Title: 2003 NELAC Standards Microbiology Checklist: How to Pass your Assessment
12003 NELAC StandardsMicrobiology Checklist
How to Pass your Assessment
- By
- Denise K. Williams
- Biological Scientist III
- Florida Department of Health
- Environmental Laboratory Certification Program
- FSEA Microbiology Workshop
- Rev. 2 5-22-07
2Do Your Homework
- Use the Microbiology Checklist!
- Read Appendix D.3 to NELAC Chapter 5
- Make sure previous deficiencies are still
corrected (if still applicable) - Review original methods and your current Standard
Operating Procedures SOP - Do internal audits of each method
- Maintain Demonstrations of Capability DOC
3Work Area
- D.3.8 (a) Are the laboratory floors and work
surfaces where Microbiology testing takes place
non-absorbent and easy to clean and disinfect? - D.3.8 (a) Are microbiology work surfaces
adequately sealed? - D.3.8 (a) Are laboratory storage spaces for
Microbiology testing sufficient, clean, and free
from accumulation of dust? - D.3.8 (a) Does the laboratory prohibit plants,
food, and drink from the Microbiology work area?
4Incubator/Water Bath Records
- D.3.8(b)(6)(i) - Does the laboratory document
temperatures of incubators water baths twice
daily, at least 4 hours apart, on each day of
use? - Are your times documented to demonstrate that
checks were performed 4 hours apart? - Temperature Units? (oC?)
5Incubator/Water Bath Records
- Do the laboratory's analytical records on strip
charts, tabular printouts, computer data files,
analytical notebooks, run logs include the
following essential information? - 5.4.12.2.5.3(f) - Analyst or operator initials/
signature(Initials?) - 5.4.12.2.5.3(c) - Instrumentation identification
instrument operating conditions/parameters (or
reference to such data)(Equipment ID?) - 5.5.5.5(b) - Manufacturer's name, type
identification, serial number or other unique
identification (Unique ID?)
6Incubator/Water Bath Records
- 5.5.5.2.1(d) - Is the support equipment
acceptability for use according to the needs of
the analysis or the application for which the
equipment is being used? - Are your incubation temperatures in the proper
range per method requirements? - (Typically 35oC/- 0.5 or 44.5oC/- 0.2)
7Incubator/Water Bath Records
- If temperatures were out of range, did you
- (1) Qualify the data? NELAC 5.5.10.3.1
- (2) Take corrective action (and document that
action)? NELAC 5.4.10.3 - (3) Result of corrective action when did
temperature become acceptable?
8Incubator/Water Bath Records
- D.3.8(b)(6)(i) - Has the laboratory established
the stability, uniformity of temperature
distribution, time to re-establish thermal
equilibrium conditions (after test sample
additions) in incubators water baths? - Two parts (1) Stability/Uniformity (2)
Equilibrium - One time study (can you find your records?)
9Refrigerator
- 5.5.5.2.1(d) - Temperatures checked once per day?
- Equipment ID? Analyst Initials? Temperature
Units? - Standard Methods for the Examination of Water and
Wastewater SM, 20th Edition specifies 1-4oC - Out of range? Need corrective action
- (Clean vents? Too close to wall?)
10Thermometers
- 5.5.5.2.1(d) - Is the following support equipment
associated with microbiological testing checked
with NIST traceable materials (where available)? - Includes Refrigerator(s) for sample storage
and/or media storage Water Baths Incubators - Traceability Do you have the NIST certificates??
11Thermometers
- D.3.8(b)(1) - Is each temperature measuring
device (e.g., liquid-in-glass thermometers,
thermocouples, platinum resistance thermometers)
calibrated at least annually to national or
international standards for temperature? - SM 20th Ed. Requires semi-annual calibration for
microbiology
12Thermometers
- D.3.8(b)(1) - Are the available temperature
monitoring devices that are used in incubators,
autoclaves, refrigerators, or other equipment
where temperature accuracy has a direct effect on
the Microbiological analysis of appropriate
quality to achieve specifications in the test
method (e.g., no separations in liquid column for
liquid-in-glass thermometers)? - LOOK at your thermometers
13Thermometers
- D.3.8(b)(1) - Is the scale of graduations for
each temperature measuring device appropriate for
the required accuracy of measurement? - LOOK at your thermometers
- 35.0oC- need at least 0.5 units
- 44.5oC- need at least 0.1 units
14Thermometers
- Recording temperatures is dependent on the
graduations of the thermometer. - For example, if your thermometer is in 0.5
increments, temperature recordings must either
end in NN.0 or NN.5.
