility for its correctness and
completeness. It is the analysts responsibility to verify that the instrument was calibrated and
was performing properly.

The chief operator looks over and double checks the bench sheets. He checks all
calculations, looks to see that all data makes sense and that the numbers were rounded properly
(section 10.5) and that proper significant numbers were recorded (section 10.4).

After transferring the data to the WR-43 report forms he checks for transcription errors.

The assistant operator performs a quick check of bench vs. WR-43 data before the report
is given to the town manager for his signature.

The town manager looks at the WR-43 report form and questions the chief operator
concerning any unusual or suspicious looking result before signing the report. As a rule the
town manager and chief operator meet to discuss the report regardless of whether or not there are
irregularities.

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Data Handling - Reduction, Validation and Reporting (continued)

10.4 Significant Digits

Parameter Significant Digits

BOD - No digit after decimal point. 28 mg/L
Chlorine Residual - Two digits after decimal point. 0.51 mg/L
Coliform - No digits after decimal point. 50/100 ml.

TKN, NH3, NO3 - One digit after decimal point. 17.6 mg/L
D.O. - Two digits after decimal point. 7.35 mg/L

Settleable Solids - One digit after decimal point. 5.1 ml/L

Metals - One digit after decimal point. 436.3 ppb
pH - Two digits after decimal point. 7.00 pH units
Suspended Solids - No digit after decimal point. 22 mg/L
Temperature - One digit after decimal point. 17.2C

10.5 Rounding Policy

All digits are used in calculations, then are rounded, using the following guidelines.
Numbers following decimals shall be rounded to the next higher or lower number based on this
method.

For example: 3.57 is rounded to 3.6

2.41 is rounded to 2.4

7.55 is rounded to 7.6

7.44 is rounded to 7.4

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11.0 Preventive Maintenance Procedures and Schedules

11.1 All laboratory equipment is serviced and professionally calibrated by QC Services on an annual
basis. A service contract is maintained to include annual equipment calibration.

11.2 Routine and professional calibration/maintenance schedules are summarized in Table 11.2.

11.3 Preventive maintenance responsibilities are assigned to specific laboratory personnel. Only the
lab supervisor is allowed to perform other than routine calibration or minor repair.

11.4 A maintenance log is kept in the lab for each instrument. All calibration, repairs and service
visits are recorded an entitled by the responsible party.

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11.0 Preventative Maintenance Procedures and Schedules Table 11.2 (Continued)

Instrument - Manufacturer - Model Calibration Maintenance Preventative

Frequency Contractor Maintenance
Schedule
Atomic Absorption, Perkin-Elmer 3030B Daily Perkin-Elmer As needed
Autoclave, Barnstead 4/year MDT Biologic Co. 4/year
Waterbath, Precision Scientific - 83 1/year QC Services 1/year
Waterbath, Blue M 1/year QC Services 1/year
Air Incubator, Boekel 1/year QC Services 1/year
pH/Millivolt Meter, Orion Model 811 Each use QC Services 1/year
pH/Millivolt Meter, Orion Model 720A Each use QC Services 1/year
Ratio Turbidmeter Hach Model 18900 Daily QC Services 1/year
Spectrophotometer, Bauch & Spectronic 100 Daily QC Services 1/year
Dissolved Oxygen Meter, YSI Model 57 Every 4 hours QC Services 1/year
Fluorometer, Turner Model 111 Yearly QC Services 1/year
COD Reactor, Hach Model Yearly QC Services 1/year
Centrifuge, International Equipment --- QC Services ---
Conductance Meter, YSI Model 32 Yearly QC Services 1/year
Chlorine Meter, Hach Model DR100 Each use QC Services ---
Balance, Mettler AE 200 Daily QC Services 1/year
Balance, OHAUS B1500D Daily QC Services 1/year

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12.0 Corrective Action Contingencies

12.1 Corrective actions are required as a result of less than acceptable Performance Evaluation study
results, or poor comparison in split sample analyses (State lab results significantly different than
facility results). The steps taken in the corrective action process include:

Identify and define the problem.

Determine who will be responsible for investigating the problem.

Find the cause of the problem.

Determine the actions needed to eliminate the problem.

Implement the corrective actions and

Establish the effectiveness of the corrective action.

Usually the lab supervisor is responsible for initiating the corrective action under these
conditions. Documentation of actions taken and their effectiveness is forwarded to the lab
supervisor for review and distribution.

12.2 Corrective actions might also be initiated by an analyst during or after sample analysis. These
actions may be necessary because of

Unacceptable blank results (BOD blank depletion >.2 mg/L)

Suspicious positive control results (every few colonies on E Coli. positive control
sample).

QC data outside the warning or control limits for precision and accuracy.

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12.0 Corrective Action Contingencies (continued)

Duplicate or replicate results are inconsistent.

Under these conditions the analyst generating the data is expected to initiate and
document corrective action.

12.3 Corrective Actions Required for Specific Problems are Listed in Table 12.4

Table 12.4

Problem Corrective Action

Blank contamination is indicated Determine cause of contamination, eliminate
cause and repeat analysis.
Unexpected, unusual results occur more than Investigate possible causes. Try to eliminate
once every 20 analyses interferences. Keep a close watch on future
analyses.
One or more data points falls outside the Determine cause eliminate it an repeat analysis.
control limit.
Two out of three successive data points fall Repeat analysis. If the next data point is less than
outside the established warning limits. warning limit continue the analyses. If the next
point exceeds warning limits discontinue analyses
and correct the problem.
4 out of 5 successive data points exceeds 1 Analyze another sample. If the next data point is
standard deviation or are in decreasing or less than 1 standard deviation or changes the order
increasing order. continue analyses. If not discontinue and correct
the problem.
Six successive sample results are Analyze another sample. If the next result is on
above/below the central line on the control the other side of the central line continue analyses.
chart. If the next point is on the same side of the central
line discontinue analyses and correct the problem.

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13.0 Quality Control Procedures (General)

This section describes the method used at this laboratory to evaluate the quality of data
generated.

13.1 Sampling Quality Control Checks

Sampling QC checks provide information regarding the precision and accuracy of the
entire process from sample collection through analyses. Included in this category are:

13.1.1 Equipment Blanks

Equipment blanks are used to determine if contamination has been introduced

through contact with sampling equipment or to verify effectiveness of equipment
cleaning procedures.

Clean laboratory water is pumped through the Isco sampler. We usually place the
sampler probe in a 4 liter jug of RO/DI water, pump for 10 - 15 seconds, purge for 5
seconds. This process is repeated 2 to 3 times before the equipment blank sample is
collected. This sample is then taken to the lab and processed along with other samples.

13.1.2 Split Samples

Split samples are replicate samples, two aliquots taken from the same sample
container. The samples are then analyzed independently by our own lab and a contract
laboratory. If significant differences are noted the cause is determined and corrected.

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13.0 Quality Control Procedures (General) (continued)

13.1.3 Duplicate Samples

Duplicate samples are samples collected at the same location at the same time.
Collection of duplicate samples serves as a check on sampling and processing technique.
Each sample is analyzed individually. Results must be within an acceptable range (10%)
or the cause must be determined and corrected.

13.1.4 Replicate Samples

Replicate sample are two aliquots taken from the same sample container that are
processed and analyzed separately. The results are used to measure analytical precision
from sample preparation through analysis. Certain analyses are run in replicate every
time the test is performed. A minimum 10% replication schedule is established for all
analyses.

13.2 Procedures Used to Assess Data Quality

13.2.1 Precision

Precision is a measure of the closeness with which multiple analyses of a sample agree
with each other. We calculate precision from results of replicates and duplicate analyses of
quality control samples.

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13.2 Procedures Used to Assess Data Quality (continued)

13.2.1 Precision (continued)

Here at the WWW Wastewater laboratory we use Relative Percent Difference (RPD) as a
measure of precision. The formula used to calculate RPD is:

(C1 & C2) x 100%

RPD '
(C1 % C2) /2

Where: RPD = relative percent difference

C1 = larger of the two observed values

C2 = smaller of the two observed values

If calculated from three or more replicates, we use relative standard deviation rather than
RPD:

RSD = (s/y) X 100%

where: RSD = relative standard deviation

s = standard deviation
y = mean of replicate analyses

Standard deviation is defined as follows:

'Xi &
('Xi)
n
SD '
n & 1

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13.2 Procedures Used to Assess Data Quality (continued)

13.2.1a Control Charts

Control charts are used to demonstrate the precision calculated from replicates
and quality control sample duplicates.

The Relative Percent Difference (RPD) values for each sample are plotted on
control charts and where upper and lower warning and control limits are depicted.

These control charts are used by the analysts to help make them aware of
suspicious or out-of-control variability at the time of analysis. The warning and control
limits are recalculated annually. An example of a control chart cal be found at the end of
the section.

13.2.1b Control Limits

Control limits are defined as the mean + / - 3 standard deviations. An RPD value
that falls outside the control limits is considered out-of-control and requires the analysts
to repeat the analysis immediately. If the repeat value is within the control limit analysis
may continue. If the repeat value exceeds the control limit analysis must stop and the
problem must be corrected.

13.2.1c Warning Limits

Warning limits are narrower than control limits and are defined as the mean + / -
2 standard deviations. An RPD value that falls outside the warning limit is considered
suspicious. If two out of three consecutive points exceed a warning limit the analyst
must calculate the RPD on another sample. If the warning limit is exceeded analysis
must stop and the problem must be corrected.

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13.2 Procedures Used to Assess Data Quality (continued

13.2.2 Accuracy

Accuracy is expressed as a percent bias or percent recovery and is determined from the
analysis of quality control reference samples or spikes. Method accuracy is calculated on a daily
basis and summarized annually in the Laboratory Quality Assurance Plan.

Percent recovery is calculated from spike results using the following equation:

S&U
% R = 100% x
Csa

where: %R = percent recovery

S = measured concentration in spike aliquot

U = measured concentration in unspiked aliquot

Csa = actual concentration of spike added

(O & T)
% B ' 100 x
T

Where: %B = percent bias

O = measured concentration of reference material

T = actual concentration of reference material

o:\home\lori\andy\labqc.mnl

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Vermont Department of Environmental Conservation
Quality Assurance Guidelines for
Wastewater Treatment Facility Laboratories

1996

Prepared by:

Andrew Fish, C.E.T.

