10.87 DETERMINATION OF CYANIDES Discussion, The theory of the titration of cyanides with silver nitrate solution has been given in Section 10.44. All silver salts except the sulphide are readily soluble in excess of a solution of an alkali cyanide, hence chloride, bromide, and iodide do not interfere. The only difficulty in obtaining a sharp end point lies in the fact that silver cyanide is often precipitated in a curdy form which does not readily re-dissolve, and, moreover, the end point is not easy to detect with accuracy. There are two methods for overcoming these disadvantages. In the first the precipitation of silver cyanoargentate at the end point can be avoided by the addition of ammonia solution, in which it is readily soluble, and if a little potassium iodide solution is added before the titration is commenced, sparingly soluble silver iodide, which is insoluble in ammonia solution, will be precipitated at the end point. The precipitation is best scen by viewing against a black background. In the second method diphenylcarbazide is employed as an adsorption indicator. The end-point is marked by the pink colour becoming pale violet (almost colourless) on the colloidal precipitate in dilute solution (ca 0.01M) before the opalescence is visible. In 0.1 M solutions, the colour change is observed on the precipitated particles of silver cyanoargentate. Procedure. NOTE: Potassium cyanide and all other cyanides are deadly poisons, and extreme care must be taken in their use. Details for the disposal of cyanides and other dangerous and toxic chemicals may be found in Refs 14 and 15. For practice in the method, the cyanide content of potassium cyanide (laboratory reagent grade) may be determined. Method A, Weigh out accurately about 3.5g of potassium cyanide from a glass-stoppered weighing bottle, dissolve it in water and make up to 250 mL in a graduated flask. Shake well. Transfer 25.0 mL of this solution by means of a burette and NOT a pipette to a 250 mL conical flask, add 75 mL water, 5—6 mL 6M ammonia solution, and 2mL 10 per cent potassium iodide solution. Place the flask on a sheet of black paper, and titrate with standard 0.1 M silver nitrate. Add the silver nitrate solution dropwise as soon as the yellow colour of silver iodide shows any signs of persisting. When one drop produces a permanent turbidity, the end-point has been reached. Method B. Prepare the solution and transfer 25 mL of it to a 250 mL conical flask as detailed under Method A. Add two to three drops of diphenylcarbazide indicator and titrate with standard 0.1 M silver nitrate solution until a permanent violet colour is just produced. The diphenyicarbazide indicator is prepared by in 100 mL of ethanol, solving 0.1 g of the solid 1 mole AgNO 2 moles CN~A simple example of the application of a compiexation reaction to a titration procedure is the titration of cyanides with silver nitrate solution. When a solution of silver nitrate is added to a solution containing cyanide ions (¢.g. an alkali cyanide) a white precipitate is formed when the two liquids first come into contact with one another, but on stirring it re-dissolves owing to the formation of a stable complex cyanide, the alkali salt of which is soluble: Ag* +2CN~ =[Ag(CN)2] When the above reaction is complete, further addition of silver nitrate solution yields the insoluble silver cyanoargentate (sometimes termed insoluble silver cyanide); the end point of the reaction is therefore indicated by the formation of a permanent precipitate or turbidity. The only difficulty in obtaining a sharp end point lies in the fact that silver cyanide, precipitated by local excess concentration of silver ion somewhat prior to the equivalence point, is very slow to re-dissolve and the titration is time-consuming. In the Dénigés modification, iodide ion (usually as KI, ca 0.01 M) is used as the indicator and aqueous ammonia (ca 0.2M) is introduced to dissolve the silver cyanide. The iodide ion and ammonia solution are added before the titration is commenced; the formation of silver iodide (as a turbidity) will indicate the end point: [Ag(NH,).]* +1> = Ag +2NHy During the titration any silver iodide which would tend to form will be kept in solution by the excess of cyanide ion always present until the equivalence point is reached Agh +2CN~ = [Ag(CN).]7 +17 The method may also be applied to the analysis of silver halides by dissolution in excess of cyanide solution and back-titration with standard silver nitrate. It can also be utilised indirectly for the determination of several metals, notably nickel, cobalt, and zinc, which form stable stoichiometric complexes with cyanide ion. Thus ifa Ni(II) salt in ammoniacal solution is heated with excess of cyanide ion, the [Ni(CN),]? ion is formed quantitatively; since it is more stable than the [AgiCN)2] ion, the excess of cyanide may be determined by the Licbig—Denigés method. The metal ion determinations are, however, more conveniently made by titration with EDTA: see the