Method

Available Cyanide by ASTM D 6888

Document #40821214

Flow Solution™ FS 3700 Automated Chemistry Analyzer

Available Cyanide by Automated Ligand Exchange
ASTM D 6888, Flow Injection Analysis (FIA)
Cartridge Part Number 330091CT

Scope and Application

This method is used for determining available cyanide in water and wastewater by automated ligand exchange, flow
injection analysis, and amperometric detection according to ASTM Method D 6888-09 (Reference 2). This method is
approved for use in the USEPA’s data gathering and monitoring programs associated with the Clean Water Act.

Cyanide ion (CN–), hydrogen cyanide in water (HCN(aq)), and cyano-complexes of zinc, copper, cadmium, mercury, nickel,
and silver can be determined by this method. The presence of polysulfides and colloidal material may prove intractable for
application of this method.

Method Performance
Range 5.0 µg/L–0.5 mg/L
Rate 30 samples/hour
Precision at 50 µg/L <2% RSD
Method Detection Limit (MDL) 1.0 µg/L

The range may be extended to analyze other concentrations by changing the size of the sample loop. The quality of the
analysis is assured through reproducible calibration and testing of the Flow Injection Analysis (FIA) system.

A general flow diagram of the Flow Injection Analysis (FIA) system is shown below (see Figure 1 for a detailed flow
diagram).

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 Available Cyanide by ASTM D 6888
Reagents and Calibrants
Chemical Name CAS # Chemical Formula Part Number
Acetic acid, glacial 64-19-7 C2H4O2
Acetone 67-64-1 C 3H 6O
Bismuth nitrate pentahydrate 10035-06-0 Bi(NO3)3 • 5H2O
Water, deionized H 2O
5-[4-(Dimethylamino) benzylidene] rhodanine 536-17-4 C12H12N2OS2
DOWFAX 2A1 12626-49-2 A000080
Ethylenediamine 107-15-3 C2 H 8 N 2
Ligand exchange reagent, available cyanide A001416
(WAD) reagent A
Ligand exchange reagent, available cyanide A001417
(WAD) reagent B
Mercury (II) cyanide 594-04-1 Hg(CN)2
Nickel potassium cyanide 14220-17-8 K2Ni(CN)4
Potassium cyanide 151-50-8 KCN
Silver nitrate 7761-58-3 AgNO3
Sodium acetate 127-09-3 C2H3O2Na
Sodium arsenite 7784-46-5 NaAsO2
Sodium hydroxide 1310-73-2 NaOH
Sulfuric acid, concentrated 7664-93-9 H2SO4
Sulfuric acidification reagent (SAR) A001872

Summary of ASTM D 6888

ASMT D 6888 summarizes the method and interferences as follows below (Reference 6).

Method

l Prior to analysis, treat the sample to remove potential interferences. Ligand exchange reagents form
thermodynamically stable complexes with the transition metal ions, releasing the cyanide ion from the cyano-complex-
es. Addition of acid converts the cyanide ion to hydrogen cyanide gas (HCN), which passes under a gas diffusion mem-
brane. The hydrogen cyanide gas diffuses through the membrane into an alkaline receiving solution where it converts
back to cyanide ion. The cyanide ion is monitored amperometrically with a silver working electrode, silver/silver chloride
reference electrode, and platinum/stainless steel counter electrode at an applied potential of zero volt. The current gen-
erated is proportional to the cyanide concentration present in the original sample.

Interferences

l Method interferences can be caused by contaminants in the reagents, reagent water, and glassware, which may bias
the results. Take care to keep all such items free of contaminants.

l Sulfide, at concentrations greater than 50 ppm, is a positive interferant in this method (Reference 11). When sulfide is
acidified, it forms hydrogen sulfide, which passes through the gas diffusion membrane and produces a signal at the
silver electrode. In addition, sulfide ion reacts with cyanide ion in solution to reduce its concentration over time.

l Treat samples containing water soluble aldehydes, such as formaldehyde or acetaldehyde, by adding ethylenediamine
solution, at the time of sampling.

l Remove oxidizing agents that decompose cyanides by adding ascorbic acid if analysis is to be within 24 hours,
otherwise use sodium arsenite.