Wastewater Laboratory Specialist
for the VT Dept. of Environmental Conservation
 PREFACE

These guidelines have been developed to assist operators and technicians at Vermont
Wastewater Treatment Facilities to prepare acceptable Quality Control Manuals specific to their
facilities. It is further hoped that the guidelines will encourage the implementation of these
procedures. The quality control procedures outlined in this manual represent the minimum
requirements considered acceptable by the Vermont Department of Environmental Conservation
for facilities regulated under the NPDES program.

1
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 The laboratory quality control manual described in this guidance document is an integral
part of the appropriate quality assurance procedures required of all NPDES regulated facilities
pursuant to Title 40 of the Code of Federal Regulations, Part 122.41 (e) any commercial
laboratory performing analyses for an NPDES regulated facility must certainly meet these
minimum requirements.

It is absolutely essential that all laboratories analyzing wastewater compliance samples

follow defined quality assurance procedures. Those procedures must be documented in a
laboratory quality control manual.

To satisfy minimum NPDES requirements a quality control manual must address the
following items. (Each of which will be discussed in detail in the following pages.)

I. Quality Assurance Program Plan Elements

II. Staff Organization and Responsibilities
III. Quality Assurance Objectives and Activities
IV. Sample Collection Procedures
V. Sample Handling and Documentation Procedures
VI. Laboratory Water Quality
VII. Standard Operating Procedure for Each Analytical Method
VIII. Reagent/Standard Quality Preparation and Traceability
IX. Calibration Procedures and Frequencies
X. Data Handling - Reduction, Validation and Reporting
XI. Preventive Maintenance, Procedures and Schedules
XII. Corrective Action Contingencies
XIII. Quality Control Procedures (General)

2
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 In order to produce a truly useful QA/QC manual a table of contents should be
included. You may also wish to include things such as laboratory utilities information,
electrical supply, compressed air source, etc. or anything that might effect the function of
the laboratory.

I. Quality Assurance Project Plan Elements

This section of the manual simply lists the topics which will be included in your
quality assurance plan. It is basically a table of contents without the page numbers. It
should be the first step in preparing the manual as it will help you to organize your
thoughts and establish distinct sections.

II. Staff Organization and Responsibilities

Laboratory analytical centers can obviously vary considerably in size, number,

complexity of analyses performed, and in the number of people employed.

This section of the manual should identify all persons responsible for laboratory
activities from sampling to preparation and signing of the Discharge Monitoring Report.
It should describe the responsibilities of each of the individuals in certain terms;
identifying each individual using that persons name (not just the position - i.e. Andy
Fish, QA coordinator not QA coordinator).

A typical municipal wastewater facilitys staff organization and responsibilities

section might include:

1). The town manager as the person responsible for signing the WR-43.
2) The chief operator as the preparer of the WR-43.
3). Assistant operator as responsible for sample collection.
4) Lab tech - responsible for performing analyses maintaining bench sheets,

3
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 assisting in data review etc.

Each individuals responsibilities and limitations should be spelled out in this

section.

As an example of limitations, an analysts responsibilities might include informing

the chief operator if accuracy values are beyond established warning and control limits.

----This clearly shows that the analysts responsibility ends with informing his
supervisor of a problem. It is presumably the chief operators responsibility to determine
the action taken from that point.

An organization chart is very useful in identifying positions (where they fit into
the scheme of things).

III. Quality Assurance Objectives

This section, besides being useful to inspectors and certification officials can be
very helpful to the person preparing the manual to figure out just what am I trying to
demonstrate in this manual? What am I really after here?

Here is an example of typical Quality Assurance Objectives.

1. To ensure data produced by the laboratory is accurate and defensible.

2. To ensure samples collected are representative of preferred universe.
3. To ensure that all laboratory procedures are EPA approved.
4. To ensure that all equipment is properly calibrated and meets EPA specifications.
5. To ensure that proper corrective actions are initiated when necessary.

III. Quality Assurance Objectives (continued)

4
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 6. To ensure that each sample is tracked from collection time until the report is
finalized - with records maintained for the required intervals. Of course in
addition to simply listing the objectives you might summarize referencing specific
sections of the manual, how these objectives will be met.

IV. Sample Collection Procedures

Careful and precise documentation of sampling procedures is absolutely

imperative to ensure that good representative samples are consistently collected. Untold
time and dollars are wasted on analysis of samples that are collected at an improper
location, time or in an unacceptable container. An exact sampling procedure should be
written for each parameter required at your facility. It is smart to include detailed
photographs and or diagrams along with sampling location descriptions.

The sampling procedures section of your QA manual should include:

1) Description of container - glass, plastic, etc.

2) Container cleaning requirements -- acid washed, DI rinsed, etc.
3) Sample preservation techniques .
4) Sample holding times.
5) Exact sampling location description.
6) Exact sampling time for most representative sample.
7) Collection method/technique - grab, composite (time-flow, auto sampler)
etc.
8) Volume of sample required for analysis.

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V. Sample Handling and Documentation Procedures

This section must include:

A written description of exactly how the samples are handled after collection.
The methods for documentation must be clearly stated. The following items must be
included in this section.

1) Documentation of exact time, date and location sample was collected.

2) Labeling of sample bottles.
3) Chain of Custody Procedures (if analysis is performed by an independent
laboratory).

a) Sample custody forms, labels, seals.

b) Sample transportation and delivery procedure.

V. Laboratory Water Quality

As many of us are painfully aware, the quality of the water we use in rinsing
glassware, preparing reagents and standards, sample dilution, and blank preparations can
be the determining factory between accurate and inaccurate analytical results.

There are rather strict monitoring requirements for laboratories analyzing

drinking water. These requirements are listed in EPA/570/9-90/008 document Manual
for the Certification of Laboratories Analyzing Drinking Water, and include:

6
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V. Laboratory Water Quality (continued)

1) General Lab Water

Parameter Limit Monitoring Frequency

Conductivity <2 umhos/cm @ 25C Monthly

2) Water for Microbiological Analyses

Tests Limit Monitoring Frequency

Conductivity <2 umhos/cm @ 25C Monthly
Total Residual Chlorine <detection limit Monthly
Heavy Metals, Single4
(Cd, Cr, Cu, Ni, Pb, Zn) <0.05 mg/L Annually
Heterotrophic Plate Count <500 CFU/ml Monthly
Water Quality Test5
(Biosuitability) 0.8 - 3.0 ratio Annually

In wastewater analysis our main concerns regarding laboratory water quality are
conductivity, chlorine and metals that might interfere with certain analyses (i.e. high
copper level can affect results in the BOD analysis).

Generally accepted monitoring frequencies and limits for those parameters are
listed below.

7
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V. Laboratory Water Quality (continued)

2) Water for Microbiological Analyses (continued)

Parameter Limit Monitoring Frequency

Conductivity <2 umhos/cm @ 25C Monthly
Total Residual Chlorine 0.0 or <detectable limit Monthly
Copper < .05 mg/L Semi-Annually
Other Metals < .05 mg/L Annually
(Cd, Cr, Ni, Pb, Zn)

* Reality Check: The Vermont Department of Environmental Conservation does

not expect a small wastewater treatment facility required to perform pH analyses
only to have their own laboratory water analyzed for all the parameters listed
above. Such a facility should consider purchasing commercially available
distilled water, where concentrations of these parameters are often listed on the
label. Larger
facilities, performing BODs and more sophisticated analyses would be expected
to verify conductivity, chlorine and copper limits. Commercial laboratories
performing numerous parameters more frequently than the average treatment
facility would be expected to perform more frequent laboratory water analyses.

VII. Standard Operating Procedures - For Each Analytical Method

This section of the manual should include a simple step - by step procedure which
details the exact method in which the analysis is performed. It should NOT simply be a
copy of an analytical method from Standard Methods but instead a practical What you
really do to perform this analysis, type of description in an easy to read format.

8
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VII. Standard Operating Procedures - For Each Analytical Method (continued)

Include specific items such as Turn the blender to setting 6 and blend for 5
minutes to get a good homogenous sample or Use only Class A volumetric glassware
from drawer #2.

Items that must be included in this section are:

1) Reference Method, number and Title

2) Description of instrumentation
3) Specific glassware and equipment cleaning procedures
4) Reagent and Standard Preparation
5) Step by Step Procedures (SOPs)
6) Calibration and Standardization procedures - specific to the analysis
7) Equations and Calculations

You may reference an SOP manual to satisfy #5 if you have a separate and
complete manual.

VIII. Reagent/Standard Quality, Preparation and Traceability

This section of the manual should describe the general procedures for preparation
of reagents and standards. The types of questions that should be addressed include:

1) Grade of materials used i.e. ASC or Analytical Reagent grade chemicals primary
standards, etc.
2) Measuring methods i.e. calibrated analytical balance, class A volumetric
glassware etc.
3) Reagent labeling: Must Include

9
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VIII. Reagent/Standard Quality, Preparation and Traceability (continued)

A. Reagent name
B. Concentration
C. Preparation and Expiration dates and/or received the dates a chemical is
received as well as the date it is opened.

IX. Calibrations Procedures and Frequencies

Documentation of all instrument or equipment calibration is essential to any good

quality assurance plan.

This section should detail the procedures, frequencies and type of calibration as
well as whether it was routine or professionally performed and finally who performed the
calibration.

Written documentation must include at a minimum:

1) Established frequency of routine calibration for each piece of equipment i.e. daily,
weekly, monthly.
2) Established frequency of professional calibrations for each piece of equipment.
3) Calibration procedures - Standards used, etc.
4) Dates and times calibrations are performed.
5) The name (* not just initials of the person who calibrated the instrument.)

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X Data Handling - Reduction Validation and Reporting

Reduction of Data

Data reduction is the process of transforming raw data into final results that are
reported in standard units to some authority. An efficient method of data reduction must
exist to reduce transcription and calculation errors. That methodology, whatever it may
entail, should be described in this section.

Validation of Data

Data validation starts with the analyst. It is usually the analysts responsibility to
ensure that instruments have been properly calibrated and are operating properly. He or
she then records the results on a bench sheet. Generally someone then transfers the data
to another form after, perhaps performing some calculations.

At this point there should be some mechanism in place to ensure that the data has
been accurately transferred and that any calculations were performed properly.

At least one other person beside the analyst should look over and double check
the bench sheet data vs. final results. One or both of these individuals must understand
the concept of significant figures and units of measures.

* More than one technical violation could have been avoided had this knowledge
been applied.

This section should describe the procedure by which data is checked. Specific
responsibilities should be spelled out here.

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X Data Handling - Reduction Validation and Reporting (continued)

Reduction of Data (continued)

Each WR-43 report form contains a certification signed by the permittees

authorized agent which states I certify under penalty of law that I have personally
examined and am familiar with the information submitted herein.

