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Development of Arduino Uno-Based TCS3200 Color Sensor and Its Application
on the Determination of Rhodamine B Level in Syrup
Muhammad Syukri Surbakti
1
, Muhammad Farhan
2
, Zakaria Zakaria
1
, Muhammad Isa
1
, Elly Sufriadi
2
,
Sagir Alva
3
, Elin Yusibani
1
, Leni Heliawati
4
, Muhammad Iqhrammullah
5
, and Khairi Suhud
2*

1
Department of Physics, Faculty of Mathematics and Natural Sciences, Universitas Syiah Kuala, Banda Aceh 23111, Indonesia
2
Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Syiah Kuala, Banda Aceh 23111, Indonesia
3
Department of Mechanical Engineering, Faculty of Engineering, Mercubuana University, Jakarta 11650, Indonesia
4
Department of Chemistry, Faculty of Mathematics and Natural Sciences, Ibnu Khaldun University, Bogor 16162, Indonesia
5
Graduate School of Mathematics and Applied Science, Universitas Syiah Kuala, Banda Aceh 23111, Indonesia
* Corresponding author:
tel: +62-85297492376
email: [email protected]
Received: September 18, 2021
Accepted: February 1, 2022
DOI: 10.22146/ijc.69214

Abstract: The use of the notorious synthetic dye, rhodamine B, in food and beverage
products has been widely reported. This application urges the need to develop an
analytical method that can provide reliable rhodamine B data with an easy operational
technique. Therefore, this research is aimed to develop an Arduino Uno-based TCS3200
color sensor and study its application to determine rhodamine B levels in syrup. The
design of the analytical instrument included TCS3200, an Arduino Uno microcomputer,
an Integrated Development Environment (IDE) software, a black box container, and a
24 × 2 matrix display screen, where samples were prepared via absorption using wool
thread. With a linear range of 1–20 mg/L, our proposed colorimetric sensor had recoveries
of 96.25–110.3%, which was better compared to that was obtained from the UV-vis (81.8–
100.6%) method. The detection and quantification limits of the sensor were 2.766 and
8.383 mg/L, respectively. The syrup samples used in this study were purchased from the
local stores in Banda Aceh. Based on the proposed TCS3200 color sensor, the highest
rhodamine B concentration from the syrup sample was 16.74 mg/L. The t-test analysis in
this study revealed that the Rhodamine B levels quantified using the newly developed
TCS3200 color sensor were not statistically or significantly different from the UV-Vis
spectrophotometer method.
Keywords: color sensor; TCS3200; rhodamine B; Arduino Uno; Zn(CNS)
2; IDE software

■ INTRODUCTION
As a form of consumer protection efforts, sensor
technology for food or beverage products has been
developed intensely [1-3]. For example, Fourier Transform
Infrared (FTIR) spectroscopy has been employed to
separate halal and non-halal meatballs [4]. The color
spectroscopy method has also reported the detection and
analysis of synthetic dye rhodamine B in chili powder [5].
Dyes used in food products are derived from natural and
artificial chemicals. Synthetic dyes are widely used
because it is more cost-efficient and available.
Additionally, the synthetic dye may give a brighter color
to the food or beverage product [6].
As one of 30 synthetic dyes available, Rhodamine
B is considered as a dangerous dye, where its use in food
or beverage products has been prohibited. Nevertheless,
Rhodamine B is often used in processing industries,
papers, and fabrics [7-8]. Moreover, it could be
employed as a ligand to bind metal ions [9]. However, in
Indonesia, rhodamine B is still very popular as a food
coloring agent, including in iced syrup. The syrup is
intentionally added with rhodamine B, so the products
obtain a more attractive appearance [10].

