Forensic Science International, 16 (1980) 43 - 52 0 Elsevier Sequoia S.A.

, Lausanne - Printed in the Netherlands

43

THE APPLICATION OF ANODIC STRIPPING VOLTAMMETRY FORENSIC SCIENCE. I. THE CONSTRUCTION OF A LOW-COST POLAROGRAPH

TO

JUEI H. LIU and WEN-FA LIN Department (U.S.A.) of Criminal Justice, University of Illinois at Chicago Circle, Chicago, Illinois

LOTHAR TAYLOR Department of Information Illinois (U.S.A.) (Received January 14, 1980; 1980) Engineering, University of Illinois at Chicago Circle, Chicago, 20, 1980; accepted February 22,

in revised form February

Summary
Intended for forensic gunshot residue analyses and other applications, a low-cost (about $250.00) polarograph was constructed. This instrument is essentially an operational amplifier version of a three-electrode potentiostat containing a voltage ramp generator, a scan amplifier, an amplifier and a current-to-voltage converter, two voltage followers and an electrolysis cell. An anodic stripping procedure was developed for the simultaneous determination of a zinc, cadmium, lead, antimony and copper mixture, and used for testing the instrument. Effects of pre-electrolysis time, concentration changes, presence of other species and overlapping were also studied.

Introduction Modern methods of analysis are often achieved by the use of expensive and sophisticated equipment. While this approach gives satisfactory results in well-funded organizations, it is usually beyond the reach of small laboratories. Considering the limited resources most forensic laboratories face, the -authors constructed a low-cost polarograph intended for anodic stripping voltammetric (ASV) analysis of forensic samples. Although ASV is widely used in environmental [l] and other trace analyses [ 2, 31, its application in forensic science is rare. Forensic toxicological uses include the determination of cadmium poisons [4] and drugs of abuse [5], and the determination of overdose levels of 1,4-benzodiazepines in tissue analysis [ 61. Compared to atomic absorption spectroscopy (another practical trace analysis method), ASV has the following advantages [ : I] (1) determination of anions and organic compounds is possible; (2) simultaneous determination of several species is common; (3) the instrument is easy to construct and is relatively inexpensive; (4) it is a non-destructive

V
20 10K 5K K Id IK IO0 Y

0
109 1 S

b ,T

Fig. 1. Scheme of the polarograph and the system lay-out. A, counter electrode; B, reference electrode; C, hanging mercury (working) electrode; D, nitrogen inlet; E, port for standard addition; F, Teflon stopper; G, rubber ring; H, sample cell; I, Teflon-coated magnetic stirring bar; Al, current to voltage converter; A2 and A5, linear ramp generator; A3, summing amplifier; A4 and A6, voltage follower; Sl, baseline compensation onoff; S2 and S7, anodic, hold, cathodic scan control; S3 and S8, scan rate (fine); S4, sensitivity; 55, working electrode on-off; S6, zero adjustment; S9, scan rate (coarse); SlO, scan enable; Sll, scan reset; Vd, signal output; E, applied voltage.

technique in the sense that the sample is retrievable; and (5) there are extensive theoretical studies in the literature which provide a profound basis for practical application. This paper describes the construction of a low-cost (about $250.00, excluding recording devices) polarograph mainly designed for ASV experiments. The instrument is tested by the simultaneous determination of trace quantities of zinc (Zn), cadmium (Cd), lead (Pb), antimony (Sb) and copper (Cu). Fundamental electroanalytical procedure and problems, such as supporting electrolytes, linearity, overlapping voltammogram and interference, are also discussed in relation to the test mixture. The subsequent paper will present the application of this instrument and analytical technique to gunshot residue analysis. Applications of this instrument to other samples of forensic interest will be investigated later.

