@pikaa-ai/pikaa 0.3.1 → 0.3.4

package/skills/analytical-method-validation/references/compendial-and-clsi.md

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-# Compendial, CLSI, and ISO Sources (No Standard Text)
-
-Research basis: **2026-07-27**. This reference identifies documents, their scope, and where to obtain
-them. **It does not reproduce their requirements, thresholds, or study designs**, because they are
-copyrighted and paywalled.
-
-## Copyright boundary
-
-USP–NF general chapters, CLSI documents, and ISO/IEC standards are copyrighted works sold by their
-publishers. Do not ask an agent to retrieve, transcribe, summarise clause-by-clause, reconstruct, or
-store their text. Vendor application notes and training decks that quote them are equally
-constrained, and a paraphrase that carries the same numbers is still a reproduction of the
-substantive content.
-
-The practical consequence: **when a numeric criterion or a study design lives in one of these
-documents, read it from the authorised copy.** An agent asked for "the USP <621> tailing factor
-limit" or "the CLSI EP15 number of days" will produce a plausible number. Plausible is not the same
-as correct, and the difference is discovered at audit.
-
-Record publisher, title, designation, edition, amendments, authorised location, access date, and
-review date in the laboratory's controlled source register.
-
-## USP–NF general chapters
-
-| Chapter | Title | Scope |
-| --- | --- | --- |
-| `<1220>` | Analytical Procedure Life Cycle | Three-stage lifecycle: procedure design (Stage 1), performance qualification (Stage 2), ongoing performance verification (Stage 3), organised around an analytical target profile. Official 1 May 2022 (incorporated into USP–NF 2022 Issue 1 on 1 Nov 2021). Integrates the concepts previously spread across `<1224>`, `<1225>`, and `<1226>`. |
-| `<1225>` | Validation of Compendial Procedures | Validation of non-compendial procedures, and of compendial procedures used outside their stated scope. Stage 2 activity under `<1220>`. |
-| `<1226>` | Verification of Compendial Procedures | Assessment of selected performance characteristics showing a compendial procedure works under actual conditions of use. **Verification is not revalidation** and does not repeat the full validation. |
-| `<1224>` | Transfer of Analytical Procedures | Transfer between laboratories. |
-| `<1010>` | Analytical Data — Interpretation and Treatment | Statistical treatment of analytical data. |
-| `<621>` | Chromatography | System suitability and chromatographic operating parameters, including the extent to which a compendial procedure may be adjusted without triggering revalidation. |
-| `<711>` / `<1092>` | Dissolution / The Dissolution Procedure | Dissolution testing and development/validation of the procedure. |
-
-Obtain from the USP–NF (<https://www.uspnf.com/>). Regional pharmacopoeias — Ph. Eur., JP, ChP —
-carry their own general chapters; check which pharmacopoeia the specification cites, because
-adjustment allowances and system suitability requirements differ between them.
-
-**The `<1226>` decision.** Verification applies when using a compendial procedure as written and
-within its scope. Two situations push you back to `<1225>` validation: using the procedure outside
-its stated scope (a different matrix, a different dosage form, a concentration range it does not
-cover), or modifying it beyond the adjustments the relevant chapter permits. Getting this wrong in
-either direction is expensive — unnecessary full validation, or an unsupported claim of verification.
-
-## CLSI EP series
-
-Designations and titles below were taken from clsi.org listings and secondary sources on the
-research date. **Editions change; confirm the current edition on <https://clsi.org/> before designing
-a study.** Marked `[confirm]` where the edition was not read from the publisher directly.
