Unitad States         Office of Water            EPA-B21-R-04-D01
Environmental Protection    (4303)
Agency                            January 2004
     Method OlA-1677, DW
     Available Cyanide by Fiow
     Injection, Ligand Exchange,
     and Amperometry

 Method OIA-1677, DW

This method was  developed by Michael Straka of OI Analytical in cooperation with Emil
Milosavljevic and Ljiljana Solujic of the University of Nevada Reno Mackay School of Mines and
guidance from William A. Telliard of the Engineering and Analysis Division (BAD) within the U.S.
Environmental Protection Agency's (EPA's) Office of Science and Technology (OST). Additional
assistance in preparing  the  method was  provided  by DynCorp  Information and Enterprise
Technology and Interface, Inc..

This Method has been reviewed and approved for publication by the Analytical Methods Staff within
EPA's Engineering and Analysis Division.  Mention of trade names or commercial products does
not constitute endorsement or recommendation for use.
Questions concerning this Method or its application should be addressed to:

W.A. Telliard
Engineering and Analysis Division (4303)
U.S. Environmental Protection Agency
Ariel Rios Building
1200 Pennsylvania Avenue, N.W.
Washington, DC  20460
Phone: 202/566-1061
Fax:  202/566-1053
                                                                            January 2004

                                                                   Method OIA-1677, DW
                               Table of Contents

   Introduction		  iv

   1.0          Scope and Application	1

   2.0          Summary of Method	.1

   3.0          Definitions  	2

   4.0          Contamination and Interferences	 2

   5.0          Safety	3

   6.0          Equipment and Supplies  	4

   7.0          Reagents and Standards	4

   8.0          Sample Collection, Preservation, and Storage	8

   9.0          Quality Control	 10

   10.0         Calibration and Standardization	15

   11.0         Procedure  	16

   12.0         Data Analysis and Calculations 	16

   13.0         Method Performance	.17

   14.0         Pollution Prevention and Waste Management	17

   15.0         References	 18

   16.0         Tables	19

   17.0         Glossary	 20
January 2004

Method OIA-1677, DW

Method OIA-1677 was developed by ALPKEM, a division of OI Analytical, in cooperation with the
University of Nevada Reno Mackay School of Mines, as a way to measure available cyanide without the
interference problems of the currently approved available cyanide methods. EPA proposed the use of
Method OIA-1677 on July 7, 1998 (63 FR 36809). EPA is approving the use of Method OIA-1677 for
compliance monitoring under Section 304(h) of the Clean Water Act. Method OIA-1677 is an additional
test procedure for measuring the same cyanide species as are measured by currently approved methods
for cyanide amenable to chlorination (CATC). In some matrices, CATC methods are subject to
significant test interferences. Method OIA-1677 has been  added to the list of approved methods because
it is more specific for available cyanide, is more rapid, measures cyanide at lower concentrations, offers
improved safety, reduces laboratory waste, and is more precise and accurate than currently approved
CATC methods.
                                             iv                                    January 2004

                                 Method OIA-1677

       Available Cyanide by Flow Injection, Ligand Exchange, and

LOScope and Application

1.1     This method is for determination of available cyanide in water and wastewater by flow
       injection, ligand exchange, and amperometric detection. The method is for use in EPA's
       data gathering and monitoring programs associated with the Clean Water Act, Resource
       Conservation and Recovery Act, Comprehensive Environmental Response,
       Compensation and Liability Act, and Safe Drinking Water Act.

1.2     Cyanide ion (CN~), hydrogen cyanide in water (HCNaq), and the cyano-complexes of
       zinc, copper, cadmium, mercury, nickel, and silver may be determined by this method
       (see Section 17.2.1).

1.3     The presence of polysulfides may prove intractable for application of this method.

1.4     The method detection limit (MDL) is 0.5  ug/L and the minimum level (ML) is 2.0 |ig/L.
       The dynamic range is approximately 2.0 |ag/L (ppb) to 5.0 mg/L (ppm) cyanide ion using
       a 200 fjiL sample loop volume.  Higher concentrations can be determined by dilution of
       the original sample or by reducing volume of the sample loop.

1.5     This method is for use by analysts experienced with flow injection equipment or under
       close supervision of such qualified persons.

2.0Summary of Method

2.1     The analytical procedure employed for determination of available cyanide is divided into
       two parts: sample pretreatment and cyanide detection. In the pretreatment step, ligand-
       exchange reagents are added at room temperature to 100 mL of a cyanide-containing
       sample. The ligand-exchange reagents form thermodynamically stable complexes with
       the transition metal ions listed in Section  1.2, resulting in the release of cyanide ion from
       the metal-cyano complexes.

       Cyanide detection is accomplished using a flow-injection analysis (FLA) system
       (Reference 15.6). A 200-jnL aliquot of the pre-treated sample is injected into the flow
       injection manifold of the system. The addition of hydrochloric acid converts cyanide ion
       to hydrogen cyanide (HCN) that passes under a gas diffusion membrane. The HCN
       diffuses through the membrane into an alkaline receiving solution where it is converted
       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 generated is
       proportional to the cyanide concentration  present in the original sample. Total analysis
       time is approximately two minutes.
 January 2004

  Method OIA-1677, DW
 2.2     The quality of the analysis is assured through reproducible calibration and testing of the
        FIA system.

