5 Colo. Code Regs. § 1005-7-7

Current through Register Vol. 47, No. 24, December 25, 2024

Rule 5 CCR 1005-7-7 - [Effective 1/14/2025] Natural Medicine Testing Facilities: Analytical Processes

7.1Method Validation and Verification. Analytical method selection, validation, and verification must ensure that the test method used is fit-for-purpose and that the Natural Medicine Testing Facility can successfully perform the testing.

7.1.1 The demonstration of testing validity must ensure consistent, accurate and reproducible analytical performance in the matrices tested by the laboratory.

7.1.2 Method performance specifications must ensure analytical tests are sufficiently sensitive for the purposes of the detectability requirements of Natural Medicine Division Rules, 1 CCR 213-1.

7.1.3 A Natural Medicine Testing Facility must validate new methodology and revalidate any changes to approved methodology prior to analyzing samples.

7.1.4 The Natural Medicine Testing Facility must implement a performance-based measurement system for the selected methodology and validate the method following good laboratory practices in accordance with AOAC International, United States Pharmacopeia (USP), United States Food and Drug Administration (FDA), United States Department of Agriculture (USDA), or other reputable validation guidelines and methodology prior to reporting results. Validation, verification, or matrix extension of other or new methodology must include when applicable, but is not limited to:

7.1.4.1 Verification of Accuracy

7.1.4.2 Verification of Precision

7.1.4.3 Verification of Analytical Sensitivity

7.1.4.4 Verification of Analytical Specificity

7.1.4.5 Verification of the LOD

7.1.4.6 Verification of the LOQ

7.1.4.7 Verification of the Reportable Range

7.1.4.8 Identification of Interfering Substances

7.1.4.9 Exclusivity, which means the specificity of the test method for validating microbial testing methods. It evaluates the ability of the method to distinguish the target organisms from similar but genetically distinct non-target organisms;

7.1.4.10 Inclusivity, which means, related to microbiological method validation, the sensitivity of the test method. It evaluates the ability of the test method to detect a wide range of target organisms by a defined relatedness;

7.1.4.11 Verification of Recovery. The laboratory shall take action to remediate recovery issues where Matrix effects affect recovery at greater than plus or minus 20% of the theoretical value;

7.1.4.12 Measurement Uncertainty.

7.1.4.12.1 Subsequent to initial validation, Measurement Uncertainty must be re-evaluated at least annually or whenever method modifications are made.

7.1.5 For qualitative methods, a minimum of three spiking/naturally occurring levels of target Analyte must be included assessing accuracy, precision and specificity per each matrix validated (microbiological methods shall be considered accurate/precise if fractional results are obtained when limits are <1 CFU preanalytical portion): one negative level in which no Analyte is present, one low level at which Analyte is present at the regulatory limit and one high level at which the Analyte is present at 10x the regulatory limit.

7.1.6 For qualitative methods, at least two levels must be included for the negative and high level spikes - at least ten replicates must be included at the fractional level, per Analyte, per matrix.

7.1.7 For quantitative methods, laboratories must include a minimum of four spiked levels or more, covering the analytical range must be included assessing accuracy, precision and specificity per each matrix validated: one negative level in which no Analyte is present, one low level at which Analyte is present at the regulatory limit, one medium level for which the Analyte is present at a concentration close to the middle calibrator and one high level at which the Analyte is present at the upper level of the calibration.

7.1.8 For quantitative methods, at least two replicates must be performed at each spiked level per each matrix validated.

7.1.9 Validation or verification of methodology must be documented in a validation report. The validation report shall include, but is not limited to, the following:

7.1.9.1 Validation plan;

7.1.9.2 Introduction and summary;

7.1.9.3 Materials, to include identification of Certified Reference Materials, and preparation methods;

7.1.9.4 Method parameters;

7.1.9.5 Raw data, including instrument raw data such as chromatograms, for each test method and each instrument, if any;

7.1.9.6 Instrument calibration data, if any;

7.1.9.7 Data, calculations, and results;

7.1.9.8 Method Acceptability Criteria performance data;

7.1.9.9 Interlaboratory Comparison

7.1.9.10 Verification of spreadsheets and/or laboratory information management system;

7.1.9.11 Conclusion and discussion; and

7.1.9.12 References.

7.1.10 Natural Medicine Testing Facilities must validate or verify instrumentation and methodology immediately and prior to use following a change in location.