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 Available Cyanide by ASTM D 6888
l High concentrations of carbonate can result in a negative response in the amperometric detector when carbon dioxide
diffuses across the gas diffusion membrane into the alkaline receiving solution, reducing its pH. Treat effluents from
high-carbonate containing wastes, such as coal gasification waste and atmospheric emission scrub water, with
hydrated lime to stabilize the sample (Reference 11).

l Tests conducted on samples containing large amounts of colloids indicate rapid cyanide losses. Filter turbid or colloidal
samples.

l Nitrate and nitrite do not interfere in this method.

l Thiocyanate, sulfite, and thiosulfate do not interfere with this method.

Safety

WARNING: Cyanide solutions may produce toxic hydrogen cyanide gas when acidified. For these reasons, any method
involving cyanide in a sample or reagent should only be performed by properly trained personnel wearing adequate
personal protective equipment.

WARNING: Two separate waste bottles should be used: one clearly marked “Cyanide Waste” and another marked “Acid
Waste”. The waste line from the amperometric flow cell must go to the Cyanide Waste bottle. The waste line from the gas
diffusion module must go to the Acid Waste bottle.

WARNING: Cyanide ion, hydrocyanic acid, all cyanide salts, and most metal-cyanide complexes are extremely danger-
ous. As a contact poison, cyanide does not need to be ingested to produce toxicity. Also, cyanide solutions produce fatally
toxic hydrogen cyanide gas when acidified. For these reasons, any method involving cyanide in a sample or reagent
should only be performed by properly trained personnel wearing adequate personal protective equipment.

1. The toxicity or carcinogenicity of each compound or reagent used in this method has not been fully established. Each
chemical should be treated as a potential health hazard. Exposure to these chemicals should be reduced to the
lowest possible level.

2. For reference purposes, a file of Material Safety Data Sheets (MSDS) for each chemical used in this method should
be available to all personnel involved in this chemical analysis. The preparation of a formal safety plan is also
advisable.

3. Chemicals used in this method may be highly toxic or hazardous and should be handled with extreme caution at all
times. Consult the appropriate MSDS before handling.

4. Unknown samples may be potentially hazardous and should be handled with extreme caution at all times.

5. Proper personal protective equipment (PPE) should be used when handling or working in the presence of chemicals.

6. This method does not address all safety issues associated with its use. The laboratory is responsible for maintaining a
safe work environment and a current awareness file of OSHA regulations regarding the safe handling of the chemicals
specified in this method.

Sample Handling and Preservation

1. Collect samples in amber glass bottles with minimal headspace that have been thoroughly cleaned and rinsed with
reagent water.

2. Ensure the volume of sample collected is sufficient to obtain a representative sample, analyze replicates, and
minimize waste disposal. 40 mL amber VOA vials have been found sufficient.

3. Add NaOH solution to pH 11 and cool samples to 2-6 °C.

4. Do not add NaOH if the cyanide concentration would change as a result of the addition.

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5. Treat samples containing sulfide ion.
a. Test samples with lead acetate test paper to determine the presence or absence of sulfide ion. Note that the lead
acetate test paper can be unreliable and is typically not usable for sulfide concentrations below approximately
50 ppm.
i. If sulfide is detected by the lead acetate test strip (sulfide > 50 ppm), dilute sample so that sulfide is below
amount detected by test strips. Record dilution factor on chain of custody/ sample bottle analyze samples
within 24 hours.
ii. Lead acetate test paper should be tested for minimum level of sulfide detection by spiking reagent water
aliquots with decreasing levels of sulfide. The spiked samples are tested with lead acetate test paper
moistened with acetate buffer. Test each new batch of test paper or acetate buffer to determine the lowest
level of sulfide ion detection prior to use.
b. If the sample contains particulate matter that would be removed with filtration, filter the sample. Available CN
species are soluble in water, so particulate matter need not be analyzed.

6. Treat samples containing or suspected to contain formaldehyde, acetaldehyde, or other water-soluble aldehydes with
20 mL of 3.5% ethylenediamine solution per liter of sample.

NOTE: Samples that have been disinfected by UV irradiation are likely to contain aldehydes.

