Subchapter D. QUALITY ASSURANCE AND QUALITY CONTROL REQUIREMENTS
Sec.
252.401. Basic requirements.
252.402. Essential quality control requirements—chemistry.
252.403. Essential quality control requirements—toxicity testing.
252.404. Essential quality control requirement—microbiology.
252.405. Essential quality control requirement—radiochemistry.§ 252.401. Basic requirements.
(a) An environmental laboratory shall develop and maintain a quality manual appropriate to the type, range and volume of testing and analysis of environmental samples. The quality manual shall be available to and used by environmental laboratory personnel. The quality manual must contain the following:
(1) The full name and physical address of the laboratory.
(2) The name, address (if different from paragraph (1)), and telephone number of the laboratory supervisors.
(3) A revision number and effective date.
(4) A table of contents, and applicable lists of references, glossaries and appendices.
(b) The quality manual must state the environmental laboratory’s policies, operational procedures, protocols and practices established to meet the requirements of this chapter. These policies and procedures must include:
(1) An ethics policy statement as specified in subsection (d).
(2) A document control system as specified in subsection (c).
(3) Recordkeeping as specified in § 252.706 (relating to recordkeeping).
(4) The procedures for termination of operations and transfer of records as specified in § 252.706.
(5) The procedures for detecting and permitting departures from established procedures as specified in subsections (i) and (h).
(6) The procedures for detecting and preventing improper practices as specified in § 252.304 (relating to personnel requirements).
(7) The sample handling and acceptance procedures as specified in subsections (f) and (g).
(8) The reporting of analytical results as specified in subsection (j).
(9) The monitoring of the quality of analysis as specified in subsection (l).
(c) An environmental laboratory shall have a document control system that provides procedures for control and maintenance of all documents. The document control system must ensure that standard operating procedures, methods, manuals or documents clearly indicate the time period during which the procedure or document was in force.
(d) An environmental laboratory shall develop and maintain an ethics policy statement relevant to the employee’s duties and responsibilities under the act.
(1) The laboratory shall implement procedures for educating and training personnel in their ethical and legal responsibilities under the act.
(2) The laboratory shall provide training in ethical and legal responsibilities within 2 months of employment to the laboratory and at least every 14 months thereafter for all employees.
(e) An environmental laboratory shall maintain records of the technical personnel, which include dates of employment, signatures, initials and a list of persons authorized to approve or release reports of testing or analysis of environmental samples.
(f) An environmental laboratory shall establish procedures for handling environmental samples.
(1) The environmental laboratory shall implement procedures for checking and verifying the condition of the sample. The results of these checks shall be recorded. The environmental laboratory shall check:
(i) The sample container and the sample preservation, both thermal and chemical, of each sample.
(ii) The sample pH for all samples to be analyzed for whole effluent toxicity and safe drinking water chemistry fields of accreditation, unless the sample is collected by the environmental laboratory performing the analysis.
(iii) The sample for the presence of residual chlorine when the presence of residual chlorine will compromise the validity of the test.
(2) The laboratory shall utilize a recordkeeping system that meets the requirements of § 252.706 to document receipt of all sample containers. The recordkeeping system must include the following:
(i) The client/project name.
(ii) The date, time and location of sample collection, name of sample collector and field identification code.
(iii) The date and time of laboratory receipt and identification of the individual receiving the sample at the laboratory.
(iv) Any comments resulting from inspection for sample rejection shall be linked to the laboratory ID code.
(v) A unique laboratory ID code that corresponds to the information required by this paragraph.
(vi) An identification of the person making the entries.
(g) An environmental laboratory shall have a sample acceptance policy that clearly outlines the circumstances under which environmental samples will be accepted or rejected. The environmental sample acceptance policy must include the following areas:
(1) Sample identification, location, date and time of collection, collector’s name, preservation type and sample type.
(2) Sample labeling.
(3) Use of appropriate containers and sample preservation method.
(4) Adherence to holding times specified in the regulation and when not specified by the regulation, adherence to the holding times specified by the method.
(5) Sufficient sample volume shall be available to perform the necessary testing and analysis, including any required quality control testing or analysis.
(6) Procedures to be used when samples show signs of damage, contamination or inadequate preservation.
(h) An environmental laboratory shall document the laboratory management’s processes and procedures for permitting departures from the method, quality manual, established policies and procedures or standard operating procedures.
(i) An environmental laboratory shall establish procedures for detecting when departures from the method or quality manual have occurred. These procedures must include the following:
(1) Identify the individuals responsible for assessing each quality control type.
(2) Identify the individuals responsible for initiating or recommending, or both, corrective actions.
(3) Define how the analyst shall treat the results of testing or analysis of environmental samples if the associated quality control measures fail to meet the requirements of the method.
(4) Specify how out-of-control situations and subsequent corrective actions are to be documented.
(5) Specify procedures for the laboratory supervisor to review corrective action reports.
(j) An environmental laboratory shall develop procedures for reporting results of testing or analysis of environmental samples. Each test report must include at least the following information, except as specified in subsection (k).
(1) The name and address of the laboratory.
(2) The total number of pages in the report, including any addendums, in the format of Page x of y.
(3) The name and address of the client.
(4) An identification of the test method used.
(5) An identification of the samples including the client identification code.
(6) The date and time of sample collection.
(7) The date of sample analysis.
(8) The date and time of sample preparation or analysis, or both, if the holding time requirement for either activity is less than or equal to 72 hours.
(9) The test results and units of measurement.
(10) The quantitation limit.
(11) The names, functions and signatures of the persons authorizing the test report.
(12) An identification of results reported on a basis other than as received (for example, dry weight).
(13) An identification of testing or analysis results not covered by the laboratory’s scope of accreditation.
(14) An identification of results that do not meet the requirements of this chapter.
(15) An identification of subcontracted results.
(16) A unique test report identifier code, such as a serial number or other unique code.
(17) An identification of amendments to the test report. The laboratory shall uniquely identify all amendments to a test report. The amended report shall be issued in the form of a further document, data transfer or completely new test report, which includes the statement ‘‘Amended’’ or ‘‘Revised’’ and the identification of the unique laboratory code that meets the requirements of paragraph (16).
(k) Tests performed by an environmental laboratory operated by a facility that provides results to the facility management for compliance purposes do not need to be reported under subsection (j) regarding procedures for reporting results, provided the information required by subsection (j) is maintained under § 252.706.
(l) An environmental laboratory shall implement procedures or practices to monitor the quality of the laboratory’s analytical activities. Examples of the procedures or practices are:
(1) Internal quality control procedures using statistical techniques.
(2) Participation in proficiency testing, other inter-laboratory comparisons or round robin testing.
(3) Analysis of split samples by different laboratories.
(4) Use of certified reference materials or in-house quality control using secondary reference materials, or both.
(5) Replicate testing using the same or different test methods.
(6) Retesting of retained samples.
(7) Correlation of results for different but related analysis of a sample (for example, total phosphorus should be greater than or equal to orthophosphate).
(m) To the extent possible, results of testing or analysis of environmental samples shall be reported only if all quality control, analytical testing and sample acceptance measures are acceptable. If a quality control, analytical testing or sample acceptance measure is found to be out of control and the results of the testing or analysis of environmental samples are to be reported, all environmental samples associated with the failed quality control measure shall be documented and the results flagged in an unambiguous manner on the sample analysis report with the appropriate data qualifiers.
(n) Policies, procedures, protocols and practices specified in this section must be in writing and be followed.
(o) The environmental laboratory shall clearly identify opinions and interpretations as opinions and interpretations on test reports. When test reports include opinions and interpretations, the laboratory shall include an explanation for the basis upon which the opinions and interpretations have been made.
Authority The provisions of this § 252.401 amended under 27 Pa.C.S. § § 4103(a), 4104 and 4105; and section 1920-A of The Administrative Code of 1929 (71 P.S. § 510-20).