-
-| Designation | Subject | Note |
-| --- | --- | --- |
-| EP05 | Evaluation of precision of quantitative measurement procedures | Establishment of precision; the multi-day/multi-run designs. `[confirm edition]` |
-| EP06 | Evaluation of linearity of quantitative measurement procedures | 2nd edition reported. `[confirm edition]` |
-| EP07 | Interference testing in clinical chemistry | Screening, quantifying and confirming interferents; verifying manufacturer interference claims. 3rd edition reported. `[confirm edition]` |
-| EP09 | Measurement procedure comparison and bias estimation using patient samples | The method-comparison document. 3rd edition reported. `[confirm edition]` |
-| EP15 | User verification of precision and estimation of bias | The short study a laboratory runs to verify a manufacturer's claims. 3rd edition reported. `[confirm edition]` |
-| EP17 | Evaluation of detection capability | Limit of blank, limit of detection, limit of quantitation; verification of manufacturer claims. `[confirm edition]` |
-| EP25 | Evaluation of stability of in vitro diagnostic reagents | `[confirm edition]` |
-| EP28 | Defining, establishing, and verifying reference intervals | Formerly designated C28. An implementation guide (EP28IG) also exists. `[confirm edition]` |
-
-**Vocabulary.** CLSI distinguishes *limit of blank*, *limit of detection*, and *limit of quantitation*
-as three separate quantities with separate protocols. This is not the same taxonomy as ICH Q2(R2)'s
-detection limit and quantitation limit, and the two should not be translated into each other
-casually — the underlying definitions and the experiments differ.
-
-**Verification versus establishment.** For an FDA-cleared or CE-marked assay used as intended, a
-laboratory *verifies* the manufacturer's performance claims — a bounded study. For a
-laboratory-developed test, or an assay used off-label, the laboratory *establishes* performance,
-which is a much larger exercise. Under CLIA the distinction has direct regulatory consequences and
-also depends on test complexity. Determine which applies before designing anything.
-
-## ISO standards
-
-| Standard | Relevance |
-| --- | --- |
-| ISO/IEC 17025:2017 | Clause 7.2 selection, verification and validation of methods; clause 7.6 measurement uncertainty. Validation "to the extent necessary" for the intended application — no characteristic list, no numeric criteria. |
-| ISO 15189 | Medical laboratories: quality and competence. The clinical-laboratory counterpart to 17025. |
-| ISO 21748 / ISO 5725 series | Using repeatability, reproducibility and trueness estimates in measurement uncertainty; accuracy of measurement methods. |
-
-Obtain from ISO (<https://www.iso.org/>) or a national member body. A laboratory is **accredited** to
-ISO/IEC 17025 by an accreditation body — it is not "17025 certified", and writing "certified" is a
-substantive error assessors notice.
-
-For accreditation readiness, the quality manual, and the surrounding management system, use this
-repository's `iso-standards-readiness` skill. This skill stays at the level of the individual
-procedure.
-
-## Environmental, food, and forensic method systems
-
-Where a prescribed method system governs — a published EPA method, an AOAC Official Method, a
-standard method for water or food analysis — the validation and quality-control requirements are
-written into the method or the programme, and they take precedence. Do not substitute a
-pharmaceutical framework. Common differences: matrix spike and duplicate requirements per batch,
-prescribed calibration-verification frequencies, method detection limit procedures that differ from
-both ICH and CLSI, and mandatory participation in proficiency testing schemes.

package/skills/analytical-method-validation/references/framework-selection.md

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-# Which Framework Governs
-
-Research basis: **2026-07-27**. Confirm every date and edition against the official source before
-relying on it; see `source-ledger.md`.
-
-Framework selection is the first decision and the one most often skipped. Getting it wrong
-invalidates the protocol regardless of how well the studies are executed, because each framework
-requires a different set of characteristics, a different study layout, and a different treatment of
-acceptance criteria.
-
-## The deciding questions, in order
-
-**1. Is the measurand a drug concentration in a biological matrix, supporting a nonclinical or
-clinical study?**
-→ **ICH M10.** This covers pharmacokinetics, toxicokinetics, and bioequivalence. M10 supplies
-explicit numeric criteria, and they differ between chromatographic assays and ligand binding
-assays. Q2(R2) does not govern here.
