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#314 QC & Validation Complete

MV: APAP HPLC-UV / ICH Q2(R1) full validation

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method-validation ICH-Q2 USP-1225 HPLC-UV acetaminophen

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This template captures a full ICH Q2(R1) / USP <1225> validation of a reverse-phase HPLC-UV assay quantifying acetaminophen (APAP) in a tablet matrix, supporting the assay method’s transfer to QC release-testing.

Goal: demonstrate that the developed HPLC-UV method is fit for its intended purpose by satisfying all eight ICH Q2(R1) validation characteristics — specificity, linearity, accuracy, repeatability, intermediate precision, LOD, LOQ, range, and robustness — within acceptance criteria.

Method conditions: Phenomenex Luna C18 250 × 4.6 mm × 5 µm column; mobile phase 75:25 water:methanol with 0.1% formic acid (isocratic); 1.0 mL/min flow; 25°C column oven; 10 µL injection; UV 243 nm detection; 6 min run time. Target concentration 100 µg/mL acetaminophen, working range 50-150 µg/mL.

Acceptance criteria (per ICH Q2(R1)): specificity — no co-eluting peaks; linearity — R² > 0.999 across 5 levels; accuracy — 98-102% recovery at 3 levels (n=3 each); repeatability — RSD < 2% (n=6); intermediate precision — RSD < 3% across 2 analysts and 2 days; LOD < 1 µg/mL, LOQ < 3 µg/mL; range — 50-150% target; robustness — pass with ±0.1 pH, ±5°C, ±2% organic, ±10 nm wavelength, ±0.1 mL/min flow.

Outcome: All eight parameters met acceptance criteria. Validation report v1.0 approved 2026-06-04. Method released to QC for routine release testing.

Last saved: 34 seconds ago

12 fields loaded from Method Validation ICH Q2 baseline (SOP v3.1)

Method

Analyte

text

Acetaminophen (APAP)

Matrix

select

tablet, suspension, plasma, urine, water

Instrument

select

HPLC-UV, LC-MS/MS, GC-FID, UV-Vis

Target conc

number

100µg/mL

Linearity

Cal range

text

50-150 µg/mL (5 levels)

R² (≥ 0.999)

number

0.9998

Slope

number

24891area/µg/mL

Intercept

number

218

Accuracy & Precision

Recovery (98-102%)

text

99.4 / 100.1 / 101.2 % (50/100/150%)

Repeatability RSD

number

0.8%

Intermediate RSD

number

1.2%

LOD / LOQ

text

0.1 / 0.3 µg/mL

validation_protocol.pdf

412 KB

validation_data.xlsx

162 KB

final_report_v1.0.pdf

688 KB

01

Review approved protocol; confirm IQ/OQ/PQ current for HPLC and column

PQ valid through 2026-09-15

0:30

02

Specificity: inject blank, placebo, APAP standard, spiked placebo

No co-eluting peaks at APAP RT 3.42 min

0:45

03

Linearity: prepare 5 levels (50, 75, 100, 125, 150 µg/mL), inject in triplicate

R² 0.9998 · slope 24891 · intercept 218

2:00

04

Accuracy: spike placebo at 50, 100, 150 µg/mL × 3 reps each; calc % recovery

99.4 / 100.1 / 101.2 %

1:30

05

Repeatability: 6 injections of 100 µg/mL standard; calc RSD

RSD 0.8% (n=6)

0:45

06

Intermediate precision: 2nd analyst, 2nd day, 6 injections; combine RSD

Combined RSD 1.2% (n=12 across 2 days)

1:30

07

LOD / LOQ: prepare serial dilutions, determine S/N = 3 (LOD) and 10 (LOQ)

LOD 0.1 µg/mL · LOQ 0.3 µg/mL

1:00

08

Robustness: ±0.1 pH, ±5°C, ±2% organic, ±10 nm λ, ±0.1 mL/min flow — 7 factor matrix

