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Title: 2003 NELAC Standards Microbiology Checklist: How to Pass your Assessment


1
2003 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

2
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

3
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?

4
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?)

5
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?)

6
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)

7
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?

8
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?)

9
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?)

10
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??

11
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

12
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

13
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

14
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.

15
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?

16
Microorganisms
  • 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?

17
Microorganisms
  • 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?

18
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
    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?

19
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.

20
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

21
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.

22
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 manufacturers
    specifications.

23
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 analysts initials
    for every cycle of autoclave operations?
  • Equipment Identification?

24
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?

25
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
    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).

26
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.

27
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, analysts initials?
  • Equipment ID?

28
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 laboratorys glassware
    used for Microbiological analysis made of
    borosilicate or other non-corrosive material,
    free of chips cracks, and have readable
    measurement marks?

29
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).

30
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.

31
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,
    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?

32
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?

33
Sample 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.

34
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?

35
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.

36
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.

37
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.

38
Sample 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

39
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?

40
Sample 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).

41
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)?

42
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?)

43
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
    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?

44
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.

45
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.

46
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

47
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?

48
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.

49
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.
  • 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.

50
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

51
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).

52
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?

53
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?
  • 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.

54
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?

55
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.

56
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).

57
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 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?

58
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

59
Media- 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

60
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

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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.

62
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?

63
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)

64
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-600/8-78-017, Part IV-A, 5.2)
  • Pay attention to the required test intervals
    and the actual data!

65
Water 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)

66
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.

67
Water 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.

68
Standard 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?

69
Standard 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

70
Standard 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

71
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

72
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?

73
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 analysts DOC information in a
    personnel file. Statements should easily link to
    the data used for the DOC.

74
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.

75
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.

76
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.

77
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?
    Dont use dilution water.
  • Are samples 100 ml if performing
    Presence/Absence?
  • Verifications not required if tests are P/A.

78
Method 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?

79
Microbiological 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

80
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