You are on page 1of 5

+Model

ACA-228212; No. of Pages 5

ARTICLE IN PRESS

Analytica Chimica Acta xxx (2007) xxxxxx

Spectrophotometric determination of aqueous cyanide using a revised phenolphthalin method


David Cacace, Heidi Ashbaugh, Naomi Kouri, Sara Bledsoe, Sean Lancaster, Stuart Chalk
Department of Chemistry and Physics, University of North Florida, 4567 St. Johns Bluff Rd S, Jacksonville, FL 32224, USA Received 31 January 2007; accepted 1 February 2007

Abstract This paper focuses on a revision of the phenolphthalin method for cyanide analysis, with the intent of producing a robust and sensitive spectrophotometric method. Limitations of the phenolphthalin chemistry were overcome by the addition of EDTA to the reagent. The revised reagent was found to have suitably fast kinetics, a linear dynamic range of 0.013.0 ppm cyanide, and a detection limit of 5 ppb. The method was tested for interferences and applied to the determination of cyanide in environmental waters. 2007 Published by Elsevier B.V.
Keywords: Cyanide determination; Copper(II)EDTA binuclear complex; Phenolphthalin; Environmental analysis

1. Introduction Cyanide is found naturally in air and water and as it is toxic to humans it is a priority pollutant worldwide [1]. In air, cyanide exists primarily as hydrogen cyanide (HCN) at concentrations in the range 1.51.7 101 ppbv, and an average residence time of 2.5 years [2]. HCN is also classied as a chemical warfare agent (CWA) [1] as it is a signicant potential threat due to its volatility, toxicity, and high odor threshold (100 times the lethal limit 300 g/L) [3]. Aqueous cyanide is also dangerous because of short and long-term health effects from ingestion. The goal of this work was to evaluate existing colorimetric chemistries for cyanide analysis in the context of developing a robust methodology for cyanide in air and water. In comparing different methodologies the reactions were evaluated based on the following criteria; speed, sensitivity, detection limit, selectivity, reproducibility and reagent stability. A literature survey revealed a variety of colorimetric chemistries for cyanide [422] using batch and ow-based methods. Of those evaluated the most suitable was determined to be the phenolphthalin method [57,23,24]. This paper reports a revision of the phenolphthalin method where EDTA is added to the reagent solution to stabilize (com-

plex) copper(II)(aq) in solution at pH 10.8 [5]. The relative concentrations of the reagent were optimized and a single stable combined reagent was formulated and tested. The method is more reproducible and sensitive than the traditional reagent formulation [7] and does not require solid phase copper(II) [5] for analytical application. A discussion of the reaction chemistry of this method is included as well as analytical gures of merit, an interference study, and analysis of environmental waters for the determination and recovery of cyanide. 2. Experimental 2.1. Apparatus pH measurements were made using a Thermo Orion Model 525 pH meter (ThermoElectron, www.thermo.com). Absorbance measurements were made with a USB4000 diodearray spectrophotometer with USB-ISS-UVvis source (Ocean Optics, www.oceanoptics.com) and a 10 mm cell. For acquisition of spectral and kinetics data, SpectraSuiteTM v5.1 (Ocean Optics) was used. 2.2. Reagents and solutions 2.2.1. Borax buffer A buffer solution was prepared by dissolving 19.068 g of sodium tetraborate decahydrate (Acros, www.shersci.com,

Corresponding author. Tel.: +1 904 620 1938; fax: +1 904 620 1989. E-mail address: schalk@unf.edu (S. Chalk).