-
-**2. Is it a quality attribute of a drug substance or drug product — assay, potency, impurity,
-identity, dissolution, content uniformity?**
-→ **ICH Q2(R2)** for validation, with **ICH Q14** for development, robustness, the analytical
-target profile, and lifecycle change management. If the procedure is compendial and being used as
-written, see question 3 first.
-
-**3. Is the procedure a compendial (pharmacopoeial) procedure?**
-→ **USP <1226> verification** if it is used as written and within its stated scope. Verification
-assesses selected characteristics to show the procedure works under actual conditions of use; it is
-not revalidation and does not repeat the full study. → **USP <1225> validation** if the procedure
-is non-compendial, or compendial but used outside its scope. Both sit inside the **USP <1220>**
-three-stage lifecycle. Regional pharmacopoeias (Ph. Eur., JP) have their own general chapters —
-check which pharmacopoeia the specification cites.
-
-**4. Is it a clinical laboratory measurement procedure reporting patient results?**
-→ **CLSI EP series**, inside a CLIA/CAP or ISO 15189 quality system. The vocabulary differs from
-pharmaceutical work: *verification* of a manufacturer's claims for an FDA-cleared assay is a much
-smaller exercise than *establishment* of performance for a laboratory-developed test, and the
-distinction is regulatory, not stylistic.
-
-**5. Is the laboratory accredited to ISO/IEC 17025 and the method non-standard, laboratory-developed,
-or a modified standard method?**
-→ **ISO/IEC 17025 clause 7.2.2** requires validation as extensive as necessary to meet the needs of
-the intended application, plus measurement uncertainty under clause 7.6. It sets no characteristic
-list and no numeric criteria; the laboratory justifies both.
-
-**6. Is it an environmental, food, or forensic method under a prescribed method system?**
-→ The method system governs (for example a published EPA method, an AOAC Official Method, or a
-regulator's prescribed procedure), usually with its own validation and QC requirements written into
-the method itself. Do not substitute a pharmaceutical framework.
-
-## More than one can apply
-
-Common and legitimate. A contract laboratory accredited to ISO/IEC 17025 running a compendial assay
-for a pharmaceutical client satisfies <1226> for the procedure and 17025 clause 7.2 for the
-accreditation scope, with the client's specification supplying the criteria. Record which framework
-each requirement traces to, so a later change can be assessed against the right one.
-
-## Do not blend them
-
-The failure mode is a protocol that mixes Q2(R1)-era characteristic names, an M10 numeric tolerance
-imported because it was memorable, and a CLSI study layout. It satisfies none of the three and is
-hard to defend because no single source can be cited for any of it. If a requirement is in the
-protocol, name the framework and section it comes from.
-
-## Where the numbers come from
-
-| Framework | Numeric acceptance criteria |
-| --- | --- |
-| ICH Q2(R2) | Almost none. Derive from the specification, the ATP, or development data, and justify. |
-| ICH Q14 | None. It supplies the ATP concept and the development/robustness framework. |
-| ICH M10 | Explicit, and modality-dependent. Use them as written. |
-| USP <1225>/<1226>/<1220> | Consult the authorised text. |
-| CLSI EP | Consult the authorised text; many EP documents supply study designs rather than limits. |
-| ISO/IEC 17025 | None. The laboratory sets and justifies them. |
-
-Q2(R2)'s reticence is deliberate: a criterion that is not tied to what the result is used for is
-arbitrary. An assay releasing product against a 95.0–105.0% specification needs different precision
-than one supporting a 70–130% content-uniformity limit. Deriving the criterion from the decision the
-result supports is the substance of the exercise, not paperwork around it.
-
-## Related skills in this repository
-
-- `iso-standards-readiness` — the surrounding quality system (ISO/IEC 17025, ISO 15189
-  accreditation readiness, quality manual, CAPA). That skill operates at the laboratory level; this
-  one operates at the level of a single procedure.
-- `statistical-analysis`, `statistical-power` — general inference and study sizing.