7. Oxidizing agents such as chlorine can destroy most of the cyanide during storage and manipulation. Destroy the
oxidizing agent at the time of sample collection so the test result reflects an accurate cyanide concentration in the
source at the time of collection. Treat samples containing chlorine, hypochlorite, or sulfite with 0.1 g sodium arsenite
per 100 mL sample. Sodium thiosulfate may be used in place of sodium arsenite.
a. Using a plastic pipette, place a drop of sample on a strip of potassium iodide-starch test paper.
b. If observing a bluish discoloration on the paper, add a few crystals of sodium arsenite to the sample with a
spatula. Continue to add crystals and testing a drop of sample on the test paper until color is no longer produced
on the test paper.

NOTE: Do not add an excess of reducing agent.

8. High concentrations of carbonate can result in a negative response in the amperometric detector when carbon dioxide
diffuses across the membrane into the alkaline acceptor reagent, reducing its pH. The high buffer capacity of the
acceptor reagent largely eliminates sensitivity to carbonate. Nevertheless, Standard Methods recommend using
hydrated lime to stabilize samples such as effluents from coal gasification wastes, atmospheric emission scrub waters,
and other high carbonate-containing wastes.
a. Slowly add hydrated lime while stirring to raise the pH of the sample to 12.0–12.5.
b. Decant the sample after the precipitate settles.

Configuration and Start-Up

Refer to the Flow Solution™ 3700 Operator Manual (Reference 3) and/or the FlowView Quick Start Reference Guide
(Reference 14) for assistance configuring the FS 3700 and FlowView.

1. Instrument Configuration

a. Verify that the cartridge is configured as illustrated in the cartridge schematic shown in Figure 1.
b. Connect the appropriate pump tubes to the cartridge and to their appropriate reagent containers according to the
flow diagram.
c. Create and Save Method in FlowView, as follows:

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Primary Chassis Pump at Start 40%

Valve Load Inject (sec) 35
Cycle Duration (sec) 120
Heater Setpoint (°C) Not Used
UV Lamp Not Used
Detector Detector Mode Amperometric
Sample Gain 1X
Applied Potential 0
Polarity Normal

Calibration Settings Calibrant Units ppm

Final Units ppm
Conversion Factor 1.00
Calibration Curve Fitting Method 2nd Order
Include Origin as a Calibration Point? No
Peak Measurement Measure Peaks by Height
Baseline Value for Peak Marking Sync Peak Starts as Baseline (Default)
Apply Carryover Correction? No
Smoothing Smoothing Mode (#points) Moving Average (5 points)
Sync-Peak Marking Recognize Sync Peak Start by a rise 100
of
Total Counts within 5 seconds
Sync Peak Ignore Time (sec) 50
Use Return to Baseline? √
Return to Baseline (%) 99
Minimum Peak Height 400
Minimum Peak Width 60
Use 1st derivative? √
Minimum Return to Baseline (%) 90
Cutoff Counter Value for End of Peak 4

2. Instrument Stabilization

a. Start the pump, allowing the solution to flow through the entire system.
b. Make sure that the flow cell of each detector is purged of all bubbles and the flow is stable and free from surging.
c. For trace level analysis, OI Analytical recommends calibrating from 2.00-100 µg/L.

3. Amperometric Cell Conditioning

Perform the following after servicing the amperometric cell, or as needed.

a. Load a 5 mg/L cyanide standard into the sampling valve and inject it into the segmented FIA system.
b. Continue to inject 5 mg/L cyanide standard until three successive peak heights or area results are within 5% RSD,
indicating a stable electrode system.
c. Following stabilization, or to demonstrate stability of the electrode system, inject the 0.5 mg/L cyanide calibrant until
three successive peak heights or area results are within 3% RSD.

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4. Start the Run

a. Select Sample Table and specify Result File prior to starting the run. For help on creating a Sample Table, refer to
the FlowView Software Quick Start Guide (Reference 14).
b. Press Start to begin to collecting baseline data. Verify that the baseline does not drift and is free from large
fluctuations. When ready, click Run.

NOTE: Very sharp fluctuations in the baseline and/or consistent drifting are typically signs of bubbles in the flowcell. The
flowcell must be free of bubbles prior to beginning analysis.