0003-2670/$ see front matter 2007 Published by Elsevier B.V. doi:10.1016/j.aca.2007.02.004

Please cite this article in press as: D. Cacace et al., Spectrophotometric determination of aqueous cyanide using a revised phenolphthalin method, Anal. Chim. Acta (2007), doi:10.1016/j.aca.2007.02.004

+Model
ACA-228212; 2 No. of Pages 5

ARTICLE IN PRESS
D. Cacace et al. / Analytica Chimica Acta xxx (2007) xxxxxx

Cat# AC41945-0010, Lot# A0169149) in 2.0 L Milli-Q water (with heating/stirring). The pH was increased to 10.8 with 0.5 M sodium hydroxide (Fisher, www.shersci.com; Cat# S320-3; Lot# 007230) prior to making the solution to volume. 2.2.2. Phenolphthalin solution A 0.0111 M stock solution was prepared by dissolving 1.777 g phenolphthalin (Acros; Cat# 417231000; Lot# A0203911001) in 500 mL of 50% (v/v) ethanol (Pharmco, Brookeld, CT; Cat# 111ACS200) in Milli-Q water. 2.2.3. Copper (II) nitrate/EDTA solution A 2.0 102 M copper(II) stock solution was prepared by dissolving 1.876 g Cu(NO3 )2 (Sigma, www.sigma.com; Cat# C2646; Lot# 73H3452) in 500 mL of Milli-Q water. Additionally, solutions with different ratios of EDTA added to copper(II) (in the range of 1:0.11:1 mole ratio Cu(II):EDTA) were made. For example, a half mole equivalent (the 1:0.5 ratio) solution was made by the addition of 1.861 g of disodium EDTA dihydrate (Sigma; Cat# E-5134; Lot# 107H093915) to a 2.0 104 M copper(II) solution in a 500 mL volumetric ask. 2.2.4. Cyanide solution A 1000 ppm cyanide (KCN) stock solution (CertiPrep, www.certiprep.com; Cat# RSCN9-2X; Lot# 33-5AS) was used to make diluted stock and working solutions. 2.2.5. Stabilizing solution A stabilizing solution was made by dissolving 1.25 g Na2 SO3 (Fisher; Cat# S430-500; Lot# 934,683) and 0.324 g triethanolamine hydrochloride (Kodak; Rochester, NY; Cat# 1135458; Lot# 1161042163) in Milli-Q water in a 100 mL volumetric ask. A separate stabilizing solution of EDTA was prepared by dissolving a 1:0.5 copper(II):EDTA mole ratio of EDTA in 500 mL Milli-Q water. 2.2.6. Interference solutions One thousand parts per million solutions were prepared from solid/liquid stocks (Fisher), which were then diluted as needed. 2.2.7. Combined reagent solution An optimized combined reagent solution was prepared by adding the reagents to a 50 mL volumetric ask in the following order. 0.2326 g copper(II) nitrate was added in 20 mL Milli-Q water followed by 0.1861 g EDTA. After the EDTA dissolved, 0.1777 g phenolphthalin was added followed by 25 mL ethanol. Once the phenolphthalin had dissolved the ask was lled to the mark with Milli-Q water. 2.3. Procedures All experiments for the optimization and evaluation of the reaction chemistry were performed with a specic reagent addition order (see Section 3). A 50 mL volumetric ask was lled with 45 mL borax buffer and the appropriate volume of cyanide stock added, followed by phenolphthalin and copper solution (or copper/complexing agent), starting the reaction. Exactly 3 min