-- `uncertainty-and-units` — unit handling and measurement uncertainty propagation, which ISO/IEC
-  17025 clause 7.6 requires alongside validation.

package/skills/analytical-method-validation/references/ich-m10-bioanalytical.md

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-# ICH M10 — Bioanalytical Criteria, by Modality
-
-Research basis: **2026-07-27**, read from the ICH Harmonised Guideline *Bioanalytical Method
-Validation and Study Sample Analysis M10*, Step 4 dated 24 May 2022. ICH licenses its documents for
-reuse with acknowledgement. Confirm the current text and your region's implementation at
-<https://database.ich.org/sites/default/files/M10_Guideline_Step4_2022_0524.pdf>.
-
-M10 harmonised what had been separate FDA and EMA bioanalytical guidance for studies in its scope:
-methods quantifying drug and metabolite concentrations in biological matrices supporting nonclinical
-and clinical studies, plus the analysis of study samples.
-
-## The distinction that matters most
-
-**Chromatographic assays (section 3) and ligand binding assays (section 4) have different numeric
-criteria throughout.** They are not stylistic variants of one set. Applying chromatographic
-tolerances to an LBA is the most common error in this area, and importing the LBA total-error
-criterion into a chromatographic method is its mirror image.
-
-| | Chromatographic | Ligand binding assay |
-| --- | --- | --- |
-| Calibration levels (minimum) | 6, including LLOQ | 6, including LLOQ |
-| Calibration standard tolerance | ±15% | ±20% |
-| … at LLOQ | ±20% | ±25% |
-| … at ULOQ | ±15% | ±25% |
-| Calibration standards that must pass | ≥75% | ≥75%, excluding anchor points |
-| Accuracy | ±15% | ±20% |
-| … at limits | ±20% at LLOQ | ±25% at LLOQ **and** ULOQ |
-| Precision (%CV) | ≤15% | ≤20% |
-| … at limits | ≤20% at LLOQ | ≤25% at LLOQ **and** ULOQ |
-| A&P QC levels | minimum 4 | 5 (LLOQ, low, medium, high, ULOQ) |
-| A&P replicates per level per run | ≥5 (within-run) | ≥3 |
-| A&P runs | ≥3 runs over ≥2 days | ≥6 runs over ≥2 days |
-| **Total error** | **no such criterion** | **≤30%; ≤40% at LLOQ and ULOQ** |
-| Routine run QC tolerance | ±15% | ±20% |
-| Routine run QC pass rule | ≥2/3 of all QCs **and** ≥50% at each level | same rule, ±20% |
-| Dilution integrity | mean within ±15% | mean within ±20% |
-| Stability | mean at each QC level within ±15% | mean within ±20% |
-| ISR agreement | within ±20% for ≥2/3 of repeats | within ±30% for ≥2/3 of repeats |
-| Selectivity sources/lots | ≥6 individual sources | ≥6 individual sources |
-| Carry-over in blank | ≤20% of LLOQ analyte response and ≤5% of IS response | per guideline |
-
-Verify any figure against the guideline before using it in a protocol; regional implementation and
-subsequent revisions can change the picture.
-
-## Chromatographic QC placement (section 3)
-
-Accuracy and precision validation QCs at a minimum of **four** concentration levels:
-
-- the **LLOQ**
-- **low QC** — within three times the LLOQ
-- **medium QC** — around 30–50% of the calibration curve range
-- **high QC** — at least 75% of the ULOQ
-
-For runs that are not accuracy-and-precision runs, low, medium and high QCs may be analysed in
-duplicate; these plus the calibration standards form the basis for accepting or rejecting the run.
-
-Calibration standards and QCs should be prepared from **separate stock solutions**, to avoid a bias
-that is not a property of the analytical performance. If a single stock must serve both, verify the
-accuracy and stability of that stock. A single source of blank matrix may be used if it is free of
-interference and matrix effects.
-
-Calibration curves for accuracy and precision assessment should use freshly spiked standards in at
-least one run; if other runs use frozen standards, demonstrate their stability.
