2003 NELAC Standards Microbiology Checklist: How to “Pass” your Assessment

2003 NELAC Standards
Microbiology 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
Do 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]
Work 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?
Incubator/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?)
Incubator/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?)
Incubator/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)
Incubator/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?
Incubator/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?)
Refrigerator
• 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?)
Thermometers
• 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??
Thermometers
• 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
Thermometers
• 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
Thermometers
• 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
Thermometers
• 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”.
Microorganisms
• 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?
Microorganisms
• D.3.7(a) - Note: Microorganisms can be singleuse 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?]
Microorganisms
• D.3.7(a) - Note: Microorganisms can be singleuse 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?
Microorganisms
• 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 refrozen & re-used?
• Are microorganism working stocks not sequentially
subcultured more than 5 times?
• Are working stocks of microorganisms not subcultured to
replace reference stocks?
Autoclave
• 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.
Autoclave
• D.3.8(b)(2)(i) - Is the autoclave capable of
meeting specified temperature
tolerances?
– Check manufacturers specifications.
– [SM 20th Ed. media: 121-124oC]
Autoclave
• 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.]
Autoclave
• 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 manufacturer’s
specifications.]
Autoclave
• 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] analyst’s initials for
every cycle of autoclave operations?
– Equipment Identification?
Autoclave
• 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?
Autoclave
• 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 sporeforming 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 manufacturer’s instructions).
UV 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.
Ovens 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, & analyst’s initials?
– Equipment ID?
Washing
• 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 laboratory’s glassware
used for Microbiological analysis made of
borosilicate or other non-corrosive material, free
of chips & cracks, and have readable
measurement marks?
Washing- 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).
Washing- 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.
Sample 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, presterilized 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?
Sample 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?
Sample Containers-Chlorine
– Requirement depends on laboratory’s 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.
Sample 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?
Sample 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.
Sample 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.
Sample 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.
Sample Receipt- Temperature
• DEP-SOP-001/01, Table FS 1000-8 :
• Drinking Water (Total coliforms, fecal
coliforms, E. coli, HPC): < 10oC
• DEP-SOP-001/01, Table FS 1000-4:
• Non-Potable Water (Total coliforms, fecal
coliforms): 4oC
• NEW: 40 CFR Part 136, Table II: </= 10oC
Sample 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?
Sample ReceiptTemperature 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).
Sample 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)?
Sample 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?)
Volumetric 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 [nondisposable] 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 tipsDELETE] once per lot?
Environmental 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.
Colony 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.
Sterility 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
Sterility 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?
Sterility 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.
Sterility 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.
– Don’t forget to check TSB itself.
– Don’t 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?
– Don’t forget HPC, PCA, etc.
Media 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
Media- 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).
Media: +/- 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?
Media: +/- 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?
– Don’t 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.]
Media- 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?
Media- 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.
Media- 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).
Media 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 laboratory’s 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?
Media-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
Media- Storage
• Storage of prepared media (SM9020B, 3h4; EPA-600/878-017, 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 < 1 mL
Dilution 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
Dilution 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.
Support 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?
Water 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)
Water 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-600/8-78017, Part IV-A, 5.2):
**Pay attention to the required test intervals and the actual
data!
Water Quality
•
•
•
•
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pH 5.5-7.5 (measured each use)
Residual Chlorine < 1.0 mg/L (monthly)
Conductivity < 2.0 umho/cm at 25oC (with each use)
Heterotrophic Plate Count < 1000 CFU/ mL (monthly)
[Bacteriological ratio 0.8 – 3.0 (annually, EPA-600/8-78017 only)*]
Cd, Cr, Cu, Ni, Pb, Zn each < 0.05 mg/L, collectively <
0.1 mg/L (annually)
NH3, Organic Nitrogen < 0.1 mg/L (monthly check)
TOC < 1 mg/L (monthly)
Student's t < 2.78 for Use Test (quarterly & for new water
source)
Water 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.
Water Quality- 3/07 Guidance
• Note: The manufacturer’s 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.
Standard Operating Procedures
• NELAC 5.5.4.1.2 (a) – Does the laboratory have an inhouse 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?
Standard Operating Procedures
NELAC 5.5.4.1.2 (b) Does each test method in the inhouse 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, preserv’n, shipment, & storage
Standard Operating Procedures
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(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
[Initial] 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
Demonstration 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?
[Initial] 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 analyst’s DOC information in a
personnel file. Statements should easily link to the
data used for the DOC.
Demonstration 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.
[Continued 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.
Method 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.
Method 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? Don’t
use dilution water.
• Are samples 100 ml if performing Presence/Absence?
• Verifications not required if tests are P/A.
Method RequirementsCommon 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?
Microbiological Sludge
• If your lab is analyzing microbiology sludge
samples…the 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
The End