All 7 factors within ±2% of nominal — PASS

3:00

09

Compile validation report; tabulate parameter / criterion / observed / status

Report v1.0 drafted

1:30

10

Reviewer + QA approver sign electronic record; release method to QC

Signed by D. Hayes (QA) 2026-06-04 16:32

0:30

	A	B	C	D
1	Parameter	Acceptance criterion	Observed	Status
2	Specificity	no co-eluting peaks	no interference at RT	PASS
3	Linearity	R² ≥ 0.999	R² = 0.9998	PASS
4	Accuracy 50%	98-102% recovery	99.4%	PASS
5	Accuracy 100%	98-102% recovery	100.1%	PASS
6	Accuracy 150%	98-102% recovery	101.2%	PASS
7	Repeatability	RSD < 2% (n=6)	RSD 0.8%	PASS
8	Intermediate prec	RSD < 3% (2 days)	RSD 1.2%	PASS
9	LOD	< 1 µg/mL	0.1 µg/mL	PASS
10	LOQ	< 3 µg/mL	0.3 µg/mL	PASS
11	Range	50-150% target	50-150% verified	PASS
12	Robustness (7 factors)	± nominal within 2%	all 7 factors ≤ 2%	PASS

EXP-2026-0421 APAP HPLC method development draft M. Reyes Apr 21

EXP-2026-0612 APAP first QC release test (post-MV) M. Reyes Jun 12

SOP-0501 Method Validation SOP v3.1 (ICH Q2) QA Team May 1

REG-ICH ICH Q2(R1) Validation of Analytical Methods Regulatory Refs Nov 5

USP-1225 USP <1225> Validation of Compendial Procedures Regulatory Refs Aug 18

Acetaminophen USP reference standard

USP 1003009 · Lot R12270

In stock

Methanol (HPLC grade)

34860 · Lot SHBT1421

In stock

Formic acid (LC-MS grade)

14265 · Lot SLBR2784

In stock

Water (LC-MS grade)

00612 · Lot SLBT6618

In stock

Phenomenex Luna C18 column (250 × 4.6 × 5)

00G-4252-E0 · SN 4521A

In use

APAP tablet sample matrix (placebo + active)

internal · Lot R&D-2026-04

In stock

APAP USP reference standard

RT · Reference Std Cabinet · Box USP-A

×1

Mobile phase aliquots (1 L, prepared)