-
-## Reporting obligations that catch people out
-
-**Report everything.** Validation data and the determination of accuracy and precision must include
-*all* results obtained, including individual QCs outside the acceptance criteria — except cases where
-errors are obvious and documented. Silently dropping an out-of-criteria QC is a data integrity
-problem, not a rounding decision.
-
-**Within-run accuracy and precision are reported per run.** If the within-run criteria are not met in
-every run, calculate an overall estimate of within-run accuracy and precision for each QC level.
-Between-run (intermediate) accuracy and precision combine data from all runs.
-
-**Trend within a run.** It is recommended to demonstrate accuracy and precision over at least one run
-sized like a prospective study-sample run, so time-dependent drift is visible.
-
-## Incurred sample reanalysis (section 5)
-
-ISR repeats the analysis of a subset of study samples in separate runs, to verify that measured
-concentrations in real samples are reproducible. It is not a substitute for QCs — QCs are spiked,
-incurred samples are not, and only incurred samples can reveal metabolite back-conversion, protein
-binding effects, or matrix instability.
-
-- The extent depends on the analyte and the samples and should be justified.
-- Objective criteria for choosing the subset should be **predefined**; selecting samples around
-  Cmax and the elimination phase is recommended.
-- **Do not pool samples** — pooling masks anomalous findings.
-- ISR samples and QCs are processed and analysed in the same manner as the original analysis.
-- Percent difference is `(repeat value - initial value) / mean value x 100` -- assessed
-  against the **mean of the two**, not against the initial value.
-- Repeats are performed within the analyte's stability window, but **not on the same day**
-  as the original analysis.
-- Acceptance: within ±20% for at least 2/3 of repeats (chromatographic), or within ±30% for at least
-  2/3 (LBA).
-
-For nonclinical studies in scope, ISR should in general be performed; the guideline notes incurred
-samples need only be included if available, so inclusion was not felt to be mandatory in every case.
-Confirm the situations requiring ISR against the guideline text for your study type.
-
-## Study sample reanalysis is a separate thing
-
-ISR is a method-reliability check. *Reanalysis of study samples* for a reportable-value decision is
-different, and the reasons for reanalysis, the number of replicates, and the criteria for selecting
-the value to report must be **predefined in the protocol, study plan, or SOP before study sample
-analysis begins.** Deciding after the fact which of two values to report is the classic finding.
-
-## Partial and cross validation
-
-M10 addresses partial validation (a change to a validated method — matrix, anticoagulant, species,
-instrument, or a range change) and cross validation (comparing data from two methods or two
-laboratories contributing to the same study). Both are scoped by the change and the risk; consult
-the guideline for what each requires. For a cross validation between sites or methods, the
-statistics in `compare_methods.py` — equivalence testing against a pre-stated margin, and a
-regression that allows error in both measurements — are the appropriate treatment.
-
-## Biomarkers and other contexts
-
-M10's scope centres on drug and metabolite concentration measurement. Biomarker assays, immunogenicity
-assays, and diagnostic measurements are addressed differently or fall outside scope; do not assume the
-concentration-assay criteria transfer. Where a biomarker assay supports a regulatory decision, the
-fit-for-purpose framework and the applicable regional guidance govern the extent of validation.

package/skills/analytical-method-validation/references/ich-q2r2.md

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-# ICH Q2(R2) — Structure and Recommended Data
-
-Research basis: **2026-07-27**, read from the ICH Harmonised Guideline *Validation of Analytical
-Procedures Q2(R2)*, Final Version adopted 1 November 2023, with the error correction dated
-30 November 2023. ICH licenses its documents for reuse with acknowledgement, so requirements are
-summarised here directly. Confirm the current text and your region's implementation date at
-<https://database.ich.org/sites/default/files/ICH_Q2%28R2%29_Guideline_2023_1130.pdf>.