15Microorganisms
- D.3.7(a) - Does the laboratory use reference
cultures of microorganisms for positive
negative controls obtained from a recognized
national collection, organization, or
manufacturer recognized by the NELAP Accrediting
Authority? - Typically ATCC
- Do you have your certificates?
- Do you use these organisms for your media QC?
- Are your reference cultures not expired?
- Traceability of use?
16Microorganisms
- D.3.7(a) - Note Microorganisms can be
single-use preparations or cultures maintained by
documented procedures that demonstrate continued
purity viability of the organism. - Documented Do you have a written procedure?
- Do you have a subculture record (traceability)?
- 5.5.4.1.1 - Does the laboratory have standard
operating procedures that accurately reflect all
phases of current laboratory activities?
17Microorganisms
- D.3.7(a) - Note Microorganisms can be
single-use preparations or cultures maintained by
documented procedures that demonstrate continued
purity viability of the organism. - Purity How do you know there is only a single
type organism? - Gram stain? Streak for isolated colony? API?
- Viability How do you know the organism is still
alive after storage under your specified
conditions? - How do you know your negative control culture is
viable?
18Microorganisms
- D.3.7(a)(1) and (2)
- Are reference cultures of microorganisms revived
(if freeze-dried) or transferred from slants
subcultured only once to provide reference
stocks? - Are reference stocks preserved by a technique
that maintains the desired characteristics of the
strain? - Are the working stocks of microorganisms for
routine work prepared from the reference stocks? - Are reference stocks that have been thawed not
re-frozen re-used? - Are microorganism working stocks not sequentially
subcultured more than 5 times? - Are working stocks of microorganisms not
subcultured to replace reference stocks?
19Autoclave
- D.3.8(b)(2)(i) - Has the laboratory evaluated the
functional properties performance (e.g., heat
distribution characteristics) for each autoclave
with respect to typical uses? - One time study- can you locate your records?
- Can be done with biological indicators or maximum
registering thermometer.
20Autoclave
- D.3.8(b)(2)(i) - Is the autoclave capable of
meeting specified temperature tolerances? - Check manufacturers specifications.
- SM 20th Ed. media 121-124oC
21Autoclave
- D.3.8(b)(2)(ii) - Does the laboratory demonstrate
sterilization temperature by using a continuous
temperature recording device or maximum
registering thermometer with each cycle? - D.3.8(b)(2)(iv) - Does the laboratory perform
autoclave maintenance annually (either internally
or by service contract) which includes a pressure
check calibration of the temperature device?
SM requires semi-annual calibration of
temperature devices.
22Autoclave
- D.3.8(b)(2)(iv) - Does the laboratory perform
autoclave maintenance annually (either internally
or by service contract) which includes a pressure
check calibration of the temperature device? - Pressure check Is there a leak around the
seal/gasket of the autoclave? Is the pressure
15-20 psi during the sterilization portion of the
autoclave cycle? Check manufacturers
specifications.
23Autoclave
- D.3.8(b)(2)(iii) - Does the laboratory record the
1 date, 2 contents, 3 maximum temperature
reached, 4 pressure, 5 time in sterilization
mode, 6 total run time (may be documented as
time in time out), and 7 analysts initials
for every cycle of autoclave operations? - Equipment Identification?
24Autoclave
- D.3.8(b)(2)(v) - Does the laboratory check the
autoclave mechanical timing device quarterly
against a stopwatch and document the actual
elapsed time? - Includes automatic autoclaves with continuous
recorders. - D.3.8(b)(2)(ii) - Does the laboratory use
temperature sensitive tape with the contents of
each autoclave run to indicate that the autoclave
contents have been processed? -Tape only
indicates that materials have been inside an
autoclave, not that autoclave conditions have
been met. - D.3.8(b)(2)(i) - Are pressure cookers not used
for sterilization of growth media?