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Methods that have been previously developed to
identify rhodamine B in food ingredients include thin-
layer chromatography (TLC) [11], voltammetry [12], and
the standard method using UV-Vis spectrophotometer
(the best option for identifying compounds with color).
However, UV-Vis spectrophotometry has several
drawbacks; not portable, complicated, and expensive [13].
Hence, this study tried to overcome the stated drawbacks
by developing a simple measurement method using a
portable sensor.
The sensor system developed in the present work
was based on the TCS3200 color sensor constructed by the
console to overcome external noise and program library
modification [14]. The TCS3200 color sensor has been
widely reported for different applications, including
measuring levels of cyanide [14], nitrogen [15], and heavy
metals [16]. In the case of colorimetric sensors, analytes
should first be reacted with a complexing agent to cause a
color change [17-18]. In this study, the sensor detects
color degradation from tissue paper that has been spiked
with reagents, so its sensitivity is specifically improved for
rhodamine B analysis. The reagent used was Zn(CNS)
2,
which can cause a color change from red to purple due to
the formation of the rhodamine B-Zn-thiocyanate ((RhB)
complex 2Zn(CNS)
4) [19].
The color intensity contributed by the presence of
rhodamine B was converted through the sensor output
pin in the form of a square signal in which its frequency
depends on the concentration. The box's signal with
varied frequency was then processed using a
microcontroller on Arduino Uno. In this processing, four
filters were used, namely green, blue, red, and no filter. In
this case, no filter was excluded because the three
parameters were sufficient to represent the color
degradation of rhodamine B in the sample [20]. Filter
settings were performed by providing low and high logic
in the Arduino IDE program, following the reported
study [21].
The distance between the sample and the 8 × 8 diode
array was set at 3 cm, following the sensor system's
geometry. The console's color was made black so the color
could be absorbed fully, and influence from the
degradation of colors that enter the diode array could be
avoided. After obtaining the concentration of
rhodamine B using the Arduino Uno-based TCS3200
color sensor, the results were compared with the
standard UV-Vis spectrophotometric method. Finally,
the analysis results were compared to obtain the data on
sensitivity and accuracy of the newly developed sensor
[22].
■ EXPERIMENTAL SECTION
Materials
The materials used were a UV-Vis
spectrophotometer (Thermo Fisher Scientific, Selangor
Malaysia), a color sensor TCS3200 (ICTAOS/AMS), a
console, and an Arduino Uno (wavgat). Syrup samples
tested for rhodamine B levels were procured from local
stores in Banda Aceh. The standard rhodamine B was
purchased from The National Agency of Drug and Food
Control of Indonesia (BPOM RI). All other chemicals
used, i.e. NH
4OH, NaOH, HCl, C2H5OH, CH3CO2Na,
ZnCl
2, CH3CO2H, and KCNS, were obtained from
Merck (Selangor, Malaysia) in analytical grade.
Hardware Design
The hardware design was initiated by developing a
console for the TCS3200 color sensor, then connecting
the output port of the color sensor via a jumper cable to
the Arduino Uno microcomputer to process frequency
data and convert it into 8-bit RGB digital data. There
were 256 color digit variations for each RGB color
component that was sortable and distinguishable by the
processing. These color digit variations were displayed
on the computer screen and converted to reduce color
variations. These color variations were also recorded in
.xls format (MS Excel) (Fig. 1).
Development of the TCS3200 Sensor Console
The TCS3200 console sensor was designed in black
to absorb all color wavelengths. The distance between
the diode array and the color object was 3 cm. The
console was arranged in such a way that light from
outside could not enter. The TCS3200 sensor was
positioned opposite the color sample, which was
absorbed into a filter paper. Four LED units with white

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Fig 1. Schematic diagram of hardware design

wavelengths would hit the filter paper, and the intensity
light reflected the diode array following the color intensity
of the sample.
Software Design
Construction of the software design was initiated
with a blink test on the Arduino Uno system to determine
the response and performance of the microcomputer. The
software used was Arduino IDE with available open-
source libraries – C programming language. The program
library was modified to enable the required color filters,
Arduino Uno pins, the required display format, and data
storage mode (Fig. 2).
Rhodamine Analysis Using TCS3200 Color Sensor
Construction of the calibration curve for rhodamine B
Briefly, the rhodamine B solution was added with
3.0 mL of Zn-thiocyanate. Then, the standard solution
(RhB)
2-Zn(CNS)4 with different concentrations measured
the RGB value with the TCS3200 color sensor and
absorbance with UV-Vis at the maximum wavelength
obtained. The solution was prepared with 100 mg/L
rhodamine B as the stock solution, which was then diluted
using distilled water into standard solutions with varying
concentrations ranging from 1 to 20 mg/L. These
solutions were prepared to determine the maximum
wavelength of rhodamine B and as a database for the
TCS3200 color sensor. Following that, a solution of 1 mL
ZnCl
2 2 M and 2 mL KCNS 2 M as a reagent was made to
detect the presence of rhodamine B, as suggested by a
previous report [23].

