UMIST

Separation of Metal Ions using Isotachophoresis

Jeff E. Prest*, Sara J. Baldock, Nihal Bektas, Peter R. Fielden and Bernard J. Treves Brown
Department of Instrumentation and Analytical Science (DIAS), UMIST, PO Box 88, Manchester M60 1QD, UK
* Jeff Prest is supported by BNFL, Research & Technology, Springfields, Salwick, Preston, Lancashire, PR4 OXJ.

Introduction
Electrophoretic separation techniques have become popular analytical tools in recent years. This has largely been in the field of molecular biology and biochemistry where they have proved useful in the analysis of things such as proteins. However some of the techniques, notably capillary zone electrophoresis and (the less commonly encountered) isotachophoresis, can also be used for the separation of ionic substances. Figure 1 gives an indication of the wide range of metallic ions which have been separated using these techniques. This poster pays particular attention to showing the usefulness of isotachophoresis for performing such separations and also the great potential for miniaturising the technique.
Key
H Li Be Na Mg B C N O

The separation of uranium from a further selection of metals can be seen in Figure 4, which shows the separation of the following
Figure 1 Metals which have been separated using electrophoretic techniques
He F Ne Al Si P S Cl Ar I Xe

E lectro ly te S y stem P a ra m eters

HCl
L ead in g E lectro lyte(1 )

N a O H / H IB A
L ead in g E lectro lyte(2 ) N a + , 0 .0 3 M H IB A , 0 .0 1 5 M P ro p io n ic acid 4 .9 H E C , 0 .1 T erm in atin g E lectro lyte C arn , 0 .0 1 M

mixture: 10 ppm Co(II), Cr(III), Cu(II), Fe(III), Mn(II), Ni(II), Zn(II) and U(VI) Na (contamination) Indeed the HCl electrolyte system allows uranium to be separated from any other metal.
io n , co n cen tra tio n co m p lex in g a g en t p H B u ffer pH a d d itiv e, (% w /w )

H + , 0 .0 1 M H E C , 0 .1

K Ca Sc Ti V Cr Mn Fe Co Ni Cu Zn Ga Ge As Se Br Kr Rb Sr Y Zr Nb Mo Tc Ru Rh Pd Ag Cd In Sn Sb Te Cs Fr
LaBa Lu Hf Ta W Re Os Ir AcRa Lr Unq Unp Unh Uns Uno Une

Pt Au Hg Tl Pb Bi Po At Rn

La Ce Pr Nd Pm Sm Eu Gd Tb Dy Ho Er Tm Yb Lu Ac Th Pa U Np Pu AmCm Bk Cf Es Fm Md No Lr

C arn ; carn itin e h yd ro ch lo rid e H E C ; h yd ro x yeth yl cellu lo se H IB A ; a -h yd ro x yiso b u tyric acid T ab le 1 E lectro lyte system s u sed fo r iso tach o p h o retic sep aratio n s

Isotachophoresis and Capillary Zone Electrophoresis Isotachophoresis Capillary Zone Electrophoresis

100 90 80

Figure 5 Calibration graph of uranium standard samples

The linearity of the isotachophoretic zones produced by uranium was assessed by calculating a standard calibration curve for a series of 7 uranium samples, of concentrations ranging from 5 to 100 ppm. As can be seen in Figure 5 good linearity is exhibited, with a correlation coefficient of 0.999 being obtained. Figure 4 also highlights why complexing agents are used to aid separations as the transition metals all co-migrate when using the HCl electrolyte system.
y = 0.9847x - 2.4351 R2 = 0.9973

ZONE LENGTH (s)