7.1.11 Any changes to the approved other or new methodology must be revalidated and documented prior to testing samples.

7.1.12 Testing and Validation of Complex Matrices. A Natural Medicine Testing Facility must include a variety of matrices as part of the validation/verification process. During method validation/verification, a Natural Medicine Testing Facility must:

7.1.12.1 Select matrices which best represent each category of products to be tested as listed in 1.3.12 "Harvest Lot" and 1.3.28 "Production Lot." The Natural Medicine Testing Facility shall independently determine the category of matrix a product falls within; properties to consider include fat content, Tryptamine content, pH, salt content, sugar content, water activity, the presence of known chemical compounds, microbial flora and antimicrobial compounds.

7.1.12.2 Perform a new Matrix validation, prior to reporting results, on matrices which are either a new category of Matrix or are considerably different from the original matrix validated within the category.

7.1.12.2.1 For example, the Natural Medicine Testing Facility intends to receive gelatin-based gummies" for testing, but previous validation studies included powdered capsules and chocolate. A new validation should be performed for the product prior to testing since salt content and other properties differ vastly from the original matrices validated.

7.1.12.3 Perform a Matrix verification (a client Matrix spike or similar consisting of the target Analyte(s) at the time of analysis) on matrices submitted for testing which differ slightly from those initially validated but which fall within a category already validated.

7.1.12.3.1 For example, the Natural Medicine Testing Facility Natural Medicine Testing Facility receives a new edible type matrix for testing (agar-based gummies) but previous validation included soft confection edibles. A spike of a portion of the submitted material must be analyzed prior to, or at the time of, sample analysis.

7.1.13 Software must be validated prior to testing samples, including but not limited to: analytical software, application programming interface(s) (APIs), laboratory information management systems (LIMS), etc., to include any calculations performed by the software.

7.1.14 Prior to use, methodology must have a Standard Operating Procedure approved and signed by the laboratory director.

7.1.15 Testing analysts must have documentation of competency assessment prior to testing Samples.

7.1.16 Any changes to the approved methodology must be revalidated and documented prior to testing Samples. The documentation of changes and revalidation must be provided to the Department prior to implementation.

7.2General Laboratory Equipment. A Natural Medicine Testing Facility must:

7.2.1 Track, verify and apply correction factors where applicable;

7.2.2 Perform annual calibration, monthly spore ampule checks and daily temperature checks of autoclaves;

7.2.3 Verify pipette calibration on a monthly basis across the volume range the pipette is used;

7.2.4 Perform annual calibration of balances and analytical weights. Balances must be verified across the appropriate mass range on each day of use;

7.2.5 Perform verification of laboratory dispensers on each day of use;

7.2.6 Perform annual gravimetric verification of volumetric glassware;

7.2.7 Calibrate pH meters on each day of use with buffers across the analytical range (i.e., 4, 7, 10);

7.2.8 Perform annual calibration of thermometers to a NIST traceable reference thermometer. Correction factors must be applied when such a factor could impact the final result;

7.2.9 Calibrate analytical timers annually.

7.3Gas Chromatography (GC). A Natural Medicine Testing Facility using GC must:

7.3.1 Document the conditions of the gas chromatograph, including the detector response;

7.3.2 Perform and document preventive maintenance as required by the manufacturer and SOPs;

7.3.3 Ensure that records are maintained and readily available to the staff operating the equipment;

7.3.4 Document the performance of new columns before use;

7.3.5 Use an internal standard for each qualitative and quantitative analysis that has similar chemical and physical properties to that of the compound identified;

7.3.6 Establish Acceptability Criteria for variances between different aliquots and different columns;

7.3.7 Document the monitoring of the response (area or peak height) of the internal standard to ensure consistency over time of the analytical system;

7.3.8 Evaluate the performance of the instrument after routine and preventive maintenance prior to analyzing subject Samples; and

7.3.9 Monitor and document the performance of the instrument each day of testing.

7.4Gas Chromatography Mass Spectrometry (GC/MS). A Natural Medicine Testing Facility using GC/MS must:

7.4.1 Perform and document preventive maintenance as required by the manufacturer and SOPs;

7.4.2 Document and maintain records when cleaning or changes in source, source conditions, column, or other routine maintenance are made to the instrument;