Fig 2. Display of the Arduino IDE Software main menu
Determination of rhodamine B level using the
TCS3200
color sensor
The standard curve of (RhB)2-Zn(CNS)4 was
obtained by measuring the RGB values of the standard
solution (RhB)
2-Zn(CNS)4 using the TCS3200 sensor.
The concentration of rhodamine B used was 1 to 20
mg/L, which were priorly reacted with reagents.
Measurements were carried out three times, and the
concentration was averaged. Thereafter, RGB values
were converted into a color index, namely Hue,
Intensity, and Saturation (HIS). Conversion of RGB
values to HIS values was carried out using the following
Equations.
R
R
Red color index (I )
RGB

 (1)

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G
G
Green color index (I )
RGB

 (2)
B
B
Blue color index (I )
RGB

 (3)
The HIS color model was designed to resemble the
perception of human vision, while the RGB values
resembled the image of the display system [20]. The
results of the calculation of the HIS value were then
plotted as the dependent variable (y-axis) to the variation
of concentration (RhB)
2-Zn(CNS)4 (x-axis).
TCS3200 color sensor method validation
Method validation included accuracy, precision,
sensitivity, and linearity, which were conducted based on
the suggestion from a previous report [22].
Syrup sample preparation
Samples of commercial red syrup were purchased
from local stores in Banda Aceh. Each sample (10 mL) was
taken and put into an Erlenmeyer which was subsequently
mixed in 20 mL of 25% ammonia solution (dissolved in
70% ethanol) for 24 h and evaporated on a hot plate. The
evaporation residue was dissolved in 10 mL distilled water
containing acid (10 mL distilled water and 5 mL acetic
acid 10%). Wool thread (15 cm) was dipped into the acid
solution and simmered for 10 min until the dye colors
appeared on the wool thread, then lifted. The wool thread
was then washed with distilled water, and the wool thread
was dissolved in ethanol 70% and heated to a boil (Fig. 3).
This solution was used as the sample, per suggestion by a
published work [24]. The wool thread was used to extract

Fig 3. The extraction of rhodamine B from commercial
red syrups using wool thread. Wool thread was dipped
into the dissolved syrup residue for 10 min (a).
Rhodamine B-containing wool thread before re-
immersed to ethanol 70% and boiled (b)
rhodamine B-containing samples in an acidic
environment. A comparative study has reported that
wool thread has the highest dye adsorption as compared
with silk and nylon [25]. Adsorption of dye analyte in
wool thread is determined by its O- and N-containing
functional groups, which has been reported in many
published papers [26-28]. The dyed wool was then
immersed in ethanol 70% and boiled until its original
color returned. The obtained solution was analyzed for
its rhodamine B levels using the TCS3200 color sensor
and a reference method – UV-Vis spectrophotometry.
Quantitative Analysis
The prepared sample was added with Zn-
thiocyanate and then dipped in filter paper. Rhodamine
B levels were measured using the TCS3200 color sensor
[29]. The concentration was obtained based on the linear
equation obtained from the calibration curve.
Method Comparison using Two-Way t-Test
Results of the samples between the TCS3200 color
sensor and the UV-Vis spectrophotometry method were
compared [30]. In addition, a two-way t-test was carried
out to see the significance between the newly studied
TCS3200 color sensor method and the reference method
using UV-Vis spectroscopy by calculating the t value for
each method and then comparing it with the t
theoretical.
■ RESULTS AND DISCUSSION
Maximum λ of Rhodamine B Complex
The complex (RhB)2-Zn(CNS)4 was produced to
give rhodamine B a specific color, allowing easier
analysis. The solution of rhodamine B, which was
initially red, turned to purple and was then measured
using a UV-Vis spectrophotometer at a wavelength
ranging from 574 to 600 nm. The UV-vis absorbance
corresponding to the (RhB)
2-Zn(CNS)4 complex
scanned from 574 to 600 nm is presented in Fig. 4.
Based on the measurement results, the UV-Vis
spectrometer spectrum of (RhB)
2-Zn(CNS)4 showed a
maximum absorption (0.442 au) at a wavelength of
590 nm. The difference in wavelength between
rhodamine B and (RhB)
2-Zn(CNS)4 is due to a shift in