 2.3     A flow diagram of the FIA system is shown in Figure 1.
 Coll    Diffusion
                        Figure 1. Row injection Manifold used In the quantification of
                                cyanide in the pretreated sample. Carrier (0.1 M
                                HO); Add (0.1 M HCI); Acceptor (0.1 M NaOH).
        Definitions for terms used in this method are given in the glossary at the end of the


4.1     Solvents, reagents, glassware, and other sample-processing hardware may yield artifacts
        that affect results.  Specific selection of reagents or purification of these reagents may be

4.2     All materials used in the analysis shall be demonstrated to be free from interferences
        under the conditions of analysis by running laboratory blanks as described in Section 9.4.

4.3     Glassware is cleaned by washing in hot water containing detergent, rinsing with tap and
        reagent water, and drying in an area free from interferences.

4.4     Interferences extracted from samples will vary considerably from source to source,
        depending upon the diversity of the site being sampled.

4.5     Sulfide is a positive interferent in this method (References 15.3 and 15.4), because an
        acidified sample containing sulfide liberates hydrogen sulfide that is passed through the
        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. To overcome this
        interference, the sulfide ion must be precipitated with lead ion immediately upon  sample
        collection.  Sulfide ion and lead sulfide react with cyanide ion to form thiocyanate which

                                             2                                    January 2004

                                                                   Method OIA-1677, DW
       is not detected in the analytical system. Tests have shown (Reference 15.7) that if lead
       carbonate is used for sulfide precipitation, the supemate containing cyanide must be
       filtered immediately to avoid loss of cyanide through reaction with precipitated lead
       sulfide (Section 8.2.1).

4.6    Though not interferences, substances that react with cyanide should also be removed from
       samples at time of collection.  These  substances include water soluble aldehydes that
       form cyanohydrins and oxidants such as hypochlorite and sulfite.  Water soluble
       aldehydes react with cyanide to form cyanohydrins that are not detected by the analytical
       system; hypochlorite and sulfite oxidize cyanide to non-volatile forms. Procedures for
       the removal of these substances are provided in Sections 8.2.2 and 8.2.3.


5.1    The toxicity or carcinogenicity of each compound or reagent used in this method has not
       been precisely determined; however,  each chemical compound should be treated as a
       potential health hazard. Exposure to these compounds should be reduced to the lowest
       possible level.

5.2    Cyanides and cyanide solutions

WARNING:   The  cyanide  ion,  hydrocyanic acid, all cyanide salts,  and
most metal-cyanide complexes are  extremely dangerous.   As a contact
poison,  cyanide  need not be  ingested to produce  toxicity.   Also,
cyanide  solutions produce  fatally toxic hydrogen cyanide gas  when
acidified.   For  these  reasons, it is mandatory that work with cyanide
be carried out in a well-ventilated hood  by properly trained  personnel
wearing  adequate protective  equipment.	

5.3    Sodium hydroxide solutions

CAUTION:   Considerable heat  is generated  upon dissolution of  sodium
hydroxide  in water.  It may  be advisable  to cool the container in an
ice bath when  preparing sodium hydroxide  solutions.	

5.4    Unknown samples may contain high  concentrations of volatile toxic compounds.  Sample
       containers should be opened in a hood and handled with gloves to prevent exposure.

5.5    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.
       A reference file of material safety data sheets (MSDSs) should be available to all
       personnel involved in these analyses. Additional information on laboratory safety can be
       found in References 15.8 and 15.9.
 January 2004

 Method OIA-1677, DW
G.OEquipment and Supplies
NOTE:   Brand  names,  suppliers,  and part numbers  are for illustrative
purposes only.   No  endorsement  is implied.   Equivalent  performance  may
be achieved using apparatus and materials  other  than those specified
here, but demonstration  of equivalent  performance  that  meets  the
requirements  of this method is  the responsibility  of the laboratory.

6.1    Flow injection analysis (HA) system—ALPKEM Model 3000 (Reference 15.5), or
       equivalent, consisting of the following:

       6.1.1  Injection valve capable of injecting 40 to 300 fJiL samples

       6.1.2  Gas diffusion manifold with a microporous Teflon® or polypropylene membrane

       6.1.3  Amperometric detection system with:

     Silver working electrode

     Ag/AgCl reference electrode

     Pt/stainless steel counter electrode

     Applied potential of 0.0 volt

6.2    Sampling equipment—Sample bottle, amber glass, 0.1-L,  with polytetrafluoroethylene
       (PTFE)-lined cap.  Clean by washing with detergent and water, rinsing with two aliquots
       of reagent water, and drying by baking at 110 -150 °C for one hour minimum.

6.3    Standard  laboratory equipment including volumetric flasks, pipettes, syringes, etc. all
       cleaned, rinsed and dried per bottle cleaning procedure in Section 6.2.

7.0Reagents  and Standards

7.1    Reagent water—Water in which cyanide and potentially interfering substances are not
       detected at the MDL of this method. It may be generated by any one of the methods listed
       below. Reagent water generated by these methods shall be tested for purity utilizing the
       procedure in Section 11.

       7.1.1   Activated carbon—Pass distilled or deionized water through an activated carbon
              bed (Calgon Filtrasorb-300 or equivalent).

       7.1.2   Water purifier—Pass distilled or deionized water through a purifier (Millipore
              Super Q, or equivalent).

7.2    Sodium hydroxide—ACS reagent  grade.

7.3    Potassium cyanide—ACS reagent  grade.

                                         4                                January 2004

                                                                      Method OIA-1677, DW
7.4    Mercury (II) cyanide, ^99% purity—Aldrich Chemical Company Catalog No. 208140, or

7.5    Potassium nickel (II) cyanide—Aldrich Chemical Company Catalog No. 415154, or

7.6    Silver nitrate—ACS reagent grade.  Aldrich Chemical Company Catalog No. 209139, or

7.7    Hydrochloric acid—approximately 37%, ACS reagent grade.

7.8    Preparation of stock solutions.  Observe the warning in Section 5.2.

       7.8.1  Silver nitrate solution, 0.0192 N—Weigh 3.27 g of AgNO3 into a 1-L volumetric
              flask and bring to the mark with reagent water.