5. Operating Notes

a. Sensitivity and performance at the low end may be enhanced by:

i. Increasing the size of the sample loop.
ii. Decreasing the range of calibrants to closer fit the concentration of the samples being run.
b. Performance may be enhanced and the range extended on the high end by:
i. Decreasing the size of the sample loop.
ii. Making the shortest possible connection between the cartridge and the detector.
iii. Increasing the cycle duration time in “Cartridge configuration”.
iv. Moving the calibrant range up so that the calibrants are closer to the concentration of the actual samples.

6. After the Run

a. Disconnect the lead from the working electrode.

b. Place acid reagent pump tubes into reagent water and pump reagent water through the system for at least
10–15 minutes. Do not pump reagent water in place of base reagent.
c. Stop the pump, and wait for flow to stop.
d. Release tension on all pump tubes.
e. Store the Gel Polymer Reference Electrode (PN 325348) in electrode storage solution when not in use with
Reference Electrode Storage Kit (PN 326133).
f. Power off the system.

NOTE: For short-term and overnight shutdowns, simply turn off the pump and release tension on the pump tubes.

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References
1. Wilmont, J.C.; Solujic, L.; Milosavljevic, E. B.; Hendrix, J.L.; Rader, W.S. Formation of Thiocyanate During Removal of
Sulfide as Lead Sulfide Prior to Cyanide Determination. Analyst 1996, 121, 799–801.

2. Standard Test Methods for Cyanides in Water. Annual Book of ASTM Standards Volume 11.02, ASTM International;
ASTM D 6888-09.

3. Flow Solution™ 3700 Operator’s Manual (part number 329998). Available from OI Analytical, P.O. Box 9010, College
Station, TX, 77842–9010.

4. Milosavljevic, E.B.; Solujic, L. How to Analyze for Cyanide. Cyanide: Social Industrial and Economic Aspects. The
Minerals, Metals, and Materials Society. 2001, 125–128.

5. Ingersol, D.; Harris, W.R.; Bomberger, D.C.; Coulson, D.M. Development and Evaluation Procedures for the Analysis
of Simple Cyanides, Total Cyanides, and Thiocyanate in Water and Waste Water: 1983; EPA-600/4-83-054; U.S.
Environmental Protection Agency, Environmental Monitoring Systems Laboratory, U.S. Government Printing Office:
Washington, DC, 1983.

6. Milosavljevic, E.B.; Solujic, L.; Hendrix, J.L. Environ. Sci. Technol. 1995, 29 (No. 2), 426–430.

7. Federal Register, CFR Title 40, Part 136, Appendix B, U.S. Government Printing Office: Washington, DC, 1994.

8. Guide to Method Flexibility and Approval of EPA Water Methods. Available from the National Technical Information
Service (PB97-117766).

9. Handbook for Analytical Quality Control in Water and Wastewater Laboratories: 1979; EPA-600/4-79-019; U.S.
Environmental Protection Agency, Environmental Monitoring Systems Laboratory, U.S. Government Printing Office:
Washington, DC, 1979.

10. Less is Better: Laboratory Chemical Management for Waste Reduction. Available from the American Chemical
Society, Department of Government Regulations and Science Policy, 1155 16th Street, NW, Washington, DC 20036.

11. ASTM D 7365-09a Standard Practice for Sampling, Preservation and Mitigating Interferences in Water Samples for
Analysis of Cyanide.

12. Report of the Draft Method OIA-1677 Interlaboratory Validation Study, March 1997. Available from OI Analytical, Box
9010, College Station, TX 77842-9010.

13. Report of the Draft Method OIA-1677 Single Laboratory Validation Study, November 1996. Available from OI
Analytical, Box 9010, College Station, TX 77842-9010.

14. Flow Solution™ 3700 ACA Flow Analyzer Software Quick Start Guide (part number 327069). Available from OI
Analytical, P.O. Box 9010, College Station, TX, 77842–9010.

