Proceedings of the International Cyanide Detection Testing Workshop

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Search for a Simplifi ed Field Test: the Soundararajan Digestion Combined with ISE or Colorimetric Methods for Detection of Cyanide Ion Concentrations in Marine Fish A new method for testing fi sh samples for the presence of cyanide was developed by Dr. Rengarajan Soundararajan in 1990 (Rubec and Soundararajan 1991). The method involves the digestion of fi sh tissues in concentrated sodium hydroxide (NaOH). The method is appealing since it is relatively simple and quick to conduct. The cyanide ion is released from the tissues into a basic, high pH solution preventing the cyanide from being lost to the atmosphere. Cyanide ion concentrations can then be measured with an ion-selective electrode (ISE). Five fi sh species obtained from Indonesia tested in the USA had cyanide ion concentrations ranging from 5.8 to 23 mg/kg (ppm) (Rubec and Soundararajan 1991). A Clown Triggerfi sh obtained from the Philippines exhibited a cyanide ion measurement of 1120 mg/kg. No cyanide was detected in two Flame Angelfi sh obtained from the Marshall Islands, for two French Angelfi sh from the Caribbean, and two species of surgeonfi sh obtained from Hawaii. Hence, cyanide was detected in fi sh from countries known to have collectors using cyanide and no cyanide was detected in fi sh obtained from countries where cyanide is not used for collecting marine aquarium fi sh. Peter J. Rubec 1 and Benita Manipula 2 International Marinelife Alliance 1. peterrubec@cs.com 2. dragon9262002@yahoo.com 150 The International Marinelife Alliance (IMA) evaluated the technique in 1991, but abandoned it after it was found to give anomalously high cyanide readings. It was suspected that the anomalously high readings found by the IMA might have occurred because of false-positive readings caused by sulfi de interference with the ISE electrode. Further research was conducted by Aquarium Systems with a small grant obtained by the IMA from the Columbus Zoo (Frakes and Studt 1996). The basic procedure was to expose the fi sh to known concentration of cyanide ion, kill the fi sh, then puree the samples in 5M-NaOH. The NaOH volume was calculated to yield a 10% by weight fi sh slurry. This slurry was allowed to settle and a clear aliquot was diluted with distilled water to produce another 10% by weight dilution. Cyanide ion (CN - ) concentrations in the fi nal solution, 1% fi sh tissue, and approximately 0.5M-NaOH were measured in millivolts (mV) recorded with an Orion CN - ISE linked to an Orion ISE meter. These readings were compared with a semi-log plot produced with known cyanide concentrations of 0.1, 1.0, and 10 mg/L
levels with mV readings recorded as each level (Frakes and Studt 1996). Lead carbonate was added to the supernatant solution to precipitate sulfi des from solution. The fi rst trial compared readings with Atlantic Blennies, which were net-caught and exposed to cyanide. Both the test and the control fi sh (not exposed to cyanide) were found to exhibit cyanide levels of about 300 mg/L (ppm). Hence, the addition of lead carbonate to the solution prior to ISE testing did not eliminate the anomalously high readings. A second experiment evaluated whether the high readings might be due to iodide interference with the ISE. Again, the experiment was inconclusive. In a third experiment, test fi sh (4 Caribbean Blue Chromis, 1 Atlantic Wrasse, and 3 goldfi sh) were exposed to cyanide concentrations ranging from 1.3 to 3.9 ppm CN - for times ranging from one minute (for the 4 chromis) to 14 minutes (for a goldfi sh). Readings for control fi sh (1 wrasse and 2 goldfi sh) not exposed to cyanide produced higher mV readings than fi sh exposed to cyanide. Recent analyses by Ms Benita Manipula using the Soundararajan procedure and cyanide measurements obtained using an Industrial Test Systems (ITS) Inc. colorimetric Cyanide Reagent Strip TM (Cyanide Test Kit 484003) have also experienced diffi culty in obtaining reliable measurements of cyanide concentrations in comparison to known concentrations of cyanide ion in sodium hydroxide solution (lacking slurry). The problem does not appear to be related to the reliability of the 151 ITS cyanide test strips, since they have been demonstrated to be sensitive (down to 0.02 mg/L) and reliable for measuring cyanide ion concentrations in water (Battelle 2005). The most likely explanation for these results is that organics in the digested tissue solutions containing sodium hydroxide produced the anomalous readings both with the ISE and the ITS test strips. Further research is needed to see whether it is possible to measure cyanide ion concentrations in solutions obtained from marine fi sh using the Soundararajan tissue digestion method. References Battelle (2005) Industrial Test Systems, Inc. Cyanide ReagentStrip TM Test Kit. Environmental Technolgy Verifi cation Report, 42 pp, under contract to U.S. Environmental Protection Agency. Frakes, T. A. and Studt, R. (1996) Evaluation of Methods for the Cyanide Detection Test for Fish Utilizing Alkaline Digestion and Cyanide Specifi c Electrode. Report by Aquarium Systems associated with IMA grant from the Columbus Zoo, 8 pp, Columbus, OH. Rubec, P.J. and Soundararajan, R. (1991) Chronic toxic effects of cyanide on tropical marine fi sh. Proceedings of the Seventeenth Annual Toxicity Workshop: November 5-7, 1990 (Vancouver, B.C.). Canadian Technical Report Fisheries and Aquatic Sciences 1:1774, 243-251.