       7.8.2  Rhodanine solution, 0.2 mg/mL in acetone—Weigh 20 mg of p-
              dimethylaminobenzalrhodanine (Aldrich Chemical Co. Catalog No. 114588, or
              equivalent) in a 100-mL volumetric flask and dilute to the mark with acetone.

       7.8.3  Potassium cyanide stock solution, 1000 mg/L

      Dissolve approximately 2 g (approximately 20 pellets) of sodium
                      hydroxide in approximately 500 mL of reagent water contained in a one
                      liter volumetric flask.  Observe the caution in Section 5.3. Add 2.51 g of
                      potassium cyanide (Aldrich Chemical Co. Catalog No. 207810, or
                      equivalent), dilute to one liter with reagent water, and mix well.  Store
                      KCN solution in an amber glass container at 0-4°C.

      Standardize the KCN solution (Section by adding 0.5 mL of
                      rhodanine solution (Section 7.8.2) to 25 mL of KCN solution and
                      titrating with AgNO3 solution (Section 7.8.1) until the color changes
                      from canary yellow to a salmon hue.  Based on the determined KCN
                      concentration, dilute the KCN solution to an appropriate volume so the
                      final concentration is 1.00 g/L, using the following equation:
 January 2004

Method OIA-1677, DW
                                    EQUATION 1

                                x * v = 1 g/L x l
                     x=concentration ofKCN solution determined from titrations
                     v=volume ofKCN solution needed to prepare 1 L of 1 g/L KCN solution
                                                                          January 2004

                                                                        Method OIA-1677, DW
                       If the concentration is not 1.00 g/L, correct the intermediate and working
                       calibration concentrations accordingly.

        7.8.4   1M sodium hydroxide—Dissolve 40 g of sodium hydroxide pellets in
               approximately 500 mL of reagent water in a 1-liter volumetric flask, observing the
               caution in Section 5.3. Dilute to one liter with reagent water.  Store in an amber
               bottle at room temperature.

7.9     Secondary standards.

        7.9.1   Cyanide, 100 mg/L—Dilute 100.0 mL of cyanide stock solution (Section
               and 10 mL of 1M sodium hydroxide (Section 7.8.4) to one liter with reagent water
               (Section 7.1). Store in an amber glass bottle at 0-4°C.

        7.9.2   Cyanide, 10 mg/L—Dilute 10.0 mL of cyanide stock solution and 10 mL of 1M
               sodium hydroxide to one liter with reagent water.  Store in an amber glass bottle
               at 0-4°C.

        7.9.3   Cyanide, 1 mg/L—Dilute 1.0 mL of cyanide stock solution and 1 mL of 1M
               sodium hydroxide to one liter with reagent water.  Store in an amber glass bottle
               at 0-4°C.

        7.9.4   Cyanide working calibration standard solutions (2 - 5000 |ig/L as
               cyanide)—Working calibration standards may be prepared to cover the desired
               calibration range by adding the appropriate volumes of secondary standards
               (Sections 7.9.1, 7.9.2, 7.9.3) to 100 mL volumetric flasks that contain 40 mL of
               reagent water (Section 7.1) and 1 mL of 1M sodium hydroxide (Section 7.8.4).
               Dilute the solutions to 100 mL with reagent water. Prepare working calibration
               standards daily.  The following table provides the quantity of secondary standard
               necessary to prepare working standards of the specified concentration.
 January 2004

 Method OIA-1677, DW

Secondary Standard Solution Volume
Secondary Standard
(Section 7.8.3)


Secondary Standard
(Section 7.8.2)

Secondary Standard
(Section 7.8.1)

        If desired, the laboratory may extend the analytical working range by using  standards that
        cover more than one calibration range, so long as the requirements of Section 10.3 are

7.10    Sample Preservation Reagents

        7.10.1  The presence of sulfide may result in the conversion of cyanide to thiocyanate.
               While lead acetate test paper has been recommended for determining the presence
               of sulfide in  samples, the test is generally unreliable and is typically not usable for
               sulfide concentrations below approximately 1 ppm.  The use of lead carbonate
               (Aldrich Chemical Co. Catalog No. 336378, or equivalent), followed by
               immediate filtration of the sample is required whenever sulfide ion is present. If
               the presence of sulfide is suspected but not verifiable from the use of lead acetate
               test paper, two samples may be collected, one without lead carbonate addition and
               another with lead carbonate addition followed by immediate filtration. Analyze
               both samples.  If sulfide is present, the preserved sample should contain higher
               levels of cyanide than the unpreserved sample. Lead acetate test paper may be
               used, but should be tested for minimum level of sulfide detection by spiking
               reagent water aliquots with decreasing levels of sulfide and determining the
               lowest level of sulfide detection attainable. The spiked samples are tested with
               lead acetate test paper moistened with acetate buffer solution. The buffer solution

                                             8                                    January 2004

                                                                      Method OIA-1677, DW
              is prepared by dissolving 146 g anhydrous sodium acetate, or 243 g sodium
              acetate trihydrate in 400 mL of reagent water, followed by addition of 480 g
              concentrated acetic acid.  Dilute the solution to 1 L with reagent water.  Each new
              batch of test paper and/or acetate buffer should be tested to determine the lowest
              level of sulfide ion detection prior to use.