after addition of copper, the stabilizing solution was added. The ask was lled to the mark with buffer and analyzed at 553 nm. When using the combined reagent solution, timing was started upon its addition. 2.3.1. Chemistry optimization Optimization of each reagent was performed independently. Initial reagent concentrations were determined based on stoichiometric amounts needed for reaction with 3 ppm cyanide. Alternating variable search was performed to nd the optimum conditions. 2.3.2. Calibration curve and linear range After reagent optimization, calibrations were performed to determine the linearity of the chemistry over a certain concentration grade. 0.0, 0.5, 1.0, 1.5, 2.0, 2.5, and 3.0 ppm cyanide standards were determined against a borax blank. 2.3.3. Detection limit The detection limit (blank + 3s of the blank) of the optimized method was estimated by analyzing the background absorbance of ve reagent blank solutions (ve times each) at 553 nm against a borax blank. The calculated absorbance was applied to the calibration curve and the resulting cyanide detection limit concentration analyzed against a borax blank. 2.3.4. Method reproducibility A study was performed to determine method precision and evaluate if EDTA improved reproducibility. Ten solutions of each of the following were made and analyzed; 2 ppm cyanide with EDTA, 2 ppm cyanide without EDTA, 0.2 ppm cyanide with EDTA and 0.2 ppm cyanide without EDTA. 2.3.5. Reagent stability A reagent stability study was performed by using fresh reagent and 1000 ppm cyanide stock solutions and performing calibrations with these solutions over time. Solutions were stored at room temperature in clear Nalgene bottles. 2.3.6. Reagent addition order The effect of the reagent addition order (copper(II)/EDTA, phenolphthalin, cyanide) was evaluated by using a 2 ppm cyanide standard and adding the reagents in the six addition order combinations. Addition of the stabilizer was started after addition of the last reagent. 2.3.7. Interferences Different concentrations of various interfering ions were added to a 50 mL volumetric ask prior to 2 ppm cyanide and reagent addition. Each solution was analyzed and compared to a 2 ppm cyanide standard. A 5% bias of the interfering ion signal was considered signicant. 2.3.8. Kinetics The kinetics of the reaction was evaluated by recording the absorbance of a 2 ppm cyanide solution over time. Reagents were added in their optimized order (without stabilizer) and the

Please cite this article in press as: D. Cacace et al., Spectrophotometric determination of aqueous cyanide using a revised phenolphthalin method, Anal. Chim. Acta (2007), doi:10.1016/j.aca.2007.02.004

+Model
ACA-228212; No. of Pages 5

ARTICLE IN PRESS
D. Cacace et al. / Analytica Chimica Acta xxx (2007) xxxxxx 3

solution was made up to volume and analyzed immediately. The absorbance at 553 nm was recorded every second for 1 h. For comparison, an identical solution was prepared with stabilizer added after 3 min of development. 2.3.9. Water samples Environmental samples were collected, ltered using a 0.45 m cellulose acetate lter, and then analyzed for cyanide. Samples spiked with 2 ppm cyanide were also analyzed and compared to evaluate cyanide recovery. 3. Results and discussion 3.1. Phenolphthalin chemistry As shown in Fig. 1(a), when cyanide is added to aqueous copper(II) it preferentially forms the tetracyanocopper(II) complex. Tetracyanocopper(II) in base is then immediately converted to the tetracyanocopper(I) complex [25], producing cyanogen ((CN)2 ). Results in this work support the hypothesis that cyanogen immediately reacts with phenolphthalin producing phenolphthalein. There are two phases to the phenolphthalein color development. Initially, there is an instantaneous color development, followed by a slow absorbance increase over time [7]. This can be explained by the regeneration of cyanide ion by the reductive splitting of cyanogen upon reaction with phenolphthalin, allowing cyanide to act as a catalyst continuously producing phenolphthalein until no phenolphthalin remains. In order to stop color development, a stabilizing solution can be added to form a more stable copper complex [6] than with cyanide. A high pH (10.8) is necessary to produce the tetracyanocopper(II) complex and convert phenolphthalein to its colored form. However, this pH affects the reproducibility of the method as any non-complexed copper(II) precipitates as Cu(OH)2 (Fig. 1(d)) which interfers with the absorbance at 553 nm. This problem gets worse at lower cyanide concentrations as more precipitate is formed. 3.2. Revised phenolphthalin chemistry A complexing agent was added to the phenolphthalin chemistry to eliminate the formation of Cu(OH)2 (s). EDTA complexation of copper(II) has been extensively studied in the
Fig. 2. Variation of absorbance for 2 ppm cyanide with changing copper(II):EDTA ratios.

