☠️ Cyanides in water

Physicochemistry PN-EN ISO 6703

Determination of total and free cyanides in water after distillation — spectrophotometric method with pyridine and barbituric acid at 578 nm.

Overview

Cyanides are among the most highly toxic water contaminants. The cyanide ion CN⁻ blocks cytochrome oxidase in mitochondria, inhibiting cellular respiration. The lethal dose of KCN for humans is only 1–3 mg/kg body weight. The permissible cyanide concentration in drinking water is 50 µg/L (Journal of Laws 2017, item 2294).

ISO 6703 standard comprises three parts: Part 1 — determination of total cyanide, Part 2 — free cyanides (easily liberatable), Part 3 — cyanogen chloride. The method includes reflux distillation in acidic environment to release HCN from cyanide complexes, absorption in NaOH, and subsequent spectrophotometric determination.

Cyanides enter waters from electroplating, coking, metallurgical wastewaters, gold extraction processes (cyanide leaching), and plastics and pesticide production. Cyanide monitoring is required in water permits of industrial facilities.

Method principle

The method comprises two stages: (1) distillation and (2) spectrophotometric determination.

Distillation stage: The sample is acidified with phosphoric acid to pH < 2, which releases hydrogen cyanide (HCN) from cyanide compounds (including complexes with metals). HCN is distilled from the sample and absorbed in NaOH solution (0.4 mol/L), where it converts to ionic CN⁻.

Determination stage: Cyanides in the distillate react with chloramine-T forming cyanogen chloride (CNCl) at pH < 8. Then CNCl reacts with pyridine-barbituric acid forming a red-violet colored complex. Absorbance is measured spectrophotometrically at 578 nm (with barbituric acid) or 620 nm (with pyridine-pyrazolone).

The photometric method allows determination of 0.002–0.025 mg CN⁻, titration method (Tyndall effect) > 0.005 mg, and titration method with indicator > 0.05 mg.

Applications

Key parameters

ParameterValue
Photometric range0.002–0.025 mg CN⁻ in sample
Titrimetric range> 0.05 mg CN⁻
Limit of quantification~5 µg/L (photometry)
Wavelength578 nm (pyridine-barbituric acid) or 620 nm (pyridine-pyrazolone)
Sample max concentration< 100 mg CN⁻/L (without dilution)
InterferencesSulfides, aldehydes (formaldehyde), nitrites, thiocyanates

Standard

Standard number
PN-EN ISO 6703:2021
Title (PL)
Jakość wody — Oznaczanie cyjanków
Title (EN)
Water quality — Determination of cyanide

Step-by-step procedure

1. Sample preservation

Acidification is FORBIDDEN (would release HCN!). Add NaOH to pH > 12. Store in dark, 4°C. Test for sulfides (lead paper) — if present, add PbCO₃.

⏱ Time: 5 min

2. Distillation

Transfer 500 mL sample to round-bottom flask. Add H₃PO₄ to pH < 2. Connect to condenser and absorber (25 mL NaOH 0.4 mol/L). Distill under reflux until ~450 mL distillate collected.

⏱ Time: 60–90 min • 🌡 Temperature: ~100°C

3. Reagent preparation

Prepare fresh chloramine-T solution (daily). Check pyridine-barbituric acid reagent (stored at 4°C, valid ~1 week).

⏱ Time: 10 min

4. Color reaction

To 50 mL distillate add 0.5 mL chloramine-T. Mix 1 min. Add 5 mL pyridine-barbituric acid solution. Mix. Wait 8 min for color development.

⏱ Time: 10 min

5. Spectrophotometric measurement

Measure absorbance at 578 nm in 1 cm cuvette (or 5 cm for low concentrations). Read concentration from calibration curve. Reading within 15 min of adding coloring reagent.

⏱ Time: 5 min

6. Calibration

KCN standard solutions: 0; 0.02; 0.05; 0.1; 0.2; 0.5 mg/L CN⁻ (distilled like samples). Calibration curve: A = f(c). R² > 0.998.

⏱ Time: 15 min

7. Quality control

Blank sample (dist. water through distillation). CRM per series. Duplicate every 10 samples. Recovery 85–115%.

⏱ Time: 5 min

Required equipment and apparatus

EquipmentExampleIndicative price
Reflux distillation apparatusGlass set for cyanide distillation (1L round-bottom flask + condenser + absorber)3 000–8 000 PLN
UV-Vis spectrophotometerHach DR6000, Shimadzu UV-1900, Thermo Evolution 30015 000–80 000 PLN
Hot plate or heating mantleIKA, Heidolph — boiling temperature control2 000–5 000 PLN
Volumetric flasks and pipettesClass A flasks 100, 250, 500 mL; automatic pipettes 1–10 mL50–500 PLN/pc
Spectrophotometric cuvettesQuartz or glass cuvettes 1 cm, 5 cm100–500 PLN/pc

Reagents, media and consumables

ReagentCASDetails
Phosphoric acid (H₃PO₄)7664-38-2For acidifying sample during distillation (HCN release)
Sodium hydroxide (NaOH)1310-73-20.4 mol/L, absorption solution for capturing HCN from distillate
Chloramine-T127-65-10.014 mol/L (0.40 g/100 mL). For CN⁻ → CNCl conversion. Prepare fresh daily
Barbituric acid67-52-715 g barbituric acid + 75 mL pyridine + 15 mL conc. HCl to 1 L. Store at 4°C
Pyridine110-86-1Component of coloring reagent, analytical grade purity
KCN standard solution151-50-81000 mg/L CN⁻. Deadly toxic! Store in NaOH, away from acids

Health and safety (OHS)

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