5 Colo. Code Regs. § 1005-5-7

Current through Register Vol. 47, No. 11, June 10, 2024

Rule 5 CCR 1005-5-7 - Hemp Testing Laboratories: 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 laboratory 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 Rules Pertaining to the Administration and Enforcement of the Industrial Hemp Regulatory Program Act, 8 CCR 1203-23 Part 4 and Department hemp product regulations.

7.1.3 To the extent practicable, laboratory test methods must meet AOAC International standard method performance requirements.

7.1.4 The laboratory must implement a performance based measurement system for the selected methodology and validate the method following good laboratory practices in accordance with AOAC, United States Pharmacopeia (USP), United States Food and Drug Administration (FDA) United States Department of Agriculture (USDA), and other reputable validation guidelines and methodology prior to reporting results. Validation, verification, or Matrix extension of 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 Verification of Recovery

7.1.4.10 Inclusivity

7.1.4.11 Exclusivity

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.

7.1.4.12.2 For GC cannabinoid methods, experimental determination of actual conversion rate of THCA to THC.

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

7.1.5.1 Validation plan;

7.1.5.2 Introduction and summary;

7.1.5.3 Materials, to include identification of certified Reference Materials, and preparation methods;

7.1.5.4 Method parameters;

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

7.1.5.6 Instrument calibration data, if any;

7.1.5.7 Data, calculations, and results;

7.1.5.8 Method Acceptability Criteria performance data;

7.1.5.9 Conclusion and discussion; and

7.1.5.10 References.

7.1.6 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.

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

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

7.1.9 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.

7.1.9.1 Laboratories must validate or verify instrumentation and methodology immediately and prior to use following a change in location.

7.2Gas Chromatography (GC). A Hemp Testing Laboratory using GC must:

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

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

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

7.2.4 Document the performance of new columns before use;

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

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

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

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

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

7.3Gas Chromatography Mass Spectrometry (GC/MS). A Hemp Testing Laboratory using GC/MS must:

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

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

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

7.3.4 Maintain records of mass spectrometric tuning;

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

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

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

7.3.8 Use selected ion monitoring within each run to assure that the laboratory compares ion ratios and retention times between calibrators, controls and Samples for identification of an Analyte;

7.3.9 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;

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

7.3.11 Define the criteria for designating qualitative results as positive;

7.3.12 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;

7.3.13 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

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

7.4Immunoassays. A Hemp Testing Laboratory using Immunoassays must:

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

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

7.4.3 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

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

7.5High Performance Liquid Chromatography (HPLC). A Hemp Testing Laboratory using HPLC 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 Monitor and document the performance of the HPLC instrument each day of testing;

7.5.4 Evaluate the performance of new columns before use;

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

7.5.6 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;

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

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

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

7.6Liquid Chromatography Mass Spectrometry (LC/MS). A Hemp Testing Laboratory using LC/MS 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 Establish written criteria for an acceptable mass-spectrometric tune;

7.6.4 Maintain records of mass spectrometric tuning;

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

7.6.6 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;

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

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

7.6.10 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

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

7.7Inductively Coupled Plasma Mass Spectrometry (ICP/MS). A Hemp Testing Laboratory using ICP 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 (counts per second) of the internal standard to ensure consistency over time of the analytical system;

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

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

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

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

7.8Microbial Assays. A Hemp Testing Facility using microbial assays 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 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;

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

7.8.5 Verify the stated detection limit of qualitative assays through "dilution to extinction" studies in which the calculated extinction dilution is corroborated with cultural data.

7.8.6 The laboratory 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;

7.8.7 For molecular methods, the laboratory 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;

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

7.8.9 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.

7.9Moisture Content Analysis. A Hemp Testing Laboratory analyzing percent moisture 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 an approved method when a Sample is not included within the types of Samples for which the method was originally validated;

7.9.4 Ensure SOPs specify all unique method parameters, such as temperature, sample surface area, etc., that prevent loss of volatile compounds, the oxidation of oils and/or the re-absorbance of water;

7.9.5 Ensure that appropriate quality assurance and Quality Control measures are performed and documented as necessary for the assay;

7.9.6 Evaluate the performance of the method after routine and preventive maintenance prior to analyzing subject Samples.

7.9.7 Establish criteria for acceptable moisture analyzer performance. It may be necessary to obtain a reference material that is tested prior to analyzing samples each day in order to ensure the acceptability of the analyzer.

7.10Other Analytical Methodology. A Hemp Testing Laboratory 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;

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

7.11Cannabinoid Methodology. At a minimum, analytical testing of Hemp for delta-9 tetrahydrocannabinol (THC) must use post-decarboxylation or other similarly reliable methods. The testing methodology must consider the potential conversion of delta-9 tetrahydrocannabinolic acid (THCA) into THC. The results reported must reflect the Total THC content.

7.11.1 The Total THC concentrations of Hemp shall be determined and reported on a Dry Weight Basis.

7.11.1.1 A Hemp Testing Laboratory must ensure reporting of Total THC includes a calculation for moisture correction based on a theoretical concentration of zero percent moisture. The following conversion formula is recommended:

Click to view image

Where:

P2 = adjusted constituent percentages at moisture M2 (percent)

M2 = moisture basis (percent, i.e., 0%)

P1 = original (as-is) constituent percentage

M1 = original moisture (percent)

7.11.1.2 The Cannabinoid concentrations of Hemp Products shall be determined and reported on an "as-is" basis (i.e., in the form submitted to the laboratory).

7.12 Testing and validation of complex matrices. a hemp testing laboratory must include a variety of matrices as part of the validation/verification process. during method validation/verification, a hemp testing laboratory must:

7.12.1 Select matrices which best represent each category of products to be tested as listed in department hemp product regulations. the laboratory shall independently determine the category of matrix a product falls within. properties to consider include fat content, cannabinoid content, pH, salt content, sugar content, water activity, the presence of known chemical compounds, microbial flora and antimicrobial compounds.

7.12.2 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.

7.12.2.1 For example, the hemp testing laboratory intends to receive the topical product "bath bombs" for testing, but previous validation studies for topical products include lotion and massage oil. 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.

7.12.3 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.

7.12.3.1 For example, the hemp testing laboratory receives a new edible type matrix for testing (snickerdoodle cookies), but previous validation included gummies and hard candy. a spike of a portion of the submitted material must be analyzed prior to, or at the time of, sample analysis.

5 CCR 1005-5-7

44 CR 06, March 25, 2021, effective 4/14/2021

46 CR 21, November 10, 2023, effective 12/15/2023