It would seem reasonable then to involve this person in the data validation
process.

It is important to include in this section.

Examples on description of

1) 8Bench sheets, lab books

2) A description of the check system used to avoid transcription and
calculation errors.
3) A procedure of how invalid data is handled on bench sheets and on the
WR-43 report form (i.e. BOD results not meeting the R1 D2) requirement
- How is this handled?
4) Rules for consistent rounding off of numerical results.

XI. Preventive Maintenance, Procedures and Schedules

Preventive maintenance is very important in order to minimize instrument Down

Time and ensure the continued accuracy of analytical results.

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XI. Preventive Maintenance, Procedures and Schedules (continued)

The old excuse I couldnt report pH results for the month of April because my
pH probe was broken, just isnt acceptable as a defense.

Proper preventative maintenance and contingency plans are essential for

uninterrupted analyses.

This section must describe procedures for routine and scheduled contract services.
Include:

1) Comprehensive routine maintenance schedule - list of instruments.

2) Professional service/maintenance schedule - list of instruments.
3) A list of employees responsible for performing maintenance.
4) A check list to ensure duties have been completed.

XI. Corrective Action

This section of the manual will outline the steps taken when any portion of the
quality assurance process become questionable or invalid.

These steps should include:

1) Identifying and defining the problem i.e. QC is outside control limits,

what caused unacceptable results.
2) A list defining need for corrective actions - i.e. (Poor blank or standards
results)
3) Actions to eliminate the problem.
4) Actions to prevent recurrence . You may include a copy of the
checklist(s)

13
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XI. Corrective Action (4) (continued)

in the manual and include a reference describing where the actual list(s) is/are
located if you desire.
5) Procedure for informing superiors of other authorities (if applicable).
6) Protocol for resampling and retesting.

XIII. Quality Control Procedures:

Quality control is defined in the 18th edition of Standard Methods for the
Examination of Water and Wastewater, 1992.

As as set of measures within a sample analysis methodology to assure that the

process is in control. This section of the manual is dedicated to describing those
measures taken to ensure process control. In it you should define terms and describe
general procedures which can be referenced in the QC portion of each analytical method.

Quality control consists of analyzing and reporting the results of quality control
standards, sample duplicates and replicates, spikes and blanks.

EPA recommends that a frequency of 10% to 20% be applied for quality control
measures. That is, for every 5 to 10 samples analyzed there should be one quality control
sample analyzed for each parameter being tested.

For commercial laboratories analyzing multiple samples, a quality control

standard or spike, a sample duplicate or replicate and a blank is expected to be run with
each batch of samples analyzed.

14
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XIII. Quality Control Procedures: (continued)

The minimum frequency for quality control analyses in laboratories performing

one sample at a time would be one-in every 10 samples. *Note: Certain analyses are
required by the Vermont Department of Environmental Conservation to be run in
duplicate 100%. (T.S.S. and E. Coli).

The Vermont Department of Environmental Conservation has the following

minimum requirements regarding QC for the common parameters listed in the Table
below:

Calibration/ QC Duplicates/
Parameter Standardization Standards Replicates Spikes Blanks

Ammonia As per method 1/10 Tests 1/10 Tests 1/Yr 1/each

BOD Meter before each use 1/10 Tests 1/10 Tests 1/Yr 1/each
IGGA/Each
if seeded

Chlorine Check standard 1/month 1/10 Tests 1/Yr 1/each

Residual Meter curve daily standard for each use

Titrimetric FAS stand 1/month 1/month 1/10 Tests 1/Yr 1/each

COD as per method 1/10 Tests 1/10 Tests 1/Yr 1/each

Cyanide as per method 1/each 1/each 1/each 1/each

E. Coli Additional QC required for E. Coli 2/Yr 1/each N/A 1/each

includes equipment sterility checks /POS
(indicator tape, Kilit ampules) control/each
each tests and pH check of test
dilution water (7>1 .2) each tests

Metals as per method 1/each 1/each 1/each 1/each

Nitrate Nitrogen as per method 1/10 Tests 1/10 Tests 1/Yr 1/each

Oil & Grease as per method 1/each 1/each 1/Yr 1/each

pH Minimum 2 point calibration each (3rd buffer)

use 1/each 1/10 Tests N/A N/A

Phosphorus 5 Standards/each 1/10 Tests 1/10 Tests 1/Yr 1/each

Settleable Solids N/A N/A 1/10 Tests N/A N/A

Total Suspended constant weights before and after 1/10 Tests 1/each N/A 1/each
Solids filtering sample

15
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XIII. Quality Control Procedures: (continued)

XIII. 1B. Spikes

Spikes are prepared by adding a predetermined quantity of a known standard to a

sample. By analyzing an unspiked sample and a spiked sample it is possible to determine
if there is a substance in the sample that interferes with test results.

From the results of analysis of a spike and unspiked sample, percent recovery can
be calculated using the equation:

S&U
% Recovery ' 100% x
Csa

Where S = Measured concentration in spiked aliquot

U = Measured concentration in unspiked aliquot
Csa = Actual concentration of spike added

If the Percent Recovery is outside the range of 80% to 100% the contents of the
sample may be interfering with the method of measurement for that analysis.

In this event the cause of the interference must be determined and corrective
actions must be initiated (i.e. change methods, etc.)

* One spike or Quality Control Standard should be run with one of every 10
samples run.

* For laboratories performing sample analysis in batches one spike or quality

control standard should be run with each batch.

16
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XIII. Quality Control Procedures: (continued)

XIII. 2B. Quality Control Standards

Quality control standards are substances of known concentration. They are used
to verify the accuracy of your analysis.

Quality control standards:

1) Must be prepared from a source different than what was used to calibrate
the instrument or prepare the standard curve.

2) The quality control standard must be specific for the parameter being
analyzed.

3) The quality control standard must approximate the concentration of the

sample and must have a value between that of the highest and lowest
calibration standards used.

For Example: If a sample is expected to have a pH of

approximately 6, calibration standards of pH 4 and 7 might be used along
with a quality control standard with a pH value of 6.

4) Remember: Quality control standards are not used to adjust the instrument
as are calibration standards. The resulted quality control standards are
simply recorded an compared to the actual known value.

17
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XIII. Quality Control Procedures: (continued)

XIII. 2B. Quality Control Standards (continued)

5) Unless control charts are used to determine the acceptable quality control
standard range. Results of the quality control standard must fall within a
range of 20% of the known value. That is the Percent Error must be
from 80% to 120% of the actual standard

Observed Value & Known Value

Percent Error ' x 100
Known Value

6) If results of a quality control standard do not fall within the acceptable

range the cause must be determined and recorded on bench sheets or other
appropriate record books. A not should be placed in the comments section
of the WR-43 report form. Corrective actions must be implemented.

III. Duplicates/Replicates
1) Duplicate samples are samples which are collected in two separate containers at
the same time and place.

2) Replicate samples are samples that are collected in a single container and are
poured off into a second container for separate analyses of the same parameter(s).

3) An absolute minimum of one duplicate or replicate per 10 samples is required.

4) The result of duplicate/replicate samples should not vary by more than 20% from
the original sample.

XIII. 2B. Quality Control Standards (continued)

18
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III. Duplicates/Replicates (continued)

5) Do not average original and duplicate/replicate results. Record the result of the
original sample. Then record on bench sheets or other appropriate record book
the Percent error for the duplicate/replicate.

Percent error is determined by applying the following equation.

Observed Value & Known Value

Percent Error ' x 100
Known Value

If the duplicate/replicate results fall outside the acceptable ( 20%) range of the
original sample a note should be made in the comments section of the WR-43
report form and on the bench sheet. Corrective action should be implemented.

IV. Blanks

1) A blank consists of either laboratory water or parameter specific dilution water.

Blanks provide a check of the laboratory water quality as well as a check of the
analysts analytical technique.

2) Blank must be analyzed with each analysis where applicable.

19
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XIII. 2B. Quality Control Standards (continued)

IV. Blanks (continued)

3) Acceptable blank values are included with each analytical method.

For example:

BOD = # 0.2 mg/L

E. Coli = 0 colonies/100 mls

If blank limit values are exceeded a note must be included in the comment section of the
WR-43 report form, and on the bench sheet. In some instances such as in the E. Coli
analysis, this invalidates the sample data. (Check the method info) In any blank
exceedance corrective action contingencies must be implemented.

20
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 The following pages contain an

Example QC Manual

Hopefully this example will help you in preparing

your own manual.

Remember: The manual must be specific to your own

facility and will remain a work in progress requiring
periodic updating and revisions.

21
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 Table of Contents
A. Title Page Page 17A
B. Introduction Page 18
1. Quality Assurance Project Plan Elements Page 19
2. Laboratory Staff Organization and Responsibilities Page 20
2.1. Town Manager Page 21
2.2 Chief Operator Page 21
2.3 Lab Supervisor Page 21
2.4 Operators Page 22
. 2.5 Skills and Training Page 22
3.0 Quality Assurance Objectives and Activities Page 23
4.0 Sample Collection Procedures Page 23
4.1 General Guidelines Page 23
4.2 Sample Type and Holding Times Page 24
4.3 Sample Preservation Page 25
5.0 Sample Handling and Documentation Procedures Pages 27 & 28
6.0 Laboratory Water Quality Page 28
6.1 Deionized Water Pages 28 & 29
7.0 Standard Operating Procedure for Each Analytical Method Pages 30
7.1 pH Pages 30 to 32
7.2 Total Residual Chlorine Pages 32 to 35
7.3 Settleable Solids Pages 35 to 36
7.4 Total Suspended Solids Pages 36 to 40
8.0 Reagent/Standard Quality, Preparation and Traceability Pages 40 to 41
9.0 Calibration Procedures and Frequencies Page 41
9.1 Instrument Calibration Procedures Pages 41 to 43
10.0 Data Handling-Reduction, Validation and Reporting Page 43
10.1 Data Reduction Pages 43 to 44
10.2 Data Validation Page 44
10.3 Reporting Pages 45 to 46
11.0 Preventive Maintenance, Procedures and Schedules Page 47
12.0 Corrective Action Contingencies Pages 48 to 49
13.0 Quality Control Procedures (General) Pages 50 to 55

22
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 INTRODUCTION

This Quality Control/Assurance Manual is written

specifically for the WWW Wastewater Facility in an attempt to
assure the continued high quality and accuracy of analytical
results. This will be accomplished by monitoring the accuracy
and precision of these results, by providing clear, concise
Standard Operating Procedures defining sampling and analytical
procedures specific to the WWW facility and by clearly defining
the responsibilities of all personnel regarding required
performance and documentation.