literature [2629] and the complex does not have a signicant spectral overlap with phenolphthalein. Fig. 1(ei) summarizes reactions of Cu/EDTA pertinent to this work. Comparing solutions of copper(II) with and without EDTA, at a 1:0.5 mole ratio of Cu:EDTA, signicantly less precipitate was observed. Fig. 2 shows that the 1:0.5 mole ratio of Cu:EDTA is optimum for use in this chemistry. Lower ratios result in more precipitation of Cu(OH)2 while ratios closer to 1:1 result in a dramatic decrease in signal. This optimum can be explained by the complexation of the copper(II) as the sparingly soluble binuclear Cu2 EDTA4H2 O(aq) [28,29]. If EDTA is present at higher ratios the sensitivity of the method decreases because of the formation of [CuEDTA]2 (aq), which is more stable then both Cu2 EDTA4H2 O(aq) and [Cu(CN)4 ]2 (aq) complexes and thus stops the reaction. The formation of the binuclear complex is key to the success of the revised reagent. The formation constant of Cu2 EDTA4H2 O(aq) is lower than the formation constant of the tetracyanocopper(II) complex (see Table 1), thus copper(II) is solubilized at pH 10.8 and available to complex cyanide. Tanaka et al. [27] report that the reaction of Cu2 EDTA4H2 O(aq) with cyanide is the rate limiting step in the formation of the tetracyanocopper(II) complex. EDTA increases the kinetics slightly in both phases of the reaction. This is because copper(II) is more available in solution via the binuclear complex. This also indicates that copper(II) is part of the rate determining step in the second phase of the reaction. Whether the kinetics of the loss of cyanogen to the atmosphere is important to this step is unknown at this point. EDTA can serve a second purpose in this methodology. As mentioned previously, the reaction can be quenched by the addition of sulte/triethanolamine, forming a Cu(NC6 H15 O3 )4 SO4 complex [6]. Replacing sulte/triethanolamine with EDTA (1.8 104 M) results in the same quenching effect. 3.3. Reagent optimization 3.3.1. Buffer type and pH A borax solution at pH 10.8 was chosen due to its high buffer capacity and non-reactivity with any of the reagents

Fig. 1. Reactions of copper(II), phenolphthalin, and EDTA.

Please cite this article in press as: D. Cacace et al., Spectrophotometric determination of aqueous cyanide using a revised phenolphthalin method, Anal. Chim. Acta (2007), doi:10.1016/j.aca.2007.02.004

+Model
ACA-228212; 4 Table 1 Physical properties of reaction species Species [Cu(CN)4 [Cu(CN)4 ]2 [Cu(CN)4 ]2 /[Cu(CN)4 ]3 Cu2 EDTA + 4CN [Cu(CN)4 ]2 + [CuEDTA]2 Cu2 EDTA4H2 O [CuEDTA]2Cu(OH)2 Cu(II)/Cu(I) Phenolphthalin (CN)2 + 2H+ + 2e 2HCN ]3 No. of Pages 5

ARTICLE IN PRESS
D. Cacace et al. / Analytica Chimica Acta xxx (2007) xxxxxx

Type Formation constant () Formation constant () Reduction potential (Eo ) Rate constant (k) Stability constant (Kstab ) Solubility product constant (Ksp ) Stability constant (Kstab ) Solubility product constant (Ksp ) Reduction potential (Eo ) Half-wave potential (E1/2 ) Standard reduction potential

Value 1022 L4 mol4 0.66 V 6.8 107 L4 mol4 s1 2.5 103 L mol1 2 105 mol2 L2 6.0 1018 L mol1 2.2 1020 mol2 L2 0.153 V 1.33 V 0.716 0.010 V 1030.53 L4 mol4

Ref. [25] [25] [25] [27] [29] [26] [31] [31] [32] [33]