       7.10.2 Ethylenediamine solution—In a 100 mL volumetric flask, dilute 3.5 mL
              pharmaceutical-grade anhydrous ethylenediamine (Aldrich Chemical Co. Catalog
              No. 240729, or equivalent) with reagent water.

       7.10.3 Ascorbic acid—Crystals—Aldrich Chemical Co. Catalog No. 268550, or

7.11   FIA Reagents.

       7.11.1 Carrier and acid reagent (0.1M hydrochloric acid)—Dilute 8 mL of concentrated
              hydrochloric acid to one liter with reagent water.

       7.11.2 Acceptor reagent (0.1M sodium hydroxide)—Dilute 100 mL of sodium hydroxide
              solution (Section 7.8.4) to 1000 mL with reagent water.

       7.11.3 Ligand-exchange reagent A-ALPKEM part number A001416, or equivalent.

       7.11.4 Ligand-exchange reagent B-ALPKEM part number A001417, or equivalent.

7.12   Quality control solutions

       7.12.1 Mercury (II) cyanide stock solution (1000 mg/L as cyanide)—Weigh 0.486 g of
              mercury (II) cyanide (Section 7.4) in a 100-mL volumetric flask. Add 10 - 20 mL
              of reagent water and 1 mL of 1M sodium hydroxide solution (Section 7.8.4).
              Swirl to mix. Dilute to the mark with reagent water.

       7.12.2 Laboratory control sample (LCS)—Place 0.20 mL of the mercury (II) cyanide
              stock solution (Section 7.12.1) in a 100-mL volumetric flask and dilute to the
              mark with reagent water to provide a final cyanide concentration of 2.00 mg/L.

S.OSample Collection, Preservation, and Storage

8.1    Sample collection and preservation—Samples are collected using manual (grab)
       techniques and are preserved immediately upon collection.

       8.1.1  Grab sampling—Collect samples in amber glass bottles with PTFE-lined caps
              cleaned according to the procedure in Section 6.2.  Immediately after collection,
              preserve the sample using any or all of the preservation techniques (Section 8.2),
              followed by adjustment of the sample pH to > 12 by addition of 1M sodium
              hydroxide and refrigeration at 0-4°C.
 January 2004

 Method OIA-1677, DW
        8.1.2   Compositing—Compositing is performed by combining aliquots of grab samples
               only. Automated compositing equipment may not be used because cyanide may
               react or degrade during the sampling period. Preserve and refrigerate each grab
               sample immediately after collection (Sections 8.1.1 and 8.2) until compositing.

        8.1.3   Shipment—If the sample will be shipped by common carrier or mail, limit the pH
               to a range of 12.0 -12.3. (See the footnote to 40 CFR 136.3(e), Table E, for the
               column headed "Preservation.")

8.2     Preservation techniques

        8.2.1   Samples containing sulfide ion

        Test the sample with  lead acetate test paper (Section 7.10.1) to determine
                       the presence or absence of sulfide ion.  If sulfide ion is present, the
                       sample must be treated immediately (within 15 minutes of collection)
                       with sufficient solid lead carbonate (Section 7.10.1) to remove sulfide
                       (as evidenced by the lead acetate test paper), and immediately filtered
                       into another sample bottle to remove precipitated lead sulfide.

        If sulfide ion is suspected to be present, but its presence is not detected
                       by the lead acetate paper test, two samples should be collected. One is
                       treated for the presence of sulfide and immediately filtered, while the
                       second is not treated for sulfide. Both samples must be analyzed.  (Tests
                       conducted prior to the interlaboratory validation of this method showed
                       significant and rapid losses of cyanides when lead sulfide was allowed to
                       remain in contact with the sample during holding times of three days or
                       less. As a result, the immediate filtration of samples preserved with lead
                       carbonate is essential (Reference 15.6)).

        If the sample contains particulate matter that would be removed upon
                       filtration, the sample must be filtered prior to treatment with lead
                       carbonate to assure that cyanides associated with the particulate matter
                       are included in the measurement. The collected particulate matter must
                       be saved and the filtrate treated using the sulfide removal procedure
                       above (Section The collected particulate and treated filtrate
                       must be recombined and homogenized, and then sent to the laboratory
                       for analysis.

       8.2.2  Samples containing water soluble aldehydes—Treat samples containing or
              suspected to contain formaldehyde, acetaldehyde, or other water soluble aldehydes
              with 20 mL of 3.5% ethylenediamine solution (Section 7.10.2) per liter of sample.

       8.2.3  Samples known or suspected to contain chlorine, hypochlorite, and/or
              sulfite—Treat with 0.6 g of ascorbic acid (Section 7.10.3) per liter of sample.
              EPA Method 330.4 or 330.5 may be used for the measurement of residual  chlorine
              (Reference 15.1).

                                           10                                  January 2004

                                                                        Method OIA-1677, DW
8.3     Sample holding time—Maximum holding time for samples preserved as above is 14
        days. Unpreserved samples must be analyzed within 24 hours, or sooner if a change in
        cyanide concentration will occur. (See the footnotes to Table n at 40 CFR 136.3(e).)

S.OQuality Control

9.1     Each laboratory that uses this method is required to operate a formal quality assurance
        program (Reference 15.9).  The minimum requirements of this program consist of an
        initial demonstration of laboratory capability, and the periodic analysis of LCSs and
        MS/MSDs as a continuing check on performance.  Laboratory performance is compared
        to established performance criteria to determine if the results of the analyses meet the
        performance characteristics of the method.