70 60 50 40 30 20

Isotachophoresis
Capillary isotachophoresis (CITP) is an electroseparation technique which utilises the electrokinetic phenomenon of electrophoresis. It is a high resolution, technically simple method which can be readily utilised for the analysis of ionic substances. The technique utilises a discontinuous electrolyte system. This consists of a leading electrolyte, which has a higher mobility than any of the ions of interest in the sample being separated; and a terminating electrolyte, which has a lower mobility than any of the ions of interest in the sample. The sample is introduced between these electrolytes. When an electric current is applied a uniform strength electric field occurs over the sample zone. Separation then occurs because one of the parameters in the equation which determines how ionic species move under an electric field: n = mE where n = electrophoretic migration velocity m = electrophoretic mobility of ion E = electric field strength of applied field is fixed. Thus as the mobilities of different species are different they will also move with different migration velocities. Hence all of the ionic species in the sample separate in order of decreasing electrophoretic mobility. Eventually a situation is reached where each zone contains only one ionic species. This is known as the steady state, where all the zones migrate at the same rate, determined by the migration velocity of the leading electrolyte, and are not subjected to a constant electric field strength. In the steady state the electric field strength applied across the zones iincreases from the leading zone to the terminating zone as the mobility decreases. This characteristic gives rise to one of the significant features of isotachophoretic systems, the self correction of zone boundaries, which enable high resolution separations to result. If an ion diffuses into a following zone its electrophoretic mobility will increase because trailing zones have a greater field strength across them. This causes the ion to run forward to its correct position. Likewise if a zone diffuses into a proceeding zone it is subjected to a lower electric field strength which retards its electrophoretic velocity allowing it to be caught up by the correct zone. A further useful feature of isotachophoresis, which makes it particularly suitable for use with dilute samples is that the concentrations of all the separated zones are determined by the concentration of the leading electrolyte, a parameter which can be easily manipulated. The actual concentration of all the separated zones is constant, with the length of the zone giving a measure of the concentration of the individual species.
Figure 2 Schematic of the ItaChrom EA101 isotachophoretic separation unit
Terminating electrolyte reservoir

Separations have also been carried out using a complexing

10 0 0 10 20 30 40 50 60 70 80

electrolyte system, the NaOH / HIBA system shown in Table 1. The migration order for a number of metallic cations with this system has been found and is shown in Figure 5.

90

100

CONCENTRATION (ppm)

Na(I)

Mg(II)

Mn(II)

Co(II)

Ni(II)

La(III)

Pr(III)

Nd(III)

Eu(III)

Cu(II)

Ho(III)

Carn U(VI)
Slower than terminating electrolyte

Ca(II)

Li(I)

Cd(II)

Cr(III)

Zn(II)

Ce(III)

Fe(II)

Sm(III)

Gd(III)

Y(III)

Yb(III)

Separations are only possible between adjacent ions if there is a colour change Figure 6 Migration order obtained using the NaOH / HIBA electrolyte system

Figure 7 Isotachopherogram of a separation of 17 metallic cations

3.14

A number of separations of complex simulated samples have been achieved using the NaOH / HIBA electrolyte system which particularly show the potential of isotachophoresis for the analysis
MAGNITUDE OF RESPONSE (V)
2.71

Yb(III)

TE

of ionic substances. Figure 7 shows a separation of the following 17 ions: 3 ppm Ca(II), Mg(II) 5 ppm Mn(II), Co(II), Ni(II), Zn(II), Cu(II) 10 ppm Y(III) 15 ppm Ce(III), Sm(III), Eu(III) 0.0001 M Pr(III), Nd(III), Gd(III), Ho(III), Yb(III) 0.0003 M La (III). achieved using the two columns of the separation device as effectively a single, long column. Of note is that this separation includes some ions which are generally regarded as being difficult to separate, such as Eu(III) and Gd(III).

Eu(III)
2.28

Cu(II) Gd(III)

Ho(III) Y(III)

Nd(III) Ce(III) Pr(III) Ni(II) Mn(II) Ca(II) Co(II) La(III) Zn(II) Sm(III)

1.85

Mg(II)

1.42

LE

0.99 2926

3030

3135

3239

3344

3449

TIME (s)
where LE = leading electrolyte TE = terminating electrolyte

Injection valve Sample syringe Buffering block Upper (preseparation) column Conductivity detector Leading electrolyte syringe Bifurcation block Lower (analytical) column Conductivity and UV detector

Capillary Scale Separations

Capillary scale separations were carried out using an ItaChrom EA101 Analyser, which is a commercially available separation system. A schematic of the device can be seen in Figure 2. The system has a power supply which can provide 0-15 kV and currents of 0-500 mA. The device utilises a valve injection system which includes a 30 ml sample loop.

The information garnered from these separations shows that the two electrolyte systems can thus be used together in a two dimensional separation. The HCl system would be used in the preseparation column and the NaOH / HIBA system in the analytical column. This would allow uranium and most other metal cations to be separated and identified in a single run.