Source The provisions of this § 252.401 amended April 9, 2010, effective April 10, 2010, 40 Pa.B. 1898; amended July 28, 2017, effective July 29, 2017, 47 Pa.B. 4085. Immediately preceding text appears at serial pages (348810) to (348813).
Cross References This section cited in 25 Pa. Code § 252.5 (relating to NELAP equivalency); 25 Pa. Code § 252.402 (relating to essential quality control requirements—chemistry); 25 Pa. Code § 252.403 (relating to essential quality control requirements—toxicity testing); 25 Pa. Code § 252.404 (relating to essential quality control requirement—microbiology); and 25 Pa. Code § 252.405 (relating to essential quality control requirement—radiochemistry).
§ 252.402. Essential quality control requirements—chemistry.
(a) In addition to the requirements of § 252.401 (relating to basic requirements), laboratories performing testing or analysis of environmental samples in the area of chemistry shall comply with this section.
(b) When the method selected by an environmental laboratory in accordance with § 252.307 (relating to methodology) contains more stringent requirements than the requirements of this section, the environmental laboratory shall follow the more stringent requirements contained in the method.
(c) Initial calibration requirements are as follows:
(1) An environmental laboratory shall follow the initial calibration requirements of the method.
(2) The results of testing or analysis of environmental samples shall be determined from an initial calibration and may not be determined from any continuing calibration verification, unless otherwise required by regulation, method or program.
(3) The details of the initial calibration procedures including calculations, integrations, acceptance criteria and associated statistics shall be included or referenced in the laboratory’s standard operating procedure.
(4) Raw data records shall be retained to permit reconstruction of the initial calibration, including identification or reference to the reagents, standards and supplies used, dates of analysis, instrument identification, results of the initial calibration, calibration criteria and analyst identification.
(5) Initial calibrations shall be verified with a standard obtained from a second manufacturer or with a standard from the same manufacturer if the verification standard is documented by the manufacturer as prepared independently of the standard used during initial calibration.
(6) The lowest standard used for initial calibration may not be below the detection limit. The lowest standard must be at or below the lower limit of the range of quantitation.
(d) Except for methods that explicitly allow initial calibration using a single concentration of standard, initial calibration shall be done using multiple concentrations of standards according to the requirements of this subsection.
(1) Unless otherwise specified in the method, the initial calibration must meet one of the following criteria:
(i) A relative standard deviation of less than 20% for the calculated response factors.
(ii) A coefficient of determination (r2) of 0.99 for a linear calibration curve.
(iii) A coefficient of determination (r2) of 0.999 for a nonlinear calibration curve determined with the use of at least 6 calibration standards or as otherwise specified by the Department.
(2) If the initial calibration fails to meet established acceptance criteria, corrective action shall be performed and all associated environmental samples shall be reanalyzed after an acceptable initial calibration is obtained. If reanalysis of the environmental samples is not possible, a new environmental sample shall be collected.
(3) If the results of testing or analysis of environmental samples that are below the initial calibration range are reported, the results shall be reported with appropriate data qualifiers.
(4) If the results of testing or analysis of environmental samples are above the initial calibration range, the environmental sample shall be diluted and reanalyzed or the results reported with appropriate data qualifiers. Sample results within the established calibration range will not require data qualifiers.
(5) The lowest calibration standard may not be below the detection limit and may not be above the MCL.
(6) If the method does not specify the number of calibration standards, the minimum number of calibration standards for a response factor or linear calibration, not including blanks or a zero standard, shall be determined as follows:
(i) For an initial calibration covering a range up to 20 times the lowest quantitation level, a minimum of three calibration standards shall be used.
(ii) For an initial calibration covering a range from greater than 20 times and up to 50 times the lowest quantitation level, a minimum of four calibration standards shall be used.
(iii) For an initial calibration covering a range greater than 50 times and up to 100 times the lowest quantitation level, a minimum of five calibration standards shall be used.
(e) For a method that explicitly allows calibration using a single concentration of a standard, not including a blank or zero concentration standard, the initial calibration shall meet the requirements of this subsection.
(1) Prior to the testing or analysis of environmental samples, the linear range of the instrument shall be established by analyzing a series of standards, one of which shall be at the lowest quantitation level.
(2) An initial calibration using a single calibration standard and a zero point shall be performed at the beginning of each analysis day.
(3) A standard corresponding to the lowest quantitation level must be analyzed with each analytical batch and must meet the acceptance criteria established by the method. When there are no established criteria in the method, an environmental laboratory shall determine internal criteria and document the procedure used to establish the acceptance limits.
(4) If the results of testing or analysis of environmental samples that are below the lowest quantitation level verification standard, specified in paragraph (3), are to be reported, the results shall be reported with appropriate data qualifiers.
(5) If the results of testing or analysis of environmental samples produce a result above the associated single point standard, the environmental laboratory shall do one of the following:
(i) Analyze a standard at or above the sample concentration that meets established acceptance criteria to validate linearity.
(ii) Dilute the sample so that the result falls below the single point calibration concentration.
(iii) Report the data with an appropriate data qualifier.
(f) Calibration verification requirements are as follows:
(1) A calibration verification standard shall be analyzed at the beginning and end of each analysis day. For methods that use an internal standard, a calibration verification standard is not required at the end of the analysis day unless specified in the method, or State or Federal law or regulation.
(2) A calibration verification standard shall be analyzed after every ten samples, unless a different frequency is specified in the method.
(3) At a minimum, the laboratory shall verify the calibration curve of each analytical batch with calibration verification standards at a low and a high level.
(i) The concentration of the low calibration verification standard shall be within the lower 20% of the calibration curve and not more than five times the lowest quantitation level.
(ii) The concentration of the high calibration verification standard shall be within the upper 20% of the calibration curve.
(4) Details of the calibration verification procedure including calculations, integrations, acceptance criteria and associated statistics shall be included or referenced in the laboratory’s standard operating procedure.
(5) Raw data records shall be retained to permit reconstruction of the calibration verification.
(6) Acceptance criteria for calibration verification standards in the method shall be followed. When there are no established criteria in the method, an environmental laboratory shall use the acceptance criteria described in an equivalent method for the same type of analysis. When an equivalent method is not available, the laboratory shall establish control charts in accordance with Standard Methods for the Examination of Water and Wastewater (available from the American Public Health Association, 800 I Street, NW, Washington, D.C. 20001) to determine internal criteria and document the procedure used to establish the acceptance limits.
(7) If a calibration verification standard fails the established acceptance criteria, an environmental laboratory shall initiate corrective actions. If the corrective actions fail to produce an immediate consecutive calibration verification standard within the acceptance criteria, a new calibration verification standard shall be prepared. If the freshly prepared calibration verification standard fails to produce a result within the established acceptance criteria, the environmental laboratory shall recalibrate the test or analysis according to the method or as set forth in subsection (c) and as set forth in either subsection (d) or (e).
(8) To the extent possible, and as provided by paragraph (1), environmental samples not bracketed by acceptable calibration verification standards shall be reanalyzed. If the calibration verification standard is found to be out of control, and the results of the testing or analysis of environmental samples are to be reported, all environmental samples associated with the failed calibration verification standard shall be documented and the results flagged in an unambiguous manner on the sample analysis report with the appropriate data qualifiers.
(g) Method blank requirements are as follows:
(1) A method blank must be processed along with and under the same conditions as the associated environmental samples including all steps of the analytical procedure.
(2) A method blank must be analyzed at a minimum of one per preparation batch. When no separate preparation method is used (example: volatiles in water), the batch shall be defined as no more than 20 environmental samples that are analyzed together using the same method, personnel and lots of reagents.
(3) A method blank must consist of a matrix that is similar to the associated environmental samples and is free of the analytes of interest. When a matrix that is similar to the associated environmental samples that is free of the analytes of interest is not available, reagent water or an artificial or simulated matrix may be used.