        9.1.1  The laboratory shall make an initial demonstration of the ability to generate
              acceptable precision and accuracy with this method.  This ability is established as
              described in Section 9.2.

9.2     Initial demonstration of laboratory capability

        9.2.1  Method Detection Limit (MDL)—To establish the ability to detect cyanide at low
              levels, the laboratory shall determine the MDL per the procedure in 40 CFR Part
               136, Appendix B (Reference 15.4) using the apparatus, reagents, and standards
              that will be used in the practice of this method. An MDL less than or equal to the
              MDL listed in Section 1.4 must be achieved prior to practice of this method.

        9.2.2  Initial Precision and Recovery (IPR)—To establish the ability to generate
              acceptable precision and accuracy, the laboratory shall perform the following

      Analyze four samples of the LCS (Section 7.12.2) according to the
                       procedure beginning in Section 10.

      Using the results of the set of four analyses, compute the average percent
                       recovery (x) and the standard deviation of the percent recovery (s) for
                       cyanide. Use Equation 2 for calculation of the standard deviation of the
                       percent recovery.
 January 2004                                 11

 Method OIA-1677, DW
                                       EQUATION 2
                              s  =
n - 1
                       n = Number of samples
                       x = Percent recovery in each sample
        9.2.3  Compare s and x with the acceptance criteria specified in Table 1. If s exceeds the
              precision limit or x falls outside the range for recovery, system performance is
              unacceptable and the problem must be found and corrected before analyses can

9.3     Matrix spike/matrix spike duplicate (MS/MSD)—The laboratory shall spike, in duplicate,
        a minimum of 10 percent of all samples (one sample in duplicate in each batch of ten
        samples) from a given discharge.

        9.3.1  The concentration of the spike in the sample shall be determined as follows:

       If, as in compliance monitoring, the concentration of cyanide in the
                       sample is being checked against a regulatory concentration limit, the
                       spiking level shall be at that limit or at 1 to 5 times higher than the
                       background concentration of the sample (determined in Section 9.3.2),
                       whichever concentration is higher.

       If the concentration of cyanide in a sample is not being checked against a
                       limit, the spike shall be at the concentration of the LCS or at 1 to 5 times
                       higher than the background concentration, whichever concentration is

        9.3.2  Analyze one sample aliquot out of each set of ten samples from each discharge
              according to the procedure beginning in Section 11 to determine the background
              concentration (B) of cyanide.

       Spike this sample with the amount of mercury (n) cyanide stock solution
                       (Section 7.12.1) necessary to produce a cyanide concentration in the
                       sample of 2 mg/L. If necessary, prepare another stock solution
                       appropriate to produce a level in the sample at the regulatory compliance
                       limit or at 1 to 5 times the background concentration (per Section 9.3.1).

       Spike two additional sample aliquots with the  spiking solution and
                       analyze these aliquots to determine the concentration after spiking (A).

                                           12                                  January 2004

                                                                       Method OIA-1677, DW
      9.3.3  Calculate the percent recovery of cyanide in each aliquot using Equation 3.

                                     EQUATION 3

                                          100  (A - B)
                                     P =
                      P = Percent recovery
                      A = Measured concentration of cyanide after spiking
                      B = Measured background concentration of cyanide
                      T = True concentration of the spike	
       9.3.4  Compare the recovery to the QC acceptance criteria in Table 1. If recovery is
             outside of the acceptance criteria, and the recovery of the LCS in the ongoing
             precision and recovery test (Section 9.6) for the analytical batch is within the
             acceptance criteria, an interference is present.  In this case, the result may not be
             reported for regulatory compliance purposes.

       9.3.5  If the results of both the MS/MSD and the LCS test fail the acceptance criteria,
             the analytical system is judged to be out of control.  In this case, the problem shall
             be identified and corrected, and the analytical batch reanalyzed.

       9.3.6  Calculate the relative percent difference (RPD) between the two spiked sample
             results (Section 9.3, not between the two percent recoveries) using Equation 4.
                                      EQUATION 4
                                        ID. - D,|
                                                   x 100
                      RPD = Relative percent difference
                      Dj = Concentration of cyanide in the spiked sample
                      D2 = Concentration of cyanide in the spiked duplicate sample
       9.3.7   Compare the precision to the RPD criteria in Table 1. If the RPD is greater than
              the acceptance criteria, the analytical system is judged to be out of control, and the
              problem must be immediately identified and corrected, and the analytical batch

       9.3.8   As part of the QC program for the laboratory, method precision and accuracy for
              samples should be assessed and records should be maintained. After the analysis
              of five spiked samples in which the recovery passes the test in Section 9.3.4,
              compute the average percent recovery (Pa) and the standard deviation of the

January 2004                                 13

 Method OIA-1677, DW
               percent recovery (sp).  Express the accuracy assessment as a percent recovery
               interval from Pa - 2sp to Pa + 2sp. For example, if Pa = 90% and sp = 10% for five
               analyses, the accuracy interval is expressed as 70 - 110%. Update the accuracy
               assessment on a regular basis (e.g., after each five to ten new accuracy

9.4     Laboratory blanks—Laboratory reagent water blanks are analyzed to demonstrate
        freedom from contamination.

        9.4.1   Analyze a reagent water blank initially (i.e., with the tests in Section 9.2) and with
               each analytical batch.  The blank must be subjected to the same procedural steps
               as a sample.

        9.4.2   If cyanide is detected in the blank at a concentration greater than the ML, analysis
               of samples is halted until the source of contamination is eliminated and a blank
               shows no evidence of contamination.

