Fluorat_Nucleic_Acids_Quantifier

URS & FS — User Requirements and Functional Specification

This document describes the controlled interface, user requirements and functional behaviour of Fluorat_Nucleic_Acids_Quantifier. The utility is intended for automated verification of laboratory, pharmacopoeial, analytical or manufacturing QC parameters using input.csv and producing a structured output.json result.

Domain limits and critical parameters

• • Concentration: above limit of detection (LOD, usually >1 ng/uL)
• • Signal-to-Background Ratio (SBR): ≥3.0
• • Calibration Curve Coefficient of Determination (R²): ≥0.99
• • Fluorometric methods using intercalating dyes (PicoGreen, SYBR Green, RiboGreen) are significantly more sensitive than UV spectrophotometry (A260).
• • Allows determination of concentrations in the picogram/nanogram range.
• • Lumex Fluorat instruments provide excitation stability and low noise levels, important for accurate quantitative analysis.
• • Dye specificity allows distinction between single-stranded and double-stranded nucleic acids.
• • Concentration must exceed the method's limit of detection (LOD).
• • Signal-to-Background Ratio (SBR) must be high to minimize pipetting error and solvent background.
• • Calibration curve must be linear in the working range (R² ≥ 0.99).
• • Avoid contamination with DNase/RNase and fluorescence inhibitors.
• • High-sensitivity DNA/RNA determination.
• • Analytical signal quality control (SBR, R²).
• • Support for Lumex Fluorat fluorometer data.
• • Concentration: > LOD (e.g., 1.0 ng/uL)
• • Signal-to-Background Ratio: >= 3.0

URS — User Requirements Specification

input.csv contract

BatchNumber,Concentration_ng_uL,SampleRFU,BlankRFU,StandardCurveR2
DNA-FLUOR-2026-001,55.0,2500.0,50.0,0.998
RNA-FLUOR-2026-001,42.5,1800.0,45.0,0.995
NA-FAIL-LOW-001,0.5,60.0,55.0,0.992

FS — Functional Specification

Example output.json

{
  "utilityId": "fluorat-nucleic-acids-quantifier",
  "utilityName": "Fluorat_Nucleic_Acids_Quantifier",
  "overallStatus": "PASS|WARNING|FAIL",
  "sourceFile": "input.csv",
  "checks": [
    {
      "parameter": "BatchNumber",
      "value": "DNA-FLUOR-2026-001",
      "status": "PASS|WARNING|FAIL",
      "message": "Rule-based check result"
    },
    {
      "parameter": "Concentration_ng_uL",
      "value": "55.0",
      "status": "PASS|WARNING|FAIL",
      "message": "Rule-based check result"
    },
    {
      "parameter": "SampleRFU",
      "value": "2500.0",
      "status": "PASS|WARNING|FAIL",
      "message": "Rule-based check result"
    },
    {
      "parameter": "BlankRFU",
      "value": "50.0",
      "status": "PASS|WARNING|FAIL",
      "message": "Rule-based check result"
    },
    {
      "parameter": "StandardCurveR2",
      "value": "0.998",
      "status": "PASS|WARNING|FAIL",
      "message": "Rule-based check result"
    }
  ],
  "criticalFindings": [],
  "warnings": [],
  "generatedFor": "QA/QC review and LIMS integration"
}

Traceability matrix

OQ/PQ test scenarios

QA/QC and change control

• Do not rename columns without updating the validator, documentation and test set.
• Retain input.csv, output.json, executable version, documentation and checksum.
• Before production use, perform IQ/OQ/PQ or equivalent CSV/CSA verification.
• Critical limits must be verified against the approved specification, local SOPs and registration dossier.