🧪 Formaldehyde in Water

Physicochemistry PN-ISO 12460

In short

Spectrophotometric determination of formaldehyde in water by acetylacetone method (Hantzsch). The test is performed according to PN-EN ISO 12460-5:2016; Measurement range: 10–5,000 µg/L (with dilution). The procedure comprises 10 steps (about 3 h 20 min in total); it is used for: Drinking water quality control — formaldehyde monitoring, Industrial wastewater testing (chemical plants, sawmills, resin production), Monitoring by-products of water ozonation.

At a glance

  • Standard: PN-EN ISO 12460-5:2016
  • Category: Physicochemistry
  • Procedure steps: 10
  • Total time of stages: 3 h 20 min
  • Measurement wavelength: 410 nm
  • Measurement range: 10–5,000 µg/L (with dilution)
  • Limit of quantification (LOQ): 10–50 µg/L (1–5 cm cuvette)

Overview

Formaldehyde (HCHO) is the simplest aldehyde, occurring in water as industrial pollution (resin, glue, textile production), by-product of water ozonation, and natural product of organic matter decomposition. It is classified by IARC as group 1 carcinogen. The permissible formaldehyde concentration in drinking water is 50 µg/L according to WHO.

The acetylacetone method (Nash/Hantzsch method) is a classic colorimetric technique based on condensation reaction of formaldehyde with acetylacetone and ammonium ion in acetic acid medium. The reaction product is 3,5-diacetyl-1,4-dihydrolutidine (DDL) — a yellow dye with absorption maximum at 410 nm. Color intensity is proportional to formaldehyde concentration in sample.

The method has good selectivity — other aldehydes (acetaldehyde, propionaldehyde) do not interfere at typical concentrations found in water. The limit of quantification is 10–50 µg/L depending on cuvette path length. The method is routinely used in water and wastewater laboratories.

Alternatively, formaldehyde in water can be determined by HPLC with DNPH derivatization (more sensitive, LOD ~1 µg/L) or enzymatic method. However, the spectrophotometric method remains the simplest and cheapest in routine laboratory practice.

Method principle

Formaldehyde reacts with acetylacetone (2,4-pentanedione) and ammonium ion (from ammonium acetate) in Hantzsch reaction, forming 3,5-diacetyl-1,4-dihydrolutidine (DDL). This is a condensation reaction leading to yellow dye with absorption maximum at λ = 410 nm. The reaction proceeds at 40°C for 30 minutes or at room temperature for 60 minutes. Absorbance is measured with UV-Vis spectrophotometer in 1 cm or 5 cm cuvette. Formaldehyde concentration is calculated from calibration curve prepared from formaldehyde standard solutions.

Applications

Key parameters

ParameterValue
Measurement wavelength410 nm
Measurement range10–5,000 µg/L (with dilution)
Limit of quantification (LOQ)10–50 µg/L (1–5 cm cuvette)
Reaction temperature40°C / 30 min or 20°C / 60 min
Precision (CV)3–8%
SelectivityHigh — no interference from C2–C4 aldehydes

Standard

Standard number
PN-EN ISO 12460-5:2016
Title (PL)
Oznaczanie formaldehydu w wodzie — Metoda spektrofotometryczna z acetyloacetonem
Title (EN)
Determination of formaldehyde in water — Spectrophotometric method with acetylacetone

Step-by-step procedure

  1. Nash reagent preparation

    Dissolve 150 g ammonium acetate in ~800 mL deionized water. Add 3 mL glacial acetic acid and 2 mL acetylacetone. Make up to 1 L. Store in dark bottle at 4°C (stable 1 month).

    ⏱ Time: 15 min

  2. Standard preparation

    From formaldehyde standard (37%) prepare working solution 100 mg/L. Prepare calibration series: 0, 0.05, 0.1, 0.25, 0.5, 1.0, 2.0, 5.0 mg/L in 25 mL volumetric flasks.

    ⏱ Time: 20 min

  3. Color reaction — standards

    To each standard (10 mL) add 2 mL Nash reagent. Mix. Place in water bath at 40°C for 30 minutes.

    ⏱ Time: 30 min • 🌡 Temperature: 40°C

  4. Cooling

    Remove tubes from water bath. Cool to room temperature in darkness (30 min). DDL dye is stable for 2 hours.

    ⏱ Time: 30 min

  5. Absorbance measurement — calibration

    Measure absorbance of standards at 410 nm against blank (water + Nash reagent). Plot calibration curve A = f(c).