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Proceedings of the International Cy
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Proceedings of the Cyanide Detectio
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ACKNOWLEDGEMENTS The Proceedings of
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PREFACE The International Cyanide D
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EXECUTIVE SUMMARY The International
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ATCA) in homogenized fi sh tissue m
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Validation of the ISE method applie
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Importing countries should also req
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INTRODUCTION Cyanide fi shing is a
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through the implementation of a net
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Working Group 2 The Export Working
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o o o o Can the U.S. require verifi
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Polymeric membrane based ion select
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Goal 2: Provide recommendations on
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hands-on training that develops a d
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Goal 3: Identify research needs to
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Working Group 1 Participants Bob KO
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estimated at roughly 3 to 5 days. H
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Goal 2 : Identify steps, agreements
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3) certifi cate, but tests positive
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Report from Working Group 3 The Par
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No. of Shipments 6,000 5,000 4,000
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Goal 3: Provide a recommendation wi
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Working Group 3 Participants Eric P
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Introduction Since the early 1960
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ainbow trout exposed to 40 mg SCN -
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Blennies, which were net-caught and
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electrical potential (similar to an
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in the Philippines under contract w
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endorsing the IMA’s use of the IS
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The intended products from the rese
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WPCF 1992, 1998). Both of these stu
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ASTM (1997) Standard test methods f
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Kuegsen, M., Kloock, J.P., Knobbe,
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Philippines. In Proceedings from th
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3) 4) 5) 6) the metabolism of cyani
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or new matrices. 5.2.3 Modifi catio
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Level Two: The method should be val
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of cyanide exposure and to identify
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Special considerations for the anal
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tissue samples indispensable. Altho
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y reduction and subsequent separati
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can also be used for analysis of cy
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thiocyanate in saliva and serum bas
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are many discrepancies in the liter
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Table 3. Analytical Methods to dete
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Poisoning, In Textbook of military
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(70) Odoul, M., Fouillet, B., Nouri
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V. (1975) Stable reagents for the c
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(159) Felscher, D. and Wulfmeyer, M
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Purpose of this Document The purpos
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day. One cyanide tablet (2 g) is mi
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Page 111 and 112: existing cyanide detection methods
Page 113 and 114: - Plasma lactate — elevated plasm
Page 115 and 116: Method Advantages Disadvantages Hig
Page 117 and 118: Paper Method Matrix Detection Limit
Page 119 and 120: Table 4 lists experts identifi ed t
Page 121 and 122: X Jerry Thomas CDC Em Steve Borron
Page 123 and 124: References Barber, C. and Pratt, V.
Page 125 and 126: Rates of Recovery. Environmental Ma
Page 127: COUNTRY REPORTS
Page 130 and 131: The first three of these destructiv
Page 132 and 133: enforcement is minimal. There is an
Page 134 and 135: 1. Management/guidance to the Minis
Page 136 and 137: in Vietnam now and requires more ef
Page 138 and 139: Figure 2: Administrative structure
Page 140 and 141: References Baquero, J. (1999) Marin
Page 142 and 143: Acronyms • CDT • Cyanide Detect
Page 144 and 145: Law/Ordinance/Decree /Decision 52/2
Page 146 and 147: of samples required for analysis. U
Page 148 and 149: Field test kit. BFAR recognizes the
Page 150 and 151: Political Will Having planted the s
Page 152 and 153: gravely by fi shing communities. Th
Page 154 and 155: ecome the partner and counterpart o
Page 157 and 158: Ion-Selective Electrodes for Cyanid
Page 159: References Alban, J., Manipula, B.E
Page 163 and 164: Trends Determined by Cyanide Testin
Page 165 and 166: Implementing agencies: • Quaranti
Page 167: APPENDIX
Page 170 and 171: 1:30 Overview of and Potential Cyan
Page 172 and 173: Kristine Johnson Director Kingfi sh
Page 174: Assessment of U.S. Role in Trade: U
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