The WWW facility performs analyses required by the

NPDES permit for discharge to the WWW River, in-house
progress control analyses, and occasionally pH and chlorine
analyses for the Water Supply Department.

Analytical results generated for NPDES permit are

submitted to the State of Vermont on a monthly basis. Results
of process control analyses are recorded on bench sheets and
maintained in-house files for a period of three years.

23
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I. Quality Assurance Project Plan Elements

The fourteen Quality Assurance Project Plan Elements addressed in this manual
are:

Title Page
Introduction
Staff Organization and Responsibilities
Quality Assurance Objectives and Activities
Sample Collection Procedures
Sample Handling and Documentation Procedures
Laboratory Water Quality
Standard Operating Procedure for Each Analytical Method
Reagent/Standard Quality Preparation and Traceability
Calibration Procedures and Frequencies
Data Handling - Reduction, Validation and Reporting
Preventive Maintenance, Procedures and Schedules
Corrective Action Contingencies
Quality Control Procedures (General)

24
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2.0 Laboratory Staff Organization and Responsibilities

The Fuzzyville Wastewater Treatment facility has 5 permanent full-time

employees. At some time, all employees are responsible for collection of samples either
on a daily basis or on a rotating weekend schedule.

One employee is dedicated to laboratory work only, while all 5 might perform
daily analyses including pH, Cl2 and settleable solids.

The organizational structure is summarized in Table I below.

Table I

25
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2.1 Town Manager

The town manager is responsible for signing the WR-43 report form.

Before signing he/she questions the chief operator or to the reports accuracy and
any special comments.

Reviews all data.

Compares WR-43 results to bench sheet results.

Provides final data check.

Sends report to the proper state authority.

Makes final decision on equipment purchases after conferring with chief operator.

Reviews and is familiar with the laboratory QC manual.

2.2 Chief Operator

The chief operator is responsible for signing the WR-43 report form as the
preparer.

Transfer all results from bench sheets to the WR-43 report form.

Compares final WR-43 results to bench sheets to guard against transcription

errors etc.

Reviews an is familiar with the laboratory QC manual.

Schedule special sampling and analytical projects.

Reviews bench sheet data to ensure proper WC was practiced.

26
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 Is responsible for purchasing laboratory equipment.

Occasionally collect and analyzes effluent samples for pH.

On occasion when pH is analyzed he/she properly calibrates the pH meter.

2.3 Lab Supervisor

Is responsible for overall technical quality of the work performed in the

laboratory.

Ensures the use of acceptable Standard analytical methods.

Provides training to all persons responsible for sampling.

Is responsible for preparation and revisions of laboratory QC manual.

Informs chief operator of any equipment needs.

Performs all instrument calibrations and reagent preparations.

Maintains proper bench sheets and equipment maintenance logs.

Responsible for maintaining current SOPs.

Collects and composites effluent samples.

27
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 2.4 Operators

Are responsible for sampling of specific parameters.

Are properly training and responsible for analysis of pH, Total Residual Chlorine
and Settleable solids when necessary.

Maintain proper bench sheets and calibration logs.

2.5 Skills and Training

EDUCATION
EXPERIENCE
Time & Type
RESPONSIBILITIES
REQUIREMENTS SPECIFIC TO A POSITION

Example:

SPECIAL
NAME POSITION EDUCATION EXPERIENCE SKILL RESPONSIBILITIES REQUIREMENTS

Kim Lab B.S. 15 yrs. as lab Phosphorus As described in None

Istry Supervisor Chemistry analyst determination organizational
using auto chart
analyzer

28
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 EXAMPLE

3.0 Quality Assurance Objectives

a. To ensure data produced by the laboratory is accurate and defensible.

b. To ensure samples collected are representative of preferred universe.

c. To ensure that all laboratory procedures are E.P.A. approved.

d. To ensure that all equipment is properly calibrated and meets E.P.A.

specifications.

e. To ensure that proper corrective actions are initiated when necessary.

f. To ensure that each sample is tracked from collection time until the report is
finalized - with records maintained for the required intervals.

4.0 Sample Collection Procedures

4.1 General Guidelines

a. Samples are collected in a well mixed area at the center of the channel avoiding
eddies, backwaters and area where settling might take place.

b. Influent samples are collected after the comminuter but above the RAS line. See
Diagram.

c. Effluent samples are collected after all treatment processes just before discharge
to the stream. See Diagram for specific parameters.

29
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4.1 General Guidelines (continued)

d. Grab samples

Grab samples are collected via a Nalgene container attached to an 8 foot

extendable aluminum pole. All grab samples are collected and immediately
returned to the lab for analysis.

*Note* E. Coli - collected directly into sterilized 250 ml. Plastic bottle containing
4 drops 10% Sodium Thiosulfate solution.

e. Composite samples

All composite samples consist of 24 disrete samples collected hourly via an Isco
automatic sampler. Each disrete sample (approximately 500 mls) is distributed
into properly cleaned, 1 liter Isco sample bottles. Composite samples are cooled
by ice packs which are placed in the sampler wells. Samples are composited at
the lab based on readings from flow charts.

30
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 SAMPLING PROCEDURES

4.2 Type of Sample and Holding Time

Type of Holding Sample

Parameter Sample Time Container Location
pH Grab 15 Minutes Plastic Bottle Refer to Diagram
Temperature Grab At Site Plastic Bottle Refer to Diagram
Dissolved Oxygen Grab 15 Minutes B.O.D. Bottle Refer to Diagram
B.O.D. Composite Flow 6 Hours Composite Refer to Diagram
Proportional Sampler Plastic
Bottles
Total Coliform Grab 1 Hour Sterile Sample Refer to Diagram
Fecal Coliform Plastic Bottle
E. Coli w/Sodium
Thiosulfate
Chlorine Residual Grab 15 Minutes Opaque B.O.D. Refer to Diagram
Glass Bottle
Total Suspended Composite Flow 6 Hours Composite Refer to Diagram
Solids Proportional Sampler Plastic
Bottles
Specific Grab 15 Minutes Glass Refer to Diagram
Conductance
Metals Grab/Comp 6 Months Amber Glass Refer to Diagram
TKN Grab 1 Hour Plastic Bottle Refer to Diagram
Settleable Solids Grab 30 Minutes Plastic Bottle Refer to Diagram

31
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 4.3 Sample Preservation

If samples are taken and not run within the time frame as in Chart 9.1 then the following
chart applies.

PRESERVATION CONDITIONS

Parameter Container Volume Preservation Holding Representative Sampling

Time Time
BOD, TSS P 1L 4 C 24 Hours 8 AM - 8 AM
TKN P .5L H2SO4 pH < 2.0 28 Days 8 AM - 8 AM
4C
Oils & Grease G 1L HCl pH < 2.0 24 Hours Between 8 AM and 12 PM
Metals GA .2L HN03 pH < 2.0 6 Months 8 AM - 8 AM
Phenols G .5L H3PO4 pH 24 Hours 8 AM - 8 AM
< 2.0 + 1t CuSO4 4C
Sulfides P .5L 2 ml Zinc 24 Hours 8 AM - 8 AM
Acetate 4C
Cyanides (T) P 1L NaOH pH > 12.0 4 C 24 Hours 2 PM
VOC V 40 ml 4C 24 Hours 2 PM

G = Glass bottle with Teflon lined lid

GA = Amber bottle with Teflon lined lid

P = Plastic Bottle

V = Approved glass vials with Teflon and pure rubber seals

Note: All samples are refrigerated at 4C after preservation.

32
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4.4 Sample Collection Location

BAD EXAMPLE : Effluent sample is collected at end of chlorine contact tank.

BETTER : The effluent grab samples for pH and Total Residual Chlorine are
collected at the outfall end of the chlorine contact tank at a point
approximately 1 foot upstream of the v-notch weir.

BEST : The effluent grab samples for pH and Total Residual Chlorine are
collected at the outfall end of the chlorine contact tank at a point
approximately 1 foot upstream of the v-notch weir, in the center of
the channel at a depth of approximately 1 foot. The open container
attached to a 6 foot aluminum pole is lowered into the waste
stream with the open end facing down stream as shown in pictures
#1 and #2.

Picture #1 Picture #2
Be specific and include
descriptions for each different type of sample (re: grab, composite)

33
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5. Sample Handling Documentation Procedures

5.1 All grab samples collected for analysis of pH, Total Residual Chlorine and
Settleable Solids are immediately taken to the lab. The pH and Chlorine analyses
are begun immediately. The settleable solids sample is poured immediately into
the Imhoff cone.

5.2 The grab sample collected for analysis of E. Coli is immediately placed into a
cooler with four blue ice packs and sealed. It is immediately transported by WWW
to WWW laboratory. The approximate travel time is 35 minutes.

Each bottle is properly labeled and accompanied with a chain of custody form.
One copy of the COC form is maintained at the facility.

5.3 Composite samples

Immediately after the last 24 disrete sample is collected the entire sampler is
brought into the laboratory where the hourly sample volumes are calculated
measured and poured into a four liter jug. Aliquots are poured from the container
for specific parameters (BOD, TSS etc.) After thorough shaking. Sample
containers are then placed into the refrigerator at 4C until the analysis is
performed.

34
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5. Sample Handling Documentation Procedures (continued)

5.4 Sample Identification/Labeling

a) All sample containers are labeled with the following information.

1) Type of sample (influent, effluent, grab, composite)

2) Parameter to be analyzed

3) Time and date collected

4) Initials of person who collected it

5) Preservation information

6) Any special instructions or remarks

b) All data from the label is transferred to bench sheets.

c) A chain of Custody form is completed. One copy accompanies the sample

to the lab. One copy is retained at the wastewater facility.

*Include a copy of the form in the manual*

d) The sample is delivered directly to the laboratory by name, assistant

operator

35
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5. Sample Handling Documentation Procedures (continued)

5.4 Sample Identification/Labeling (continued)

e) When the sample is handed over to the commercial laboratory the

recipient signs the chain of custody form indicating the time the sample
was received.

f) A copy of the chain of custody form is returned to the wastewater facility

along with analytical results.

6.0 Laboratory Water Quality

This example was borrowed from a QC manual (prepared by personnel of the

Nashua NH Wastewater Treatment Facility (copied here with their kind permission).