25Autoclave
- D.3.8(b)(2)(ii) - Does the laboratory use
appropriate biological indicators once per month
if in use to determine effective sterilization? - -Biological indicator contains thermophilic
spore-forming organism, typically Bacillus
stearothermophilus - -Have you ever processed a non-autoclaved
control? (See NELAC 5.4.6.2) - -Do you keep records? Optimally, these records
would include the incubator ID, incubation
period, traceability to lot of spores used,
retention of spore certificates, analyst
initials? - -Do you incubate at the proper temperature?
Typically, 55-60oC (see manufacturers
instructions).
26UV Sterilization
- D.3.8(b)(4) - If used for sanitation, are UV
instruments tested quarterly for effectiveness
with an appropriate UV light meter or by plate
count agar spread plates? - Note UV bulbs must be replaced if output is
less than 70 of the original for light tests or
if count reduction is less than 99 for a plate
containing 200-300 organisms.
27Ovens for Sterilization
- D.3.8(b)(6)(ii) - Are ovens used for
sterilization checked for sterilization
effectiveness monthly with appropriate biological
indicators? - D.3.8(b)(6)(ii) - Does the laboratory maintain
records of each sterilization cycle for the oven
that include date, cycle time, temperature,
contents, analysts initials? - Equipment ID?
28Washing
- D.3.8(b)(7)(i) - Does the laboratory have a
documented written procedure for washing
labware? - D.3.8(b)(7)(i) - Does the laboratory use
detergents Alconox, Liquinox designed for
laboratory use for washing labware? - D.3.8(b)(7)(ii) - Is the laboratorys glassware
used for Microbiological analysis made of
borosilicate or other non-corrosive material,
free of chips cracks, and have readable
measurement marks?
29Washing- IRT
- IRT- Inhibitory Residue Test
- D.3.8(b)(7)(iii) - Is labware that is washed
reused tested for possible presence of residues
which may inhibit or promote growth of
microorganisms by performing the Inhibitory
Residue Test annually? - D.3.8(b)(7)(iii) - Does the laboratory perform
the Inhibitory Residue Test each time it changes
the lot of detergent or washing procedures? - Either current (annual) certificate or annual
test is OK. - Procedure is in SM 20th Ed, 9020 B, 4(a)(2).
30Washing- pH test
- D.3.8(b)(7)(iv) - Does the laboratory test washed
labware at least once daily, each day of washing,
for possible acid or alkaline residues by
testing at least one piece of labware with a
suitable pH indicator such as bromothymol blue? - Note Records of these tests must be maintained.
31Sample Containers- Sterility
- D.3.1(a)(4) - Does the laboratory perform sample
container sterility checks on at least one
container for each lot of purchased,
pre-sterilized containers, or on one container
per sterilized batch for containers prepared
sterilized in the laboratory, with nonselective
growth media such as TSB. - -Includes sample bottles, sample bags,
Quantitray. - -Did you record the lot number?
32Sample Containers- Volume
- Non-disposable
- D.3.8(b)(3)(ii) - Does the laboratory calibrate
volumetric equipment such as filter funnels,
bottles, non-Class A glassware, other marked
containers once per lot prior to first use? - Disposable
- D.3.8(b)(3)(iii) - Does the laboratory check the
volume of disposable volumetric equipment such as
sample bottles, disposable pipettes,
micropipette tips-DELETE once per lot? - Did you record the batch number (sterilization
date) or lot number?
33Sample Containers-Chlorine
- Requirement depends on laboratorys procedures
for sample receipt but checks are typically
required. - 5.5.8.3.1(a)(2) - Has the laboratory checked
samples for proper preservation (e.g. pH, absence
of free chlorine, temperature) prior to or during
sample preparation or analysis? - Note These checks are not required for
chlorinated water systems as long as The
laboratory must have records showing that
Chlorine was measured in the field the actual
concentration is documented AND - The laboratory must check one sample container
from each commercial lot or prepared batch (for
adequate Na2S2O3), to prove that 5 mg/L Chlorine
in Drinking Water 15 mg/L Chlorine in
Non-Potable Water can be neutralized.