7.4.3 Ensure that records are maintained and readily available to the staff operating the equipment;

7.4.4 Maintain records of mass spectrometric tuning;

7.4.5 Establish written criteria for acceptable mass-spectrometric tuning parameters;

7.4.6 Document corrective actions if a mass-spectrometric tune is unacceptable;

7.4.7 Monitor analytic analyses to check for contamination and carry-over;

7.4.8 Use selected ion monitoring within each run to assure that the Natural Medicine Testing Facility compares ion ratios and retention times between calibrators, controls and Samples for identification of an Analyte;

7.4.9 Use an internal standard for qualitative and quantitative analysis that has similar chemical and physical properties to that of the compound identified and is isotopically labeled when available or appropriate for the assay;

7.4.10 Document the monitoring of the response (area or peak height) for the internal standard to ensure consistency over time of the analytical system;

7.4.11 Define the criteria for designating qualitative results as positive;

7.4.12 When a library is used to qualitatively identify an Analyte, the identity of the Analyte must be confirmed before reporting results by comparing the relative retention time and mass spectrum to that of a known standard or control run on the same system;

7.4.13 Evaluate the performance of the instrument after routine and preventive maintenance (e.g. clipping or replacing the column or cleaning the source) prior to analyzing subject Samples; and

7.4.14 Monitor and document the performance of the instrument each day of testing.

7.4.15 Monitor gas phase standards frequently by comparison to the initial calibration curve. Fresh standards must be prepared if this check exceeds 20% drift. Gas standards must be replaced after one week or as recommended by the standard manufacturer. Alternatively, standards must be replaced after an established period of time for which the Natural Medicine Testing Facility has demonstrated the stability of the standard..

7.4.16 Monitor non-gas standards frequently by comparison to the initial calibration. Fresh standards must be prepared if this check exceeds 20% drift. Non-gas standards must be replaced after one month for working standards and three months for opened stocks or as recommended by the standard manufacturer. Alternatively, standards must be replaced after an established period of time for which the Natural Medicine Testing Facility has demonstrated the stability of the standard; and

7.4.17 Monitor and document the performance of the instrument each day of testing.

7.5Immunoassays. A Natural Medicine Testing Facility using Immunoassays must:

7.5.1 Perform and document preventive maintenance as required by the manufacturer and SOPs;

7.5.2 Ensure that records are maintained and readily available to the staff operating the equipment;

7.5.3 Validate any changes or modifications to a manufacturer's approved assays or testing methods when a Sample is not included within the types of Samples approved by the manufacturer; and

7.5.4 Define acceptable separation or measurement units (absorbance intensity or counts per minute) for each assay, which must be consistent with manufacturer's instructions.

7.6High Performance Liquid Chromatography (HPLC). A Natural Medicine Testing Facility using HPLC must:

7.6.1 Perform and document preventive maintenance as required by the manufacturer and SOPs;

7.6.2 Ensure that records are maintained and readily available to the staff operating the equipment;

7.6.3 Monitor and document the performance of the HPLC instrument each day of testing;

7.6.4 Evaluate the performance of new columns before use;

7.6.5 Create written standards for acceptability when eluting solvents are recycled;

7.6.6 Use an internal standard for each qualitative and quantitative analysis that has similar chemical and physical properties to that of the compound identified when available or appropriate for the assay;

7.6.7 Document the monitoring of the response (area or peak height) of the internal standard to ensure consistency over time of the analytical system;

7.6.8 Evaluate the performance of the instrument after routine and preventive maintenance prior to analyzing subject Samples; and

7.6.9 Monitor and document the performance of the instrument each day of testing.

7.7Liquid Chromatography Mass Spectrometry (LC/MS). A Natural Medicine Testing Facility using LC/MS must:

7.7.1 Perform and document preventive maintenance as required by the manufacturer and SOPs;

7.7.2 Ensure that records are maintained and readily available to the staff operating the equipment;

7.7.3 Establish written criteria for an acceptable mass-spectrometric tune;

7.7.4 Maintain records of mass spectrometric tuning;

7.7.5 Document Corrective Actions if a mass-spectrometric tune is unacceptable;

7.7.6 Use an internal standard with each qualitative and quantitative analysis that has similar chemical and physical properties to that of the compound identified and is isotopically labeled when available or appropriate for the assay;

7.7.7 Document the monitoring of the response (area or peak height) of the internal standard to ensure consistency over time of the analytical system;

7.7.8 Compare two transitions and retention times between calibrators, controls and Samples within each run;

7.7.9 Ensure ion ratios do not exceed +/- 30% relative to the average of calibration standards from the same sequence.

7.7.10 Document and maintain records when changes or cleaning in source, source conditions, eluent, or column are made to the instrument;

