Accepted Manuscript

A novel self-powered time-temperature integrator (TTI) using modified biofuel cell for
food quality monitoring

Min Jung Kim, Hye Won Shin, Seung Ju Lee

PII: S0956-7135(16)30265-1
DOI: 10.1016/j.foodcont.2016.05.031
Reference: JFCO 5048

To appear in: Food Control

Received Date: 11 January 2016

Revised Date: 30 April 2016
Accepted Date: 21 May 2016

Please cite this article as: Kim M.J., Shin H.W. & Lee S.J., A novel self-powered time-temperature
integrator (TTI) using modified biofuel cell for food quality monitoring, Food Control (2016), doi: 10.1016/
j.foodcont.2016.05.031.

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 ACCEPTED MANUSCRIPT

1 A novel self-powered time-temperature integrator (TTI) using

2 modified biofuel cell for food quality monitoring
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4 Min Jung Kim, Hye Won Shin, Seung Ju Lee*

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6 Center for Intelligent Agro-Food Packaging (CIFP), Department of Food Science and
7 Biotechnology, Dongguk University-Seoul, Seoul 100-715, Korea

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9 * Corresponding author at: Department of Food Science and Biotechnology, Dongguk

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10 University-Seoul, Seoul 100-715, Korea. Tel: 82-2-2260-3372, Fax: 82-2-2260-3372,
11 E-mail: Lseungju@dongguk.edu

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12
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13 ABSTRACT
14 A self-powered time–temperature integrator (TTI) was developed by modifying a biofuel cell and
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15 characterised in terms of kinetics and temperature dependency. The TTI was composed of two
16 immobilised enzyme electrodes, (+) and (-), and a cavity containing substrates. The enzymes (laccase
17 and glucose oxidase), mediators (methylene blue, 8-hydroxyquinoline-5-sulfonic acid hydrate,
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18 ferrocenium hexafluorophosphate, 4,4-sulfonydiphnol) and stabilisers (pyrrole) were immobilised on

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19 a glassy carbon electrode by electrodeposition by applying a square wave (10 s at 4 V then 3 s at 0.5
20 V) for 180 cycles. The primarily measured output voltage, which corresponds to a rate of food
21 quality change, was mathematically converted to a secondary variable, its integral value,
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22 representing the food quality level. The temperature dependency of the self-powered TTI was
23 modified by using sodium azide, and the shelf-life of TTI could be adjusted to match that of the food
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24 product by changing glucose concentrations. The optimum Arrhenius activation energy was found to
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25 be 78.22 kJ/mol with a sodium azide concentration of 0.02 M and the glucose concentration at the
26 maximum (5.05 M), it used continuously for 7 days at 25°C. Through a case study with milk, the
27 novel TTI was found to accurately detect milk quality changes. The self-powered TTI can be utilised
28 as an intelligent versatile sensor in the food distribution system with an appropriate IT platform. In
29 particular, if the TTI is incorporated with RFID tag, we will see a new generation of smart RFID tag
30 for intelligent food packaging.

31 Keywords : Time-temperature integrator, Biofuel cell, Arrhenius activation energy, Electro-deposition

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32 1. Introduction
33
34 The time–temperature integrator (TTI) is a small label which is attached on food packages. The
35 TTI colour and food quality change by experiencing a time–temperature history, and the food quality
36 level can be predicted by the TTI colour. TTIs play an important role in improving the efficiency of
37 the food distribution system by minimising economic losses caused by food quality changes (Tsoka,

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38 Taoukis, Christakopoulos, Kekos, & Macris, 1998; Simpson et al., 2012). As the food distribution
39 system has become broader, IT has become an integral part of the system. If a TTI has an electrical

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40 sensor function as well as a colour-changing indicator, it could be linked with IT technology, such as
41 radio frequency identification (RFID tag)/ubiquitous sensor network (USN) (Wan & Knoll, 2016).