4 °C · Fridge 4 · Shelf 2 · Box MOBILE-APAP

×3

Validation worksheets + signed report

Document Mgmt · Validation/2026/MV-APAP-001

×1

Visibility

QC Validation GroupM. Reyes

Can write

M. ReyesT. HayesD. Hayes

Color

Stroke width

Eraser

Use Ctrl + click to add text

{
    "experiment": {
        "id": 314,
        "custom_id": "MV-APAP-HPLC-2026-04",
        "title": "MV: APAP HPLC-UV / ICH Q2(R1) full validation",
        "template": "Method Validation",
        "template_version": "v3.2",
        "category": "QC & Validation",
        "status": "Complete",
        "started_on": "2026-07-21",
        "tags": [
            "method-validation",
            "ICH-Q2",
            "USP-1225",
            "HPLC-UV",
            "acetaminophen"
        ]
    },
    "main_text": "This template captures a full ICH Q2(R1) / USP <1225> validation of a reverse-phase HPLC-UV assay quantifying acetaminophen (APAP) in a tablet matrix, supporting the assay method’s transfer to QC release-testing.\nGoal: demonstrate that the developed HPLC-UV method is fit for its intended purpose by satisfying all eight ICH Q2(R1) validation characteristics — specificity, linearity, accuracy, repeatability, intermediate precision, LOD, LOQ, range, and robustness — within acceptance criteria.\nMethod conditions: Phenomenex Luna C18 250 × 4.6 mm × 5 µm column; mobile phase 75:25 water:methanol with 0.1% formic acid (isocratic); 1.0 mL/min flow; 25°C column oven; 10 µL injection; UV 243 nm detection; 6 min run time. Target concentration 100 µg/mL acetaminophen, working range 50-150 µg/mL.\nAcceptance criteria (per ICH Q2(R1)): specificity — no co-eluting peaks; linearity — R² > 0.999 across 5 levels; accuracy — 98-102% recovery at 3 levels (n=3 each); repeatability — RSD < 2% (n=6); intermediate precision — RSD < 3% across 2 analysts and 2 days; LOD < 1 µg/mL, LOQ < 3 µg/mL; range — 50-150% target; robustness — pass with ±0.1 pH, ±5°C, ±2% organic, ±10 nm wavelength, ±0.1 mL/min flow.\nOutcome: All eight parameters met acceptance criteria. Validation report v1.0 approved 2026-06-04. Method released to QC for routine release testing.",
    "extra_fields": {
        "Method": {
            "Analyte": {
                "type": "text",
                "value": "Acetaminophen (APAP)"
            },
            "Matrix": {
                "type": "select",
                "value": "tablet, suspension, plasma, urine, water"
            },
            "Instrument": {
                "type": "select",
                "value": "HPLC-UV, LC-MS/MS, GC-FID, UV-Vis"
            },
            "Target conc": {
                "type": "number",
                "value": "100 µg/mL"
            }
        },
        "Linearity": {
            "Cal range": {
                "type": "text",
                "value": "50-150 µg/mL (5 levels)"
            },
            "R² (≥ 0.999)": {
                "type": "number",
                "value": "0.9998"
            },
            "Slope": {
                "type": "number",
                "value": "24891 area/µg/mL"
            },
            "Intercept": {
                "type": "number",
                "value": "218"
            }
        },
        "Accuracy & Precision": {
            "Recovery (98-102%)": {
                "type": "text",
                "value": "99.4 / 100.1 / 101.2 % (50/100/150%)"
            },
            "Repeatability RSD": {
                "type": "number",
                "value": "0.8 %"
            },
            "Intermediate RSD": {
                "type": "number",
                "value": "1.2 %"
            },
            "LOD / LOQ": {
                "type": "text",
                "value": "0.1 / 0.3 µg/mL"
            }
        }
    },
    "attached_files": [
        {
            "filename": "validation_protocol.pdf",
            "type": "pdf",
            "size": "412 KB"
        },
        {
            "filename": "validation_data.xlsx",
            "type": "xlsx",
            "size": "162 KB"
        },
        {
            "filename": "final_report_v1.0.pdf",
            "type": "pdf",
            "size": "688 KB"
        }
    ],
    "steps": [
        {
            "n": 1,
            "description": "Review approved protocol; confirm IQ/OQ/PQ current for HPLC and column",
            "duration": "0:30",
            "complete": true,
            "note": "PQ valid through 2026-09-15"
        },
        {
            "n": 2,
            "description": "Specificity: inject blank, placebo, APAP standard, spiked placebo",
            "duration": "0:45",
            "complete": true,
            "note": "No co-eluting peaks at APAP RT 3.42 min"
        },
        {
            "n": 3,
            "description": "Linearity: prepare 5 levels (50, 75, 100, 125, 150 µg/mL), inject in triplicate",
            "duration": "2:00",
            "complete": true,
            "note": "R² 0.9998 · slope 24891 · intercept 218"
        },
        {
            "n": 4,
            "description": "Accuracy: spike placebo at 50, 100, 150 µg/mL × 3 reps each; calc % recovery",
            "duration": "1:30",
            "complete": true,
            "note": "99.4 / 100.1 / 101.2 %"
        },
        {
            "n": 5,
            "description": "Repeatability: 6 injections of 100 µg/mL standard; calc RSD",
            "duration": "0:45",
            "complete": true,
            "note": "RSD 0.8% (n=6)"
        },
        {
            "n": 6,
            "description": "Intermediate precision: 2nd analyst, 2nd day, 6 injections; combine RSD",
            "duration": "1:30",
            "complete": true,
            "note": "Combined RSD 1.2% (n=12 across 2 days)"
        },
        {
            "n": 7,
            "description": "LOD / LOQ: prepare serial dilutions, determine S/N = 3 (LOD) and 10 (LOQ)",