6.1 Deionized Water

R/O Deionized water is used in the laboratory for reagent preparation, glassware
rinsing, and BOD dilution water.

System includes: Millipore R/O system followed by a Milli-Q deionization

system and a 0.02 micron final filter.

TYPE: I : R/O, Deionization and 0.02 um. filter.

TYPE: II: R/O only

36
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6.1 Deionized Water (continued)

Quality Control Tests:

Parameter Requirements Frequency

(Required) Conductivity > 0.5 megohms resistance or Once Per Month
< 2 microhms/cm at 25C
(Suggested) Total Chlorine Non-detectable Once Per Month
Residual
(Suggested) Biosuitability of Ratio of 0.8 - 3.0 Semi-annually
Laboratory Pure (April & Oct.)
Water
(Suggested) Metals Lead, Nickel, Zinc, Copper, Semi-Annually
Cadmium, Chromium, Iron, (April & Oct.)
Silver
(Suggested) Total Plate Count < 200 colonies/100 ml. Once Per Month

* All QC data generated is recorded in the bound QC Manual.

Other Quality Control Measures

R/O system is back flushed daily for one half of an hour. Water for BOD dilution
water and reagent dilution is made fresh after morning flush cycle.
Tubing and associated hardware (if in contact with the water) are replaced as needed.

6.2 Distilled Water

Occasionally bottled distilled water is purchased (Poland Springs) for

preparation of reagents, BOD dilution water and glassware rinsing.

37
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6.1 Deionized Water (continued)

Other Quality Control Tests: (continued)

The quality specifications for this bottled water are included on the product label
and meets all requirements for analytical use.

7. Standard Operating Procedures for Each Analytical Method

7.1 pH

7.1 A. Method Reference

Electrometric Method #4500 - HB. 18th Edition Standard Methods pages

for the Examination of Water and Wastewater 4 - 65 ... 4 - 69.

7.1 B. Apparatus
pH meter (make and model)
pH temperature probe (type)
Buffer solutions Fisher, NIST certified, 4.0, 7.0, 10.0
Glass beakers (50 ml)
Magnetic stirrer and stir bars
Safety Glasses, gloves
Squeeze bottle for RO/DI water

7.1 C. Standardization

- The meter is standardized before each use with fresh pH 4.0 and 10.0
buffer solutions.

38
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7.1 C. Standardization (continued)

- The pH of a fresh pH 7.0 buffer is checked and the result is recorded in the
QC log.

- If the result of the checked buffer varies by more than .1 standard unit.
The meter is restandardized and the problem is corrected.

- The Fill hole and electrode cap are removed during use and replaced
after each use.

7.1 D. Interferences

- It is important, even with temperature compensation capabilities to

approximate as closely as possible the sample temperature when
standardizing the meter. To accomplish this , buffers are stored in the
refrigerator during winter months and at room temperature during summer
months.

- Sodium error is not a problem at this facility.

- Oil material, especially from influent testing can cause development of a

film on the electrode causing poor response. This can be avoided by
periodically washing the probe with a mild detergent followed through
rinses with RO/DI water.

- pH Electrodes are replaced if proper standardization cannot be

accomplished.

39
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7. Standard Operating Procedures for Each Analytical Method (continued)

7.1 E. Sample Preparation and Preservation

A minimum 25 ml sample is collected in a glass container. The

sample is taken immediately to the lab for analysis.

7.1 F. Procedure

Place pH and temperature probes into the sample. Allow meter to

equilibrate. Then record the reading in pH units at temp C.

7.1 G. Calculation

The pH meter reads directly in a pH standard units pH is reported

to the nearest 0.1 unit at a given temperature.

7.1 H. Quality Control

1. All required calibrations and other preventative maintenance on

instrumentation and equipment is performed regularly and
recorded on bench sheets and calibration logs.

2. Buffers are NIST Traceable, are changed daily and recorded.

3. 10% Duplication and Replication.

40
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7. Standard Operating Procedures for Each Analytical Method (continued)

7.1 H. Quality Control (continued)

4. The pH meter will be calibrated before each use, and separate

standard will be processed as a control buffer to validate the
calibration, pH 10 buffer will be used as a control.

5. Grab samples will be delivered immediately to the lab for analysis.

6. Glassware will be thoroughly rinsed with purified water.

7. Temperature probe will be calibrated against a NIST traceable

thermometer.

8. Calibration sheet will include: date and time of calibration, buffers

used and order of use, exact meter results, analyst performing
calibration, buffer and sample temperature.

9. Meter range 0-14, accuracy 0.1 pH, repeatability 0.1 pH,

temperature compensation and capable of 2 point calibration.

7.2. Total Residual Chlorine

7.2 A. Method Reference

Hach DR 700 method which is based on the DPD colorimetric

method #4500-CL G 18th Edition Standard Methods pages 4-45 and 4-46.

41
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7.2. Total Residual Chlorine (continued)

7.2 B. Apparatus

Spectro Photometer Model DR 700

DPD Total Residual Chlorine powder pillow/packets
Vials 10 ml - marked - dedicated currettes
Kimwipe Tissues
Safety glasses, gloves
Scissors or clippers

7.2 C. Standardization

The meter has an internal calibration which is automatically

enabled for each use.

In addition a Hach brand Voluette ampule certified standard is

used daily. Results are recorded and standard curve is plotted.

A blank consisting of the raw sample is used to zero the instrument

each use.

7.2 D. Interference

Color and turbidity interferences are reduced by the use of an

untreated sample as a blank to zero the device.

Organic contaminants may interfere but can usually be spotted by

the strange coloration - yellowish or greenish.

42
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7.2 D. Interference (continued)

Oxidized manganese because of its orange color can create a

problem but is at least partially compensated for by use of a blank.

7.2 E. Sample Preparation and Preservation

Because of the unstable characteristics of chlorine in an aqueous

solution it is important to take great care in the sample collection not to
alter the sample chlorine concentration.

The sample is collected into an airtight BOD bottle which has been
covered with black electrical tape. The bottle is filled well into the neck
so as to overflow when the ground glass stopper is inserted. Make sure
there are no bubbles in the bottle. It is then taken to the dab for immediate
analysis (within 5 minutes from collection to analysis).

7.2 F. Procedure

Rinse the sample cells and caps thoroughly with RO/DI.

Pour sample into 2 sample cells to the 10 ml mark.

Empty the contents of one Total Cl2 powder pillow or packet into
one of the sample cells. Cap and invert 10 - 15 minutes to mix.

Wait 3 minutes.

43
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7.2 F. Procedure (continued)

Using the up and down arrows set the device to mode 5207 (515
nanometers).

Wipe both sample cells carefully with a Kimwipe being careful to

remove any fingerprints or other marks.

Place the cell containing untreated sample (Blank) into the cell
compartment. Close the light shield and press the zero button.
The meter should read 0.00 mg/L.

Place the cell containing treated sample into the cell compartment
and push the read button.

Record the result in mg/L.

7.2 G. Calculations

The DR700 reads concentration of Total Residual Chlorine in

mg/L. Results are recorded directly to bench sheets.

Standard curves are plotted in mg/L.

7.2 H. Quality Control

1. All reagents are NIST traceable and preparation and expiration dates are
recorded.

44
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7.2 H. Quality Control (continued)

2. The Hach DR700 colorimeter is maintained by QC Services with regularly

scheduled service every 6 months.

3. Glassware is thoroughly washed with a 2% solution of Micro brand

detergent, and rinsed with RO deionized water. Separate cells for
duplicate total chlorine determinations.

4. Hach Volette Ampule Certified Standards are used daily, and are recorded
as a standard curve for a control chart.

5. Duplicate samples are analyzed once per week.

6. Grab samples are recorded at the exact time of sampling and are analyzed
immediately after entering the laboratory. The time of analysis and date
are recorded.
NO HOLD TIME

Calculation for Voluette Ampule Standards:

ml of standard addition
x Chlorine conc. of Vol. Ampule
ml of standard additional % ml of sample

= Concentration of chlorine added to the sample.

45
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7.3. Settleable Solids

7.3 A. Method Reference

Volumetric Method #2540 F 18th Edition Standard Methods for the

Examination of Water and Wastewater pages 2-57.

7.3 B. Apparatus

1 Liter Imhoff Cones

7.3 C. Standardization N/A

7.3 D. Interferences N/A

7.3 E. Sample Preparation and Preservation

Mix the sample thoroughly before pouring off.

7.3 F. Procedure

Place one liter of well mixed sample into a clean Imhoff cone.

Allow the sample to settle for 45 minutes.

Rotate the cone to dislodge solids on the side of the cone.

Allow to settle for an additional 15 minutes.

46
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7.3 F. Procedure (continued)

After a total of elapsed time of 60 minutes from the time the sample was
poured. The settled solids are read in ml/liter and recorded.

7.3 G. Calculations

Results are recorded as ml/L.

7.3 H. Quality Control

Duplicate samples are collected and analyzed weekly.

7.4 Total Suspended Solids

7.4 A. Method Reference

Total Suspended Solids Dried at 103 - 105C Method #2540 D.

18th Edition Standard Methods for the Examination of Waste and

Wastewater pages 2-56.

7.4 B. Apparatus

Drying Oven 104 C ( 1C)

Filter Paper, glass fiber 934AH

Buchner Funnel

47
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 7.4 B. Apparatus (continued)

Rubber stopper (to fit Buchner funnel to 1 liter side arm flask)

(2) 1 foot sections of rubber hose (one to connect side arm to

filtration flask to safety trap) One to connect trap to vacuum pump.

(2) 1 liter side arm flasks - 1 filtration 1 safety vacuum pump.

Graduated Cylinders

Desiccator - fresh desiccant

Forceps

Safety glasses and gloves

Aluminum weighing tray

(4) Place analytical balance squeeze bottle for RO/DI water.

7.4. C. Standardization (or more accurately pre-test preparation)

The balance must be checked to be sure it is level and then

properly zeroed after brushing any debris off the pan.

48
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7.4. C. Standardization (or more accurately pre-test preparation)
(continued)

Filters must be rinsed with RO/DI water dried at 104C for

approximately 15 minutes, weighed and then redried, redesiccated and
reweighed to establish constant weight ( 0.5 mg) before the analysis is
performed.

- Make sure the drying oven is maintained at a constant temperature

of 104C 1.

7.4. D. Interferences

Water with every high mineral content might need to be dried

longer, desiccated and weighed quickly as moisture will be quickly
absorbed from the atmosphere. This affect can be reduced by placing
desiccant in a container inside the balance chamber.