34Sample Receipt- Chlorine
- 5.5.8.3.1(a)(2) - Has the laboratory checked
samples for proper preservation (e.g., pH,
absence of free chlorine, temperature) prior to
or during sample preparation or analysis? - Note These checks are not required for
chlorinated water systems as long as - Sufficient Na2S2O3 was added to each sample
container to dechlorinate at least 5 mg/L
Chlorine in Drinking Water samples at least 15
mg/L Chlorine in Non-Potable Water samples. - -How do you demonstrate this?
35Sample Receipt- Chlorine
- (1) The laboratory must have records showing
that Chlorine was measured in the field the
actual concentration is documented AND - (2) The laboratory must check one sample
container from each commercial lot or prepared
batch (for adequate Na2S2O3), to prove that 5
mg/L Chlorine in Drinking Water 15 mg/L
Chlorine in Non-Potable Water can be neutralized.
36Sample Receipt- Chlorine
- If no field chlorine level is documented for the
sample, the laboratory must check the sample for
chlorine. - The laboratory may check each sample for chlorine
and not check each lot/batch of sample containers
for adequate Na2S2O3. - Even sources known not to contain chlorine must
have chlorine checks.
37Sample Receipt- Temperature
- 5.5.8.3.1(a)(2) - Has the laboratory checked
samples for proper preservation (e.g. pH, absence
of free chlorine, temperature) prior to or during
sample preparation or analysis? - DEP-SOP-001/01, Table FS 1000-4 However, even
if ice is present when the samples arrive, it is
necessary to immediately measure the temperature
of the samples (document actual temperature) - Lab must also document if sample was received on
ice or not on ice.
38Sample Receipt- Temperature
- DEP-SOP-001/01, Table FS 1000-8
- Drinking Water (Total coliforms, fecal coliforms,
E. coli, HPC) - DEP-SOP-001/01, Table FS 1000-4
- Non-Potable Water (Total coliforms, fecal
coliforms) 4oC - NEW 40 CFR Part 136, Table II
39Sample Receipt- Temperature
- How do I take sample temperature?
- Temperature of ice water bath in cooler must not
be used for sample temperature. - Can take temperature of each sample,
representative sample, or dummy sample per
cooler. - Lab needs to have a procedure!
- If using non-invasive temperature measuring
device (IR gun), have you calibrated it?
40Sample Receipt- Temperature Acceptability
- 5.5.8.3.1(a)(1) - For samples that require
thermal preservation, does the laboratory
consider acceptable only those samples where the
arrival temperature is within 2oC of the required
temperature or method-specified range OR is
within 0-6oC (where the specified temperature is
4oC). - Note For samples hand-delivered to the
laboratory on the same day calendar date that
they are collected, samples are considered
acceptable if there is evidence that the chilling
process has begun (e.g., arrival on ice).
41Sample Receipt- Holding Times
- DEP-SOP-001/01, Table FS 1000-8
- Drinking Water (Total coliforms, fecal coliforms,
E. coli) 30 hrs (HPC) 8 hrs - DEP-SOP-001/01, Table FS 1000-4
- Non-Potable Water (Total and fecal coliforms) 6
hrs - NEW 40 CFR Part 136, Table II 6 hrs transport
with 2 hours to begin analysis after receipt - Hold time Time between collection and analysis
- Is the sample time and date documented?
- Is the analysis time and date documented?
- Do you reject the sample or qualify the data if
holding times are exceeded (procedure should be
in corrective action/contingency plan)?
42Sample Receipt- pH
- 5.5.8.3.1(a)(2) - Has the laboratory checked
samples for proper preservation (e.g., pH,
absence of free chlorine, temperature) prior to
or during sample preparation or analysis? - pH not typically required for microbiology
samples (virus method?)
