EndotoxinLALChecker

URS & FS — User Requirements and Functional Specification

This document describes the controlled interface and behaviour of EndotoxinLALChecker for Endotoxin LAL Checker.

Domain limits and critical parameters

• Endotoxin LAL Checker — Control of Bacterial Endotoxins (LAL Test)
• ℹ️ Utility checks bacterial endotoxin content using Limulus Amebocyte Lysate (LAL) according to Ph. Eur. 2.6.14 and USP <85>:
• • Endotoxin Concentration (EU/mL) — main safety parameter
• • Calculation of individual limit using K/M formula (where K=5.0 EU/kg for parenterals)
• • Precision (CV): ≤ 25% for replicates
• ⚠️ CRITICAL: If Spike Recovery is outside 50-200%, the test is considered INVALID!
• EndotoxinLALChecker.exe → demo mode (console output)
• EndotoxinLALChecker.exe input.csv output.json → evaluate your data
• BatchNumber,ProductName,EndotoxinConcentration_EU_ml,DilutionFactor,MaxDose_ml_per_kg,K_Factor,RecoveryPercent,CV_Percent,MethodType,NC_Response,PC_Response
• Bacterial endotoxins (lipopolysaccharides of Gram-negative bacteria) cause pyrogenic reactions in patients.
• ⚠️ CRITICAL:
• • Endotoxin Limit (K): Calculated as K/M (where M is maximum drug dose per kg body weight per hour).
• • Dilution: If interference is detected, the sample must be diluted until recovery is 50-200%.
• • Specificity: LAL reacts only with endotoxins (unlike old methods), but may give false positives in presence of (1→3)-β-D-glucans (unless glucan-blocking reagent is used).
• • Endotoxin limit compliance assessment
• • Compliance with GMP and Pharmacopoeia requirements (Ph. Eur. 2.6.14)
• Critical parameters:
• • Endotoxin: ≤ specified limit (K/M)

URS — User Requirements Specification

input.csv contract

BatchNumber,ProductName,EndotoxinConcentration_EU_ml,DilutionFactor,MaxDose_ml_per_kg,K_Factor,RecoveryPercent,CV_Percent,MethodType,NC_Response,PC_Response
LAL-2026-WFI-001,Water for Injection (WFI),0.03,1.0,10.0,5.0,98.0,2.5,Chromogenic,0.005,0.450
LAL-2026-API-002,Active Substance X,0.15,10.0,0.5,5.0,110.0,4.0,GelClot,0.002,0.500
LAL-2026-FAIL-003,Complex Formulation Y,0.80,5.0,1.0,5.0,40.0,3.0,Turbidimetric,0.004,0.480

Input validation rules

FS — Functional Specification

Example output.json

{
  "utilityId": "endotoxinlalchecker",
  "utilityFolder": "EndotoxinLALChecker",
  "package": "Vet",
  "overallStatus": "PASS|WARNING|FAIL",
  "sourceFile": "input.csv",
  "processedAtUtc": "2026-06-10T00:00:00Z",
  "checks": [
    {
      "parameter": "BatchNumber",
      "value": "LAL-2026-WFI-001",
      "status": "PASS|WARNING|FAIL",
      "message": "Deterministic rule-based check result",
      "ruleReference": "FS-RULE-001"
    },
    {
      "parameter": "ProductName",
      "value": "Water for Injection (WFI)",
      "status": "PASS|WARNING|FAIL",
      "message": "Deterministic rule-based check result",
      "ruleReference": "FS-RULE-002"
    },
    {
      "parameter": "EndotoxinConcentration_EU_ml",
      "value": "0.03",
      "status": "PASS|WARNING|FAIL",
      "message": "Deterministic rule-based check result",
      "ruleReference": "FS-RULE-003"
    },
    {
      "parameter": "DilutionFactor",
      "value": "1.0",
      "status": "PASS|WARNING|FAIL",
      "message": "Deterministic rule-based check result",
      "ruleReference": "FS-RULE-004"
    },
    {
      "parameter": "MaxDose_ml_per_kg",
      "value": "10.0",
      "status": "PASS|WARNING|FAIL",
      "message": "Deterministic rule-based check result",
      "ruleReference": "FS-RULE-005"
    },
    {
      "parameter": "K_Factor",
      "value": "5.0",
      "status": "PASS|WARNING|FAIL",
      "message": "Deterministic rule-based check result",
      "ruleReference": "FS-RULE-006"
    },
    {
      "parameter": "RecoveryPercent",
      "value": "98.0",
      "status": "PASS|WARNING|FAIL",
      "message": "Deterministic rule-based check result",
      "ruleReference": "FS-RULE-007"
    },
    {
      "parameter": "CV_Percent",
      "value": "2.5",
      "status": "PASS|WARNING|FAIL",
      "message": "Deterministic rule-based check result",
      "ruleReference": "FS-RULE-008"
    },
    {
      "parameter": "MethodType",
      "value": "Chromogenic",
      "status": "PASS|WARNING|FAIL",
      "message": "Deterministic rule-based check result",
      "ruleReference": "FS-RULE-009"
    },
    {
      "parameter": "NC_Response",
      "value": "0.005",
      "status": "PASS|WARNING|FAIL",
      "message": "Deterministic rule-based check result",
      "ruleReference": "FS-RULE-010"
    },
    {
      "parameter": "PC_Response",
      "value": "0.450",
      "status": "PASS|WARNING|FAIL",
      "message": "Deterministic rule-based check result",
      "ruleReference": "FS-RULE-011"
    }
  ],
  "criticalFindings": [],
  "warnings": [],
  "audit": {
    "inputHash": "sha256:<calculated at runtime>",
    "rulesVersion": "<utility executable version>",
    "documentation": "EndotoxinLALChecker.documentation.html"
  }
}

Traceability matrix

IQ/OQ/PQ test scenarios

QA/QC and change control

• Do not rename columns without updating validator, documentation and test set.
• Retain input.csv, output.json, executable version and checksum.
• Before production use, perform IQ/OQ/PQ or equivalent CSV/CSA verification.
• Critical limits shall be verified against the approved specification, registration dossier and local SOPs.
• The utility provides structured QC decision support; final release decision remains with QA/QP and approved procedures.