-
-## Document history that matters
-
-| Version | Date | Note |
-| --- | --- | --- |
-| Q2A | Oct 1994 | Text |
-| Q2B | Nov 1996 | Methodology |
-| Q2(R1) | Nov 2005 | Q2B merged into the parent guideline |
-| Q2(R2) | 1 Nov 2023 | Complete revision, aligned with the new Q14 |
-| Q2(R2) correction | 30 Nov 2023 | Table 5 reportable-range linearity formulae; Tables 6–11 |
-
-If a protocol cites "ICH Q2(R1)" or lists characteristics in the R1 order, it is working from the
-superseded structure. The error correction is easy to miss and applies to the dissolution example
-and to Annex 2 Tables 6–11.
-
-## The restructure
-
-Q2(R1) presented a flat list. Q2(R2) groups methodology under section 3 by performance
-characteristic:
-
-```
-3.1  Specificity/Selectivity
-       3.1.1 General considerations (absence of interference, orthogonal comparison,
-             technology-inherent justification)
-3.2  Range                                    <-- parent characteristic
-       3.2.2 Response
-               3.2.2.1 Linear response        <-- what R1 called "linearity"
-               3.2.2.2 Non-linear response
-               3.2.2.3 Multivariate calibration
-       3.2.3 Validation of lower range limits <-- what R1 called LOD and LOQ
-3.3  Accuracy and Precision
-       3.3.1 Accuracy
-       3.3.2 Precision (repeatability, intermediate precision, reproducibility)
-       3.3.3 Combined approaches for accuracy and precision   <-- new
-3.4  Robustness  --> largely a development activity, see ICH Q14
-```
-
-Section 2 carries the general considerations, including two concepts absent from R1: **reportable
-range** (2.3) and **considerations for multivariate analytical procedures** (2.5).
-
-## Table 1 — which tests for which measured attribute
-
-Required tests follow the *measured quality attribute*, not the instrument.
-
-| Characteristic | Identity | Impurity: quantitative | Impurity: limit test | Assay (content/potency) |
-| --- | --- | --- | --- | --- |
-| Specificity test | yes | yes | yes | yes |
-| Response (calibration model) | no | yes | no | yes |
-| Lower range limit | no | QL† | DL | no |
-| Accuracy test | no | yes | no | yes |
-| Repeatability test | no | yes | no | yes |
-| Intermediate precision test | no | yes‡ | no | yes‡ |
-
-† In some complex cases DL may also be evaluated.
-‡ Not required independently where reproducibility has been performed and intermediate precision
-can be derived from that dataset.
-
-Further notes from Table 1: other quantitative measurements follow the impurity scheme when the
-range limit is close to DL/QL, and the assay scheme when it is not. Some characteristics may be
-substituted by technology-inherent justification for physicochemical properties. Lack of specificity
-in one procedure should be compensated by one or more supporting procedures unless justified.
-
-## Table 2 — reportable range examples
-
-The reportable range derives from the specification and must include the upper and lower
-specification or reporting limits. Other ranges are acceptable if justified; at low amounts a wider
-upper range may be more practical.
-
-| Use | Low end | High end |
-| --- | --- | --- |
-| Assay of a product | 80% of declared content, or 80% of the lower specification limit | 120% of declared content, or 120% of the upper specification limit |
-| Potency | lowest specification limit −20% | highest specification limit +20% |
-| Content uniformity | 70% of declared content | 130% of declared content |
-| Dissolution, IR, one point | Q − 45% of the lowest strength specification | per specification |
-| Dissolution, IR, multi-point | lower limit as justified, or QL | 130% of declared content of the highest strength |
-| Dissolution, modified release | lower limit as justified, or QL | per specification |
-| Impurity | reporting threshold | 120% of the specification limit |
-| Purity (area %) | 80% of the lower specification limit | upper specification limit, or 100% |
-
-Where assay and impurity run as a single test with one standard, linearity must be shown both at the
-impurity reporting level and up to 120% of the assay specification limit.
