28 AUTOMATIC CONTROL OF FOOD MANUFACTURING PROCESSES depth below the surface. These enzyme electrodes were derived from immobilised enzyme glucose sensors developed for medical applications. 2.4 Size, colour and turbidity ‘There are a group of requirements for optical inspection which can be met using compact and relatively low cost electro-optic equipment. These include monitoring trends in height or length and changes in depth of colour. The turbidity of beverages can be measured using similar techniques. 2.4.1 Height and length The length and width of pieces lying on a conveyor can be measured conveniently by applying counting and timing logic to the video output of a conventional monochrome video camera. The dimensional stability of the imaging does not compare well with other optical measuring systems, but repeatability to within 0.5% has been consistently achieved with arrange- ments of this type used on biscuit production lines. This degree of precision is sufficient in many process control applications. Greater speed and accuracy can be obtained using light emitting diode (LED) or laser sources and linear photodiode arrays. The arrays have excellent dimensional stability and up to 4096 diodes on one chip. Interpolation between diode counts can be made from repeated scanning. An array of 128 diodes was scanned at 500 Hz and used to measure the thickness of dark chocolate to within 1 part in 500. Two alternative arrangements of triangulation for thickness gauging using solid-state components are shown in Figure 2.5. One method (Figure 2.5(a)) using a single light source embodies 3 ‘xo ‘COED wht | ’ ) to ® " 4 a X % a Figure 2.5 Optical thickness gauging with solid-state lectron () see text cs. For explanation of parts (a) andIN-LINE SENSORS 2» the principle of the Heightscan gauge, introduced by Integrated Photomatrix of Dorchester, UK, in 1976. Figure 2.5(b) shows the inverse arrangement, with an array of LED sources, which has also been used successfully in a sensor which has the added facility of measuring depth of colour (McFarlane, 19906). The advantage of LED sources is that they can be modulated at high frequency, and synchronous demodulation of the detector output applied to make the system immune to changes in ambient illumination 2.4.2 Colour The various mechanisms causing colour changes in food materials are described by Nassau (1983). The yellow and orange carotenoid colouring of carrot, corn, pumpkin, peach and other vegetables and fruits arises from the permitted energy transitions where there are non-cyclic double bonds in a chain of carbon atoms. Closely related to beta-carotene is the structure of rhodopsin, the main agent in the eye’s perception of light. Carotenoid colours are very stable to heat. The acid-base colour change of anthocyanins provide the red, blue and purple colours of beets, red cabbage and many berries. A blue colour may be produced even in an acid solution by traces of iron (hence the need for a protective inner layer in fruit juice cans). A similar reaction occurs with the closely related anthoxanin pigments which provide the creamy white colour of onion and cauliflower. The red colour of meat is derived from myoglobin, and freshly cut meat quickly develops the bright red of oxygenated myoglobin on exposure to air. The browning of raw fruits and vegetables when exposed to air is the result of an enzyme-activated oxidation producing melanin. All foods char if overheated, but the browning of meats, bread, cake and fried potato is the result of the Maillard reaction between certain sugars and amino acids. The effect of particle size on diffuse reflectance of caramel is described by Saguy and Graf (1991). When caramel particle size was reduced from 68 to 28 jim, the lightness increased, as predicted by theory; values of L* increased from 42 to 46, When the caramel was diluted with finely powdered (125 zm) sucrose the effect was reversed; 28 yan caramel had L* value of $5 compared with 65 for 68 um particles. Finally, when the sucrose particle size was further reduced, in steps to 115, 95, 80, 65, 58 and 45 wm, the caramel colourant of smaller particle size again gave the lighter readings, but the colour was masked by the sucrose powders, and all the Z* readings were in the range 58-88. Saguy and Graf comment that to manipulate the colour values of dry foods with caramel colouring it is necessary to take account of the particle sizes of other ingredients. Colour measuring equipment usually describes colour in one of the tristimulus colour scale values, and conversion between scales is easily carried out using standard formulae. Hunter and Harold (1987) identify several single number scales adopted for rating the colour value of specific food materials,IN-LINE SENSORS 2s essential oils can be identified from milligram samples by analysis of the carbon-13 NMR spectrum ~ but low resolution NMR is more likely to find a place in food processing. Low resolution NMR utilises the magnetic properties of protons (hydrogen nucleii) using magnetic fields much lower in strength than are required for high resolution NMR. An early application for pulsed NMR described by Brosio et al. (1982) was the determination of the solid fat index in margarine, based on the longitudinal magnetisation decay curve of 1 gsamples of oil/water emulsions. Other applications, including on- line equipment, have been mainly to distinguish solid from liquid phases. Padua et al. (1991) report the measurement of three water states ~ polymer, capillary and free in following the hydration of whey protein, with relaxation rates of 28, 2.75 and 0.4 s~! respectively. An on-line sampling system and automatic calibration are features of a compact pulsed NMR system used for measurements on ground corn, reported by Pearson et al. (1987). The NMR signals from the moisture and oil in the material are separated by using two pulses, 2 ms apart. The second pulse is said to refocus the NMR signal from the oil in the sample, and an echo is, detected after a further 2ms. 24s after the first pulse, the signal is proportional to the total hydrogen (chemically bound, and in oil and water) but after 64 is the chemically bound hydrogen signal has decayed, leaving a signal proportional to the oil and water hydrogen. From the three ‘measurements, oil and water content can be deduced and corrected for sample density. Moisture predictions were within 0.6% standard error. Bellon et al. (1992) showed that NMR is more suecessful than surface NIR for assessing the ripeness of some thick-skinned fruit such as cherries and grapes. Another application for nuclear magnetic resonance has been magnetic resonance imaging (MRI) involving magnetic field gradients and examination of magnetic echoes. The equipment is cumbersome, but Sun et al. (1993) were able to map two-dimensional temperature gradients in 25mm diameter cylindrical capsules of food gels, and the technique offers a method for observing, for example, progress of crystallisation in chocolate. 