45

Materials and methods

Construction of the polarograph

The instrument consists of a power supply and five fast-precision FET operational amplifiers (models 2203 and 1027, respectively, Teledyne Philbrick, Dedham, MA) as shown in Fig. 1. These operational amplifiers were used because of their high slew rate and fast settling time. They also use FET inputs so that input loading is virtually non-existent (manufacturer s specifications should be consulted for detailed characteristics of these components). The operational amplifiers were externally zero-trimmed with 25-turn 1 K potentiometers. To minimize the noise level, appropriate capacitors were incorporated into the power supply, and operational amplifiers Al and As ; shielded cables were also used as indicated in the diagram (Fig. 1). All of the resistors used were 0.5 W with 1% precision. All components were mounted on a circuit board and housed inside a metal cabinet. Rating of each component is available upon request.
Simultaneous Apparatus determination of zinc, cadmium, lead, antimony and copper

All experiments were run on the instrument constructed. The electrolysis cell was made from a Pyrex tube with Teflon cap. Holes were drilled on the Teflon cap to accommodate the platinum counter electrode, saturated calomel reference electrode (SCE), nitrogen inlet port and the hanging mercury drop working electrode (HMDE). Platinum was used as the support for the HMDE and constructed according to the procedure described in the literature [ 8 - lo] . The solution was stirred, whenever needed, with a Teflon-coated magnetic bar. The stripping rate was controlled by a magnetic stirrer. A Honeywell Electronik 194 strip-chart recorder was used for recording voltammograms. Voltage scanning was monitored by a homeassembled digital multimeter from Heath Company, Benton Harbor, Michigan.
Reagent and sample preparation

A 0.1 M acetate buffer (pH 5.9 + 0.1) was used as supporting electrolyte. A 1 M stock buffer solution was prepared by dissolving 129.2 g of CHsCOONa*3HzO, adding 2.4 ml of 99.7% acetic acid and diluting to 1 liter with deionized-distilled water. The buffer solution was passed through Chelex 100 (Bio-Rad Labs., Richmond, CA) to remove traces of copper. Stock solutions (1 X 10M3M) of zinc, cadmium, lead and copper were prepared by dissolving appropriate quantities of reagent grade zinc acetate, cadmium nitrate, lead nitrate and copper acetate in the 0.1 M acetate buffer. A 1 X 10e3M stock solution of antimony was prepared by dissolving the appropriate quantity of antimony trioxide in 4 M hydrochloric acid and diluting to 1 M. Working solutions were prepared fresh by diluting stock

46

0.1 uA

Fig. 2. A typical anodic stripping voltammogram of zinc, cadmium, lead, antimony and copper mixture in 0.1 M acetate buffer. The concentration of each species is 4 X 10d6 M; preelectrolysis time is five minutes.

solutions in 0.1 M acetate buffer which also served as the supporting electrolyte. Working procedure One drop of mercury was collected from a capillary tube and served as the working electrode. The radius and surface area were 0.0490 cm and 0.0302 cm respectively. A typical experiment included a ten-minute , deaeration of the solution. ASV runs were carried out in a standard procedure, pre-electrolysis of the desired duration being followed by the anodic run. The pre-electrolyses were generally done at -1.25 V (us. SCE) for ten minutes with stirring and 1 minute without stirring. Anodic runs (0.25 V/min) were generally terminated at 0.04 V (US. SCE) and held for two minutes when the HMDE was to be used in the same solution again. In cases where the HMDE was to be used for the second solution, it was first cleaned in a buffer solution for 5 minutes at 0.04 V (us. SCE). A typical voltammogram is shown in Fig. 2. To minimize the variations in experimental conditions, concentrations were changed by the addition of concentrated standard solutions.

47

Standard solutions containing known quantities of individual species (cadmium and antimony; zinc, lead and copper; and zinc, cadmium, lead and copper) were used for this purpose.