43Volumetric Equipment
- D.3.8(b)(3)(i) - Does the laboratory calibrate
volumetric equipment with movable parts, such as
automatic dispensers, dispensers/diluters,
mechanical hand pipettes quarterly? - D.3.8(b)(3)(ii) - Does the laboratory calibrate
non-disposable volumetric equipment such as
filter funnels, bottles, non-Class A glassware,
other marked containers once per lot prior to
first use? - D.3.8(b)(3)(iii) - Does the laboratory check the
volume of disposable volumetric equipment such as
sample bottles, disposable pipettes,
micropipette tips- DELETE once per lot?
44Environmental Conditions
- NELAC 5.5.3.2 The laboratory does not provide
for effective monitoring, control, and recording
of appropriate environmental conditions (such as
biological sterility, dust, electromagnetic
interference, humidity, mains voltage,
temperature, and sound and vibration levels). - Note SM 9020 B, 2(e) requires monthly air
monitoring to not exceed 15 colonies/plate/15
minutes.
45Colony counts- Reproducibility
- D.3.2 - If the test method specifies colony
counts (e.g., membrane filtration, HPC), the
laboratory does not verify the ability of
individual analysts to count colonies at least
once per month by having two or more analysts
count colonies from the same plate. - Note Counts must be within 10 between multiple
analysts to be acceptable. - Note An analyst in a 1-person laboratory may do
repetitive counting on the same plate, with no
more than 5 difference between the counts.
46Sterility Checks- Misc.
- D.3.1(a) - Does the laboratory demonstrate that
filtration equipment filters, sample
containers, media, reagents have not been
contaminated through improper handling or
preparation, inadequate sterilization, or
environmental exposure? Petri dishes? Blender?
Pipettes? Quantitray? - D.3.1(a)(4) - Does the laboratory perform sample
container sterility checks on at least one
container for each lot of purchased,
pre-sterilized containers, or on one container
per sterilized batch for containers prepared
sterilized in the laboratory, with nonselective
growth media? - Non-selective growth media typically TSB
47Sterility Checks- Misc.
- D.3.1(a)(5) - Does the laboratory perform a
sterility blank on each batch of dilution water
prepared in the laboratory, on each batch of
prepared, ready-to-use dilution water, with
nonselective growth media? (may use double
strength TSB) - D.3.1(a)(6) - Does the laboratory check at least
one filter from each new lot of membrane filters
for sterility with non-selective growth media?
48Sterility Checks- Membrane Filtration Method
- D.3.1 (a)(2) Is one beginning and one ending
sterility check conducted for each laboratory
sterilized unit used in a filtration series? Is
a sterility check conducted once per lot for
pre-sterilized single-use funnels? Note The
filtration series may include single or multiple
filtration units that have been sterilized prior
to beginning the series. - D.3.1 (a)(2) Is the membrane filtration series
ended when more than 30 minutes elapses between
successive filtrations? - D.3.1 (a)(2) Is a sterility blank analyzed
every 10 samples (unless filtration units are
sanitized by UV light after each filtration)? - Note During a filtration series filter funnels
must be rinsed with three 20-30 ml portions of
sterile rinse water after each sample filtration.
49Sterility Checks- Media
- D.3.1(a)(1) - Is a sterility blank analyzed for
each lot of pre-prepared, ready-to-use medium
for each batch of medium prepared in the
laboratory? - Note This blank must be analyzed prior to
first use of the medium. - Dont forget to check TSB itself.
- Dont dilute other media with TSB.
- D.3.1(a)(3) - For pour-plate technique does the
laboratory make a sterility blank of the medium
by pouring at least one uninoculated plate for
each lot of prepared, ready-to-use media for
each batch of medium prepared in the laboratory? - Dont forget HPC, PCA, etc.
50Media QC
- TESTS
- (1) Sterility
- (2) pH
- (3) Control
- (4) Control (if applicable)
- Buy ready to use test per lot
- Lab must check even if certificate was supplied.
- Make in lab test per batch
51Media- pH
- Does the pH measured meet method (or sometimes,
manufacturer) specifications? - Have you checked the pH of Colilert and other
similar media? (Suspend in reagent water and test
pH).