(4) A method blank is considered contaminated if one of the following applies:
(i) The concentration of a target analyte in the method blank is at or above the reporting limit established by the method, by the laboratory or by regulation.
(ii) The contamination in the method blank otherwise affects the environmental sample results as described in the method or in individual project data quality objectives.
(5) If a contaminant is detected in the method blank, the source of contamination shall be investigated and measures shall be taken to minimize or eliminate the problem.
(6) Raw data records shall be retained to permit reconstruction of the method blank.
(7) To the extent possible, any environmental samples associated with a contaminated method blank shall be reprocessed for analysis. If a contaminated method blank is found to be out of control, and the results of the testing or analysis of environmental samples are to be reported, all environmental samples associated with the contaminated method blank shall be documented and the results flagged in an unambiguous manner on the sample analysis report with the appropriate data qualifiers.
(h) Laboratory control sample requirements are as follows:
(1) A laboratory control sample must be processed along with and under the same conditions as the associated environmental samples, including all steps of the preparation and analytical procedure.
(2) A laboratory control sample must consist of a matrix that is similar to the associated environmental samples and is free of the analytes of interest. When a matrix that is similar to the associated environmental samples that is free of the analytes of interest is not available, reagent water or an artificial or simulated matrix may be used.
(3) An environmental laboratory shall analyze a laboratory control sample at a minimum of one per preparation batch. When no separate preparation method is used, for example volatiles in water, the batch shall be defined as no more than 20 environmental samples that are analyzed together with the same method, personnel and lots of reagents.
(4) All analyte concentrations in the laboratory control sample must be within the calibration range of the method and at or below the maximum contaminant level.
(5) The components to be spiked into the laboratory control sample must be as specified by the method or other regulatory requirement. In the absence of specified components, the environmental laboratory shall use the following:
(i) For those components that interfere with an accurate assessment, such as spiking simultaneously with technical chlordane, toxaphene and PCBs, the laboratory control sample must represent the chemistries and elution patterns of the components to be reported.
(ii) For methods with more than ten analytes, a representative number may be chosen. The analytes selected shall be representative of all chemistries and analytes reported and shall be chosen using the following criteria:
(A) Targeted components shall be included in the laboratory control sample over a 2-year period.
(B) For methods that include one to ten components, the laboratory control sample must contain all components.
(C) For methods that include 11—20 components, the laboratory control sample must contain at least 10 components or 80%, whichever is greater.
(D) For methods with more than 20 components, the laboratory control samples must contain at least 16 components.
(6) Each individual laboratory control sample shall be compared to the acceptance criteria in the method. When there are no established criteria in the method, an environmental laboratory shall use the acceptance criteria described in an equivalent method for the same type of analysis. When an equivalent method is not available, the laboratory shall establish control charts in accordance with Standard Methods for the Examination of Water and Wastewater (available from the American Public Health Association, 800 I Street, NW, Washington, D.C. 20001) to determine internal criteria and document the procedure used to establish the limits.
(7) Raw data records shall be retained to permit reconstruction of the laboratory control sample.
(8) Environmental samples associated with an out of control laboratory control sample shall be reprocessed and reanalyzed from the beginning of the method or the results reported with the appropriate data qualifiers.
(i) Sample duplicate requirements are as follows:
(1) A sample duplicate or matrix spike duplicate must be processed along with and under the same conditions as the associated environmental samples, including all steps of the preparation and analytical procedure.
(2) A sample duplicate or matrix spike duplicate shall be analyzed at a minimum of one per preparation batch. When no separate preparation method is used, for example volatiles in water, the batch shall be defined as no more than 20 environmental samples that are analyzed together using the same method, personnel and lots of reagents.
(3) An environmental laboratory shall document the calculations used for determining the relative percent difference or other statistical method for evaluation of the duplicate pairs.
(4) Each duplicate relative percent difference shall be compared to the acceptance criteria in the method. When there are no established criteria in the method, an environmental laboratory shall use the acceptance criteria described in an equivalent method for the same type of analysis. When an equivalent method is not available, the laboratory shall establish control charts in accordance with Standard Methods for the Examination of Water and Wastewater (available from the American Public Health Association, 800 I Street, NW, Washington, D.C. 20001) to determine internal criteria and document the procedure used to establish the acceptance limits.
(5) For duplicate results outside established criteria, corrective action shall be documented and the data reported with appropriate data qualifiers.
(j) Surrogate spike requirements are as follows:
(1) Surrogate compounds, when commercially available, shall be added to all samples, standards and blanks for all organic chromatography test methods.
(2) Surrogate compounds shall be chosen to represent the various chemistries of the target analytes in the method.
(3) The results of the surrogate spike shall be compared to the acceptance criteria published in the method. When there are no established acceptance criteria for surrogate recovery in the method, the environmental laboratory shall use the acceptance criteria described in an equivalent method for the same type of analysis. When an equivalent method is not available, the laboratory shall establish control charts in accordance with Standard Methods for the Examination of Water and Wastewater (available from the American Public Health Association, 800 I Street, NW, Washington, D.C. 20001) to establish internal criteria and document the method used to establish the acceptance limits.
(4) For surrogate spike results outside established criteria, corrective action shall be documented and the data reported with appropriate data qualifiers.
(k) Detection limit requirements are as follows:
(1) A detection limit shall be determined by the protocol in the method or regulation. If the protocol for determining detection limits is not specified in the method or regulation, the environmental laboratory shall select a procedure that reflects instrument limitations and the intended application of the method.
(2) A detection limit study is not required for any component for which spiking solutions or quality control samples are not available, such as temperature. A detection limit study is not required for testing or analysis where the results are logarithmic, such as pH, or when the results are expressed as presence or absence.
(3) A detection limit shall be initially determined for the compounds of interest in each method in a matrix in which neither the target analyte nor interferences are at a concentration that would impact the results. The detection limit shall be determined in the matrix of interest.
(4) A detection limit shall be determined each time there is a change in the method that affects how the test is performed or that affects the sensitivity of the analysis.
(5) The sample processing steps of the method shall be included in the determination of the detection limit.
(6) Supporting data shall be retained to permit reconstruction of the detection limit study.
(7) An environmental laboratory shall have an established procedure to relate detection limits with quantitation limits.
(8) The method’s lower limit of quantitation shall be established and shall be above the detection limit.
(l) When retention times are used for the identification of an analyte, an environmental laboratory shall develop and document acceptance criteria for retention time windows. The laboratory shall document acceptance criteria for mass spectral tuning.
(m) When manual integrations are performed for chromatography methods, the laboratory shall have written procedures for manual integrations and instrument manipulations.
(1) The manual integration procedures must detail the steps taken to perform the integrations and define proper and improper integrations.
(2) The laboratory shall document manual integrations with the reason for the integration and the initials of the individual performing the integration.
(3) The laboratory shall retain a copy of the data before and after manual integration.
(n) The laboratory shall employ confirmation techniques to verify the compound identification when positive results are detected on a sample from a location that has not been previously tested by the laboratory or for a sample location that has not previously yielded detectable results for a particular compound.
(1) The confirmations shall be performed when analysis involves the use of an organic chromatography method not utilizing a mass spectrometer.
(2) The confirmations shall be documented.
(o) Records of all equipment, reference materials, reagents, and supplies shall be maintained in accordance with § 252.306 (relating to equipment, supplies and reference materials).
Authority The provisions of this § 252.402 amended under 27 Pa.C.S. § § 4103(a), 4104 and 4105; and section 1920-A of The Administrative Code of 1929 (71 P.S. § 510-20).
Source The provisions of this § 252.402 amended April 9, 2010, effective April 10, 2010, 40 Pa.B. 1898; amended July 28, 2017, effective July 29, 2017, 47 Pa.B. 4085. Immediately preceding text appears at serial pages (348814) to (348820).