    ⏱ Time: 15 min

  6. Sample preparation

    To 10 mL water sample add 2 mL Nash reagent. Mix. Incubate 40°C / 30 min, cool 30 min (same as standards).

    ⏱ Time: 60 min • 🌡 Temperature: 40°C

  7. Absorbance measurement — samples

    Measure sample absorbance at 410 nm in 1 cm cuvette (or 5 cm for low concentrations). Read concentration from calibration curve.

    ⏱ Time: 5 min

  8. Calculations

    HCHO concentration [µg/L] = reading from curve × dilution factor. Account for blank and control sample correction.

    ⏱ Time: 5 min

  9. Quality control

    Control sample (CRM or spike 0.5 mg/L) — recovery 90–110%. Blank < LOQ. Duplicate: RSD < 10%.

    ⏱ Time: 10 min

  10. Cleaning

    Rinse cuvettes with deionized water and ethanol. Store Nash reagent at 4°C in darkness. Collect waste in organic waste container.

    ⏱ Time: 10 min

Required equipment and apparatus

EquipmentExampleIndicative price
UV-Vis spectrophotometerHach DR 6000, Shimadzu UV-1900i, Agilent Cary 6015,000–80,000 PLN
Glass/quartz cuvettes 1–5 cmHellma, Starna, borosilicate/quartz optical cuvettes100–500 PLN/pc
Water bathMemmert WNB 10, Julabo TW 12, PolyScience WB053,000–10,000 PLN
Automatic pipettesEppendorf Research Plus 100–1000 µL, 1–5 mL800–2,000 PLN
Class A volumetric flasks25, 50, 100, 250 mL, borosilicate glass class A30–80 PLN/pc
Analytical balanceMettler Toledo XPR205, Sartorius Quintix 2248,000–25,000 PLN

Reagents, media and consumables

ReagentCASDetails
Acetylacetone (2,4-pentanedione)123-54-6Analytical grade, 2 mL per 1 L Nash reagent
Ammonium acetate (CH₃COONH₄)631-61-8150 g per 1 L Nash reagent — NH₄⁺ ion source
Glacial acetic acid (CH₃COOH)64-19-73 mL per 1 L Nash reagent — pH adjustment ~6
Formaldehyde standard solution50-00-037% formaldehyde stabilized with methanol, for calibration curve preparation
Deionized waterResistivity >18.2 MΩ·cm, for dilutions and blanks

Health and safety (OHS)

Frequently asked questions

Which standard describes this test?

The test is performed according to PN-EN ISO 12460-5:2016 — “Determination of formaldehyde in water — Spectrophotometric method with acetylacetone”.

How does this method work?

Formaldehyde reacts with acetylacetone (2,4-pentanedione) and ammonium ion (from ammonium acetate) in Hantzsch reaction, forming 3,5-diacetyl-1,4-dihydrolutidine (DDL). This is a condensation reaction leading to yellow dye with absorption maximum at λ = 410 nm.

What is the measuring range and accuracy?

Measurement wavelength: 410 nm; Measurement range: 10–5,000 µg/L (with dilution); Limit of quantification (LOQ): 10–50 µg/L (1–5 cm cuvette); Reaction temperature: 40°C / 30 min or 20°C / 60 min.

How long does the test take?

The times given for the individual stages add up to approximately 3 h 20 min. The procedure comprises 10 steps.

What equipment is required?

UV-Vis spectrophotometer, Glass/quartz cuvettes 1–5 cm, Water bath, Automatic pipettes, Class A volumetric flasks, Analytical balance. Indicative cost of the core instrument (UV-Vis spectrophotometer): 15,000–80,000 PLN.

Where is this test used?

Drinking water quality control — formaldehyde monitoring; Industrial wastewater testing (chemical plants, sawmills, resin production); Monitoring by-products of water ozonation; Swimming pool water control; Surface water testing around industrial facilities; Process water analysis in textile and paper industry.

What safety precautions apply?

Formaldehyde 37% — carcinogenic (cat. 1), mutagenic, sensitizing — work exclusively in fume hood!; Acetylacetone — flammable, irritant to respiratory tract, use under fume hood; Glacial acetic acid — corrosive, causes skin and eye burns, acid-resistant gloves; Safety glasses, lab coat, and nitrile gloves mandatory; Dispose formaldehyde waste as hazardous waste.

Which laboratory can perform this test?

The test is performed by laboratories accredited to ISO/IEC 17025. On LabCoda you can find them by the standard number PN-ISO 12460.

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