7.7.11 Evaluate and document the performance of the instrument after routine and preventative maintenance and when changes in: source, source conditions, eluent, or column are made prior to reporting test results; and

7.7.12 Monitor and document the performance of the instrument each day of testing.

7.8Inductively Coupled Plasma Mass Spectrometry (ICP/MS). A Natural Medicine Testing Facility using ICP must:

7.8.1 Perform and document preventive maintenance as required by the manufacturer and SOPs;

7.8.2 Ensure that records are maintained and readily available to the staff operating the equipment;

7.8.3 Establish written criteria for an acceptable mass-spectrometric tune;

7.8.4 Maintain records of mass spectrometric tuning;

7.8.5 Document Corrective Actions if a mass-spectrometric tune is unacceptable;

7.8.6 Use an internal standard with each qualitative and quantitative analysis that has similar chemical and physical properties to that of the compound identified and is isotopically labeled when available or appropriate for the assay;

7.8.7 Document the monitoring of the response (counts per second) of the internal standard to ensure consistency over time of the analytical system;

7.8.8 Compare mass-to-charge ratios between calibrators, controls and Samples within each run;

7.8.9 Monitor analyses to check for contamination and carry-over;

7.8.10 Evaluate and document the performance of the instrument after routine and preventative maintenance and when changes in: source, conditions, or detector are made prior to reporting test results; and

7.8.11 Monitor and document the performance of the instrument each day of testing.

7.9Microbial Assays. A Natural Medicine Testing Facility using microbial assays must:

7.9.1 Perform and document preventive maintenance as required by the manufacturer and SOPs;

7.9.2 Ensure that records are maintained and readily available to the staff operating the equipment;

7.9.3 Validate any changes or modifications to a manufacturer's approved assays or testing methods when a Sample is not included within the types of Samples approved by the manufacturer;

7.9.4 Verify the method at the Action Levels for each Analyte. Verification at the qualitative presence/absence limit shall include a fractional recovery study;

7.9.5 The Natural Medicine Testing Facility shall include controls for each set of Samples. Quantitative microbial methods shall use controls of a specific known value or set of values that lies within the quantifiable range of the method;

7.9.6 For molecular methods, the Natural Medicine Testing Facility shall include controls for each individual analytical run. Quantitative molecular methods shall use controls of a specific known value or set of values that lies within the quantifiable range of the method;

7.9.7 PCR-based and qPCR-based methods must include validated internal amplification controls; and

7.9.8 Microbial methods must include steps to confirm presumptive positive results; confirmation methods may be molecular or cultural or both. Where applicable, confirmation of viability must be performed.

7.9.9 Verify the stated Limit of Detection of qualitative assays through "dilution to extinction" studies in which the calculated extinction dilution is corroborated with cultural data.

7.10Other Analytical Methodology. A Natural Medicine Testing Facility using any other analytical methodology must:

7.10.1 Perform and document preventive maintenance as required by the manufacturer or SOP;

7.10.2 Ensure that records are maintained and readily available to the staff operating the equipment;

7.10.3 Ensure that appropriate quality assurance and Quality Control measures are performed and documented as necessary for the specific methodology; and

7.10.4 Evaluate the performance of the instrument after routine and preventive maintenance prior to analyzing subject Samples.

7.11General Quantitative Quality Parameters: A Natural Medicine Testing Facility must meet the following criteria for quantitative method controls.

7.11.1 Chemical Quality Controls must not exceed plus or minus 10% recovery of the target value when Analytes are greater than or equal to 100 parts per million (ppm).

7.11.2 Chemical Quality Controls must not exceed plus or minus 20% recovery of the target value when Analytes are greater than or equal to 10 ppm.

7.11.3 Chemical Quality Controls must not exceed plus or minus 20% recovery of the target value > when Analytes are greater than or equal to 1 ppm.

7.11.4 Chemical Quality Controls must not exceed plus or minus 20% recovery of the target value when Analytes are greater than or equal to 100 parts per billion (ppb).

7.11.5 Chemical Quality Controls must not exceed plus or minus 40% recovery of the target value when Analytes are greater than or equal to 10 ppb.

7.11.6 Chemical Quality Controls must not exceed plus or minus 60% recovery of the target value when Analytes are greater than or equal to 1 ppb.

7.11.7 Microbiology Quality Controls must not exceed plus or minus 20% recovery of the target value.

5 CCR 1005-7-7

47 CR 24, December 25, 2024, effective 1/14/2025