Dont include large floating particles in the sample unless they are
truly representative of the actual sample conditions.

7.4 E. Sample Preparation and Preservation

Collect a sample volume of at least 2 liters.

Keep samples refrigerated at 4C up until the time of analysis to

reduce the microbiological decomposition of solids.

49
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7.4 E. Sample Preparation and Preservation (continued)

Try to perform the analysis within 24 hours (7 days absolute

maximum)

Bring the sample to room temperature before beginning analysis.

7.4 F. Procedure

1. Prepare the funnel and vacuum apparatus. Make sure the vacuum
is not excessive so as to rip the filter.

2. Put the pre-weighed filters into the funnel wrinkled side

3. Seat the filter by rinsing the funnel and filter with about 25 mls of
RO/DI water. Turn on the vacuum pump until DI water is drawn
through the filter.

4. Shake sample thoroughly, then pour off 1 liter of effluent (100 ml

influent) into the graduated cylinder.

5. Pour the sample slowly into the center of the filter with the
vacuum on.

- Rinse the graduated cylinder with at least 3 successive 20

ml DI water rinses turn the graduated cylinder while
pouring rinsewater into funnel to insure that all solids are
rinsed out.

50
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7.4 F. Procedure (continued)

- Wash down the sides of the funnel into the filter.

- Remove the filter from the funnel with the forceps.

- Place the filter into an aluminum weighing pan and place onto the
center rack of the drying oven at 104C for 2 - 3 hours (preferably
overnight).

- Cool to room temperature by putting the filter into the desiccator

for 15 to 30 minutes.

- Weigh (if the aluminum pan was included in the initial weighing it
must certainly be included in all weighings.)

- Redry at 104C for at least 1 hour.

Redesiccate for 15 to 30 minutes.

- Reweigh

- Repeat drying, desiccating and weighing if necessary until

constant weight is achieved (<.5 mg difference)

7.4 G. Calculations

A & B x 100,000
TSS in mg/L =
C

51
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7.4 G. Calculations (continued)

Where A = weight of filter and residual (in grams)

B = weight of filter (in grams)

C = volume of sample filtered (in milliliters)

7.4 H. Quality Control

1. The oven temperatures are closely monitored.

Temperatures are recorded every four hours when in use.

2. The analytical balance is checked for level, auto calibrated

before each use and professionally calibrated every 6
months.

3. The temperature of the sample storage refrigerator is

checked and recorded every 4 hours when in use.

4. A replicate sample is analyzed each time the analysis is

performed. If results are not within a specified range
(determined by control chart) corrective actions are
initiated.

5. One duplicate sample is analyzed for every ten TSS

analyses performed. Results are recorded in QC log and
are plotted on control charts.

52
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 7.4 H. Quality Control (continued)

6. A RO/DI water blank is run each time the analysis is

performed. Results are plotted on a blank control chart.

7. Sufficient sample, up to one liter, is filtered to produce at

least 2.5 mg residual.

8. Results are recorded in QC log and bench sheets.

7.5 5 Day BOD

7.5 A. Method Reference: 5 Day BOD Method 521 OB 18th Edition

Standard Methods for Examination of Water
and Wastewater pages 5-2 through 5-6.
7.5 B. Apparatus

Dissolved Oxygen Bottles

Overcaps
One Liter Graduated Cylinders
Assorted Sizes of Graduated Cylinders
Beakers, Assorted Sizes
Pipettes
Pipette Bulb
Carboy
Siphon
Propeller Mixer
DO Meter
Thermometer

53
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7.5 B. Apparatus (continued)

Air Pump, Tubing, Filter

Stand
pH Meter
Incubator
Refrigerator
DO Bottles with Auto Pipettes
500 ml Erlenmeyer Flasks
25 ml Burette and Burette Stand
Dropping Bottles

54
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7.5 C. Reagents:

Reagent Recipe Hold Time

Phosphate Buffer - 8.5 KH2PO4 - Indefinite @ 4 C
(Dilution water - 21.75g K2HPO4 - Throw out if biological growth
component.) - 33.4g Na2HPO4 .7H2O appears.
- 1.7g NH4Cl
- Dilute to 1R DI water.
- Autoclave 45 minutes.
Magnesium Sulfate - 22.5g Mg SO4.7H2O in 1R DI water - Indefinite @ 4 C
- Autoclave 45 minutes. - Throw out if biological growth
appears.
Calcium Chloride - 27.5g CaCl2 in 1R DI water. - Indefinite @ 4 C
- Autoclave 45 minutes. - Throwout if biological growth
appears.
Ferric Chloride - 0.25g FeCl3 .6H2O in 1R DI water. - Indefinite @ 4 C
(Dilution water - Autoclave 45 minutes. - Throw out if biological growth
component) appears.
1N Sulfuric Acid - 28 ml conc. H2SO4 to 1R DI water.
(To adjust pH sample)
1N Sodium Hydroxide - 40g NaOH to 1R DI water.
(To adjust pH sample)
GGA ampules Order from: Hach Company
- 300 mg/l glucose PO Box 907
- 300 mg/l glutanic acid Ames, Iowa
Polyseed Order from: Polybac Corp. 1 year at 20 C
3894 Courtney Street
Bethlehem, PA
Sodium Sulfite - Prepare small vials each containing 1 - 2 hours
(Dechlorination reagent) 0.79g of Na2SO3
- Dissolve 1 vial in 500 ml DI water
prior to use.

55
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 7.5 C. Reagents (continued)

Reagent Recipe Hold Time

Phosphate Buffer - 24g Na2HPO4 - Indefinite at 4 C
(For dechlorination) - 46g KH2PO4 - Throw out if biological growth
- 800mg disodium EDTA dihydrate appears.
into 100ml DI water.
- All of above into 1R DI water.
- 20mg H2Cl2 preservative.
- Autoclave 45 minutes.
DPD Indicator - 1g N,N-Diethyl-p-phenylenediamine - Discard if pink color appears.
oxalate - Store in dark bottle @ 4 C.
- 8ml 1 +3 H2SO4
- 200mg disodium EDTA dihydrate
- Dissolve all of the above in 1R DI
water
0.25N FAS titrant - 49g Fe (NH4)2(SO4)2.6H2O - 3 months @ 4 C
- 10ml H2SO4 - Must be standardized.
- Dissolve above in 1R boiled and
cooled DI water.
1 + 3 sulfuric acid - one part H2SO4 to three parts distilled
water.
0.25N potassium - 12.26g, K2Cr2O7 to 1R DI water Indefinite at 4 C
dichromate
Potassium iodide crystals - Prepare small vials each containing Indefinite
1g KI

7.5 D. DO Meter Standardization

1. Turn DO meter onto RED LINE, allow to warm up for 15 minutes.

2. Fill 1R graduated Cylinder with DI water and siphon into 3 BOD bottles,
without aerating water.

56
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7.5 D. DO Meter Standardization (continued)

3. To two of the bottles add 2 mls of DO reagent #1 and 2 mls of DO

reagent #2 below surface.

4. Invert bottles until uniform consistency. Let stand until floc settles to
50% bottle volume. Invert a second time and let stand again.

5. Add 2 mls of DO #3, cap and invert until floc is complete gone.

6. Pour contents into 50 ml wide-mouth flask and titrate using .0375N

Na2S2O3 until pale yellow. Add 1 ml of starch indicator (1/2 dropper
full and swirl. Titrate slowly until blue becomes clear.

7. Add as many drops of black titrant (potassium bi-iodate) as are necessary

to bring blue color back. If one drop does this, record the reading on
the buret. If it takes more than 1 drop, multiply the number of drops
by 0.05 mls and subtract from buret reading.

Reading on buret is in milliliters and is equivalent to oxygen in

milligrams per liter. Report readings to nearest 0.05 ml.

8. Average two titrations and record results in record book.

9. Place DO probe into third bottle making sure there are no air bubbles
trapped. Turn knob to zero and adjust until zeroed.

57
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7.5 D. DO Meter Standardization (continued)

10. Turn knob to calibrate 1-10, turn on stirrer.

11. Let sit 2 minutes and adjust until meter reads average of two Winkler
titrations. If the average DO of the Winkler Titrations and the DO from
the meter differ by more than .5, rerun Winkler. Run samples within 4
hours of calibration.

E. Preparation of seed - do earlier on day samples arrive.

1. Empty one capsule of Polyseed BOD seed inoculum into 500 ml of

dilution water.

7.5 E. Interferences

Presence of copper and other metals in the sample can adversely affect results.

See Sample Preparation for Treatment of samples for other interferences.

7.5 F. Sample Preparation and Preservation

24 Hour Composite Samples -

Sample is refrigerated at 4 C from the time the first sample is collected

until one hour before analysis is begun.

It is placed in a warm water bath before analysis until the sample temp
reaches 20 C. The sample is thoroughly mixed just before being poured

58
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7.5 F. Sample Preparation and Preservation (continued)

24 Hour Composite Samples - (continued)

off. Dechlorination: Samples are dechlorinated using sodium sulfite.

Dechlorination is described in the Procedure section (7.5 G).

Seeding - All samples are seeded. The process is described in the Procedure
section 7.5 G.

pH: The sample pH is adjusted to between 6.5 to 7.5 if necessary using sulfuric
acid or sodium hydroxide (usually not necessary).

7.5 G. Procedure

A. Dilution Water Prep - 24 hours before use.

1. Wash 20 R carboy and siphon tubing with micro and rinse very
well with DI water.

2. Fill carboy with DI water filtered through 0.2 Fm filter.

-10R for first sample

-3R for each sample thereafter.

ex: 7 samples would require 28R (10 for the first then 3 X 6 = 18
for rest.)

59
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7.5 G. Procedure (continued)

3. Add 1 ml per R for each:

MgSO4

CaCl2

FeCl3

4. Aerate for about 15 minutes.

5. Store in 20 C incubator for at least 24 hours and no longer than

5 days.

6. Just prior to use add 1 ml phosphate buffer per liter; aerate 15

minutes; let sit for 1 hour, then use.

7. Be sure temp. Is 20 to 23 C.

8. Be sure DO is between 7.5 and 8.5 mg/1.

a) If >8.5 mg/l, shake closed container vigorously, open and

let sit.

b) If <7.5 mg/l, aerate some more.