used. The concentration of the borax buffer was maximized (2.5 102 M) so that solutions retained their high pH during analysis. 3.3.2. Phenolphthalin Ethanol was used to dissolve phenolphthalin due to its very low solubility in water, ethyl acetate, 2-propanol, and acetonitrile. The ethanol content and phenolphthalin concentration were optimized for low levels (<3 ppm) of cyanide, with the highest signal found at 0.9% (v/v) ethanol and 2.0 104 M phenolphthalin. 3.3.3. Copper(II) The optimized concentration of copper(II) was found to be 3.6 104 M. At low concentrations of copper(II), the formation of the tetracyanocopper(II) complex caused a lower absorbance. At higher concentrations the formation of copper hydroxide precipitate interfered with the detection. 3.3.4. Copper(II)/EDTA ratio The absorbance obtained by reaction of copper(II) with varying amounts of EDTA (1:01:1 mole ratios) at 553 nm is shown in Fig. 2. Addition of EDTA to solutions of copper(II) either slowed or eliminated the formation of Cu(OH)2 in basic solution is due to the formation of the Cu2 EDTA complex. The optimized ratio was found to be 1:0.5 mol copper(II):EDTA (3.6 104 M copper(II):1.8 104 M EDTA). 3.3.5. Order of reagent addition Fig. 3 shows the absorbance of a 2 ppm cyanide solution developed after three and 5 min using each of the six possible combinations of reactant addition. When cyanide and copper(II)/EDTA are added prior to phenolphthalin, the signal is signicantly decreased due to cyanogen volatilization that occurs before phenolphthalin addition. 3.4. Analytical parameters 3.4.1. Calibration curve, detection limit, and linear range Over the range 03 ppm cyanide, the calibration curve of the optimized method was absorbance = 0.3041[CN ] + 0.0236 (R2 = 0.9988) with a limit of linearity of 3 ppm. The detection

limit of the method was determined to be 5 ppb (blank + 3s of the blank). 3.4.2. Reproducibility Using the EDTA complexing reagent, 10 replicate 2 ppm cyanide and 0.2 ppm cyanide standards were found to have RSD values of 0.48 and 1.93% respectively. This compares to 2.08 and 6.45% for the same concentrations without EDTA, showing that addition of EDTA causes a signicant improvement in the method. 3.4.3. Reagent stability Over a period of 2 weeks (day 1, 4, 8, and 14), four 0.53 ppm cyanide calibrations were run. The calibration slopes were found to be 0.3013, 0.2972, 0.2944, and 0.3006, respectively (1% RSD). An additional calibration was run using the same stock cyanide/reagent solutions after 4 months (119 days) and the calibration slope was 0.2926 ppm1 (R2 = 0.9997). This shows that the combined reagent is very stable as long as the bottle is tightly closed, preventing evaporation of ethanol. 3.4.4. Interference study Carbonate, calcium, sulte, sulfate, uoride, chlorate, phosphate, and sulde ions were tested as possible interferents. Overall, the most signicant interference was sulde, which reacts with cyanide to form polysuldes and thiocyanate [30].

Fig. 3. Absorbance of 2 ppm cyanide with reagent addition order (Phe, phenolphthalin; CuEDTA, copper(II) EDTA complex; CN, cyanide).

Please cite this article in press as: D. Cacace et al., Spectrophotometric determination of aqueous cyanide using a revised phenolphthalin method, Anal. Chim. Acta (2007), doi:10.1016/j.aca.2007.02.004