9.5     Calibration verification—Verify calibration of the analytical equipment before and after
        each analytical batch of 14 or fewer measurements. (The 14 measurements will normally
        be 10  samples, 1 reagent blank, 1 LCS, 1 MS, and 1 MSD). Verification is accomplished
        by analyzing the mid-range calibration standard and verifying that it is within the QC
        acceptance criteria for recovery in Table 1. (The concentration of the calibration
        verification depends on the calibration range being used.) Failure to verify calibration
        within the acceptance criteria requires recalibration of the analysis system.

9.6     Laboratory control sample (LCS)—To demonstrate that the analytical system is in
        control, and acceptable precision and accuracy is being maintained with each analytical
        batch, the laboratory shall perform the following operations.

        9.6.1   Analyze a LCS (Section 7.12.2) with each analytical batch according to the
               procedure in Section 10.

        9.6.2   If the results for the LCS are within the acceptance criteria specified in Table 1,
               analysis of the batch may continue. If, however, the concentration is not within
               this range, the analytical process is not in control. In this event, correct the
               problem, repeat the LCS test, and reanalyze the batch.

        9.6.3   The laboratory should add results that pass the specification in Section 9.6.2 to
               IPR and previous LCS data and update QC charts to form a graphic representation
               of continued laboratory performance.  The laboratory should also develop a
               statement of laboratory data quality for cyanide by calculating the average percent
               recovery (R) and the standard deviation  of the percent recovery (sr). Express the
               accuracy as a recovery interval from R - 2sr to R + 2sr  For example, if R = 95%
               and sr = 5%, the accuracy is 85% to 105%.

9.7     Reference Sample—To demonstrate that the analytical system is in control, the laboratory
        should periodically test an external reference sample, such as a Standard Reference

                                            14                                  January 2004

                                                                        Method OIA-1677, DW
        Material (SRM) if an SRM is available from the National Institutes of Standards and
        Technology (NIST). The reference sample should be analyzed quarterly, at a minimum.
        Corrective action should be taken if the measured concentration significantly differs from
        the stated concentration.

10.0   Calibration and Standardization

This section describes the procedure to calibrate and standardize the FIA system prior to cyanide

10.1    Instrument setup

        10.1.1  Set up the FIA system and establish initial operating conditions necessary for
               determination of cyanide. If the FIA system is computerized, establish a method
               for multi-point calibration and for determining the cyanide concentration in each

        10.1.2  Verify that the reagents are flowing smoothly through the FIA system and that the
               flow cell is purged of air bubbles.

10.2    Instrument Stabilization

        10.2.1  Load a 10 mg/L KCN standard (Section 7.8.3) into the sampling valve and inject
               into the FIA system.

        10.2.2  Continue to inject 10 mg/L KCN standards  until 3 successive peak height or area
               results are within 2% RSD, indicating that the electrode system is stabilized.

        10.2.3  Following stabilization, inject the highest concentration calibration standard until
               3 successive peak height or area results are  within 2% RSD indicating
               stabilization at the top of the calibration range.

10.3    External standard calibration

        10.3.1  Inject each of a minimum of 3 calibration standards. One of the standards should
               be at the minimum  level (ML) unless measurements are to be made at higher
               levels. The other concentrations should correspond to the expected range of
               concentrations found in samples or should define the working range of the FIA

        10.3.2  Using injections of a constant volume, analyze each calibration standard
               according to Section 11 and record peak height or area responses against the
               concentration. The results can be used to prepare a calibration curve.
               Alternatively, if the ratio of response to amount injected (calibration factor) is
               constant over the working range (<10% RSD), linearity through the origin can be
               assumed and the averaged calibration factor (area/concentration) can be used in
               place of a calibration curve.

 January 2004                                  15

 Method OIA-1677, DW
11.0  Procedure

This section describes the procedure for determination of available cyanide using the FIA system.

11.1   Analysis of standards, samples, and blanks

       11.1.1 Ligand-exchange reagent treatment of standards, samples, and blanks.

       11.1.2 To 100-mL of cyanide-containing sample (or standard or blank) at pH of
             approximately 12, add 100 uL of ligand-exchange reagent Part B (Section 7.11.5),
             50uL of ligand-exchange reagent Part A (Section 7.11.4), and mix thoroughly.
             Load the sample, standard, or blank into the sample loop.
NOTE:   The  ligand-exchange reagents, when added  to 100 mL  of sample at
the  specified volume,  will liberate cyanide from metal complexes  of
intermediate  stability up to 5  mg/L cyanide ion.   If higher
concentrations are anticipated,  add additional ligand-exchange reagent,
as appropriate,  or dilute the sample.-  The ligand-exchange reagents
have an approximate lifetime of 6 months  after opening. The reagents
should be stored in a  refrigerator at 4°C.  Samples should be analyzed
within 2 hours of adding the ligand-exchange reagents.  The reagents
should always be used  in solutions similar to cyanide samples (pH 12
adjusted).  It is recommended that the ligands be checked  monthly.
This can be done by preparing pH 12 adjusted 2 mg/1 solutions of
mercury(II) cyanide  (Section 7.4)  and of  potassium nickel(II) cyanide
(Section 7.5).   Add ligand-exchange reagent B to the mercury(II)
standard and  ligand-exchange reagent A to the potassium nickel(II)
cyanide standard and confirm cyanide recovery.	

       11.1.3 Inject the sample and begin data collection. When data collection is complete,
             analyze the next sample, standard or blank in the batch until analyses of all
             samples in the batch are completed.