23.3 pH and ion-selective electrodes ‘The measurement of pH of process fluids is usually made with a pH-sensitive glass electrode, monitoring the concentration-dependent energy change associated with the ionisation of water at the gel-like hydrated surfaces of glass;a second electrode, not sensitive to pH, is required to measure the output Of the glass electrode, Hulley (1986) reviews the use of industrial pH sensors, and notes that the reference electrode has been the subject of more development than the measuring electrode. The main problem of the standard sealed reference electrode was that process liquids either diffused or were thermally pumped into electrode, poisoning it. Ways to overcome this include pressurising the electrolyte in the reference electrode, interposing an26 AUTOMATIC CONTROL OF FOOD MANUFACTURING PROCESSES intermediate liquid between the sensitive element and the process fluid or, as in the system of Great Lakes Inc., using a pH electrode in conjunction with a buffer solution intermediate between the electrode and the process fluid to ensure a stable output even in the event of the ingress of traces of process liquor. Solid-state electrodes are also available, with the sensitive electrode let into a body of sintered PTFE and potassium chloride saturated solution, more resistant to fouling Electrode cleaning is achieved by various methods, including use of brushing, jet or ultrasonic cleaning and abrasion with PTFE pellets. ‘Temperature compensation is required, which must be proportional not only to temperature change but also to the difference of pH from the isopotential pH. Very high impedances exist in the measuring circuit, and it is essential to keep electrical connections clean and dry, and to earth any screens. Some electrodes exhibit flowrate sensitivity. In a few applications electrodes have indefinite life, but in many others deterioration of response occurs within days or hours. Nylander (1985) notes that by making membranes of different glasses electrodes for other ions such as sodium or potassium can be manufactured; and that various metal salts can be used as membranes in ion-selective electrodes in most cases with high enough electrical conductivity to be mounted directly onto a metal electrode. Another large group of electrodes have a liquid or polymer membrane with incorporated ion exchangers or neutral ion carriers; the liquid can be in gel form, or supported by an acetate membrane. Bergveld (1989) describes progress with ion-selective field effect transistors (ISFETs). An ion-sensitive film is deposited over the transistor gate; ions diffusing into the gate region polarise the interface between the gate and the fluid. While in the case of a glass membrane electrode only a small offset occurs, the ISFET sensor inevitably exhibits a large offset, related to the manufacturing process. Moreover, in the case ofan ISFET with two dielectric layers, additional charges may collect at the interface between the two insulators, while it isalso reported that the top layers show polarisation which will result in long-term drift. These effects do not exist in glass membrane electrodes because the conducting properties of the glass membrane prevent the occurrence of an electric field. Schemes have been proposed to null the offset of the basic ISFET sensor, and with further development semiconductor sensors may be expected to overtake glass membrane electrodes in performance. 2.3.4 Immobilised enzyme sensors Biosensors can be constructed by allowing an enzyme to produce ionic products from an organic species to be analysed. Product concentration is then measurable by ion-selective electrode. For example, by adding theIN-LINE SENSORS, 27 enzyme urease to urea solution, ammonium ions are released and concentra- tion can be determined with an ammonium-selective sensor. The selectivity of the enzyme is high, but the selectivity of the electrode is limited. Nylander (1985) points out that instead of adding enzyme to the analyte, itis preferable to immobilise the enzyme in or on supporting material for re-use, or for use in aflow through cell. Immobilised enzymes tend to be more stable than enzymes in solution, and the lifetime of some enzyme reactors for amino acids can be up to several months, Figure 2.4 shows how a potentiometric enzyme electrode can be based on a gas-sensitive electrode which in turn is based on an ion- selective electrode; the high selectivity of the gas-permeable membrane prevents interference from ionic species in the analyte. Initial applications have been for medical purposes and for environmental monitoring. Gaisford and Rawson (1989) report the use of whole cell biosensors for monitoring the effect of toxicants on biological oxygen demand (BOD) in waste water. Various systems based on oxygen electrodes have been investigated, but an alternative approach to monitoring the status of the bacterial cell is by accessing the electron transport systems involved in respiration or photosynthesis using a chemical mediator. By selection of suitable catalysts and immobilisation at the surface of a graphiteclectrode ina flowing solution, disturbance of the metabolic state of the organism can be detected by monitoring current pulses generated as a result of pulsed photosynthetic stimulation. Gaisford and Rawson quote close correlation of microbial electrode output with BOD for effluents containing starch and palm oil. The electrodes are low cost and may be treated as disposable items. Kress-Rogers et al, (1993) describe the development of biosensor array for meat freshness measurement, In meat, the depletion of glucose near the surface relative to the bulk provides an indication of microbial activity. A blade was construcied holding disposable sensor electrodes which penetrate to selected ‘Ag/AgCl WIRE ee HCL NH CI ‘4 ION SELECTIVE MEMBRANE GAS PERMEABLE MEMBRANE ENZYMES DIALYSIS MEMBRANE Figure 2.4 Enzyme electrode on gas-sensitive electrode on ion-selective membrane. (After Nylander, 1985, with permission of the Institute of Physics.)