-
-**Reportable range vs working range.** The reportable range is the interval of *reported results*.
-A working range is what is presented to the instrument, and may differ because of dilution or other
-sample preparation. They can be identical. Mathematical calculation normally links the two.
-
-## Recommended data, by characteristic
-
-**Specificity (3.1).** Demonstrate absence of relevant interference, or compare against an
-orthogonal procedure, or justify from the technology. For a stability-indicating claim (2.4),
-include samples containing relevant degradation products: spiked with target analytes and known
-interferences, stressed physically and chemically, and aged or stress-stored product samples.
-
-**Response — linear (3.2.2.1).** Evaluate across the range. **A minimum of five concentrations,
-appropriately distributed, is recommended.** Report the plot, the correlation coefficient or
-coefficient of determination, the y-intercept, the slope, and *an analysis of the deviation of the
-actual data points from the regression line* — for a linear response, assess the impact of any
-non-random pattern in the residual plot. Data may be transformed (for example logarithmically) if
-necessary. Other approaches require justification.
-
-**Response — non-linear (3.2.2.2).** Some procedures are legitimately non-linear; immunoassays and
-cell-based assays commonly give an S-shaped curve, typically modelled with four- or five-parameter
-logistic functions. For these, **linearity of the concentration–response relationship is not
-required.** Assess the model by non-linear regression, and evaluate whether results are proportional
-to the true values across the range.
-
-**Response — multivariate (3.2.2.3).** Algorithms may be linear or non-linear. Accuracy depends on
-the distribution of calibration samples across the range and on the reference procedure's error.
-Assess how the residuals change across the calibration range, graphically.
-
-**Lower range limits (3.2.3).** Four approaches:
-
-| Approach | DL | QL |
-| --- | --- | --- |
-| Visual evaluation (3.2.3.1) | lowest reliably detected | lowest reliably quantitated |
-| Signal-to-noise (3.2.3.2) | S/N 3:1 generally acceptable | S/N at least 10:1 |
-| SD of response and slope (3.2.3.3) | 3.3σ / S | 10σ / S |
-| Accuracy and precision at the limit (3.2.3.4) | — | validated directly, not estimated |
-
-σ may come from the SD of blank responses, the residual SD of the regression line, or the SD of
-y-intercepts of regression lines. S is the calibration slope. Signal-to-noise applies only where
-there is baseline noise, and the noise region should sit around where the peak would appear.
-
-Reporting (3.2.3.5): give the limit **and the approach used**. An estimated limit should then be
-validated by analysing a suitable number of samples at or near it. **For impurity tests the QL must
-be at or below the reporting threshold.** Where the QL is well below the reporting limit — roughly
-ten times lower — the confirmatory validation may be omitted with justification.
-
-**Accuracy (3.3.1).** Establish across the reportable range under regular test conditions, including
-the sample matrix and the described preparation steps. Three routes: comparison against a reference
-material of known purity, a spiking study into matrix, or comparison against an orthogonal
-procedure. Accuracy can be inferred once precision, response within the range, and specificity are
-established.
-
-Recommended data (3.3.1.4): an appropriate number of determinations and levels across the reportable
-range — **for example 3 concentrations × 3 replicates of the full procedure.** Report as mean percent
-recovery of a known added amount, or as the difference between the mean and the accepted true value,
-**together with an appropriate 100(1−α)% confidence interval** or justified alternative interval. The
-observed interval should be compatible with the accuracy criterion. For impurities, state whether
-the determination is weight/weight or area percent. For quantitative multivariate procedures use
-RMSEP, compared against an acceptable RMSEC.
-
-**Precision (3.3.2).** Use authentic homogeneous samples, or artificially prepared ones if
-unavailable.
-
-- *Repeatability (3.3.2.1)*: **a minimum of 9 determinations covering the reportable range** (for
-  example 3 concentrations × 3 replicates), **or a minimum of 6 determinations at 100% of the test
-  concentration.**
-- *Intermediate precision (3.3.2.2)*: establish the effects of random events — typically different
-  days, environmental conditions, analysts, and equipment. **Studying these effects individually is
-  not necessary**, and design of experiments is encouraged. The extent should be justified from
-  development understanding and risk assessment (ICH Q14).