§ 252.403. Essential quality control requirements—toxicity testing.
(a) In addition to the requirements of § 252.401 (relating to basic requirements), an environmental laboratory that measures the toxicity or bioaccumulation of contaminants, including testing of effluents, receiving waters, sediments, elutriates, leachates and soils shall comply with this section.
(b) When the method selected by an environmental laboratory in accordance with § 252.307 (relating to methodology) contains more stringent requirements than the requirements of this section, the environmental laboratory shall follow the more stringent requirements contained in the method.
(c) An environmental laboratory that measures toxicity or bioaccumulation of contaminants shall comply with Chapter 16 (relating to water quality toxics management strategy—statement of policy) regarding counting of neonates, algae cells and weighing of fish for selected endpoints.
(d) Negative control requirements are as follows:
(1) In addition to the negative controls specified by the method, permit or regulation, additional negative controls shall be included when sample adjustments (for example, pH adjustments or dechlorination) or solvent carriers are used in the test.
(2) The results of the negative controls shall be compared to the acceptance criteria published in the method. When there are no established acceptance criteria for the negative control in the method, the environmental laboratory shall establish internal criteria and document the method used to establish the acceptance limits.
(3) The test acceptability criteria for negative controls as specified in the method must be achieved for both the reference toxicant and the environmental sample toxicity test.
(e) The requirements for reference toxicants are as follows:
(1) The environmental laboratory shall demonstrate the ability to obtain consistent results with reference toxicants before performing toxicity tests on environmental samples.
(i) Intralaboratory precision shall be determined by performing a minimum of five acceptable reference toxicant tests for each method and species using different batches of organisms and negative controls (water, sediment or soil) before performing testing or analysis on environmental samples.
(ii) An environmental laboratory shall maintain control charts for the control performance and reference toxicant statistical endpoint (such as NOEC or ECp) and evaluate the intralaboratory variability with a specific reference toxicant for each method.
(iii) The results of the toxicant test shall be compared to the acceptance criteria published in the method. When there are no established acceptance criteria for the toxicant test in the method, the environmental laboratory shall establish internal criteria and document the method used to establish the acceptance limits.
(2) The following minimum frequency of reference toxicant testing shall be met:
(i) Each batch of test organisms obtained from an outside source, field collection or from laboratory spawning of field-collected species not amenable to routine laboratory culture shall be evaluated with a reference toxicant test of the same type as the environmental toxicity test within 7 days preceding the test or concurrently with the test.
(ii) Test organisms obtained from in-house laboratory cultures shall be tested with reference toxicant tests at least once each month for each method.
(iii) If a species produced by in-house laboratory cultures is used less than once per month, a reference toxicant test of the same type shall be performed with each environmental toxicity test.
(iv) When methods and species commonly used in the laboratory are only tested on a seasonal basis, reference toxicant tests shall be conducted each month the method is in use.
(3) Ongoing environmental laboratory performance shall be documented by maintaining laboratory quality control charts that meet the following requirements:
(i) For endpoints that are point estimates (ICp, ECp), control charts shall be constructed by plotting the cumulative geometric mean and the limits that consist of the upper and lower 95% confidence limits (+2 standard deviations).
(ii) For endpoints from hypothesis tests (NOEC, NOAEC), control charts shall be constructed by plotting the values directly and the control limits shall consist of one concentration interval above and below the concentration representing central tendency or the mode.
(iii) After 20 data points are collected for a method and species, the control charts shall be maintained by using only the most recent 20 data points.
(iv) Test results that fall outside of control chart limits at a frequency of 5% or less shall be retested and confirmed before reporting and all results shall be documented in the report of the testing and analysis.
(v) The endpoint shall be compared to the acceptance criteria published in the method.
(vi) When there are no established acceptance criteria for the endpoint in the method, the environmental laboratory shall establish internal criteria and document the method used to establish the acceptance limits.
(vii) If the reference toxicant fails to meet acceptance criteria, the results of environmental toxicity tests conducted during the affected period shall be examined for defects and the test repeated using a different batch of organisms or the results shall be reported with appropriate data qualifiers.
(4) Reference toxicant tests conducted for a method and species must use the same reference toxicant, test concentrations, dilution water and data analysis method as the environmental toxicity tests for which the precision is being evaluated unless otherwise specified in the method.
(5) The test duration, dilution or control water, feeding, organism age, age range and density, test volumes, renewal frequency, water quality measurements, number of test concentrations, replicates and organisms per replicate must be the same as the environmental toxicity test. A dilution factor of greater than 0.5 shall be used for both acute and chronic tests.
(f) Sensitivity requirements are as follows:
(1) If the Dunnett’s procedure or hypothesis test (NOEC, NOAEC) is used, the statistical minimum significant difference (SMSD) by species shall be calculated according to the formula specified by the method and reported with the test results. The SMSD must be estimated for nonnormal distribution or heterogeneous variances, or both.
(2) Confidence intervals for point estimates (LCp, ICp or ECp) shall be reported as a measure of the precision around the point estimate value.
(g) When required, the data shall be plotted in the form of a curve relating the dose of the chemical or concentration of sample to cumulative percentage of test organisms demonstrating a response, such as death.
(h) At least once every 30 days, an environmental laboratory shall verify and document that the reagent grade water meets the following criteria:
(1) Conductivity must be less than 0.1 µmhos/cm or resistance greater than 17 megohms at 25°C.
(2) pH must be between 5.5 to 7.5.
(3) Total residual chlorine must be nondetectable.
(i) Reagent water used for culturing and testing shall be analyzed for toxic metals and organics whenever the minimum acceptability criteria for control survival, growth or reproduction are not met and no other cause can be identified.
(j) An environmental laboratory shall demonstrate that any analyte at a measured concentration or the reported detection limit does not exceed 1/10 the expected chronic value for the most sensitive species tested or cultured.
(k) Air used for aeration of test solutions, dilution waters and cultures must be free of oil and fumes.
(l) The requirements for test organisms are as follows:
(1) An environmental laboratory shall positively identify test organisms to species on an annual basis. The taxonomic reference (citation and pages) and the names of the taxonomic experts shall be documented. When organisms are obtained from an outside source, an environmental laboratory shall obtain the information from the supplier.
(i) Organisms used for a test must be from the same source. When available, certified seeds shall be used for soil tests.
(ii) Organisms used in tests or as brood stock to produce neonate test organisms must appear healthy, show no signs of stress or disease and exhibit survival of greater than 90% during the 24-hour period immediately preceding use in tests.
(iii) An environmental laboratory shall document the health and culturing conditions of all organisms used for testing. The documentation must include culture conditions and observations of any stress, disease or mortality.
(iv) When organisms are obtained from an outside source, the laboratory shall obtain written documentation of the water quality parameters and biological observations for each lot of organisms received.
(v) An environmental laboratory shall record the water quality parameters and the biological observations when the organisms arrive at the environmental laboratory.
(vi) Supporting information such as hatch dates and times, times of brood releases and metrics (for example, chironomid head capsule width) shall be documented.
(vii) Organisms obtained from an outside source may not be from different batches.
(viii) The control population of Ceriodaphnia in chronic effluent or receiving water tests may not contain more than 10% males.
(ix) Test soils and sediments must be within the geochemical tolerance range of the test organism.
(2) The requirements for feeding of test organisms are as follows:
(i) For each new batch of laboratory-prepared food or lot of commercial food used by the environmental laboratory, the performance of organisms fed with the new food shall be compared with the performance of organisms fed with a food of known quality. The suitability of food used for culturing shall be determined using a measure that evaluates the effect of food quality on survival and growth or reproduction of each of the relevant test species.
(ii) Foods used only in chronic toxicity tests shall be evaluated using the reference toxicant employed in the environmental laboratory quality assurance program, and shall be compared with results of previous tests using a food of known quality.