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42 TTI production is based on physical, chemical, microbial or biological mechanisms, such as
43 diffusion–migration, polymerisation of monomers, microbial reactions or enzymatic reactions
44 (Taoukis & Labuza, 1989; Lee & Shin, 2012). Among these, the biological method is most suitable

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45 for adding an electrical function to TTI as biologically produced TTI is based on an oxidation reaction
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46 by the relevant enzyme. The oxidation transfers electrons from electron donors to electron receptors,
47 hence, it causes an electric current or voltage. Kim, Kim, and Lee (2012) developed a TTI based on an
48 enzymatic oxidation reaction, in which laccase and 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulphonic
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49 acid (ABTS) were used as an enzyme and substrate, respectively. Laccase is usually available in the
50 oxidised form and is able to oxidise the ABTS, thus producing an electron and changing the substrate
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51 colour.
52 A biofuel cell is a kind of battery that converts chemical energy produced by a redox reaction to
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53 electrical energy. A biofuel cell consists of two poles, the anode and the cathode, and glucose oxidase
54 and laccase with some mediators are immobilised to the anode and cathode, respectively (Sassolas,
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55 Blum & Leca-Bouvier, 2012). As mentioned above, laccase is commonly used both in TTIs and
56 biofuel cells (Chen et al, 2001; Brunel et al., 2007). In the biofuel cell, both oxidation and reduction
57 reactions are involved. The components of the cell system relay the electrons derived from the redox
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58 reactions, causing the electrons to flow from the anode to the cathode. A particular circuit is created to
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59 utilise the produced electron and in this way it can act as a battery. The components are redox
60 enzymes, redox mediators and substrates, such as laccase, glucose oxidase, alcohol dehydrogenase;
61 ABTS, osmium; and glucose or alcohol (electron donor) and oxygen (electron receptor), respectively.
62 Likewise, the material formulation of TTI is similar to that of a biofuel cell, and therefore it
63 could be hypothesised that a biofuel cell could be applied as a TTI. However, it should be
64 remembered that the power of a biofuel cell and colour change of a TTI owing to electron transfer are
65 different quantitatively. The electrical power reflects the rate of electron transfer, whereas any changes

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66 based on oxidation reflect the amount or concentration of electron transfer. Therefore, some
67 manipulation would be required to relate the biofuel cell power to the TTI response.
68 If a TTI is developed to have an electrical output like that of a biofuel cell, i.e. a self-powered
69 TTI, as mentioned above, innovative applications would be available as follows. Recently, USN has
70 been used for platforms of food distribution systems, in which RFID tag is employed as a sensor
71 (Jones, Clarke-Hill, Comfort, Hillier, & Shears, 2005; Regattieri, Gamberi, & Manzini, 2007). If a

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72 self-powered TTI is connected to the RFID tag, real-time food quality information can be provided to
73 optimise the management of food distribution. Actually, a smart RFID tag with a temperature sensor is
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available, but its function is totally different from that of RFID tag with a self-powered TTI. The
75 smart RFID tag just measures the temperature, but an RFID tag with a self-powered TTI measures the
76 food quality level exactly like a conventional TTI. Another merit of the new TTI is to be able to

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77 provide electrical power. This aspect is very important when any devices are connected to RFID tag,
78 because RFID tag may need the additional electrical power for the operations. Biofuel cells have been

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79 reported as a power source for active RFID tags.
80 In this study, a self-powered TTI was created by using a formulation modified from that of
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81 common biofuel cells. The modification was done in order to adjust two essential functions (self-life
82 and temperature dependency) required for TTI. Electrochemical immobilisation was employed to
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83 build the anode and cathode with enzymes and mediators. Substrates (different amounts of glucose
84 and sodium azide) were formulated to adjust the self-life and temperature dependency. The primary
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85 variable, TTI voltage output, was mathematically manipulated to estimate a secondary variable
86 appropriate for indicating food quality. The reaction kinetics and temperature dependency of the self-
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87 powered TTI were investigated in a case study.