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 Available Cyanide by ASTM D 6888
Figures
Figure 1. Diagram of 330091CT Available Cyanide Cartridge

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Figure 2. Available Cyanide Calibration Series

Figure 3. Calibration Curve and Statistics

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Figure 4. Method MDL, Accuracy & Precision

Calibrant Calibrant Calibrant Calibrant

0.002 mg/L 0.010 mg/L 0.050 mg/L 0.100 mg/L
Replicate 1 0.0014 0.0108 0.0532 0.1044
Replicate 2 0.0016 0.0108 0.0520 0.1044
Replicate 3 0.0016 0.0107 0.0517 0.1036
Replicate 4 0.0013 0.0106 0.0515 0.1037
Replicate 5 0.0010 0.0105 0.0514 0.1032
Replicate 6 0.0013 0.0104 0.0514 0.1033
Replicate 7 0.0015 0.0102 0.0510 0.1027
Replicate 8 0.0014 0.0108 0.0513 0.1029
Replicate 9 — 0.0104 0.0514 0.1035
Replicate 10 — 0.0102 0.0508 0.1039
Mean 0.0014 0.0105 0.0516 0.1036
Standard Deviation 0.000196 0.000237 0.000669 0.000570
%RSD 14.12% 2.25% 1.30% 0.55%
%Accuracy — — — —
MDL 0.000587 — — —

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Appendix A - Prep Guide

Reagent Preparation

NOTE: For best results, filter and degas all reagents prior to use.

WARNING: Do not add surfactants to reagents as their presence could severely hinder or impede proper
diffusion of cyanide thorugh the gas diffusion membrane.

Reagent Water
1. Prepare degassed and deionized reagent water using one of the following methods:
2. Place distilled/deionized water under a strong vacuum for 15–20 minutes. Magnetic stirring or sonification
aids in the degassing process.
3. Purge distilled/deionized water with nitrogen gas (or other inert gas) through a glass frit for approximately
5 minutes.
4. Boil distilled/deionized water in an Erlenmeyer flask for 15–20 minutes. Remove the flask from the heat
source, cover it with an inverted beaker, and allow it to cool to room temperature. After preparing degassed
reagent water, store it in a tightly-sealed container to protect it from reabsorbing atmospheric gases. For best
results, store degassed reagent water under a slight vacuum when not used.

Stock base reagent, 10 M sodium hydroxide

1. While stirring, carefully add 400 g of sodium hydroxide in 700 mL of reagent water in a 1 L volumetric
flask.
2. Cool to room temperature.
3. Dilute to volume with reagent water and mix well.
4. Store in an amber bottle at room temperature.

NOTE: CO2 from atmosphere absorbs into NaOH solution. Do not shake or mix stock reagent prior to pipeting.
Always pipet from top.

Sodium Hydroxide, 1 M
1. While stirring, carefully add 40 g of sodium hydroxide in 700 mL of reagent water in a 1 L volumetric
flask.
2. Cool to room temperature.
3. Dilute to volume with reagent water and mix well.
4. Store at room temperature.

Sample Preservation Reagents

a. Acetate Buffer
1. Dissolve 146 g of sodium acetate in 400 mL of reagent water in a 1 L volumetric flask.
2. Add 480 g of glacial acetic acid.
3. Dilute to volume with reagent water and mix well.

b. Ethylenediamine Solution, 3.5% (v/v)

1. Add 3.5 mL of ethylenediamine to a 100 mL volumetric flask.
2. Dilute to volume with reagent water and mix well.

c. Ascorbic Acid—Crystals

d. Sodium Arsenite—Crystals

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FIA Reagents
Carrier - Reagent Water

Sulfuric Acidification Reagent SAR

1. Weigh 1.00 g of bismuth nitrate pentahydrate into a 1 L volumetric flask.
2. Add 55 mL of reagent water.
3. Carefully add 55 mL of concentrated sulfuric acid. Gently swirl the flask to dissolve the bismuth nitrate
pentahydrate.
4. Dilute to volume with reagent water and mix well.

NOTE: Directions must be followed exactly or the bismuth nitrate will not dissolve.

Base reagent, 0.1 M sodium hydroxide

1. Dilute 10 mL of acceptor stock solution to 1 L in a volumetric flask. and mix well.
2. Dilute to volume with reagent water and mix well.