(iii) In the case of algae, rotifers or other cultured foods, which are collected as a continuous batch, the quality of the food shall be assessed as described in subparagraphs (i) and (ii) each time new nutrient stocks are prepared, a new starter culture is employed or when a significant change in culture conditions occurs.
(iv) The environmental laboratory shall have written procedures for the statistical evaluation of food acceptability.
(v) Food used to culture organisms used in bioaccumulation tests shall be analyzed for the compounds to be measured in the bioaccumulation tests.
(m) Equipment requirements are as follows:
(1) If closed incubators are used, culturing and testing of organisms shall be separated to avoid loss of cultures due to cross-contamination.
(2) Temperature control equipment must be adequate to maintain the required test temperature. The average daily temperature of the test solutions shall be maintained within 1°C of the selected test temperature for the duration of the test. Temperature measurements shall be made at least once per 24-hour period. The test temperature for continuous-flow toxicity tests shall be monitored and recorded continuously.
(3) The test chambers used in a test must be identical.
(4) Materials used for test chambers and any material coming in contact with test samples, solutions, control water, sediment, soil or food must be nontoxic and cleaned according to the method. Materials may not add to nor reduce sample toxicity.
(5) Light intensity shall be maintained as specified in the method. Measurements shall be made and recorded at least once per 12 months.
(6) The photoperiod shall be maintained as specified in the method and be documented at least once every 90 days.
(7) For algal and plant tests, the light intensity shall be measured and recorded at the start of each test.
(n) The requirements for sample holding times and conditions are as follows:
(1) The sample holding time may not exceed 36 hours.
(2) The last use of the sample in renewal tests may not exceed 72 hours unless specifically approved by the Department.
(3) Samples shall be chilled to 4°C during or immediately after collection and held at that temperature until time of analysis.
(o) Chronic tests must have a minimum of four replicates per treatment.
(p) The requirements for testing conditions are as follows:
(1) Dissolved oxygen and pH in aquatic tests must be within acceptable ranges published in the method. When there are no established acceptance criteria in the method, the environmental laboratory shall establish internal criteria and document the method used to establish the acceptance limits.
(2) During aquatic chronic testing, dissolved oxygen and pH shall be measured daily in at least one replicate of each concentration.
(3) In static-renewal tests, dissolved oxygen shall be measured at both the beginning and end of each 24-hour exposure period.
(4) The pH shall be measured at the end of each exposure period after organism transfer.
(5) Minimal aeration may be provided to tests only if acceptable dissolved oxygen concentrations cannot be otherwise maintained or if specified by the method.
(q) Records of all equipment, reference materials, reagents and supplies shall be maintained in accordance with § 252.306 (relating to equipment supplies and reference materials).
Authority The provisions of this § 252.403 amended under 27 Pa.C.S. § § 4103(a), 4104 and 4105; and section 1920-A of The Administrative Code of 1929 (71 P. S. § 510-20).
Source The provisions of this § 252.403 amended April 9, 2010, effective April 10, 2010, 40 Pa.B. 1898. Immediately preceding text appears at serial pages (317279) to (317284).
§ 252.404. Essential quality control requirement—microbiology.
(a) In addition to the requirements of § 252.401 (relating to basic requirements), environmental laboratories performing testing or analysis in the area of microbiology shall comply with this section.
(b) When the method selected by an environmental laboratory in accordance with § 252.307 (relating to methodology) contains more stringent requirements than the requirements of this section, the environmental laboratory shall follow the more stringent requirements contained in the method.
(c) The following pieces of equipment shall be maintained according to this subsection:
(1) Autoclave.
(i) An environmental laboratory shall use autoclaves that meet specified temperature tolerances of the method. Pressure cookers may not be used.
(ii) A continuous temperature-recording device or a maximum-temperature-registering thermometer shall be used during each autoclave cycle.
(iii) An environmental laboratory shall verify the sterilization capability of each autoclave by utilizing appropriate biological indicators (for example, spore strips or ampoules) once a month. Records of biological indicator tests shall be maintained in a laboratory notebook and include the autoclave identification, date, incubation time and temperature, results and initials of the responsible individual.
(iv) An environmental laboratory shall verify the mechanical timing device, if used, for each autoclave every 3 months. Records of mechanical timer verification shall be maintained in a laboratory notebook and include the autoclave identification, date, mechanical timing device time, actual time and initials of the responsible individual. Correction factors shall be documented and used.
(v) Autoclaves shall be properly cleaned and maintained. Copies of service contracts or internal maintenance protocols and maintenance records shall be kept.
(vi) Required times for autoclaving items at 121°C are set forth in this subparagraph. The following items must be at temperature for the required amount of time. Except for membrane filters and pads and carbohydrate-containing media, indicated times are minimum times and may necessitate adjustment depending upon volumes, containers and loads. For autoclave runs that include membrane filters and pads and media, the total cycle time may not exceed 45 minutes. Autoclaved membrane filters and pads and media shall be removed immediately after completion of the autoclave cycle.
(A) Membrane filters and pads 10 minutes (B) Carbohydrate-containing media 12-15 minutes (C) Contaminated test materials 30 minutes (D) Membrane filtration units 15 minutes (E) Sample containers 15 minutes (F) Individual glassware 15 minutes (G) Dilution water 15 minutes (H) Rinse water 15-30 minutes (vii) Records of each autoclave run shall be maintained in a laboratory notebook and include the date, contents, sterilization time and temperature, total cycle time (recorded as time in and time out) and initials of the responsible individual.
(viii) If an autoclave cycle fails to meet any requirement, corrective action shall be documented. Media may not be reautoclaved.
(2) Hot air oven.
(i) An environmental laboratory shall maintain a thermometer, graduated in 10°C increments or less with the bulb placed in sand, in each hot air oven.
(ii) An environmental laboratory shall verify the sterilization capability of each hot air oven by utilizing appropriate biological indicators (for example, spore strips) once a month. Records of biological indicator tests shall be maintained in a laboratory notebook and include the hot air oven identification, date, incubation time and temperature, results and initials of the responsible individual.
(iii) An environmental laboratory shall sterilize items in a hot air oven maintaining a temperature of 170°—180°C for a minimum of 2 hours. Only dry items may be sterilized in a hot air oven.
(iv) Records of each hot air oven operation shall be maintained and include the date, contents, sterilization time and temperature, and initials of the responsible individual.
(3) Inoculating equipment.
(i) An environmental laboratory shall use appropriate sterile inoculating equipment.
(ii) Metal loops and needles must be made of nickel alloy or platinum.
(iii) Wooden applicator sticks must be sterilized using dry heat.
(iv) For oxidase tests, nickel alloy loops may not be used.
(4) Membrane filtration equipment.
(i) Membrane filtration funnels must be stainless steel, glass, porcelain or autoclaveable or presterilized plastic. Membrane filtration funnels may not be scratched or corroded and may not leak.
(ii) Membrane filtration units shall be sterilized before the beginning of a filtration series. A filtration series ends when 30 minutes or longer elapses after a sample is filtered.
(iii) Forceps must be blunt and smooth-tipped without corrugations on the inner sides of tips.
(iv) Membrane filters must meet the following requirements:
(A) Membrane filters must be made of cellulose ester, white, grid marked, 47 mm diameter and 0.45-µm pore size unless otherwise specified by the method.
(B) Membrane filters must be either purchased presterilized or autoclaved for 10 minutes at 121°C before use. Membrane filters may not be brittle or distorted.
(C) Membrane filters must be approved (based upon manufacturer data from tests for toxicity, recovery, retention and absence of growth-promoting substances) for the specified analysis for which they are to be used.
(v) An environmental laboratory using an ultraviolet sanitation lamp to sanitize filtration funnels between successive filtrations shall test the ultraviolet sanitation lamp every 3 months for effectiveness with an appropriate UV light meter or by plate count agar spread plates. Records of ultraviolet lamp tests shall be maintained and bulbs shall be replaced if output is less than 70% of original for light tests or if count reduction is less than 99% for a plate containing 200 to 300 organisms.