-- *Reproducibility (3.3.2.3)*: an inter-laboratory trial. **Usually not required for a regulatory
-  submission**, but consider it for pharmacopoeial standardisation or multi-site procedures.
-
-Recommended data (3.3.2.4): report the standard deviation, the relative standard deviation, and an
-appropriate 100(1−α)% confidence interval.
-
-**Combined accuracy and precision (3.3.3).** Instead of separate criteria, assess total impact
-against a single combined criterion, using a prediction interval, a tolerance interval, or a
-confidence interval. Report the combined value, describe the approach, and supply the individual
-results as supplemental information where they help justify suitability.
-
-**Robustness (3.4).** Deliberate variation of procedure parameters, plus stability of sample
-preparations and reagents over the duration of the procedure. Considered during development; may be
-submitted as development data case-by-case or made available on request. See ICH Q14 section 5.
-
-## Lifecycle, transfer, and prior knowledge
-
-Section 2.1 permits suitable development data (ICH Q14) to form part of the validation data, and
-allows abbreviated validation testing for an established platform procedure used for a new purpose,
-with scientific justification. A validation protocol must exist before the study, stating the
-intended purpose, the characteristics to be validated, and the associated criteria; where prior
-knowledge is used, justify it. Results are summarised in a validation report.
-
-The experimental design should reflect the number of replicates used in routine analysis to generate
-a reportable result, unless a different number is justified.
-
-Section 2.2 covers change: partial or full revalidation may be needed, decided on science and risk,
-and scoped to the characteristics the change affects. **Transfer** to another laboratory calls for
-partial or full revalidation and/or comparative analysis of representative samples; not performing
-transfer experiments requires justification. **Co-validation** across multiple sites can demonstrate
-the criteria are met and can simultaneously satisfy transfer at the participating sites.
-
-## Annex 2 — illustrative technique examples
-
-Non-mandatory worked examples, useful as a starting point for the robustness parameter list:
-
-| Table | Technique |
-| --- | --- |
-| 3 | Quantitative separation techniques (HPLC, GC, CE) for impurities or assay, and relative-area quantitation |
-| 4 | Elemental impurities by ICP-OES or ICP-MS |
-| 5 | Dissolution with HPLC as product performance test (corrected 30 Nov 2023) |
-| 6 | Quantitative ¹H-NMR for assay of a drug substance |
-| 7 | Biological assays |
-| 8 | Quantitative PCR |
-| 9 | Particle size measurement |
-| 10 | NIR analytical procedure |
-| 11 | Quantitative LC/MS |
-
-From Table 3, a detail worth carrying forward: **relative response factors.** Where the analyte
-responds differently from the reference material, calculate the RRF from the appropriate ratio of
-responses under final procedure conditions and document it. **If the RRF falls outside 0.8–1.2,
-apply a correction factor.** Where an impurity is overestimated, omitting the correction may be
-acceptable.
-
-## Multivariate procedures (2.5)
-
-Results come from a model relating many input variables to the property of interest. Validate in two
-phases:
-
-1. **Model development** — calibration plus internal testing. Test data may be a separate set or
-   part of the calibration set used rotationally, and are used to estimate performance and tune
-   parameters such as the number of PLS latent variables. See ICH Q14.
-2. **Model validation** — an independent validation set. For identification libraries, analyse
-   challenge samples *not* represented in the library to demonstrate discriminative ability.
-
-Samples need reference values or categories, normally from a validated or pharmacopoeial reference
-procedure whose performance **equals or exceeds** the expected performance of the multivariate
-procedure. Reference measurement and multivariate data collection should be on the same samples
-within a period short enough to assure sample and measurement stability. Describe any correlation or
-unit conversion, and any assumptions.