52Media /- Controls
- /- Controls Use pure, known cultures
- Note These culture controls must be analyzed
prior to first use of the medium and test
organisms need to respond in an acceptable
predictable manner. - Colilert and other media may have more controls.
- Do records show traceability to media lot number?
If media is prepared, do records show
traceability to preparation record? Do records
show traceability to reference stock and/or
working stock ID?
53Media /- Controls
- D.3.1(b), D.3.4(a) - Does the laboratory test
each lot of prepared, ready-to-use medium each
batch of medium prepared in the laboratory with
at least one pure culture of a known positive
reaction? - Dont forget TSB and HPC, PCA.
- D.3.1(c) - Does the laboratory test each lot of
prepared, ready-to-use medium each batch of
medium prepared in the laboratory with at least
one or more known negative culture controls
(non-target organisms) as appropriate to the
method? - N/A for general purpose (non-selective) media
- such as TSB, HPC, PCA, etc.
54Media- General
- D.3.6 - Does the laboratory ensure that the
quality of reagents media is appropriate for
the test concerned? - D.3.6(a) - Does the laboratory only use culture
media from commercial dehydrated powders or
purchased ready-to-use?
55Media- Original Expiration Date
- D.3.6(b) - Does the laboratory use reagents,
commercial dehydrated powders, media within the
shelf-life of the product? (Not expired?) - D.3.6(b) - Are all original containers of
reagents media labeled with an expiration
date? see also 5.5.6.4(b) - Assign expiration date if not assigned by
manufacturer.
56Media- SM Requirements
- SM 20th Ed. 9020 B, 4 (i)
- Store opened bottles in a desiccator.
- (Is your desiccant still working?)
- Use opened bottles of media within 6 months
(assign new expiration date after opening).
57Media Records
- D.3.6(d) - Does the laboratory have records on
media preparation in the laboratory that includes
the (1) date of preparation, (2) preparer's
initials, (3) type (4) amount of media
prepared, (5) manufacturer, (6) lot number, (7)
final pH of the media, (8) expiration date of
the prepared media? - D.3.6(d) - Does the laboratorys documentation on
media purchased pre-prepared, ready-to-use
include (1) manufacturer, (2) lot number, (3)
type (4) amount of media received, (5) date of
receipt, (6) pH of the media even Colilert!,
and (7) expiration date?
58Media-Misc.
- See checklist and method requirements.
- D.3.6(d) - Are the media, solutions, reagents
prepared, used, stored according to a
documented procedure that follows the
manufacturer's instructions or the test method? - HPC/PCA- Sterile agar medium melted not more than
once Melted agar used within 3 hours, agar
tempered at 44-46oC before pouring. - Media preparation- Is media autoclaved or brought
just to boiling point? - Colilert, Colilert-18, etc. Protect from light
- m-Endo Preparation Ethanol used is NOT denatured
59Media- Storage
- Storage of prepared media (SM9020B, 3h4
EPA-600/8-78-017, Part IV-A, 7.9 EPA 9131,
8.3.7) - Membrane Filter broth 4C 96 hours
- Membrane Filter agar plates 4C 2 weeks
- Media (loose-fitting closures) 4C 2 weeks
- HPC plates (sealed in plastic bags) 4C 2 weeks
- Broth media (with screw caps) 4C 3 months
- HPC agar (screw-cap container) 4C 3 months
- Refrigerated fermentation tube media incubated
overnight prior to use media indicating growth
not used OR - Fermentation tube media stored at 25oC used
within 1 week, evaporative losses
60Dilution Water
- Laboratory Prepared
- Water quality records
- Preparation record see 5.5.6.4(d)
- Sterility check (double strength TSB) see NELAC
D.3.1(a)(5) - pH check see D.3.6(d)
- Purchased Ready To Use
- Water quality records
- Sterility check
- pH check
- Precipitate check
61Dilution Water- 3/07 Guidance
- Laboratory prepared Sterility check per batch
as per Appendix D.3.1(a)(5) to NELAC Chapter 5.