(5) Culture dishes.
(i) Culture dishes must be presterilized plastic or sterilizable glass and of appropriate size for the method.
(ii) Stainless steel canisters, aluminum canisters or a wrap of heavy aluminum foil or char-resistant paper, shall be used for autoclave sterilization of glass culture dishes.
(iii) Loose-lid culture dishes shall be incubated in a tight fitting container containing a moistened paper towel.
(iv) Opened packs of disposable culture dishes shall be resealed between use periods.
(6) Culture tubes and closures. Culture tubes and containers must be of sufficient size to contain medium and sample without being more than three quarters full. Tube closures must be stainless steel, aluminum, plastic or a screw cap with a nontoxic liner.
(7) Pipettes.
(i) Pipettes must have legible markings and may not be chipped or etched and must be accurate to within 2.5% tolerance.
(ii) Stainless steel canisters, aluminum canisters or a wrap of heavy aluminum foil or char-resistant paper shall be used for autoclave sterilization of pipettes.
(iii) Opened packs of disposable sterile pipettes shall be resealed between use periods.
(8) Sample containers.
(i) Sample containers must be sterile plastic bags or wide-mouth plastic or noncorrosive glass bottles with nonleaking ground glass stoppers or caps with nontoxic liners that can withstand repeated sterilization. Sample containers must be capable of holding sufficient volume of sample for all required tests while maintaining adequate air space for mixing.
(ii) Glass stoppers must be covered with aluminum foil or char-resistant paper for sterilization.
(iii) Glass and plastic bottles that have not been presterilized shall be sterilized by autoclaving. Glass bottles may be sterilized by dry heat. Empty containers shall be moistened with several drops of water prior to autoclaving.
(9) Plastic and glassware washing procedure.
(i) Prior to the initial use of a lot of detergent or washing procedure, an environmental laboratory shall perform an inhibitory residue test utilizing the method described in the currently approved editions of Standard Methods for the Examination of Water and Wastewater (available from the American Public Health Association, 800 I Street, NW, Washington, D.C. 20001). Records of inhibitory residue tests shall be maintained and include the detergent identification, date, calculations, results and initials of responsible individual.
(ii) Washed plastic and glassware shall be tested at least once each month for possible acid or alkaline residue by testing at least one piece of plastic and glassware with a suitable pH indicator such as 0.04% bromothymol blue. Records of pH tests shall be maintained and include the date, results and identification of the responsible individual.
(10) Ultraviolet lamp. An environmental laboratory shall use a 365-nm, 6-watt ultraviolet lamp in a darkened room to view sample fluorescence.
(11) Quanti-TrayTM Sealer.
(i) An environmental laboratory shall perform a sealer check on each Quanti-Tray Sealer once a month by adding a dye to a water sample and performing the sealing procedure.
(ii) Records of the sealer check shall be maintained and include the sealer identification, date, results and initials of responsible individual. If dye is observed outside the wells, the Quanti-Tray Sealer may not be used.
(d) The requirements for reagent water are as follows:
(1) An environmental laboratory shall use reagent water in the preparation of media, solutions and buffers.
(2) An environmental laboratory shall demonstrate that reagent water meets the following criteria on a monthly basis or whenever maintenance is performed on the water treatment system or at startup after a period of nonuse longer than 1 month:
(i) Total chlorine residual must be less than 0.1 mg/L.
(ii) Conductivity must be less than 2.0 µmhos/cm or resistance greater than 0.5 megohms at 25°C.
(iii) Heterotrophic plate count must be less than 500 CFU/mL.
(3) An environmental laboratory shall demonstrate that reagent water meets the following criteria every 12 months:
(i) The individual concentration of lead, cadmium, chromium, copper, nickel and zinc must be less than 0.05 mg/L.
(ii) The total concentration of lead, cadmium, chromium, copper, nickel and zinc must be less than 0.1 mg/L.
(iii) Except as provided in subsection (d)(6), the bacteriological water quality test ratio must be between 0.8 and 3.0. The bacteriological water quality test shall be performed according to the currently approved editions of Standard Methods for the Examination of Water and Wastewater (available from the American Public Health Association, 800 I Street, NW, Washington, D.C. 20001).
(4) The metals analyses may only be performed by an environmental laboratory accredited under this chapter for those fields of accreditation.
(5) Results of the monthly and annual reagent water analysis shall be maintained and include the date, type of test, results and initials of responsible individual. Reagent water that does not meet the required criteria may not be used.
(6) The bacteriological water quality test need not be performed if the environmental laboratory can supply documentation to show that their laboratory pure water or reagent water meets the criteria, as specified in section 1080 of the currently approved editions of Standard Methods for the Examination of Water and Wastewater (available from the American Public Health Association, 800 I Street, NW, Washington, D.C. 20001), for Type I (high-quality) or Type II (medium-quality) reagent water.
(7) The heterotrophic plate count and bacteriological water quality test ratio analyses described in paragraphs (2) and (3) shall be performed by an environmental laboratory accredited under this chapter for the appropriate field of accreditation.
(e) The requirements for dilution/rinse water are as follows:
(1) Stock buffer solution or peptone water shall be prepared as specified in the currently approved editions of Standard Methods for the Examination of Water and Wastewater (available from the American Public Health Association, 800 I Street, NW, Washington, D.C. 20001).
(2) Stock buffers shall be autoclaved or filter-sterilized. Stock buffers shall be refrigerated and must be free from turbidity.
(3) Dilution/rinse water solutions shall be prepared as specified in the currently approved editions of Standard Methods for the Examination of Water and Wastewater (available from the American Public Health Association, 800 I Street, NW, Washington, D.C. 20001).
(f) The requirements for media are as follows:
(1) An environmental laboratory shall use dehydrated or commercially manufactured prepared media. Dehydrated media shall be stored in a cool, dry location. Caked or discolored dehydrated media shall be discarded.
(2) An environmental laboratory that prepares media from dehydrated stock shall follow method specifications.
(3) Media may not be reautoclaved.
(4) After preparation, media shall be stored and maintained as follows:
(i) Stored away from sources of direct light.
(ii) Prepared plates shall be stored in sealed plastic bags or containers.
(iii) Each bag, container or rack of broth or agar media shall be labeled with the date prepared or expiration date.
(iv) Fermentation media stored in a refrigerator shall be brought to room temperature before use. Media that shows growth or false positive results may not be used.
(v) Prepared liquid media shall be discarded if evaporation exceeds 10% of the original volume.
(vi) Poured agar plates and broth in tubes, bottles or flasks with loose-fitting closures shall be discarded if not used within 2 weeks of sterilization unless otherwise specified by the method.
(vii) Broth in tightly closed screw-cap tubes, bottles or flasks shall be discarded if not used within 3 months of sterilization unless otherwise specified by the method.
(g) An environmental laboratory shall demonstrate that the filtration equipment and filters, sample containers, media and reagents have not been contaminated through improper handling or preparation, inadequate sterilization or environmental exposure as follows:
(1) A sterility blank shall be analyzed for each lot of preprepared, ready-to-use medium and for each batch of medium prepared in the laboratory prior to first use of the medium. Records shall be maintained and include media identification, date and time of the start and end of incubation, results and initials of the responsible individuals. If sterility blank indicates contamination, the media may not be used.
(i) For chromogenic/fluorogenic media, add single-strength media to sterile reagent water and incubate at the appropriate temperature and time.
(ii) For all other media, incubate uninoculated, single-strength at the appropriate temperature and time.