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89 2. Material and methods

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91 2.1. Materials
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93 The materials were purchased from Sigma-Aldrich: laccase (Lac) from Trametes versicolor (1
94 units/mg), glucose oxidase (GOx) from Aspergillus niger (5.6 units/mg), methylene blue, 4,4-
95 sulfonydiphnol (SDP), 8-hydroxyquinoline-5-sulfonic acid hydrate (HQS), ferrocenium
96 hexafluorophosphate (FHFP), pyrrole, glucose and 0.01 M phosphate-buffered saline (pH 7.4). The
97 grade of water used was deionised water (≥18.2 MΩ · cm).

98

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99 2.2. Fabrication of a self-powered TTI based on a biofuel cell

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101 Through electrochemical immobilisation, an anode and a cathode with redox enzymes and
102 mediators fixed on their surface were produced. Pyrrole was used to fix the materials on the pole
103 surface through polymerisation by electrical treatment. As an electrical pole, glassy carbon (diameter
104 3 mm) was used. The enzymes and mediators for the immobilisation were laccase and glucose oxidase,

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105 and HQS, FHFP and methylene blue, respectively.
106 First, the anode of a carbon pole was produced by arranging a three-electrode cell with

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107 potentiostat (VSP-300, Bio-logic, France) as follows: a working electrode (carbon pole), reference
108 electrode (Ag/AgCl) and counter electrode (Pt). Then, the materials (GOx, methylene blue, SDP, HQS,

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109 FHFP and pyrrole) were placed in the bottle with the electrodes. Voltage was applied to the three-
110 electrode cell in square wave mode (180 cycles of 4 V for 10 sec and 0.5 V for 3 sec, consecutively)
111 (Fig. 1) or continuous polarisation mode (4 V for 30 min). After the immobilisation, the pole was

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112 cleaned with distilled water and dried at room temperature, then kept in phosphate-buffered saline (pH
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113 7.4) before use (Malika & Jan, 2010). In the same way, the cathode was made with laccase, methylene
114 blue, SDP, HQS, FHFP, pyrrole and sodium azide. The sodium azide is used to modify the common
115 temperature dependency of the biofuel cell to that of the TTI (Park, Kim, & Lee, 2013; Chang, Moon,
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116 Jang, & Kim, 2005). The electrodes of the TTI were immersed in substrate solution [0.3–5.5 M
117 glucose and 0.01 M PBS buffer (pH 7.4)] in an airtight 10-mL beaker.
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119
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2.3. Derivation of TTI response variable

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121 The voltage of the biofuel cell and the colour change of the redox TTI (e.g. laccase TTI) owing to
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122 electron transfer are different quantitatively. The electrical voltage reflects the rate of electron transfer
123 or current (the rate of one coulomb per second), whereas any changes based on oxidation reflect the
124
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amount or concentration of electron transfer. Therefore, the voltage output of biofuel cell was
125 mathematically manipulated to be appropriate as a TTI response variable:
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127 V ∝ dX/dt (1)
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129 where V is the output voltage and X is the concentration of electrons or substrates involved in the
130 redox reactions. It is X that is directly related to the accompanied food qualities, so Eq. (1) was
131 integrated to have X as the variable, in which a proportional constant was neglected in the derive Eq.
132 (2).
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134 X =  V dt (2)
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137 2.4. Analysis of kinetic and Arrhenius parameters of the TTI
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139 The voltage was used as a reaction rate to establish Arrhenius relations for temperature
140 dependency:

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142 V = k exp(   ) (3)
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144 where k0 is the Arrhenius constant, Ea is the activation energy (kJ/mol), R is the ideal gas constant
145 (8.314 × 10 kJ/K mol), and T is temperature (K). A system of measuring the TTI performance was

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146 created by connecting the two poles of the TTI to a potentiometer (recorder type, EYELA, Japan).
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The voltage output was measured during storage in a water bath under isothermal conditions at 5, 15
148 and 25°C.
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150 2.5. Prediction of food quality from TTI variable