+Model
ACA-228212; No. of Pages 5

ARTICLE IN PRESS
D. Cacace et al. / Analytica Chimica Acta xxx (2007) xxxxxx 5 [5] [6] [7] [8] [9] [10] [11] [12] [13] [14] [15] [16] [17] [18] [19] [20] [21] [22] [23] [24] [25] [26] A.T. Haj-Hussein, Talanta 44 (1997) 545. R.I. Nicholson, Analyst 66 (1941) 189. R.L. Maute, M.L. Owens Jr., Anal. Chem. 26 (1954) 1723. P.E. Kriedeman, Analyst 89 (1964) 145. G.D. Lander, A.E. Walden, Analyst 36 (1911) 266. D.D. Clyde, M.A. Warner, Analyst 103 (1978) 648. K. Groningsson, Analyst 104 (1979) 367. E. Casassas, R. Rubio, G. Rauret, Analyst 109 (1984) 1159. S. Upadhyay, V.K. Gupta, Analyst 109 (1984) 1619. T. Fonong, Analyst 112 (1987) 1033. P. Kaur, S. Upadhyay, V.K. Gupta, Analyst 112 (1987) 1681. F.J.M. de Villena Rueda, L.M. Polo Diez, R. Perez Perez, Analyst 113 (1988) 573. M. Drikas, B.I. Routley, Analyst 113 (1988) 1273. T. Ohno, Analyst 114 (1989) 857. J.C.L. Meeussen, E.J.M. Temminghoff, M.G. Keizer, I. Novozamsky, Analyst 114 (1989) 959. V. Kuban, P.K. Dasgupta, Anal. Chem. 64 (1992) 1106. J.O. Egekeze, T.M. Dowling, N. Grinberg, H.J. Perpall, G.R. Bicker, Talanta 44 (1997) 1203. H. Sulistyarti, T.J. Cardwell, S.D. Kolev, Anal. Chim. Acta 357 (1997) 103. J.M. Kolthoff, Z. Anal. Chem. 57 (1918) 1. W.A. Robbie, Arch. Biochem. 5 (1944) 49. A. Katagiri, S. Yoshimura, S. Yoshizawa, Inorg. Chem. 20 (1981) 4143. S. Aksu, F.M. Doyle, Potential-pH diagrams for copper in aqueous solutions of various organic complexing agents, in: R. Woods, F.M. Doyle (Eds.), Electrochemistry in Mineral and Metal Processing, The Electrochemical Society, Pennington, NJ, 2000, p. 258-269. N. Tanaka, M. Kamada, T. Murayama, Bull. Chem. Soc. Jpn. 31 (1958) 895. O. Gyliene, J. Aikaite, O. Nivinskiene, J. Haz. Mater. B116 (2004) 119. O. Gyliene, J. Aikaite, Pol. J. Chem. 77 (2003) 99. R.G. Luthy, S.G. Bruce Jr., Environ. Sci. Technol. 13 (1979) 1481. D.R. Lide, Handbook of Chemistry and Physics, 1991 71st edition CRC Press, Boca Raton, FL, p. 8-17. I.M. Kolthoff, D.J. Lehmicke, J. Am. Chem. Soc. 70 (1948) 1879. V.I. Belevantsev, B.I. Peshchevitskii, L.D. Tsvelodub, Russ. J. Inorg. Chem. 30 (1985) 2231.

3.4.5. Water samples Analysis of three separate environmental samples (pondwater, oceanwater, and riverwater) indicated that cyanide present in these samples were below the detection limit. Spike recoveries of 2 ppm cyanide were 96.3, 102, and 94.6% respectively. The proposed methodology therefore gives excellent recovery of cyanide even when the sample matrix is 40% (v/v) of the analysis solution (10 mL in 25 mL). 4. Conclusion The revised copper(II)/phenolphthalin method for the determination of cyanide is fast, sensitive, and applicable over a large concentration range. The addition of EDTA to the reagent has also signicantly improved the reliability of the method and produced a very stable reagent. In a subsequent paper, this chemistry will be applied to a ow injection analysis system for rapid, on site cyanide determination. Acknowledgment This research was funded through U.S. Department of Defense Contracts W911SR-04-C-0072 and W911SR-05-C0043. References
[1] The Organization for the Prohibition of Chemical Weapons, Hydrogen Cyanide, http://www.opcw.nl/resp/html/hcn.html (accessed 08/21/06). [2] R.J. Circerone, R. Zellner, J. Geophys. Res. 88 (1983) 10689. [3] World Health Organization, Hydrogen Cyanide and Cyanides: Human Health Aspects, http://www.who.int/ipcs/publications/cicad/en/ cicad61.pdf (accessed 8/21/06). [4] G. Gumus, B. Demirata, R. Apak, Talanta 53 (2000) 305.

[27] [28] [29] [30] [31] [32] [33]

Please cite this article in press as: D. Cacace et al., Spectrophotometric determination of aqueous cyanide using a revised phenolphthalin method, Anal. Chim. Acta (2007), doi:10.1016/j.aca.2007.02.004

You might also like