Recommendations from the Global Bioanalysis Consortium Team A8: Documentation

Abstract

The A8 Global Harmonization Team focused on the documentation needed to support both small and large molecule bioanalysis. Current regulatory requirements were compiled and compared. The scope of the team’s discussions included the validation report, the bioanalytical report, study plans, raw data, and bioanalytical summaries for the common technical document (CTD). A common high-level table of contents for method validation and sample analysis reports is proposed. Suggestions have been made as to how the CTD can be standardized to improve usability and review. Additional comments have been made on reports of failure investigations, study plans, and raw data documentation. The recommendation is that no prescriptive guidelines are required in these areas but should be led by good scientific practices subject to particular circumstances.

Appendices

Appendix 1: Table of Contents Validation Report

Signature Page

Statement of Regulatory Compliance

For validations in preclinical species, if performed under GLP, the compliance statement is straightforward. However, for validations in man, one should refrain from claiming GLP. A proposed wording could be: “The report and related raw data were inspected and verified for compliance to applicable governmental regulations and implemented internal standard operating procedures.”

Method Validation Summary Table

This summary table is considered very useful in facilitating the review. A template for this table is available in the EMA guideline on the investigation of bioequivalence (12). This summary table could also be included as an addendum to the analytical report (addendum 5 in Appendix 2).

Key Study Information (experiment dates, personnel, archiving)

The start and end date of the experiments could be included here. However, more detailed information should be included in Table 6.1 at the end of the report.

For validations that are not carried out under GLP, the names of the responsible personnel are considered non-essential as the signature page indicates the key contributors to the study. However, for GLP validations this is essential. According to FDA GLP regulations (13), the name of the study director, the names of other scientists or professionals, and the names of all supervisory personnel, involved in the study should be in the report.

The archiving location for the data cited should be documented in the report for GLP studies. For validations not conducted under GLP, the team recommends this be included as well to facilitate raw data retrieval.

1. Introduction

2. Methods and materials

2.1 Analytical Method Information

This section contains a high-level description of the assay (with a reference to a more detailed assay description in the addendum) and the regression model used. For large molecule bioanalysis, mention should be made of the platform employed, a high-level description of the ligand binding method, and the minimum required dilution (MRD).

2.2 Reference Compounds

2.3 Calibration Standards and QCs Preparation

2.4 Data Acquisition and Processing

A listing of the validated software that was used to acquire and process the data is necessary. A listing of calculations used may also be valuable in this section or can be described in the respective sections below.

2.5 Calculations

3. Validation Experiments and Results

3.1 Regression Model and Formula

3.2 Lower and Upper Limits of Quantification (LLOQ and ULOQ)

3.3 Accuracy (% Bias) and Precision (% CV) (including the formula used)

3.4 Carry-Over (if applicable)

3.5 Selectivity

3.6 Matrix Effect (specificity and MRD in case of LBAs)

3.7 Dilution of Samples (dilutional linearity and hook effect in case of LBAs)

3.8 Extraction Recovery (for chromatographic methods)

3.9 Incurred Sample Reproducibility (ISR) (if applicable)

4. Stability

4.1 Stability in Stock Solutions and Working Solutions

4.2 Processed Sample Stability

4.3 Stability in Matrix (short and long term, freeze/thaw)

5. Conclusions

6. Tables

6.1 Analytical Batch Overview (table of runs and analysis dates, passed or failed, link with Watson (or relevant LIMS) run number)

6.2 Back-Calculated Values Calibration Curves

6.3 Calibration Curve Parameters

6.4 Accuracy and Precision from QCs

6.5 Evaluation of Carry-Over

6.6 Evaluation of Selectivity

6.7 Matrix Effect

6.8 Dilution of Samples

6.9 Extraction Recovery

6.10 ISR

6.11 Stability Data

7. Addenda

Addendum 1 Chemical structures (for small molecule assays)

Addendum 2 Certificates of analysis

Addendum 3 Assay description

Addendum 4 Chromatograms (if applicable)

The proposal is to include only a very limited number of chromatograms in the report: a blank, a zero, an LLOQ and a ULOQ standard.

Appendix 2: Table of Contents Analytical Report

Signature Page

Statement of Regulatory Compliance

For preclinical safety studies performed under GLP, the compliance statement is straightforward. However, for clinical studies, one should refrain from claiming GLP. A proposed wording could be: “The report and related raw data were inspected and verified for compliance to applicable governmental regulations and implemented internal standard operating procedures.”

Key Study Information (sample receipt and analysis dates, personnel, archiving)

For sample receipt, especially when multiple shipments have occurred, only the date of the first and last sample reception should be sufficient (see also under 3.1.)

The start and end date of subject sample analysis could be included here. However, more detailed information should be included in the tables at the end of the report (see under 3.2)

For clinical study reports, the names of the responsible personnel are considered non-essential as the signature page indicates the key contributors to the study. However, for non-clinical GLP studies this is essential. According to FDA GLP regulations (13) the name of the study director, the names of other scientists or professionals, and the names of all supervisory personnel, involved in the study should be part of the report of non-clinical laboratory study results.