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152 Usually in the prediction procedures, Teff is mathematically estimated from TTI response
153 variables, and then food quality is predicted at the Teff for a given time (Taoukis, Koutsoumanis &
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154 Nychas, 1999; Marco Riva, Piergiovanni, & Schiraldi, 2001; Shim, Jung, & Lee, 2013). However, in
155 this study, a relation established at a reference temperature was used as the calibration curve. This
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156 method was more practical, because all the kinetic and Arrhenius equations should be available for the
157 mathematical prediction procedure.
158 As aforementioned, a calibration curve was experimentally prepared by relating X of TTI to food
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159 quality as follows:

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161 Y= f (X) (4)
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163 where Y is a quality of the food accompanied with TTI, and f(X) is a calibration function between X
164 and Y (Kim, Choi, Kim, Kim, & Lee 2013).
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166 2.6. Case study: milk storage

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167
168 A case study to evaluate the TTI performance on food quality monitoring was executed with milk.
169 Low Temperature Long Time (LTLT) pasteurized milk, purchased 7 days before the expiration date
170 from L mart (Seoul, Korea), was used for test. The 200-mL carton-packaged milk and TTI were stored
171 in a lab incubator under dynamic conditions. A time–temperature history scenario was assumed for the
172 storage test. A potentiometer outside the incubator was wired to the TTI electrodes for voltage

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173 measurement. The milk packages were sampled periodically, and aerobic mesophilic bacteria (AMB),
174 known to have Ea = 110.35 (Kim, Kim, & Lee, 2012), were evaluated according to a standard
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microbial assay (KFDA, 2000; Rosmini, Signorini, Schneider, & Bonazza, 2004). AMB was
176 considered as quality index of milk (KAPQA, 2012). To perform microbiological assay, decimally
177 diluted milk samples were tested with PetrifilmTM aerobic count plate (ACP; 3M, St. Paul, MN, USA)

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178 and the microbial counts were expressed in CFU/mL.
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180 2.7. Statistical analysis
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182 The microbial data were statically treated to show their variances. To complete the kinetic and
183 Arrhenius models, regression analysis was performed. MS Excel was used as a computation tool.
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185 3. Results and discussion

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186
187 3.1. Output of self-powered TTI
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188
189 The self-powered TTI generated a constant voltage with respect to time under isothermal
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190 conditions. Two modes of electrochemical treatments were used, but the levels of voltage were
191 different (Fig 2). The square wave mode was more efficient at producing a higher voltage than the
192 continuous polarisation mode. It is because the use of a square wave mode instead of a continuous
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193 mode considerably decreases the electrolysis of water, improving the stability of the TTI (Malika &
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194 Jan, 2010).

195 The shelf-life of a TTI means the length of time from TTI activation to termination of TTI
196 response, normally colour, and should correspond to that of the accompanied food. In the self-
197 powered TTI, the TTI generated a constant voltage at a constant temperature (Fig. 2) and the shelf-life
198 of the new TTI was regarded as the length of time for which the output voltage is maintained at a
199 constant level.
200 Fig. 3 shows that the TTI shelf-life increases with glucose concentration. When the concentration

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201 of added glucose was 300 mM, 1 M and 2.5 M, the TTI shelf-life was 2.8, 6.5 and 76 hours,
202 respectively, and when the concentration of added glucose was 5.05 M, the shelf-life was over 168
203 hours (over 7 days). The maximum amount of dissolved glucose, 5.05 M, is determined by
204 considering the solubility of glucose, which is 909 mg/mL at 25 ℃.
205 Thus, the TTI shelf-life can be adjusted by varying the amount of glucose in the formulation.
206 After the self-powered TTI passes its shelf-life, the substrate, glucose, gradually diminishes at the

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207 anode of the TTI, leading to decreased output voltage.
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209 3.2. TTI response variable in the type of electrical signal
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211 If the primary variable, voltage output, is directly used as a variable of TTI response, its changes
212 cannot reflect an accompanied food quality change. Unless the self-powered TTI has a meaningful
213 output as the TTI response, the self-powered TTI cannot be a true TTI. To derive another TTI variable

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214 that corresponds to food quality changes, the voltage output was mathematically manipulated as
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215 follows:
216 In Eq. (1), V is the output voltage and X is the concentration of electrons or a reactant involved in
217 redox reactions taking place on both the anode and the cathode. The formula below corresponds to the
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218 redox reactions on the anode.