following sections.Fluorescence is caused by the absorption of radiant energy and the re-emi of some of this energy in the form of light. The light emitted is almost always of higher wavelength than that absorbed (Stokes’ Law). In true fluorescence the absorption and emission take place in a short but measurable time — of the order of 10~!7— 107° second. If the light is emitted with a time delay (> 107* second) the phenomenon is known as phosphorescence; this time delay may range from a fraction of a second to several weeks, so that the difference between the two phenomena may be regarded as one of degree only. Both fluorescence and phosphorescence are designated by the term photoluminescence; the latter is therefore the general term applicd to the process of absorption and re-emission of light energy. At present the most widely used type of photoluminescence in analytical chemistry is fluorescence, which is distinguished from other forms of photo- Juminescence by the fact that the excited molecule returns to the ground state immediately after excitation. When a molecule absorbs a photon of ultraviolet radiation it undergoes a transition to an excited electronic state and one of its electrons is promoted to an orbital of higher energy. There are two important types of transition for organic molecules (a) n—>x*, in which an electron in a non-bonding orbital is promoted to a n-antibonding orbital: (6) x—n*, in which an clectron in a 2-bonding orbital is raised to a x-antibonding orbital. It is the n+n* type of excitation which leads to significant fluorescence, the n—n* type producing only a weak fluorescence. The electronic transitions corresponding to charge-transfer bands also lead to strong fluorescence. The electronic energy is not, however, the only type of energy affected when a molecule absorbs a photon of UV radiation. Organic molecules have a large number of vibrations and cach of these contributes a series of nearly equally spaced vibrational levels to each of the electronic states. The various energy states available to a molecule may be represented by means of an energy-level diagram. For details of such diagrams and the allowed energy transitions an appropriate textbook should be consulted (see Bibliography, Section 21.27; see also Section 17.13 for the origins of absorption spectra). It is clear that before a molecule can emit radiation by the mechanism offluorescence it must first be able to absorb radiation. Not all molecules which absorb UV or visible radiation are, however, fluorescent and it is useful to quantify the extent to which a particular molecule fluoresces. This is done by means of the quantum yield, $,, which is defined as the fraction of the incident radiation which is re-emitted as fluorescence. df <1) = Ne-ef photons emitted __ Quantity of light emitted *<"*~ No. of photons absorbed ~ Quantity of light absorbed A proportion of the excited molecules may lose their excess energy by undergoing bond dissociation leading to a photochemical reaction or may teturn to the ground state by other mechanisms. Clearly the quantum efficiency will then be less than unity and may be extremely small. The value of @, is an inherent property of a molecule and is determined to a large extent by its structure. In general a high value of 6, is associated with molecules possessing an extensive system of conjugated double bonds with a relatively rigid structure due to ring formation. This is illustrated by the intense fluorescence exhibited by organic molecules such as anthracene, fluorescein and other condensed-ring aromatic structures. The number of simple inorganic species which are fluorescent is much more limited, the chief examples being lanthanide and actinide compounds. In the case of metal ions this limitation may be overcome, however, by the formation of a complex with an appropriate organic ligand; for example, many of the metal-ion complexes formed with the complexing agent 8-hydroxyquinoline are fluorescent. Quantitative aspects. The total fluorescence intensity, F, is given by the equation F =1,, where J, is the intensity of light absorption and @, the quantum efficiency of fluorescence. ‘Since I) =1, +1, Where I = intensity of incident light and 1, = intensity of transmitted light, then F Io —T be and since I, = [ge ~*" (Beer~Lambert Law) F=Ig(l-e “by qa) For weakly absorbing solutions, when eci is small, the equation becomes F = 19.23 ecloy Q) so that for very dilute solutions (a few ppm, or less) the total fluorescence intensity F is proportional to both the concentration of the sample and the intensity of the excitation energy. Tt is instructive to compare the sensitivity which may be achieved by absorption and fluorescence methods. The overall precision with which absorbance can be measured is certainly not better than 0.001 units using a 1om cell. Since for most molecules the value of émax is rarely greater than 10°, then on the basis of the Beer-Lambert Law the minimum detectable concentration is given by