Miniaturised Separations
The miniaturised separation device is formed of two pieces of silicone rubber elastomer Sylgard 184. One of these pieces is cast molded using a copper template and the other is a plain piece of polymer. The two pieces of polymer are adhered together to form the flow channel containing chip. A schematic of the device is shown in Figure 8. The separation channel is 56 mm deep and 324 mm wide at half depth. This device is fitted with a single electrode on-column conductivity detector. This is of a novel design which offers simple fabrication and the prospect of higher resolution than is possible with detectors with two or more electrodes. The detection electrode is a piece of 25 mm platinum/iridium wire placed between the two pieces of polymer before they are fully bonded. This electrode is positioned 5 mm along the separation channel from well A.
TO DATA ACQUISITION COMPUTER HIGH VOLTAGE PROBE MEASURING ELECTRODE HIGH VOLTAGE POWER SUPPLY

Polymer chip

Waste

Leading electrolyte reservoirs

The device is made up of two fluorinated ethylene-propylene co-polymer (FEP) separation columns: a 90 mm long 0.8 mm internal diameter preseparation column and a 160 mm long 0.3 mm internal diameter analytical column. The arrangement allows for column switching to take place thus allowing more flexibility in separations, and making it easier for minor components of samples to be identified as major components can be discarded after they have passed through the preseparation column. Each column is fitted with an oncolumn conductivity detector. These detectors operate in ac mode and use transformer coupling for isolation from the high strength electric field applied to drive the separation. The analytical column is also fitted with a UV absorbance detector which was not used in this work.
Figure 3 Isotachopherogram of a separation of 5 metal cations

B
SEPARATION CAPILLARY

Figure 8 Schematic of the miniaturised separation device with a single electrode conductivity detector

Results
MAGNITUDE OF RESPONSE (V)

Terminating electrolyte
0.980 0.895 0.809 0.724 0.639 0.553 0.468
0.12 0.11
TERMINATING ELECTROLYTE

Cationic metal separations are generally carried out using electrolyte systems which contain complexing agents such as a These are added to aid the hydroxyisobutyric acid (HIBA).

An example of a separation carried out using one of these devices can be seen in Figure 9. This shows a separation of a 100 ppm U sample from sodium and potassium contamination present in water, achieved
U Na K

Uranium

separations by increasing effective mobility differences between the ions. In this work it was found that if attempts were made to separate uranium using an electrolyte system containing HIBA the resulting uranyl complex had a lower effective mobility than any terminating electrolyte and the substance did not migrate isotachophoretically. Thus the HCl electrolyte system, shown in Table 1, which does not contain a complexing agent, was used, successfully, for uranium separations. Figure 3 shows an example of the separation of the following mixture: 10 ppm K, Cd, Pb 30 ppm U Na (contamination). This result clearly shows the characteristic stepwise output which is an unusual feature of isotachopherograms.

ELECTRIC POTENTIAL (V)

Cadmium Sodium Lead Potassium

0.10 0.09 0.08 0.07 0.06 0.05 LEADING ELECTROLYTE 0.04 0.03 0.02 0.01 0.00 0 50 100 150
Channel length: 120mm Channel width: 0.324mm Channel depth: 0.056mm

using the HCl electrolyte system given in Table 1. The isotachopherogram shown is very different to those generally produced due to the operation of the detector. With the single electrode detector, which simply measures potential at a point, different components produce zones of different gradients rather than steps. However if the zones are extremely long, such as that for sodium, steps do occur.

Leading 0.383 electrolyte

0.297

0.212 936

1057

1177

1298

TIME (s)

Figure 4 Isotachopherogram of the separation of uranium from transition metals

1.654 Terminating electrolyte

200

250

MAGNITUDE OF RESPONSE (V)

1.489 1.325 1.160 0.995 0.831 0.666 0.501 0.336 0.172 1248 Leading electrolyte Sodium, Chromium(III), Manganese(II), Iron(III), Cobalt(II), Nickel(II), Copper(II), and Zinc(II) Uranium

TIME (s)

Figure 9 Separation of uranium using a miniaturised planar separation device

Conclusions
Isotachophoresis has been found to be a powerful analytical method for the separation of metallic cations, with the separation of up to 17 different ions simultaneously being found to be possible. The technique has also been found to offer great promise for use in the
1576 1905 2233

TIME (s)

developing field of miniaturisation.