12.0  Data Analysis and Calculations

12.1    Calculate the concentration of material in the sample, standard or blank from the peak
       height or area using the calibration curve or calibration factor determined in Section 10.3.

12.2    Reporting

       12.2.1 Samples—Report results to three significant figures for cyanide concentrations
             found above the ML (Section 1.4) in all samples. Report results below the ML as
             <2 /ig/L, or as required by the permitting authority or permit.

       12.2.2 Blanks—Report results to three significant figures for cyanide concentrations
             found above the MDL (Section 1.4). Do not report results below the MDL unless
             required by the permitting  authority or in the permit.
                                       16                               January 2004

                                                                      Method OIA-1677, DW
13.0  Method Performance

13.1   Method detection limit (MDL)—MDLs from nine laboratories were pooled to develop
       the MDL of 0.5 ju,g/L given in Section 1.4 (Reference 15.12).

13.2   Data obtained from single laboratory testing of the method are summarized in Table 2
       and show recoveries and reproducibility for "free" forms of cyanide, including the
       recovery and reproducibility of silver, nickel, and mercury cyanide species.
       Determination of these species tends to be problematic with other methods for the
       determination of available cyanide.  As it is the case with other methods used for
       available cyanide, iron cyanide species were not recovered and recoveries for gold and
       cobalt species were zero or very low.  The complete results from the single laboratory
       study are available in the Report of the Draft OIA Method 1677  Single Laboratory
       Validation Study (Reference  15.11).

13.3   Listed in Table 1 are the QC acceptance criteria developed from an interlaboratory
       validation study of this method.  This study was conducted following procedures
       specified in the Guide to Method Flexibility and Approval of EPA Water Methods
       (Reference 15.10). In this study, a total of nine laboratories performed analyses for
       various water matrices. Table 3 shows a summary of the interlaboratory results which
       include the accuracy and precision data as % recoveries and relative standard deviations.
       In addition to spikes of easily dissociable cyanides, some samples contained known
       amounts of cyanides that are not recoverable (e.g., Pt and Fe complexes) and thiocyanate
       was spiked to one sample to investigate the potential for interference.  The complete
       study results are available in the Report of the Draft OIA Method 1677 Interlaboratory
       Validation Study (Reference  15.12).

14.0  Pollution  Prevention  and Waste Management

14.1   The laboratory is responsible for complying with all Federal, State, and local regulations
       governing waste management, particularly hazardous waste identification rules and land
       disposal restrictions, and for protecting the air, water, and land by minimizing and
       controlling all releases from fume hoods and bench operations.  Compliance with all
       sewage discharge permits and regulations is also required. An overview  of requirements
       can be found in Environmental Management Guide for Small Laboratories (EPA 233-B-

14.2   Samples containing cyanide,  certain metals, and acids at a pH of less than 2 are hazardous
       and must be treated before being poured down a drain or must be handled as hazardous

14.3   For further information on waste management, consult Less is Better:  Laboratory
       Chemical Management for Waste Reduction, Reference  15.8.
 January 2004                                 17

 Method OIA-1677, DW
15.0   References

15.1    Environmental Monitoring Systems Laboratory. EPA Method 335.1. In: Methods for the
        Chemical Analysis of Water and Wastes (EPA/600/4-79-020).  Environmental Protection
        Agency, Cincinnati, Ohio. Revised March 1983.

15.2    American Public Health Association, American Waterworks Association, Water
        Pollution Control Board. Methods Section 4500-CN in Standard Methods for the
        Examination of Water and Wastewater, 19th Edition. American Public Health
        Association, Washington, DC. 1995.

15.3    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 (EPA-600/4-83-054), 1983.

15.4    Code of Federal Regulations, Title 40, Part 136, Appendix B.  U.S. Government Printing
        Office, Washington, D.C., 1994.

15.5    ALPKEM CNSolution Model 3000 Manual.  Available from ALPKEM / OI Analytical,
        Box 9010, College Station, TX 77842-9010.

15.6    Milosavljevic, E.B.; Solujic, L.; Hendrix, J.L. Environmental Science and Technology,
        Vol. 29, No. 2,1995, pp 426-430. Rapid Distillationless "Free Cyanide" Determination
        by a Flow Injection Ligand Exchange Method.

15.7    Wilmont, J.C.; Solujic, L.; Milosavljevic, E. B.; Hendrix, J.L.; Reader, W.S. Analyst,
        June 1996, Vol. 121, pp  799-801. Formation of Thiocyanate During Removal of Sulfide
        as Lead Sulfide Prior to Cyanide Determination.

15.8    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.

15.9    Handbook for Analytical Quality Control in Water and Wastewater Laboratories (EPA-
        600/4-79-019), USEPA, NERL, Cincinnati, Ohio 45268 (March 1979).