The archiving location for the data cited should be documented in the report for GLP studies. For clinical studies the team recommends this be included as well to facilitate raw data retrieval.

1. Introduction

2. Materials and Methods

2.1 Analytical Method Information

This section contains a high-level description of the assay (with a reference to a more detailed assay description in the addendum) and the regression model used. For large molecule bioanalysis, mention should be made of the platform employed, a high-level description of the ligand binding method, and the MRD,

2.2 Reference Compounds

2.3 Calibration Standards and QCs Preparation

The actual date of preparation of the QCs and standards need not be listed here but a statement should be added that QCs and standards were used within the proven long-term stability period.

2.4 Data Acquisition and Processing

A listing of the validated software that was used to acquire and process the data is necessary. A listing of calculations used may also be valuable in this section or can be described in the respective sections below.

3. Results

3.1 Sample Receipt and Storage

A general statement like “samples and QCs were stored in a monitored freezer with a set temperature of −20°C” should be sufficient unless there were any deviations which should be documented here along with an evaluation of the impact on the study results. More specific information should be kept in the raw data. Care should be taken that all samples are analyzed within the window of demonstrated stabilities, e.g., long-term storage stability. This can be documented by a statement that “long-term stability data cover the age of the samples and include the longest period of sample storage.”

3.2 Sample Analysis (batch overview, acceptance criteria)

Any laboratory information management system (LIMS) should be able to generate a tabular overview of all batches within a study, including the failed runs, along with the analysis date (refer to Table 5.1). This should help reviewers assess the quality of the method. The high-level reason for failure should also be described in the report, e.g., “batch failed because too many QCs outside of acceptance criterion” is considered sufficiently detailed.

For chromatography methods, ANVISA is requesting clock times of first and last injection in each run but this seems excessive, since this will be in the raw data. However, we do recommend that a good scientific practice would be to identify those runs where the analysis was interrupted due to technical issues. Also, the grouping of the subjects’ samples within the run is available in the raw data and should not be described in detail in the report.

The acceptance criteria can be described in the text or the specific SOP can be included as an attachment.

3.3 Linearity

Discussion on back-calculated values and regression parameters for calibration lines referring to Table 5.2 and 5.3.

3.4 Accuracy

Calculation of the within- and between-run QCs accuracy with reference to the data in Table 5.4.

3.5 Precision

Calculation of the within- and between-run QCs precision with reference to the data in Table 5.4.

3.6 IS evaluation (if applicable)

If an IS evaluation was done in the study the criteria should be listed here.

3.7 Repeat analysis

List the repeated samples in a table (Table 5.5) with the initial value, the reason for the repeat, the repeat value, and the reason for acceptance. This should only be done for samples that were part of accepted batches. Canada, Brazil, and the EMA request the number of repeats as a percentage of the total number of samples. There was no consensus on how exactly this number should be calculated nor on the need to include this in a report. If the purpose of this number is to assess the ruggedness of the assay, then we suggest using the percentage of failed batches as the value, although it can be argued that this is “de facto” obtained from the data in the report.

3.8 Incurred Sample Reproducibility (ISR) (if applicable)

If ISR was performed as part of the study the criteria should be listed here.

3.9 Failed Run Investigation (if applicable)

3.10 SOP/Assay Method Deviations (if applicable)

Include any deviation from the validated assay here and describe the impact on the study results.

4. Conclusions

5. Tables

5.1 Analytical Batch Overview

This is a table listing all of the analytical runs with corresponding analysis dates and indication if the run passed or failed

5.2 Back-Calculated Values Calibration Standards

5.3 Calibration Curve Parameters

These parameters include slope, intercept, and correlation coefficients. For large molecule bioanalysis, the asymptotes can also be included. Performing statistics on these parameters does not have added value and can be omitted.

5.4 Accuracy and Precision from QCs

The table should display the QC results from all accepted batches, outliers should be flagged and both intra- and inter-batch accuracy and precision should be calculated.

5.5 Reanalyzed Individual Samples

This table should trace the reanalyzed samples to a valid reported value, and include the reason for the failure.

5.6 ISR

6. Addenda

Addendum 1 Certificates of analysis for the analyte(s)

Addendum 2 Assay description

Addendum 3 Results Report

The final results are listed here in tabular format. However, results are usually listed in a specific format to fit with a database, especially in case of clinical studies. This often makes these tables very difficult to read and may therefore be of limited added value. It is therefore suggested to only list these columns in the table which allow identification of the sample along with the corresponding concentration.

Addendum 4 Chromatograms (for chromatographic methods)

Most regulators ask for 5 or 20% of the subjects with calibration samples and QCs from these runs, and only for clinical studies. It is best to define which chromatograms will be included in the report a priori in an SOP, e.g., the first 5 (or 20)% of subjects analyzed to avoid cherry picking the best looking chromatograms.

Addendum 5 Method validation summary (from validation report); optional