(2) For each reusable membrane filtration unit used during a filtration series, the laboratory shall prepare at least one sterility blank at the beginning and at the end of the series. A series is considered ended when more than 30 minutes elapses between filtrations. The laboratory shall insert a sterility blank after every ten sample aliquots filtered through each membrane filtration unit or sanitize filtration units by UV light after each sample filtration in addition to the regular rinsing procedure. Records of sterility blank results shall be maintained in the same manner as the associated sample and include the date and time of the start and end of the incubation, results and initials of the responsible individuals. If sterility blanks indicate contamination, the laboratory must treat each affected sample according to program requirements.
(3) For presterilized single use filtration funnel units, a sterility check shall be performed on one funnel unit per lot.
(4) Sterility checks on sample containers shall be performed on at least one container for each lot of purchased, presterilized containers with an appropriate nonselective growth media. For containers prepared and sterilized in the laboratory, a sterility check shall be performed on one container per sterilized batch with an appropriate nonselective growth media. Results shall be maintained and include sample container identification, date and time of the start and end of incubation, results and initials of responsible individuals. If sample container sterility check indicates contamination, the affected sample container may not be used.
(5) A sterility blank shall be performed on each batch of dilution/rinse water prepared in the laboratory and on each batch of preprepared, ready-to-use dilution water with an appropriate nonselective growth media. The concentration of media shall be single strength after addition of dilution water. Results shall be maintained and include dilution/rinse water identification, date and time of the start and end of incubation, results and initials of the responsible individuals. If dilution/rinse water sterility check indicates contamination, the affected dilution water may not be used.
(6) At least one filter from each new lot of membrane filters shall be checked for sterility with an appropriate nonselective growth media. Results shall be maintained and include membrane filter identification, date and time of the start and end of incubation, results and initials of the responsible individuals. If the membrane filter sterility check indicates contamination, the affected membrane filters may not be used.
(7) Sterility checks on Quanti-TrayTM sample trays shall be performed on at least one sample tray for each lot of purchased presterilized sample trays with an appropriate nonselective growth media. Results shall be maintained and include sample tray identification, date and time of the start and end of incubation, results and initials of the responsible individuals. If the sample tray sterility check indicates contamination, the affected lot of sample trays may not be used.
(h) The requirements for positive and negative culture control checks are as follows:
(1) Each preprepared, ready-to-use lot of medium and each batch of medium prepared in the laboratory shall be tested by the laboratory with at least one pure culture of a known positive reaction prior to first use of the medium. Records shall be maintained and include the date and time of the start and end of incubation, media lot or batch number, type of media, positive culture control organism identification, results and initials of the responsible individuals. If positive culture control checks do not meet expected results, the affected media may not be used.
(2) Each preprepared, ready-to-use lot of selective medium and each batch of selective medium prepared in the laboratory shall be tested by the laboratory with at least one pure culture of a known negative reaction prior to first use of the medium. Records shall be maintained and include the date and time of the start and end of incubation, media lot or batch number, type of media, negative culture control organism identification, results and initials of the responsible individuals. If negative culture control checks do not meet expected results, the affected media may not be used.
(3) An environmental laboratory shall use stock positive and negative culture controls that are known and traceable to a recognized National collection. Documentation of traceability shall be maintained.
(4) Stock positive and negative culture controls shall be discarded after the manufacturer’s expiration date.
(5) Culture controls may be single use or cultures maintained by the laboratory using a documented procedure that maintains the purity and viability of the organisms.
(6) For cultures maintained by the laboratory, the following criteria must be met:
(i) Reference control cultures may be revived and subcultured once to provide reference stocks.
(ii) Reference stocks shall be preserved using a method which maintains the characteristics of the organism strains. If reference stocks are thawed, they may not be refrozen and reused.
(iii) Working stocks shall be prepared from reference stocks for routine laboratory work.
(iv) If the laboratory sequentially cultures working stocks, the laboratory shall prepare a second working stock. Sequential culturing may not be performed from a working stock that has been used for routine laboratory work.
(v) Working stocks may not be used for more than 30 days.
(vi) Working stocks may not be sequentially cultured more than five times and may not be subcultured to replace reference stocks.
(vii) Secondary working stocks shall be used to prepare sequential working stocks.
(7) Positive and negative controls must be processed under the same conditions and using the same equipment as routine environmental samples, including all steps of the preparation and analytical procedure.
(i) For test methods that specify colony counts, duplicate counts shall be performed monthly on one positive sample for each month that the test is performed. If the laboratory has two or more analysts, each analyst shall count typical colonies on the same plate. Counts may not differ by more than 10%. In an environmental laboratory with only one analyst, the analyst shall count the same plate twice. Counts may not differ by more than 5%.
(j) Quality control checks, including sterility checks and positive and negative controls, shall be conducted after the laboratory receives the material or supply and before or during first use. These checks shall be performed by an environmental laboratory accredited under this chapter and utilizing the same supplies, reagents and media to be used during laboratory analysis of environmental samples. Certificates of analysis from a manufacturer may not be used to demonstrate compliance with the requirements of this subsection.
(k) Records of all equipment, reference materials, reagents, media and supplies shall be maintained in accordance with § 252.306 (relating to equipment, supplies and reference materials).
Authority The provisions of this § 252.404 amended under 27 Pa.C.S. § § 4103(a), 4104 and 4105; and section 1920-A of The Administrative Code of 1929 (71 P.S. § 510-20).
Source The provisions of this § 252.404 amended April 9, 2010, effective April 10, 2010, 40 Pa.B. 1898; amended July 28, 2017, effective July 29, 2017, 47 Pa.B. 4085. Immediately preceding text appears at serial pages (348826) to (348834).
§ 252.405. Essential quality control requirement—radiochemistry.
(a) In addition to the requirements of § 252.401 (relating to basic requirements), laboratories performing testing or analysis of environmental samples in the area of radiochemistry shall comply with this section.
(b) When the method selected by an environmental laboratory in accordance with § 252.307 (relating to methodology) contains more stringent requirements than the requirements of this section, the environmental laboratory shall follow the more stringent requirements contained in the method.
(c) The requirements for initial calibration are as follows:
(1) An environmental laboratory shall follow the initial calibration requirements of the method or regulation.
(2) Initial calibrations shall be performed using calibration standards that have the same general characteristics as the associated environmental samples, for example geometry, homogeneity and density.
(3) The initial calibration must include, when applicable, determination of instrument background, efficiency, mass attenuation and energy calibration.
(4) The results of testing or analysis of environmental samples shall be determined from an initial calibration that is not more than 12 months old and may not be determined from any continuing calibration verification, unless otherwise required by regulation, method or program.
(5) The details of the initial calibration procedures including calculations, integrations, acceptance criteria and associated statistics shall be included or referenced in the laboratory’s standard operating procedures.
(6) Raw data records shall be retained to permit reconstruction of the initial calibration.
(d) The requirements for an instrument suitability verification are as follows:
(1) An instrument suitability verification standard shall be analyzed at the beginning of each analysis day, unless a higher frequency is required in the method or regulation.
(2) The instrument suitability verification standard shall be a check source that provides adequate counting statistics for a relatively short count time and is sealed or encapsulated to prevent loss of activity and contamination of the instrument and laboratory personnel.
(3) For alpha and gamma spectroscopy systems, the instrument suitability verification standard must include determination of instrument counting efficiency, energy calibration and peak resolution.
(4) For gas-proportional and liquid scintillation counters, the instrument suitability verification standard must include determination of instrument counting efficiency.
(5) For scintillation counters, the instrument suitability verification standard must include determination of instrument counting efficiency.
(6) Details of the instrument suitability verification procedure including calculations, integrations, acceptance criteria and associated statistics shall be included or referenced in the laboratory’s standard operating procedures.
(7) Raw data records shall be retained to permit reconstruction of the instrument suitability verification.
(8) Acceptance criteria for instrument suitability verification standards in the method or regulation shall be followed. When there are no established criteria in the method or regulation, an environmental laboratory shall determine internal criteria and document the procedure used to establish the criteria.