Results and discussion Circuit description of the polarograph The circuitry is a result of significant modifications on the basic design published in the literature [ 11, 12). Basically, this is an automatic threeelectrode instrument. While the basic instrumentation of the three-electrode design is well documented in the literature [ 13 3 , the function of each operational amplifier used in our circuitry is described as follows: A,: linear ramp generator As is connected as an integrator. It integrates a constant current to generate a ramping voltage of either (+) or (-) slope. This ramping voltage is the scanning voltage used to change the working electrode voltage linearly. As: summing am$ifier As adds the voltage from A5 and A4 and outputs a voltage to the counter electrode to force the working electrode voltage to equal the scanning voltage of A+ As is unique in that its feedback loop consists of the reference electrode, amplifier A4 and a resistor. This configuration is used to accurately measure and control the voltage of the solution-working electrode interface. Aq: voltage follower A4 is used to measure the working electrode voltage through the reference electrode with very low current flow (input impedance of A4 = 1012 Ohm). Because As uses A4 as part of its feedback loop, the output of A4 is the working electrode voltage and can be monitored at the terminal E. Al: current to voltage converter Ai converts the actual current flow between the counter and indicator electrodes to a voltage, V,, that can be measured once it is buffered through Aa. A zero adjustment circuit is also included in the noninverting input of Al. As : voltage follower A6 is used as a buffer between A1 and any recording device connected to the output .terminal V,. This isolation is necessary should the recorder input impedance be less than ideal (one mega-ohm s minimum). As : linear ramp generator Az and its associate circuit are used to compensate [ 131 the rising baseline observed in anodic stripping voltammograms.

48

Performance test in the simultaneous antimony and copper

determination

of zinc, cadmium,

lead,

To test the performance of the polarograph and to demonstrate the possibility and limitation in simultaneous determination of several species by anodic stripping voltammetric method, control samples containing zinc, cadmium, lead, antimony and copper were analyzed. The discussion followed includes the choice of supporting electrolyte, data evaluation approaches and the possible deviation from the ideal linear relationship between the signal and the species concentration.
electrolyte

Various supporting electrolytes have been used for the simultaneous determination of zinc, cadmium, lead and copper 131. However, most antimony analyses have been conducted under acidic conditions, using hydrochloric acid [I4 - 201, sulfuric acid [16, 17, 19, 201 or nitric acid [16, 171. Occasionally, acetate buffer and suitable complexing agents were used to achieve better resolution when simultaneous analyses of antimony and other species such as copper, bismuth and lead [16,17] were needed. A 0.1 M acetate buffer (pH 5.9 + 0.1) was chosen in our experiments so that the pH of the solution could be controlled and resolution of the five species of interest achieved.
Data analysis

Supporting

Baselines were easily established for voltammograms containing a single species. For simplicity, peak currents were measured. Table 1 lists the concentrations, peak currents and peak potentials of the five sets of solutions analyzed. Within the concentration range analyzed, linearity is observed for all species. However, it was observed that at higher concentrations, the peak current of antimony did not increase linearly. This might be attributed to the rather low solubility of antimony in mercury [ 31. The voltammograms of mixtures are much more difficult to measure. Standard procedure [22] involves the extrapolation of the preceding peak and determination of the peak height from this extrapolated baseline. Sophisticated measurements [ 23 ] used an on-line computer. In this method an empirical equation was first developed which described the general voltammogram for a wide variety of electroactive species. The function was fitted to a number of standard voltammograms, and the constants of the function, as specifically determined for each species, were stored in a library. Upon analysis of an unknown mixture, these constants were used to regenerate the standard curves, a composite of which was then fitted to the unknown signal. It was claimed that species with peak potentials separated by less than 40 mV could be resolved under favorable conditions. While this approach is very effective it is beyond the reach of average laboratories. A simple yet effective method is developed for solving overlapping problems as described elsewhere [ 24 J .

TABLE 1

Anodic stripping voltammetry data for zinc, cadmium, lead, antimony and copper in 0.1 M acetate buffer in the presence of individual species
Peak cm& (0.1 !JA) Correlation and S.D. 0.0 0.153 0.999 0.00189 0.0362 1.20 2.50 3.60 4.76 0.00633 and S.D. Slope Intercept Statistical informatione

Species

Concentmtiona

Peak potenti&

(lO- M)b

(V, vs. SCE)

Zn

0.0

8.0 15.8

-0.86

23.5 31.1

-0.86 -0.86

-0.86

Cd 0.252 0.999 0.00260 0.0498 0.0 2.06 4.08 6.00 7.80 0.0442 0.0 2.04 0.219 0.996 0.0112 0.214 0.199 3.80 5.62 6.72