The laboratory must also have source/reagent
water tests. - Laboratory purchased per lot Sterility check
per batch as per Appendix D.3.1(a)(5) to NELAC
Chapter 5. QC per SM 9020B, 4, (c) check pH
and check for precipitate. Also, obtain
source/reagent water QC per SM 9020B, Table II
for the lot. - Note Any unused dilution water in a container
opened 1 month shall be tested for sterility
before further use in order to satisfy the
requirements of Appendix D.3.1(a) to NELAC
Chapter 5.
62Support Equipment
- 5.5.5.2.1(d) - Is the following support equipment
associated with microbiological testing checked
with NIST traceable materials (where available) - pH meter retain buffer certificate?,
Balance(s), Conductivity meter, Chlorine meter,
Refrigerator(s), Water Baths, Incubators - D.3.8(b)(5) - Are conductivity meters, oxygen
meters, pH meters, hygrometers, other support
equipment calibrated according to the
method-specified requirements?
63Water Quality
- D.3.6(c) - Is the laboratory reagent water used
in the preparation of media solutions buffers
free from bactericidal inhibitory substances? - D.3.6(c) - Is the laboratory reagent water tested
monthly, when maintenance is performed on the
water treatment system, or at start-up when the
period of disuse exceeds one month, for 1
chlorine residual, 2 specific conductance,
3 HPC? - D.3.6(c) - Does the laboratory test its
Microbiology reagent water annually for 1 toxic
metals 2 Bacteriological Water Quality?
Note The Bacteriological Water Quality Test
Ratio is not required for laboratories that
have documentation to show that their water
source is Type I or Type II reagent water. (See
SM 1080 C)
64Water Quality
- D.3.6(c) - Does the laboratory maintain records
on all water quality checks (for 5 years) meet
the following criteria for acceptance (SM9020B,
4d EPA-600/8-78-017, Part IV-A, 5.2) - Pay attention to the required test intervals
and the actual data!
65Water Quality
- pH 5.5-7.5 (measured each use)
- Residual Chlorine
- Conductivity use)
- Heterotrophic Plate Count (monthly)
- Bacteriological ratio 0.8 3.0 (annually,
EPA-600/8-78-017 only) - Cd, Cr, Cu, Ni, Pb, Zn each collectively
- NH3, Organic Nitrogen
- TOC
- Student's t new water source)
66Water Quality- 3/07 Guidance
- A. Laboratory continuous preparation QC per
Standard Methods (SM) 9020B, Table II, including
frequencies. - B. Laboratory purchased per lot QC per SM 9020B,
Table II, including frequencies. The
laboratories may obtain source/reagent water QC
per SM 9020B, Table II from the manufacturer or
arrange for the required testing.
67Water Quality- 3/07 Guidance
- Note The manufacturers certificate of testing
must only be used for first month after receipt
of the lot of reagent water (except for
conductivity and pH which have different test
frequencies the laboratory must have additional
test records for days of use). - Reagent water lots stored at the laboratory 1
month must be tested at the required frequencies.
Laboratories may obtain the same or different
lots of water from the manufacturer on a monthly
basis, along with the relevant, current
certificates, to avoid having to perform
additional tests other than pH and conductivity.
68Standard Operating Procedures
- NELAC 5.5.4.1.2 (a) Does the laboratory have an
in-house methods manual for each accredited
analyte or method? Note This manual may
consist of copies of published or referenced test
methods. - NELAC 5.5.4.1.2 (b) Does the laboratory clearly
indicate in its methods manual any modifications
made to the referenced test method and describe
any changes or clarifications where the
referenced test method is ambiguous or provides
insufficient detail? - Appendix D Does the laboratory ensure that the
essential standards outlined in Appendix D are
incorporated into the method manuals and/or
Quality Manual?