Cmnin > 10°72 /105 = 107° M.With fluorescence, however, the sensitivity is limited in principle only by the maximum intensity of the exciting light source so that under ideal conditions, Conin = 10-17 M In general the limit of detection of the fluorescence technique is of the order of 10% times lower than that for UV absorption spectrometry. Selectivity may also be superior using fluorescence methods since (a) not all absorbing species fluoresce, and (6) the analyst can select two wavelengths (excitation and emission) as compared with one for absorption methods. This inherent selectivity may be inadequate, however, and must often be enhanced by chemical separation, e.g. solvent extraction (see Chapter 6). Improved selectivity can also be achieved using derivative techniques, ic. by basing the measurement of a sample component on the derivative spectrum rather than the original fluorescence emission spectrum: thus weak shoulders on the latter convert into casily quantified peaks in the derivative spectrum. It is important to distinguish between the emission and excitation fluorescence spectra. In the former, excitation is carried out at a fixed wavelength and the emission intensity recorded as a function of wavelength. Excitation spectra, on the other hand, are obtained by measuring the fluorescence intensity at a fixed wavelength while the wavelength of the exciting radiation is varied. Factors such as dissociation, association, or solvation, which result in deviation from the Beer-Lambert law, can be expected to have a similar effect in fluorescence. Any material that causes the intensity of fluorescence to be less than the expected value given by equation (2) is known as a quencher, and the effect is termed quenching; it is normally caused by the presence of foreign ions or molecules. Fluorescence is affected by the pH of the solution, by the nature of the solvent, the concentration of the reagent which is added in the determination of inorganic ions, and, in some cases, by temperature. The time taken to reach the maximum intensity of fluorescence varies considerably with the reaction. ‘An important aspect of quenching in analysis is that the fluorescence exhibited by the analyte may be quenched by the molecules of some compound present in the sample, i.e. this is an example of a matrix effect. If the concentration of the quenching species is constant this may be allowed for by using suitable standards (ie. containing the same concentration of quenching species), but difficulties occur when there is an unpredictable variation in the concentration of quenching species. 18.2 INSTRUMENTS FOR FLUORIMETRIC ANALYSIS Instruments for the measurement of fluorescence are known as fluorimeters or spectrofluorimeters. The essential parts of a simple fluorimeter are shown in Fig. 18.1. The light from a mercury-vapour lamp (or other source of ultraviolet light)* is passed through a condensing lens, a primary filter (to permit the light band required for excitation to pass), a sample coniainer,} a secondary filter (selected to absorb the primary radiant energy but transmit the fluorescentMercury Condensing Primary Sample vapour lens filter container lamp CE Seeondary rot filter rot | Photocell or photomultiplier Galvanometer radiation), a receiving photocell placed in a position at right angles to the incident beam (in order that it may not be affected by the primary radiation), and a sensitive galvanometer or other device for measuring the output of the photocell. Since fluorescence intensity is proportional to the intensity of irradiation, the light source must be very stable if fluctuations in its intensity are not compensated for. It is usual, therefore, to employ a tWo-cell instrument; the galvanometer is used as a null instrument, and readings are taken on a potentiometer used in balancing the photocells against each other. Since the two photocells are selected so as to be similar in spectral response, it is assumed that fluctuations in the intensity of the light source are minimised. ‘The simpler fluorimeters are manual instruments operating only at a single selected wavelength at any one time. Despite this they are perfectly suitable for quantitative measurements, as these are almost always carried out at a fixed wavelength. The experiments listed at the end of this chapter have all been carried out at single fixed wavelengths. ‘The more advanced spectrofluorimeters are capable of automatically scanning fluorescent spectra between about 200 and 900 nm and produce a chart record of the spectrum obtained. These can also operate at a fixed wavelength and are equally suitable for carrying out quantitative work; their main application tends to be for the detection and determination of small concentrations of organic substances, Some commercial spectrophotometers have fluorescence attachments which allow the sample to be irradiated from an ancillary source and the resulting fluoresence to pass through the monochromator for spectral analysis.