219
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220 Glucose + Mox → Gluconic acid + Mred

221 Mred → Mox + e-
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222
223 where M is the mediator and e- is the concentration of electrons, X. The produced electrons are
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224 supplied from the anode, flowing to the cathode. In the regular interpretation of TTI colour response,
225 the substrate or product concentrations in the TTI formulations are essentially the colour origins. X is
226 one of the products and therefore is directly related to the associated food quality. Consequently, X is
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227 a true variable of TTI response rather than the primary variable, V. The output voltage was
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228 mathematically converted to X through integration. Fig. 4 shows X (an integral of V) with respect to
229 time. Unlike V which are constant, the integrals of V are shown to change with time as food quality
230 changes during storage.
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232 3.3 Kinetic and Arrhenius parameters

233 Fig. 4 shows that the output voltage is temperature-dependent, increasing with temperature. To
234 quantify the temperature dependency, the Arrhenius activation energy (Ea) was estimated from the

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235 relationship between 1/T and ln V. Fig. 5 shows the data from the TTI without a modification from
236 that of the common biofuel cell. As seen in Table 1, Ea of the unmodified cell is too low to be used
237 with any foods. Therefore, the formulation of the TTI was modified by using sodium azide, which is
238 known as an additive for adjusting Ea of TTI (Park, Kim, & Lee, 2013).
239 The output voltage increased with the sodium azide concentration (Table 1). It was speculated that
240 as sodium azide is a salt it dissociated in the solution, increasing the electrical conductivity and thus

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241 facilitating the redox reaction. The Ea increased with the concentration of sodium azide in the initial
242 stage, but decreased above 0.02 M of sodium azide. Park, Kim and Lee (2013) also reported a similar
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result, that the addition of sodium azide could change the Ea of TTI significantly.
244 The Ea values ranged from 52.4 to 78.2 kJ/mol. When compared to typical Ea values for food
245 quality losses (Labuza 1982; Labuza & Kamman, 1983), the activation energy of this novel TTI can

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246 cover the quality losses that occur due to enzymatic changes (41.84–62.76 kJ/mol), hydrolysis (62.76
247 kJ/mol), or lipid oxidation (41.84–104.6 kJ/mol). The activation energy of this TTI was comparable to

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248 a commercially available enzymatic TTI (Vitsab AB, Malmö, Sweden) type C2–15d (Ea = 50.2
249 kJ/mol) (Bobelyn, Hertog & Nicolaï, 2006) and a diffusion based TTI (Ea = 33–50 kJ/mol) (Poças,
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250 Delgado & Oliveira, 2008). In addition, the allowable difference in activation energy between the
251 food and the TTIs must be within ± 25 kJ/mol (Taoukis, 2001) to predict food quality with less than
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252 a 15% error.

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254
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255 3.4. Case study under dynamic conditions