60
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7.5 G. Procedure (continued)

B. Glassware prep. - Label all ahead of time.

Bottles 1000 ml Graduates

Blank 3 1
GGA 3 1
Seed 9 (3/dilution) 3 (1/dilution)
Sample 1 9 (3/dilution) 3 (1/dilution)
Sample 2 9 (3/dilution) 3 (1/dilution)

C. Dechlorination of samples.

1. Just prior to use, prepare 0.78g Na2SO3 to 500 mls DI water.

2. To a 500 ml wide-mouth flask, add:

- 100 ml sample

- 5 mls DPD indicator

- 5 mls PO4 buffer

- 1 g KI crystals (may be pre-weighed in small vials)

3. A faint pink-red color after 2 minutes indicates chlorine is

present. Stop here if no color.

61
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7.5 G. Procedure (continued)
4. Using standardized 0.00282N FAS, titrate to clear endpoint. Ml
FAS used equals mg/l Cl2.

5. Dechlorinate with Na2SO3 using in ml 5 times the amount of

chlorine per liter.

Example: Cl2mg/l = FAS used

2.2 mls FAS used = 2.2 mg/l Cl2

2R sample: 2R x 2.2 x 5 = 22 ml Na2SO3

Add 22 mls Na2SO3 to 2R sample to dechlorinate,

shake sample. _________________________

6. After 15 minutes repeat steps 2 and 3. If no pink color, then

sample is

2. Aerate and stir for at least 60 minutes.

3. When pipetting be sure flakes of bran do not get into pipette.

62
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7.5 G. Procedure (continued)

F. Seeding Chart

Expected Seed BOD mg/l Seed Dilution (%)* Seed (ml) to dilutions +
50 (Polyseed range) 2, 5, 10 20
75 2, 5, 10 15
100 1, 2, 5 8
125 1, 2, 5 8
150 1, 2, 5 6
175 1, 2, 5 6
200 0.51, 2, 5
225 0.5, 1, 2 5
250-400 0.5, 1,2 3

* Seed dilutions calculated for an approximate DO depletion of 3 mg/l

+ calculated for a DO depletion of approximately 1 mg/l

2. Seed Calculation:
0.8 mg/l ml seed used in samples % GGA
x 1000 '
Expected seed BOD mg/l

63
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7.5 G. Procedure (continued)

G. Dilution Technique.

1. BOD rules:

a) Blank depletion must be <0.2 mg/l.

b) Want a dilution such that residual DO is at least 1.0 mg/l.

c) Want a dilution such that depletion is at least 2.0 mg/l..

2. Calculation for dilution:

2 mg/l x 1000 ml
' mls sample to dilute 1R
Expected sample BOD

Example: Expected BOD is about 15 mg/l, you want 2 mg/l depletion.

2 mg/l x 1000 ml
' 133 ml to 1R or 13.3%
15 mg/l

3. General ranges for dilutions:

0.0 - 1.0 % for strong industrial wastes

1 - 5% for raw and settled wastewater
5 - 25% for biologically treated effluent
25 - 50% for polluted river waters

H. Actual procedure on day samples started.

7.5 G. Procedure (continued)

64
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1. Add 10 ml GGA ampule to 1R graduated cylinder (gives 2%
GGA).

2. Add seed (usually 20 ml for polyseed) to GGA and sample

cylinders.

3. Prepare dilution of samples in graduated cylinders (be sure

samples at 20 C).

4. Set up sample sheet to receive data (see sample data sheet).

5. Transfer sample dilutions, GGA, blank and seed dilutions to BOD

bottles (3 bottles per dilution).

6. Put plastic caps on 2 bottles for each dilution and place in 20

incubator for 5 days 3 hours.

7. Read DO of third bottle for each group and record as Initial DO

(see sample data sheet).

8. Read DO final after 5 days 3 hours and record both values for
each dilution.

65
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7.5 G. Procedure (continued)

Calculations:

BOD of seed x ml seed used

A. Seed factor:
1000 ml

40 mg/l x 15 ml
Example: ' 0.6 mg/l
1000 ml

B. DO depletion:

DOI & DOF ' DO depletion

DO1' Initial DO

DOF ' average final DO

Example: 8.20 mg/l & 5.4 mg/l ' 2.8 mg/l (depletion)

DO depletion & Seed Factor

C. BOD mg/l:
% Dilution

2.8 mg/l & 0.6 mg/l

Example: ' 4.4 mg/l
.5 (50% dilution)

66
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7.5 G. Procedure (continued)

Quality Control:

1. Blanks are run as a bottle and dilution water check. Values #0.2 mg/l are
acceptable.

2. GGA should run 198 30.5 mg/l.

3. A duplicate should be run in each batch.

67
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 5 Day BOD Data Sheet
Sample Locations:Sample Data Sheet
Lab ID Numbers: 96274 99385
Date & Time Sampled: 10-21-92 10:00 a.m. Date & Time Sample arrived at Lab: 10-
12-92 1:00 pm
Date & Time BOD Setup: 10-21-92 3:00 p.m. Date & Time Final DO Read: 10-26-92

1:00 p.m.
Analyst:

Sample % Dil ml Seed DOI DOF DOF Avg. DO Seed DO BOD

DOF Depl. Factor Depl. mg/l

Blank --- --- 8.80 8.75 8.65 8.70 0.10 --- --- ---
GGA 2 20 8.75 4.30 4.25 4.28 4.47 0.46 4.01 200.5
S1 2 20 8.75 8.20 8.20 8.20 0.55 --- --- <2
Depl.
S2 5 50 8.75 7.70 7.65 7.68 1.07 --- --- <2
Depl.
S3 10 100 8.80 6.60 6.40 6.50 2.30 --- --- 23
96274 A 5 20 8.75 6.00 6.05 6.07 2.68 0.46 2.22 44.4
B 10 20 8.75 3.65 3.70 3.67 5.08 0.46 4.62 46.2
C 30 80 <1 <1 --- --- --- --- --- ---

99385 D 5 20 8.75 6.55 6.65 6.60 2.15 0.46 1.69 <2

Depl.
E 10 20 8.75 5.10 4.95 5.03 3.72 0.46 3.26 32.6
F 30 20 8.80 <1 <1 --- --- --- ---

68
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 Sample % Dil ml Seed DOI DOF DOF Avg. DO Seed DO BOD
DOF Depl. Factor Depl. mg/l

Notes and Calculations:

23 mg/l x 20 mg/l
Seed Factor: ' 0.46 mg/l
1000 ml

69
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 5 Day BOD Data Sheet - Continued

Sample % Dil ml Seed DOI DOF DOF Avg. DO Seed DO BOD

DOF Depl. Factor Depl. mg/l

Notes and Calculations:

70
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8.0 Reagent/Standard Quality, Preparation and Traceability

8.1 All standards and reagents are prepared from reagent grade materials, primary standards or they
are purchased from reputable vendors. Reagents are stored according to manufacturers
instructions and discarded upon expiration. Standards and reagents are prepared using bala
nces that are calibrated daily, Class A volumetric glassware and ASTM Type II reagent water.

Once a solution is prepared it is labeled with the solution name or description, storage
requirements, concentration or normality, preparation and expiration dates and initials of
preparer. Expiration dates for standards and reagents are specified in methods that are adhered
to unless degradation prior to this date is observed. Log books are utilized to record the
preparation of standards.

Calibration standards (working standards) are dilutions or mixtures of stock standards

used to calibrate an instrument. These standards are prepared or restandardized frequently as
specified in Laboratory Standard Operating Procedures (SOPs). New standards are checked
again old standards to insure there has not been an error in preparation.

Quality control reference samples are analyzed along with most analytes, depending upon
availability to validate standards, technique and methodology. Quality control reference samples
are prepared from a different source than that used in the preparation of standards for use in the
standard curve and are US EPA certified, if possible.

9.0 Calibration Procedures and Frequencies

71
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9.1 Instrument Calibration Procedures

All instruments and equipment used are routinely calibrated by laboratory personnel or by
external calibration agencies or equipment manufacturers. Maintenance schedules can be found in the
Preventative Maintenance Section of this manual (Section 1.0). Instrument calibration procedures,
frequencies, standards and traceability are summarized in Table 9.2 To insure that instruments have
performed adequately throughout the analysis, it is laboratory practice to run a standard or quality
control reference sample at the end of an extended run.

72
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9.1 Instrument Calibration Procedures (continued)

Table 9.2

Instrument/ Calibration Procedure Calibration Quality Control

Analytes Frequency Standard Standard
(Traceability) (Traceability)
pH/Millivolt Meter Daily 2 point Certified buffers EPA Reference
- pH calibration (NIST traceable) Sample (US EPA)
bracketing
sample
Conductance meter Daily .01M Potassium Primary Grade Conductivity
- conductivity Chloride Reagents Standards (NIST)
Analytical Balances Daily Calibrated Calibration Weights NBS
according to verified using Traceable
manufacturers Class -S weights
directions.
Spectrophotometer Daily 3 - 5 Point Primary Grade EPA Reference
- COD Calibration Reagents Sample (US EPA)
Turbidity Meter Daily Calibrated AMCO Sealed AMCO Sealed
- turbidity According to Primary Secondary Standards
AMCO Calibration
instructions Standards
Dissolved Oxygen 4 Hours Winkler Primary Grade (NIST traceable)
Meter Titration Reagents
- BOD
- DO
Cloroimeter (Fixed Each use Internal NIST SMR 930 EPA Reference
Photometer) instrument S/M99 Sample (US EPA)
- Chlorine calibration and
Hach Spec T
Color Standard
COD Reactor Daily 5 Point Primary Grade EPA Reference
- COD Calibration Reagents Sample (US EPA)
Millivolt Meter Daily 4 Point Primary Grade EPA Reference
- TKN Calibration Reagents Sample (US EPA)

2 Point
Calibration
Probe

73
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 9.1 Instrument Calibration Procedures (continued)

Table 9.2

Instrument/ Calibration Procedure Calibration Quality Control

Analytes Frequency Standard Standard
(Traceability) (Traceability)
Thermometers Annually/ Accuracy Thermometer
- incubators Semi- checked over NIST
- pH Annually range to be used.
- conductivity (bacteriology) Deviations
recorded on
thermometer.
Incubators Twice Daily Temperature Thermometer
- bacteriology Check NIST
Autoclave Weekly Sterility Check Spore Strips
- bacteriology Temperature Maximum
Check Thermometer
NIST
Refrigeration Units Daily Temperature Thermometer
- reagent sample storage Check (NBS)

10.0 Data Handling - Reduction, Validation and Reporting

10.1 Data Reduction

Raw data is recorded directly onto bench sheets by the technician who actually
performed the analysis.