(9) If an instrument suitability verification standard fails the acceptance criteria, an environmental laboratory shall initiate corrective actions.
(10) Environmental samples not bracketed by acceptable instrument suitability verification standards shall be reanalyzed.
(e) The requirements for an instrument background measurement are as follows:
(1) An instrument background check shall be analyzed every analysis day.
(2) Instrument background values shall be subtracted from the total measured activity in the determination of the sample activity.
(3) Each individual background check shall be compared to the acceptance criteria in the method or regulation. When there are no established criteria in the method or regulation, an environmental laboratory shall determine internal criteria and document the procedure used to establish the limits.
(4) Environmental samples associated with an out of control instrument background check shall be reprocessed and reanalyzed from the beginning of the method or the results reported with the appropriate data qualifiers.
(f) The requirements for a method blank are as follows:
(1) A method blank shall be processed along with and under the same conditions as the associated samples including all steps of the preparation and analytical procedure.
(2) A method blank shall be analyzed at a minimum of one per preparation batch. When no separate preparation method is used, such as gamma analysis in water, the batch shall be defined as no more than 20 environmental samples that are analyzed together using the same method, personnel and lots of reagents.
(3) A method blank must consist of a matrix that is similar to the associated environmental samples and is free of the isotopes of interest. When a matrix that is similar to the associated environmental samples that is free of the analytes of interest does not exist and cannot be prepared, reagent water or an artificial or simulated matrix may be used.
(4) When an environmental sample is analyzed by gamma spectrometry by placing the sample matrix into a calibrated counting geometry, the method blank must consist of a similar counting geometry that is filled to a similar volume with reagent water to partially simulate gamma attenuation due to a sample matrix.
(5) The method blank result may not be subtracted from the sample results in the associated preparation or analytical batch unless permitted by the method or regulation.
(6) The method blank shall be prepared with similar aliquot size to that of the routine samples for analysis. The method blank result and acceptance criteria shall be calculated in a manner that compensates for sample results based upon differing aliquot size.
(7) If a contaminant is detected in the method blank, the source of contamination shall be investigated and measures shall be taken to minimize or eliminate the contamination. A method blank is considered contaminated if one of the following applies:
(i) The activity of a target isotope in the method blank is at or above the reporting limit established by the method or by regulation.
(ii) The contamination in the method blank otherwise affects the environmental sample results as described in the method, regulation or in individual project data quality objectives.
(8) Environmental samples associated with a contaminated method blank shall be reprocessed for analysis or the results reported with the appropriate data qualifiers.
(g) The requirements for a laboratory control sample are as follows:
(1) A laboratory control sample must be processed along with and under the same conditions as the associated environmental samples, including all steps of the preparation and analytical procedure.
(2) The laboratory control sample must consist of a defined matrix containing known and verified activities of isotopes. When a matrix that is similar to the associated environmental samples that is free of the analytes of interest is not available, reagent water or an artificial or simulated matrix may be used.
(3) A laboratory control sample must be analyzed at a minimum of one per preparation batch. When no separate preparation method is used, such as gamma analysis in water, the batch shall be defined as no more than 20 environmental samples that are analyzed together with the same method, personnel and lots of reagents.
(4) The activity of the laboratory control sample must be within the calibration range of the method and one of the following:
(i) Two to ten times the detection limit.
(ii) At an activity level comparable to that of the environmental samples being tested or analyzed, if the sample activities are expected to exceed ten times the detection limit.
(5) The standard used to prepare the laboratory control sample must be from a source independent of the standards used for initial calibration.
(6) When a radiochemical method, other than gamma spectroscopy, has more than one reportable isotope, for example, plutonium, Pu 238 and Pu 239, using alpha spectrometry, only one of the isotopes shall be included in the laboratory control sample. When more than one isotope is present above the specified detection limit, each isotope shall be assessed against the acceptance criteria.
(7) When gamma spectrometry is used to identify and quantitate more than one isotope, the laboratory control sample must contain isotopes that represent the low, for example americium-241, medium, for example cesium-137, and high, for example cobalt-60, energy range of the analyzed gamma spectra. The isotopes need not exactly bracket the calibrated energy range or the range over which isotopes are identified and quantitated.
(8) Each individual laboratory control sample must be compared to the acceptance criteria in the method or regulation. When there are no established criteria in the method or regulation, an environmental laboratory shall determine internal criteria and document the procedure used to establish the limits.
(9) Environmental samples associated with an out of control laboratory control sample shall be reprocessed and reanalyzed from the beginning of the method or the results reported with the appropriate data qualifiers.
(h) The requirements for sample duplicates are as follows:
(1) A sample duplicate shall be analyzed at a minimum of one per preparation batch. When no separate preparation method is used, for example gamma analysis in water, the batch shall be defined as no more than 20 environmental samples that are analyzed together using the same method, personnel and lots of reagents.
(2) An environmental laboratory shall document the calculations used for determining the relative percent difference or other statistical method for evaluation of the sample duplicate pairs.
(3) Each sample duplicate relative percent difference shall be compared to the acceptance criteria in the method or regulation. When there are no established criteria in the method or regulation, an environmental laboratory shall determine internal criteria and document the procedure used to establish the acceptance limits.
(4) For sample duplicate results outside established criteria, corrective action shall be documented and the affected data reported with appropriate data qualifiers.
(i) Tracer requirements are as follows:
(1) For those methods that utilize a tracer or internal standard, each sample result must have an associated tracer or internal standard recovery calculated and reported.
(2) The tracer or internal standard recovery shall be assessed against the acceptance criteria specified in the method or regulation. When there are no established criteria in the method or regulation, an environmental laboratory shall determine internal criteria and document the procedure used to establish the acceptance limits.
(3) For tracer or internal standard recovery outside established criteria, corrective action shall be documented and the data reported with appropriate data qualifiers.
(j) Carrier requirements are as follows:
(1) For those methods that utilize a carrier, each sample must have an associated carrier recovery calculated and reported.
(2) The carrier recovery for each sample shall be assessed against the acceptance criteria specified in the method or regulation. When there are no established criteria in the method or regulation, an environmental laboratory shall determine internal criteria and document the procedure used to establish the acceptance limits.
(3) For carrier recovery outside established criteria, corrective action shall be documented and the data reported with appropriate data qualifiers.
(k) The requirements for detection limits are as follows:
(1) A detection limit shall be determined by the protocol in the method or regulation. If the protocol for determining detection limits is not specified in the method or regulation, the environmental laboratory shall select a procedure that reflects instrument limitations and the intended application of the method.
(2) A detection limit shall be initially determined for the isotopes of interest in each method in a matrix in which neither the target isotope nor interferences are at a concentration that would impact the results. The detection limit shall be determined in the matrix of interest.
(3) A detection limit shall be determined each time there is a change in the method that affects how the test is performed or that affects the sensitivity of the analysis.
(4) The sample processing steps of the method shall be included in the determination of the detection limit.
(5) Supporting data shall be retained to permit reconstruction of the detection limit determination.
(6) An environmental laboratory shall have a written procedure to relate detection limits with quantitation limits.
(7) The method’s lower limit of quantitation shall be established and must be above the detection limit.
(l) Each result shall be reported with the associated measurement uncertainty. The procedures for determining the measurement uncertainty shall be documented and be consistent with the method and regulation.
(m) Records of all equipment, reference materials, reagents, and supplies shall be maintained in accordance with § 252.306 (relating to equipment, supplies and reference materials).
Authority The provisions of this § 252.405 amended under 27 Pa.C.S. § § 4103(a), 4104 and 4105; and section 1920-A of The Administrative Code of 1929 (71 P. S. § 510-20).
Source The provisions of this § 252.405 amended April 9, 2010, effective April 10, 2010, 40 Pa.B. 1898. Immediately preceding text appears at serial pages (317293) to (317298).
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