69Standard Operating Procedures
- NELAC 5.5.4.1.2 (b) Does each test method in the
in-house methods manual include or reference - (1) Identification of the test method
- (2) Applicable matrix or matrices
- (3) Method Detection Limit
- (4) Scope application, with components to be
analyzed - (5) Summary of the test method
- (6) Definitions
- (7) Interferences
- (8) Safety
- (9) Equipment supplies
- (10) Reagents standards
- (11) Sample collection, preservn, shipment,
storage
70Standard Operating Procedures
- (12) Quality control
- (13) Calibration standardization
- (14) Procedure
- (15) Calculations
- (16) Method performance
- (17) Pollution prevention
- (18) Data assessment acceptance criteria for QC
- (19) Corrective actions for out-of-control data
- (20) Contingencies for handling out-of-control or
unacceptable data - (21) Waste management
- (22) References
- (23) Tables, diagrams, flowcharts, validation data
71Initial Demonstration of Capability
- D.3.3 (a) Has the laboratory demonstrated
proficiency with the test method prior to its
first use? - 10 spiked samples of typical matrix passing one
PT comparison to an accredited method. - 5.5.4.2.2 (a) and C.1 Did the laboratory
perform a satisfactory demonstration of method
capability prior to the acceptance institution
of this test method? - (See also Appendix D.3.3(a)) Analysts must not
process samples before having an acceptable DOC
72Demonstration of Capability
- C.1 Does the laboratory document in its Quality
Manual other adequate approaches to Demonstration
of Capability if this procedure is not required
by the mandated test method or regulation and if
the laboratory elects not to perform this
procedure? - If your method or regulation does not describe
how your lab will perform the DOC, then the lab
must define this procedure and place it into the
QM. - If there is no proficiency test available, how do
you demonstrate capability?
73Initial Demonstration of Capability
- 5.5.4.2.2 (d) and C.2 Does the laboratory use
the NELAC-specified certification statement to
document the completion of each Demonstration of
Capability (initial)? - The only time NELAC prescribes a form- USE IT.
- Form not required for continuing DOC but is
helpful. - C.2 Are copies of certification statements
retained in the personnel records of each
employee performing the test method? - Organize your analysts DOC information in a
personnel file. Statements should easily link to
the data used for the DOC.
74Demonstration of Capability
- NELAC 5.5.4.2.2 (d) Does the laboratory retain
all associated supporting data necessary to
reproduce the analytical results summarized in
the appropriate certification statement? - A logical place to link or have a copy of these
records are in a personnel DOC file. - NELAC 5.5.4.2.2 (e) and Appendix C.1 The
laboratory does not complete a demonstration of
capability each time there is a change in
instrument type, personnel, or test method.
75Continued Proficiency Demonstration of
Capability
- NELAC 5.5.2.6 (c)(3) - Each analyst does not have
documentation of continued cDOC proficiency by
at least one of the following once per year - (1) Acceptable performance of a blind sample
(single blind to the analyst). - (2) Another demonstration of capability.
- (3) Successful performance of a blind performance
sample on a similar test method using the same
technology. - (4) At least 4 consecutive laboratory control
samples with acceptable levels of precision
accuracy. - (5) Analysis of authentic samples that have been
analyzed by another trained analyst with
statistically identical results.
76Method Requirements
- Review actual method and its Quality Control, as
well as any additional Quality Controls that may
be a NELAC requirement. - Review checklists for some method requirements.
A checklist is a tool and guide only.
77Method Requirements SM 9223 B
- SM 9223 B certification includes ONLY
- Colilert and Colilert-18
- SM 9223 B certification does not include
- Readycult Verifications no longer required
- Colisure
- Quantitray
- Lab must apply for certification (equivalent
technology PTs, iDOC, SOP, etc.) - Do you use reagent water for Colilert controls?
Dont use dilution water. - Are samples 100 ml if performing
Presence/Absence? - Verifications not required if tests are P/A.
78Method Requirements- Common Problems
- Drinking Water- 100 ml sample
- Is your sample 100 ml (/- 2.5 ml)?
- How do you subsample appropriately?
- Membrane Filtrations
- Do dilutions yield the appropriate number of
colonies? - Do you verify typical AND atypical colonies, if
required by the method?
79Microbiological Sludge
- If your lab is analyzing microbiology sludge
samplesthe lab must have certification in the
Solid and Chemical Materials matrix. - DEP QA Rule 62-160.120 (16)(c), the Solid and
Chemical Materials (SCM) matrix includes sludges
biosolids are solids. - DEP-SOP-001/01, Table FS 1000-9 preserve Cool
4oC Holding time 24 hrs
80The End