If more than one analyst is involved in setting up an analysis or reading results (Ex. BOD
initials DCs determined by one person and final DOs by another) both analysts initial the bench
sheet. All calculations are included on bench sheets.

Only the chief operator can transfer information from the bench sheets to WR-43 report
forms.

74
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Data Handling - Reduction, Validation and Reporting (continued)

10.2 Data Validation

The analyst who generates the data has the prime responsibility for its correctness and
completeness. It is the analysts responsibility to verify that the instrument was calibrated and
was performing properly.

The chief operator looks over and double checks the bench sheets. He checks all
calculations, looks to see that all data makes sense and that the numbers were rounded properly
(section 10.5) and that proper significant numbers were recorded (section 10.4).

After transferring the data to the WR-43 report forms he checks for transcription errors.

The assistant operator performs a quick check of bench vs. WR-43 data before the report
is given to the town manager for his signature.

The town manager looks at the WR-43 report form and questions the chief operator
concerning any unusual or suspicious looking result before signing the report. As a rule the
town manager and chief operator meet to discuss the report regardless of whether or not there are
irregularities.

75
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Data Handling - Reduction, Validation and Reporting (continued)

10.4 Significant Digits

Parameter Significant Digits

BOD - No digit after decimal point. 28 mg/L
Chlorine Residual - Two digits after decimal point. 0.51 mg/L
Coliform - No digits after decimal point. 50/100 ml.

TKN, NH3, NO3 - One digit after decimal point. 17.6 mg/L
D.O. - Two digits after decimal point. 7.35 mg/L

Settleable Solids - One digit after decimal point. 5.1 ml/L

Metals - One digit after decimal point. 436.3 ppb
pH - Two digits after decimal point. 7.00 pH units
Suspended Solids - No digit after decimal point. 22 mg/L
Temperature - One digit after decimal point. 17.2C

10.5 Rounding Policy

All digits are used in calculations, then are rounded, using the following guidelines.
Numbers following decimals shall be rounded to the next higher or lower number based on this
method.

For example: 3.57 is rounded to 3.6

2.41 is rounded to 2.4

7.55 is rounded to 7.6

7.44 is rounded to 7.4

76
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11.0 Preventive Maintenance Procedures and Schedules

11.1 All laboratory equipment is serviced and professionally calibrated by QC Services on an annual
basis. A service contract is maintained to include annual equipment calibration.

11.2 Routine and professional calibration/maintenance schedules are summarized in Table 11.2.

11.3 Preventive maintenance responsibilities are assigned to specific laboratory personnel. Only the
lab supervisor is allowed to perform other than routine calibration or minor repair.

11.4 A maintenance log is kept in the lab for each instrument. All calibration, repairs and service
visits are recorded an entitled by the responsible party.

77
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11.0 Preventative Maintenance Procedures and Schedules Table 11.2 (Continued)

Instrument - Manufacturer - Model Calibration Maintenance Preventative

Frequency Contractor Maintenance
Schedule
Atomic Absorption, Perkin-Elmer 3030B Daily Perkin-Elmer As needed
Autoclave, Barnstead 4/year MDT Biologic Co. 4/year
Waterbath, Precision Scientific - 83 1/year QC Services 1/year
Waterbath, Blue M 1/year QC Services 1/year
Air Incubator, Boekel 1/year QC Services 1/year
pH/Millivolt Meter, Orion Model 811 Each use QC Services 1/year
pH/Millivolt Meter, Orion Model 720A Each use QC Services 1/year
Ratio Turbidmeter Hach Model 18900 Daily QC Services 1/year
Spectrophotometer, Bauch & Spectronic 100 Daily QC Services 1/year
Dissolved Oxygen Meter, YSI Model 57 Every 4 hours QC Services 1/year
Fluorometer, Turner Model 111 Yearly QC Services 1/year
COD Reactor, Hach Model Yearly QC Services 1/year
Centrifuge, International Equipment --- QC Services ---
Conductance Meter, YSI Model 32 Yearly QC Services 1/year
Chlorine Meter, Hach Model DR100 Each use QC Services ---
Balance, Mettler AE 200 Daily QC Services 1/year
Balance, OHAUS B1500D Daily QC Services 1/year

78
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12.0 Corrective Action Contingencies

12.1 Corrective actions are required as a result of less than acceptable Performance Evaluation study
results, or poor comparison in split sample analyses (State lab results significantly different than
facility results). The steps taken in the corrective action process include:

Identify and define the problem.

Determine who will be responsible for investigating the problem.

Find the cause of the problem.

Determine the actions needed to eliminate the problem.

Implement the corrective actions and

Establish the effectiveness of the corrective action.

Usually the lab supervisor is responsible for initiating the corrective action under these
conditions. Documentation of actions taken and their effectiveness is forwarded to the lab
supervisor for review and distribution.

12.2 Corrective actions might also be initiated by an analyst during or after sample analysis. These
actions may be necessary because of

Unacceptable blank results (BOD blank depletion >.2 mg/L)

Suspicious positive control results (every few colonies on E Coli. positive control
sample).

QC data outside the warning or control limits for precision and accuracy.

79
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12.0 Corrective Action Contingencies (continued)

Duplicate or replicate results are inconsistent.

Under these conditions the analyst generating the data is expected to initiate and
document corrective action.

12.3 Corrective Actions Required for Specific Problems are Listed in Table 12.4

Table 12.4

Problem Corrective Action

Blank contamination is indicated Determine cause of contamination, eliminate
cause and repeat analysis.
Unexpected, unusual results occur more than Investigate possible causes. Try to eliminate
once every 20 analyses interferences. Keep a close watch on future
analyses.
One or more data points falls outside the Determine cause eliminate it an repeat analysis.
control limit.
Two out of three successive data points fall Repeat analysis. If the next data point is less than
outside the established warning limits. warning limit continue the analyses. If the next
point exceeds warning limits discontinue analyses
and correct the problem.
4 out of 5 successive data points exceeds 1 Analyze another sample. If the next data point is
standard deviation or are in decreasing or less than 1 standard deviation or changes the order
increasing order. continue analyses. If not discontinue and correct
the problem.
Six successive sample results are Analyze another sample. If the next result is on
above/below the central line on the control the other side of the central line continue analyses.
chart. If the next point is on the same side of the central
line discontinue analyses and correct the problem.

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13.0 Quality Control Procedures (General)

This section describes the method used at this laboratory to evaluate the quality of data
generated.

13.1 Sampling Quality Control Checks

Sampling QC checks provide information regarding the precision and accuracy of the
entire process from sample collection through analyses. Included in this category are:

13.1.1 Equipment Blanks

Equipment blanks are used to determine if contamination has been introduced

through contact with sampling equipment or to verify effectiveness of equipment
cleaning procedures.

Clean laboratory water is pumped through the Isco sampler. We usually place the
sampler probe in a 4 liter jug of RO/DI water, pump for 10 - 15 seconds, purge for 5
seconds. This process is repeated 2 to 3 times before the equipment blank sample is
collected. This sample is then taken to the lab and processed along with other samples.

13.1.2 Split Samples

Split samples are replicate samples, two aliquots taken from the same sample
container. The samples are then analyzed independently by our own lab and a contract
laboratory. If significant differences are noted the cause is determined and corrected.

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13.0 Quality Control Procedures (General) (continued)

13.1.3 Duplicate Samples

Duplicate samples are samples collected at the same location at the same time.
Collection of duplicate samples serves as a check on sampling and processing technique.
Each sample is analyzed individually. Results must be within an acceptable range (10%)
or the cause must be determined and corrected.

13.1.4 Replicate Samples

Replicate sample are two aliquots taken from the same sample container that are
processed and analyzed separately. The results are used to measure analytical precision
from sample preparation through analysis. Certain analyses are run in replicate every
time the test is performed. A minimum 10% replication schedule is established for all
analyses.

13.2 Procedures Used to Assess Data Quality

13.2.1 Precision

Precision is a measure of the closeness with which multiple analyses of a sample agree
with each other. We calculate precision from results of replicates and duplicate analyses of
quality control samples.

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13.2 Procedures Used to Assess Data Quality (continued)

13.2.1 Precision (continued)

Here at the WWW Wastewater laboratory we use Relative Percent Difference (RPD) as a
measure of precision. The formula used to calculate RPD is:

(C1 & C2) x 100%

RPD '
(C1 % C2) /2

Where: RPD = relative percent difference

C1 = larger of the two observed values

C2 = smaller of the two observed values

If calculated from three or more replicates, we use relative standard deviation rather than
RPD:

RSD = (s/y) X 100%

where: RSD = relative standard deviation

s = standard deviation
y = mean of replicate analyses

Standard deviation is defined as follows:

'Xi &
('Xi)
n
SD '
n & 1

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13.2 Procedures Used to Assess Data Quality (continued)

13.2.1a Control Charts

Control charts are used to demonstrate the precision calculated from replicates
and quality control sample duplicates.

The Relative Percent Difference (RPD) values for each sample are plotted on
control charts and where upper and lower warning and control limits are depicted.

These control charts are used by the analysts to help make them aware of
suspicious or out-of-control variability at the time of analysis. The warning and control
limits are recalculated annually. An example of a control chart cal be found at the end of
the section.

13.2.1b Control Limits

Control limits are defined as the mean + / - 3 standard deviations. An RPD value
that falls outside the control limits is considered out-of-control and requires the analysts
to repeat the analysis immediately. If the repeat value is within the control limit analysis
may continue. If the repeat value exceeds the control limit analysis must stop and the
problem must be corrected.

13.2.1c Warning Limits

Warning limits are narrower than control limits and are defined as the mean + / -
2 standard deviations. An RPD value that falls outside the warning limit is considered
suspicious. If two out of three consecutive points exceed a warning limit the analyst
must calculate the RPD on another sample. If the warning limit is exceeded analysis
must stop and the problem must be corrected.

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13.2 Procedures Used to Assess Data Quality (continued

13.2.2 Accuracy

Accuracy is expressed as a percent bias or percent recovery and is determined from the
analysis of quality control reference samples or spikes. Method accuracy is calculated on a daily
basis and summarized annually in the Laboratory Quality Assurance Plan.

Percent recovery is calculated from spike results using the following equation:

S&U
% R = 100% x
Csa

where: %R = percent recovery

S = measured concentration in spike aliquot

U = measured concentration in unspiked aliquot

Csa = actual concentration of spike added

(O & T)
% B ' 100 x
T

Where: %B = percent bias

O = measured concentration of reference material

T = actual concentration of reference material

o:\home\lori\andy\labqc.mnl

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