0.0 8.0 15.8 23.5 31.1

-0.55 -0.55 -0.56 4.55

Pb

0.0 8.0

-0.39

15.8 23.5

-0.39 -0.39

31.1

--a.39

Sb 0.0 1.20 0.193 0.998 0.00627 2.80 4.40 5.90

0.0

8.0 15.8

-0.11

-0.165 0.120

23.5 31.1

-0.11 -0.11 4.11

CU
0.0 1.50 0.999 3.38 4.80 6.40

010

8.0 15.8

0.01 0.01

0.207 0.00455

-0.0330 0.0870

23.5 31.1

0.01 0.01

%oncentration variations were obtained by the addition of standard solutions. bl X 10-l M of Zn, Cd, Pb, Sb and Cu are equivalent to 6.54, 11.2, 20.7, 12.2 and 6.35 ppb, respectively. Precision of these measurements is about f 0.01 V. dPrecision of these measurements is about ~0.003 PA. This information was computed with formulae given in ref. 21. A computer program was written for processing and plotting experimental data. This program is available upon request.

TIME

(MINUTE)

Fig. 3. Anodic stripping voltammetry data for zinc, cadmium, lead, antimony and copper mixture in 0.1 M acetate buffer. The concentration of each species in the solution is 4 x lo* M. One minute of rest time before anodic run is not included in the time axis.

A solution containing five species was analyzed. The concentrations of cadmium and antimony were increased stepwise by the addition of a standard solution including these two species. At the end of these additions, the concentrations of zinc, lead and copper were increased simultaneously by the same procedure. Results are presented in Fig. 3. The high peak currents measured for lead and copper are results of measurements from the extrapolated baselines of zinc and lead, respectively. These measurements eliminate the uncertainties in extrapolating cadmium and antimony voltammograms.
Linearity

As in any quantitative method of analysis, a linear relationship between the signal measured and the concentration is most desirable. When each species was studied individually, a linear relationship was established in the concentration range studied as shown in Table 1. In the mixture of five species, antimony and copper deviate from linearity at the same concentration range. With mixtures of high concentration, which we used to investigate the relationship between peak height and pre-electrolysis time, complete deviation from linearity was observed for both antimony and copper (Fig. 3). However, by working at a lower concentration range and shorter preelectrolysis time, a linear relationship was established for all five species in the mixture (Fig. 4). The non-linearity observed at higher concentrations is

51

8 8.00

0.50

1.00

1.50 CONCENTRATION ADDED

2.00 (X 1O-7 M)

2.50

3.00

Fig. 4. Anodic stripping voltammetry data for zinc, cadmium, lead, antimony and copper mixture in 0.1 M acetate buffer. Lines are regression results; points are experimental data. Preelectrolysis time: 3 minutes with stirring, 1 minute without stirring.

probably due to the low solubility of antimony in mercury [3] and to the limited surface available. Under these conditions the formation of CuSb and CuZn intermetallic compounds [3,25] may have occurred. This study demonstrates the applicability of constructing a low-cost polarograph for the simultaneous determination of elements commonly encountered in forensic analysis. However, due to the limited surface area of the working mercury electrode and the possibility of forming inter-metallic compounds, precautions should be taken in cases where the time-saving simultaneous determination approach is used. References
1 W. P. Ferren, Analyses of environmental samples by means of anodic stripping voltammetry. Am. Lab., 10 (1978) 52 - 58. 2 A. A. Kaplin, A. G. Stromberg and N. P. Pikula, Use of the method of polarography with accumulation (inversion voltamperometry) in the analysis of macro- and microobjects (review). Ind. Lab. (USSR), 43 (1977) 463 - 480.