            "duration": "1:00",
            "complete": true,
            "note": "LOD 0.1 µg/mL · LOQ 0.3 µg/mL"
        },
        {
            "n": 8,
            "description": "Robustness: ±0.1 pH, ±5°C, ±2% organic, ±10 nm λ, ±0.1 mL/min flow — 7 factor matrix",
            "duration": "3:00",
            "complete": true,
            "note": "All 7 factors within ±2% of nominal — PASS"
        },
        {
            "n": 9,
            "description": "Compile validation report; tabulate parameter / criterion / observed / status",
            "duration": "1:30",
            "complete": true,
            "note": "Report v1.0 drafted"
        },
        {
            "n": 10,
            "description": "Reviewer + QA approver sign electronic record; release method to QC",
            "duration": "0:30",
            "complete": true,
            "note": "Signed by D. Hayes (QA) 2026-06-04 16:32"
        }
    ],
    "spreadsheet": [
        [
            "Parameter",
            "Acceptance criterion",
            "Observed",
            "Status"
        ],
        [
            "Specificity",
            "no co-eluting peaks",
            "no interference at RT",
            "PASS"
        ],
        [
            "Linearity",
            "R² ≥ 0.999",
            "R² = 0.9998",
            "PASS"
        ],
        [
            "Accuracy 50%",
            "98-102% recovery",
            "99.4%",
            "PASS"
        ],
        [
            "Accuracy 100%",
            "98-102% recovery",
            "100.1%",
            "PASS"
        ],
        [
            "Accuracy 150%",
            "98-102% recovery",
            "101.2%",
            "PASS"
        ],
        [
            "Repeatability",
            "RSD < 2% (n=6)",
            "RSD 0.8%",
            "PASS"
        ],
        [
            "Intermediate prec",
            "RSD < 3% (2 days)",
            "RSD 1.2%",
            "PASS"
        ],
        [
            "LOD",
            "< 1 µg/mL",
            "0.1 µg/mL",
            "PASS"
        ],
        [
            "LOQ",
            "< 3 µg/mL",
            "0.3 µg/mL",
            "PASS"
        ],
        [
            "Range",
            "50-150% target",
            "50-150% verified",
            "PASS"
        ],
        [
            "Robustness (7 factors)",
            "± nominal within 2%",
            "all 7 factors ≤ 2%",
            "PASS"
        ]
    ],
    "links": {
        "experiments": [
            {
                "ref_id": "EXP-2026-0421",
                "title": "APAP HPLC method development draft",
                "owner": "M. Reyes",
                "date": "Apr 21"
            },
            {
                "ref_id": "EXP-2026-0612",
                "title": "APAP first QC release test (post-MV)",
                "owner": "M. Reyes",
                "date": "Jun 12"
            }
        ],
        "resources": [
            {
                "ref_id": "SOP-0501",
                "title": "Method Validation SOP v3.1 (ICH Q2)",
                "owner": "QA Team",
                "date": "May 1"
            },
            {
                "ref_id": "REG-ICH",
                "title": "ICH Q2(R1) Validation of Analytical Methods",
                "owner": "Regulatory Refs",
                "date": "Nov 5"
            },
            {
                "ref_id": "USP-1225",
                "title": "USP <1225> Validation of Compendial Procedures",
                "owner": "Regulatory Refs",
                "date": "Aug 18"
            }
        ]
    },
    "compounds": [
        {
            "name": "Acetaminophen USP reference standard",
            "catalog_lot": "USP 1003009 · Lot R12270",
            "stock_status": "In stock"
        },
        {
            "name": "Methanol (HPLC grade)",
            "catalog_lot": "34860 · Lot SHBT1421",
            "stock_status": "In stock"
        },
        {
            "name": "Formic acid (LC-MS grade)",
            "catalog_lot": "14265 · Lot SLBR2784",
            "stock_status": "In stock"
        },
        {
            "name": "Water (LC-MS grade)",
            "catalog_lot": "00612 · Lot SLBT6618",
            "stock_status": "In stock"
        },
        {
            "name": "Phenomenex Luna C18 column (250 × 4.6 × 5)",
            "catalog_lot": "00G-4252-E0 · SN 4521A",
            "stock_status": "In use"
        },
        {
            "name": "APAP tablet sample matrix (placebo + active)",
            "catalog_lot": "internal · Lot R&D-2026-04",
            "stock_status": "In stock"
        }
    ],
    "storage": [
        {
            "name": "APAP USP reference standard",
            "location": "RT · Reference Std Cabinet · Box USP-A",
            "count": "×1"
        },
        {
            "name": "Mobile phase aliquots (1 L, prepared)",
            "location": "4 °C · Fridge 4 · Shelf 2 · Box MOBILE-APAP",
            "count": "×3"
        },
        {
            "name": "Validation worksheets + signed report",
            "location": "Document Mgmt · Validation/2026/MV-APAP-001",
            "count": "×1"
        }
    ],
    "permissions": {
        "visibility": [
            {
                "type": "Team",
                "name": "QC Validation Group"
            },
            {
                "type": "Person",
                "name": "M. Reyes"
            }
        ],
        "can_write": [
            {
                "type": "Person",
                "name": "M. Reyes"
            },
            {
                "type": "Person",
                "name": "T. Hayes"
            },
            {
                "type": "Person",
                "name": "D. Hayes"
            }
        ]
    }
}