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257 The newly created self-powered TTI, which is based on a biofuel cell in principle, was evaluated
258 with an application to milk packaging. As a routine procedure, the similarity in temperature
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259 dependency between TTI and milk was examined. To quantify the temperature dependency of AMB
260 in milk, Ea was estimated from the regression analysis between temperature (1/T) and reaction rate (ln
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261 k). The Ea of AMB was found to be 101 kJ/mol. AMB in another food matrix, beef, was reported to
262 be 110 kJ/mol (Kim, Choi, Kim, Kim, & Lee 2013). TTI with the allowable Ea (Ea difference from
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263 milk <25 kJ/mol) was found to be at 0.02 M sodium azide (Table 1).
264 A relation established at a reference temperature (7°C) between the integral of the output voltage,
265 X of TTI and AMB was illustrated as a calibration curve (Fig. 6). 7°C was considered as a standard
266 temperature for the calibration curve. It was because refrigeration of pasteurized milk at or less than
267 7.2°C is regarded as “practical storage conditions”, the conditions that can still retain an acceptable
268 quality of pasteurized milk (DPC, 2003). This calibration curve was used when reading the AMB
269 level at a given X from the experiments to calculate the expected levels of AMB.
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270 Fig. 7 shows the time course of the TTI output voltage and temperature under dynamic
271 conditions in the case study. The output voltage was converted to its integral (Fig. 8), which was
272 substituted into the calibration curve. In Fig. 9, the predicted and experimental AMB show the similar
273 tendency to increase with storage time.
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275 4. Conclusion

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276
277 A self-powered TTI, which outputs an electrical signal (voltage), was developed by modification

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278 of a biofuel cell (BFC). The material compositions of BFC and redox enzyme-based TTIs are similar,
279 which led to the development of a self-powered TTI from a BFC. However, the characteristics of the

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280 BFC, such as temperature dependency and shelf-life, limit its application as a TTI for food. Moreover,
281 it was doubted that the electrical signal of the new TTI could represent food quality. Through some
282 modifications, these flaws were addressed. The temperature dependency of the BFC was modified by

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283 using sodium azide, and the shelf life was prolonged to the appropriate levels for TTI use by adjusting
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284 the substrate concentration. The primary output voltage was mathematically converted to be a
285 secondary variable representing food quality. Through a case study, the novel TTI could successfully
286 predict milk quality changes. Actually, the shelf life and temperature dependency tested in this paper
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287 would be limited to cover a variety of foods. So the further work on varying them would be necessary
288 in the future study. The self-powered TTI would be beneficial as a sensor which can be incorporated
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289 with RFID tag, a new generation of smart RFID tag for intelligent food packaging.
290
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291 Acknowledgment
292
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293 This research was supported by the R&D Convergence Center Support Program (710003-03) of
294 the Ministry for Food, Agriculture, Forestry and Fisheries, Republic of Korea.
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296
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363 Wan, X., & Knoll, M. (2016). A new type of TTI based on an electrochemical pseudo transistor.
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364 Journal of Food Engineering, 168, 79–83.

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11
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Table 1
Arrhenius parameters of output voltage of the self-powered TTI with different sodium azide
concentrations.
Sodium azide concentration Temperature Voltage Ea
R2
(M) (℃) (mV) (kJ/mol)

5 0.0008

PT
0 15 0.0018 52.756 0.9998
25 0.0037

RI
5 0.0008
0.002 15 0.0019 53.664 1.0000
25 0.0038

SC
5 0.0025
0.004 15 0.0055 56.726 0.9980
25 0.013

5
U 0.0035
AN
0.006 15 0.009 57.406 0.9967
25 0.0185
M

5 0.0042
0.008 15 0.0094 57.689 0.9960
25 0.024
D

5 0.0058
TE

0.01 15 0.0155 61.697 0.9985

25 0.0345

5 0.00615
EP

0.02 15 0.0357 78.222 0.9174

25 0.0588
C

5 0.006
0.04 15 0.018 65.850 0.9956
AC

25 0.0405

5 0.007
0.06 15 0.017 52.421 0.9942
25 0.032
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Figure Captions

Fig. 1. (A) Square wave voltage–time used for the electrodeposition of the enzyme–
monomer–redox mediators mixture from ultrapure water. (B) Continuous at 4 V vs.
Ag/AgCl for comparison.

PT
Fig. 2. The output voltages of the self-powered TTI at different temperatures and the effects
of potentiostat direct current potential wave types used in producing the TTI.

RI
Fig. 3. Shelf life of the self-powered TTI with different concentrations of glucose. The shelf

SC
life means the length of time for which the output voltage is maintained at a constant
level.