52 3 F. Vydra, K. Stulik and E. Julakova, Electrochemical Stripping Analysis, Halsted Press, New York, 1976. 4 R. N. Phillips and E. Spratt, Direct analysis of cadmium in blood by anodic stripping voltammetry with no sample pretreatment. 30th Annual Meeting of the American St. Louis, 1978, p. 119. Academy of Forensic Sciences, 5 M. Feldstein, Advances in polarography as applied to toxicological analysis. In A. Stolman (ed.), Progress in Chemical Toxicology, Vol. 5, Academic Press, New York, 1974, pp. 369 - 380. 6 J. M. Clifford and W. F. Smyth, The determination of some 1,4-benzodiazepines and their metabolites in body fluids -a review. Analyst, 99 (1974) 241 - 272. 7 F. P. J. Cahill and G. W. VanLoon, Trace analysis by atomic absorption spectroscopy and anodic stripping voltammetry. Am. Lab., 8, (1976) 11 - 15. 8 J. W. Ross, R. D. DeMars and I. Shain, Analytical applications of the hanging mercury drop electrode. Anal. Chem., 28 (1956) 1768 - 1771. 9 L. Meites, Polarographic Techniques, 2nd edn., Interscience, New York, 1967, pp. 455 - 456. 10 W. L. Underkofler and I. Shain, Microcell for voltammetry with the hanging mercury drip electrode. Anal. Chem., 33 (1961) 1966 - 1967. 11 W. L. Underkofler and I. Shain, A multipurpose operational amplifier instrument for electroanalytical studies. Anal. Chem., 35 (1963) 1778 - 1783. 12 S. C. Creason and R. F. Nelson, A low cost, versatile electrochemical instrument. J. Chem. Educ., 48 (1971) 775 - 779. 13 H. H. Bauer, G. D. Christian and J. E. O Reilly, Instrumental Analysis, Allyn and Bacon, Boston, 1978, p. 63. 14 T. R. Gilbert and D. N. Hume, Direct determination of bismuth and antimony in sea water by anodic stripping voltammetry. Anal. Chim. Acta, 65 (1973) 451 - 459. 15 N. K. Konanur and G. W. VanLoon, Determination of lead and antimony in firearm discharge residue on hands by anodic stripping voltammetry. Talanta, 24 (1977) 184 187. 16 V. F. Yankaukas, V. M. Pichugina and A. A. Kaplin, Amalgam polarographic determination of traces of copper, bismuth and antimony in high-purity lead. Znd. Lab. (USSR), 35 (1969) 1748 - 1750. 17 D. D. Lyungrin, V. V. Pnev and M. S. Xakharou, Anodic peaks of antimony electrodeposited on a vitreous-carbon electrode. J. Anal. Chem. USSR, 30 (1975) 1032 1034. 18 T. D. Gornostaeva and V. A. Pronin, Determination of antimony, bismuth and USSR, 30 tellurium in refined silver by inverse voltammetry. J. Anal. Chem. (1975) 958 - 961. 19 E. N. Vinogradova and A. I. Kamenev, Determination of ultrasmall amounts of bismuth and antimony in germanium and germanium tetrachloride by anodic voltamperometry on a stationary mercury electrode. J. Anal. Chem. USSR, 30 (1975) 165 - 168. 20 E. N. Vinogradova and L. N. Vasil eva, Determination of ultrasmall amounts of tin, bismuth and antimony in high-purity aluminum by anodic voltammetry on a stationary mercury electrode. J. Anal. Chem. USSR, 17 (1962) 576 - 582. 21 E. L. Bauer, A Statistical Manual for Chemists, 2nd edn., Academic Press, New York, 1971, pp. 109 - 130. 22 P. Delahay, New Instrumental Methods in Electrochemistry, Intersciences, New York, 1954, pp. 129 - 130. 23 W. F. Gutknecht and S. P. Perone, Numerical deconvolution of overlapping stationary electrode polarographic curves with an on-line digital computer. Anal. Chem., 42 (1970) 906 - 917. 24 J. H. Liu, A new approach in the quantitative analysis of overlapped anodic stripping voltammograms. Analyst, in press. 25 T. M. Florence, Determination of trace metals in marine samples by anodic stripping voltammetry. Electroanal. Chem. Znterfac. Electrochem., 35 (1972) 237 - 245.