15.10   Guide to Method Flexibility and Approval of EPA Water Methods, December, 1996,
        (EPA-821-D-96-004). Available from the National Technical Information Service

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

15.12   Report of the Draft OIA Method 1677  Interlaboratory Validation Study, March 1997.
        Available from ALPKEM / OI Analytical, Box 9010, College Station, TX 77842-9010.
                                         18                                January 2004

                                                                       Method OIA-1677, DW
16.0  Tables
Table 1. Quality Control Acceptance Criteria
Initial Precision and Recovery
Ongoing Precision and Recovery
(Laboratory Control Sample)
Calibration Verification
Matrix Spike/Matrix Spike
Required Recovery Range
92 - 122
82 - 132
82 - 130
Table 2. Species-Dependent Cyanide Recoveries Using Draft Method 1677(1)
97.4 (0.7)
100.0 (0.8)
100.9 (1.3)
101.8 (0.9)
104.3 (0.2)
100.0 (0.6)
103.4 (0.4)
1.3(2) (0.0)
2.9(2) (0.0)
2.00 ng/mL CN-
98.5 (0.7)
100.0 (0.2)
99.0 (0.6)
100.0 (0.5)
103.0 (0.5)
99.0 (0.3)
98.0 (0.3)
2.0(2) (0.0)
1 Values are % recoveries; numbers in parentheses are percent relative standard deviations.
2 Commercial product contains some free cyanide.
 January 2004

 Method OIA-1677, DW
Table 3. Cyanide Recoveries From Various Aqueous Matrices
Reagent water w/O.OlM
POTW secondary effluent
Petroleum Refinery
Secondary Effluent
Coke Plant Secondary
Rolling Mill Direct Filter
Metals Finishing Indirect
Primary Effluent
Reagent water w/O.OlM
Reagent water w/O.OlM
Mining Tailing Pond Effluent
Sample CN
3.0 \igfL
9.9 jig/L
14.0 ng/L
Added CN(I)
2 mg/L as KCN;
5 mg/L as [FeCCNy4"
50 ng/L as KCN
200 [igfL as KCN;
2 mg/L as KSCN
200 jig/L as KCN
10 mg/L as KCN;
10 mg/L as [Pt(CN)6]"-
4 mg/L as KCN
Average %
1    Cyano-complexes of Pt and Fe were added to the POTW and petroleum
    refinery effluents, respectively; and thiocyanate was added to the metals
    finishing effluent to demonstrate that the FI/LE system does not determine
    these forms of cyanide.

17.0  Glossary of Definitions and Purposes

The definitions and purposes are specific to this method but have been conformed to common
usage as much as possible.

17.1   Units of weights and measures and their abbreviations

       17.1.1 Symbols
              °C      degrees Celsius
              %      percent
              ±       plus or minus
              >       greater than or equal to

       17.1.2 Alphabetical characters
              g       gram
              L       liter
              mg      milligram
              mg/L    milligram per liter
              ptg      microgram
              ptg/L    microgram per liter
January 2004

                                                                        Method OIA-1677, DW
              mL      milliliter
              ppm     parts per million
              ppb      parts per billion
              M       molar solution
17.2   Definitions
        17.2.1 Available cyanide consists of cyanide ion (CN~), hydrogen cyanide in water
              (HCNaq) and the cyano-complexes of zinc, copper, cadmium, mercury, nickel, and

        17.2.2 Calibration blank—A 100 mL volume of reagent water treated with the ligand-
              exchange reagents and analyzed using the FIA procedure.

        17.2.3 Calibration standard (CAL)—A solution prepared from the dilution of stock
              standard solutions. A 100 mL aliquot of each of the CALs are subjected to the
              analysis procedure.  The resulting observations are used to calibrate the
              instrument response with respect to the analyte concentration.

        17.2.4 Discharge—Specific discharge (also known as "matrix type") means a sample
              medium with common characteristics across a given industrial category or
              industrial subcategory. Examples include: C-stage effluents from chlorine bleach
              mills in the Pulp, Paper, and Paperboard industrial category; effluent from the
              continuous casting subcategory of the Iron and Steel industrial category; publicly
              owned treatment work (POTW) sludge; and in-process streams in the  Atlantic and
              Gulf Coast Hand-shucked Oyster Processing subcategory.

              Specific discharge also means a discharge with characteristics different from other
              discharges.  Therefore, if there are multiple discharges from a facility  all with the
              same characteristics, these are the same discharge for the purpose of
              demonstrating equivalency of a method modification.  In this context,
              "characteristics" means that results of the matrix spike and matrix spike duplicate
              (MS/MSD) tests with the unmodified method meet the QC acceptance criteria for
              recovery and relative percent difference (RPD).

        17.2.5 Initial precision and recovery (DPR)—Four aliquots of the LRB spiked with the
              analytes of interest and used to establish the ability to generate acceptable
              precision  and accuracy.  An IPR is performed the first time this method is used
              and any time the method or instrumentation is modified.

        17.2.6 Laboratory control sample (LCS)—An aliquot of LRB to which a quantity of
              mercury (II) cyanide stock solution is added in the laboratory. The LCS is
              analyzed like a sample.  Its purpose is to determine whether the methodology is in
              control  and whether the laboratory is capable of making accurate and precise
 January 2004                                 21

Method OIA-1677, DW
       17.2.7 Laboratory reagent blank (LRB)—An aliquot of reagent water that is treated like a
             sample including exposure to all glassware, equipment, and reagents that are used
             with other samples. The LRB is used to determine if the method analyte or other
             interferences are present in the laboratory environment, reagents, or apparatus.

       17.2.8 Matrix spike/matrix spike duplicate (MS/MSD)—An aliquot of an environmental
             sample to which a quantity of the method analyte is added in the laboratory.
             MS/MSDs are analyzed like a sample. Their purpose is to determine whether the
             sample matrix contributes bias to the analytical results. The background
             concentration of the analyte in the sample matrix must be determined in a separate
             aliquot and the measured values in the MS/MSD corrected for the background

       17.2.9 Minimum level (ML)—The level at which the entire analytical system shall give a
             recognizable signal and acceptable calibration point, taking into account method
             specific sample and injection volumes.

       17.2.10  Ongoing precision and recovery (OPR)—See Laboratory control sample
                                          22                                  January 2004