Mixed ligand exchange reagent, available cyanide

1. Transfer 0.125 mL of (WAD) reagent A and 0.250 mL of (WAD) reagent B into a 100 mL volumetric
flask containing about 50 mL of reagent water.
2. Dilute to volume with reagent water and mix well.
3. Store at 4 °C. Keep the bottle closed, unless in use. If stored properly, available cyanide Mixed Ligand
Reagent is stable for approximately six months.
4. Check available cyanide mixed ligand reagent monthly after preparation, by adjusting the mixed
D 6888 LCS solution.

Calibrant Preparation

Stock Solutions

Standard Silver Nitrate Solution, 0.0192 N

a. Dissolve 3.27 g of silver nitrate in 700 mL of reagent water in a 1 L volumetric flask.
b. Dilute to volume with reagent water and mix well.
c. Store in an amber bottle at room temperature.

Rhodanine Solution, 0.2 mg/mL in Acetone

a. Dissolve 20 mg of 5-[4-(dimethylamino)benzylidene]rhodanine in 80 mL of acetone in a 100 mL volumetric
flask.
b. Dilute to volume with acetone and mix well.

Potassium Cyanide Stock Solution, 1,000 mg/mL

a. Dissolve 2 g of sodium hydroxide in 500 mL of reagent water in a 1 L volumetric flask.
b. Add 2.51 g of potassium cyanide.
c. Dilute to volume with reagent water and mix well.
d. Store in an amber bottle at 4 °C. If stored properly, this reagent is typically stable for two months.

NOTE: This stock solution can be purchased from a reputable supplier.

Standardized Potassium Cyanide Stock Solution

a. Add 0.5 mL of rhodanine solution to 25 mL of potassium cyanide stock solution.
b. Titrate with silver nitrate solution until the color changes from canary yellow to a salmon hue.
c. Based on the determined potassium cyanide concentration, dilute the potassium cyanide stock solution to
a final concentration of 1.00 g/L. If the concentration is not 1.00 g/L, correct the intermediate and working
calibration concentrations accordingly.
d. Store in an amber bottle at 4 °C. If stored properly, this reagent is typically stable for two months.

Stock Nickel (II) cyanide stock solution, 1,000 mg/L

1. Dissolve 0.2488 g of potassium tetracyanonickelate in 25 mL of reagent water and 1 mL of 1 M sodium
hydroxide in a 100 mL volumetric flask.
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 2. Dilute to volume with reagent water and mix well.
Stock Mercury (II) cyanide solution, 1,000 mg/L
1. Dissolve 0.486 g of mercury (II) cyanide in 25 mL of reagent water and 1 mL of 1 M sodium hydroxide
in a 100 mL volumetric flask.
2. Dilute to volume with reagent water and mix well.

Mixed D 6888 LCS Solution (0.2 mg/L CN)

1. Pipet 0.1 mL of Nickel (II) cyanide and mercury (II) cyanide intermediate solution into 25 mL of reagent
water and 1 mL of 1M sodium hydroxide in a 100 mL volumetric flask.
2. Dilute to volume with reagent water and mix well.
3. Store in brown glass or plastic at 2-6 °C.
4. The mixed D 6888 LCS solution evaluates the efficiency of the mixed ligand reagent addition. To
evaluate efficiency of each ligand, prepare separate nickel (II) cyanide and mercury (II) cyanide LCD solutions.

Intermediate Standards – 100 mg/L

1. Using a volumetric pipette, transfer 10 mL of stock standard to a 100 mL volumetric flask that contains
1 mL of 1 M sodium hydroxide and 40 mL of reagent water.
2. Dilute to volume with reagent water and mix well.
3. Store in an amber bottle at 4 °C. If stored properly, this reagent is typically stable for one month.

Calibration series
1. A calibration series may be prepared in 100 mL volumetric flasks containing 1 mL of 1 M sodium hydroxide
using the table below.
2. Dilute and mix well.
3. Store in an amber bottle at 4 °C.
4. Prepare fresh calibrants daily.

Final Concentration (mg/L) 0.01 0.10 0.25 0.50 1.00 5.00

Volume of Intermediate Standard to Transfer (µL) 100 1000 2500 5000 10000 5000

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