U
Fig. 4. TTI response variable (X, integral of V) with respect to time. It was created by
integration of the output voltage (V). X changes linearly at each temperature,
AN
indicating a zero-order reaction.
:5 , : 15 , : 25 .
M

Fig. 5. Arrhenius plot of 1/T vs. ln V of TTI.

D

Fig. 6. Relation, a calibration curve, between TTI response variable (integral of voltage
TE

output) and aerobic mesophilic bacteria (AMB) concentration in milk at 7 . Error

bars represent standard deviations (n=3).

EP

Fig. 7. Change in the voltage output of TTI under dynamic temperature conditions for a case
study.
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: voltage, : temperature.
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Fig. 8. Change in the integral of voltage output of TTI under dynamic temperature conditions
for a case study.

: integral of voltage, : temperature.

Fig. 9. Comparison between experimental and TTI-predicted AMB levels of milk in a case
study (Fig. 6). Error bars represent standard deviations (n=3).

: predicted levels, : experimental levels.

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4.5 4.5
4 4
3.5 3.5
3 3
Voltage (V)

Voltage (V)
2.5 2.5

PT
2 2
1.5 1.5

RI
1 1
0.5 0.5

SC
0 0
0 20 40 60 80 100 0 5 10 15
Time (sec) Time (sec)

(A)
U (B)
AN
Fig. 1.
M

0.045
D

0.04

0.035
TE

0.03
voltage (V)

EP

0.025

0.02
C

0.015
AC

0.01

0.005

0
5℃
℃ 15℃
℃ 25℃
℃

Square wave Direct current

Fig. 2.
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0.0228

0.0226

0.0224
Voltage (V)

PT
0.0222

RI
SC
0.0220

U
0.0218
0 20 40 60 80 100 120 140 160
AN
Time (hr)

300 mM 1M 2.5 M 5.05 M

M

Fig. 3.
D
TE
C EP
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0.045

0.04

0.035
Voltage (V)

0.03

PT
0.025

0.02

RI
0.015

0.01

SC
0.005

U
0
0 10 20 30 40 50 60 70
AN
Time (hr)
3
M

2.5
D
Voltage (V)

2
TE

1.5
EP

1
C

0.5
AC

0
0 10 20 30 40 50 60 70
Time (hr)

Fig. 4.
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-3.1
0.0033 0.00335 0.0034 0.00345 0.0035 0.00355 0.0036 0.00365
-3.2

-3.3

-3.4

PT
ln V

y = -1983.4x + 3.412
-3.5 R² = 0.9596

RI
-3.6

SC
-3.7

-3.8
1/T

U
AN
Fig. 5.
M

7
D

6
TE

5
AMB (Log CFU/mL)

EP

4
y = 0.7356x + 1.906
3 R² = 0.9823
C
AC

0
0 1 2 3 4 5 6
TTI

Fig. 6.
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0.045 30

0.04
25
0.035

0.03 20

℃)
Temperature (℃
Voltage (V)

PT
0.025
15
0.02

RI
0.015 10

SC
0.01
5
0.005

U
AN
0 0
0 20 40 60 80 100 120 140 160
Time (hr)
M

Fig. 7.
5 30
D

4.5
TE

25
4

3.5

℃)
Temperatrue (℃
EP

20
3
Axis Title

2.5 15
C

2
AC

10
1.5

1
5
0.5

0 0
0 20 40 60 80 100 120 140 160
Time (hr)

Fig. 8.
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4.5

4
AMB (Log CFU/mL)

3.5

PT
3

2.5

RI
2

SC
1.5

0.5
U
AN
0
0 10 20 30 40 50 60 70 80 90 100
M

Time (hr)

Fig. 9.
D
TE
C EP
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Highlights

- A novel self-powered time-temperature integrator (TTI).

- The novel TTI outputs an electrical signal instead of color change.

- The novel TTI was created by modifying a biofuel cell.

PT
- The novel TTI performance was validated in a case study with milk deterioration.

RI
U SC
AN
M
D
TE
C EP
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