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Fig 4. UV-Vis spectrometer spectrum of (RhB)
2-Zn(CNS)4 showing a maximum wavelength at 590 nm

wavelength towards the bathochromic direction caused
by substitution, solvent effects, and the influence of the
chromophore group [31]. The successful formation of the
(RhB)
2-Zn(CNS)4 complex was indicated by a color
change from red to purple and a shift in wavelength. The
equation for the reaction between rhodamine B and
Zn(CNS)
2 can be seen in Fig. 5.
Based on the graph, we can see three regression
equations obtained from each RGB index value, namely I
R
y = 0.0028x + 0.3411; I
G y = 0.0032x + 0.3513 and IB y = -
0.0058x + 0.3059. The values of the determination
coefficient (R
2
) were 0.9792,0.9700, and 0.9729
respectively. The R index had the best determination
coefficient (R
2
) of 0.9792. Therefore, the regression
equation for the R index was used to determine the
concentration of rhodamine B in the sample.
Measurement using UV-Vis Spectrophotometer
The standard curve of (RhB)2-Zn(CNS)4 was
measured at a wavelength of 590 nm by a UV-Vis
spectrophotometer. The concentration of rhodamine B
that was used ranged from 1 to 20 mg/L, which was
priorly reacted with reagents. Measurements were
carried out three times and averaged for each
concentration. The absorbance measurements can be
seen in Fig. 6. The regression equation y = 0.0023x +
0.0773 had a
determination coefficient (R
2
) of 0.9927.
Hence, it can be concluded that the concentration was

Fig 5. Calibration curve for (RhB)
2-Zn(CNS)4

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directly proportional to the absorbance, meaning that the
absorbance for the complex (RhB)
2-Zn(CNS)4 was
dependent on rhodamine B concentration.
Method Validation
Accuracy
The accuracy of the proposed sensor method was
based on the recovery (%), representing the value
proximity of the standard concentration solution to the
actual concentration. The concentrations of (RhB)
2-
Zn(CNS)
4 used were 1, 10, and 20 mg/L for the analysis
with TCS3200 and UV-Vis color sensors. The actual
concentration values and the percent recovery values
from each method can be seen in Table 1. The recovery %
calculation for the TCS3200 color sensor was still within
the allowable error range of 90–110% [32]. However, at a
concentration of 1 mg/L, UV-Vis had a recovery value
below the permissible range (81.8%). Therefore, our
proposed method was suggested to have better accuracy
for determining rhodamine B levels at a low
concentration (1 mg/L).
Precision
The precision was determined to see the proximity
of the value changes in the repetition process. The
precision value was derived from the standard curve
with a respective concentration of (RhB)
2-Zn(CNS)4 (1,
10, and 20 mg/L), expressed by the variation coefficient
(VC). The precision values for both methods based on
intra-day and inter-day repetition are presented in Table
2. The variation coefficient value obtained by the two
measurements increased with the decrement in the
concentration of the standard solution. The method is
accurate if it provides a variation coefficient value of less
than 2% [32]. Nonetheless, inter-day repetition yielded
higher variation coefficient, especially when rhodamine
B concentration was 1 mg/L.
Linearity
Linearity is the functional area of sample
measurement. The linearity of measurements using the
TCS3200 color sensor and UV-Vis spectrophotometer
for a concentration range of 1–20 mg/L is depicted in
Fig. 5 and 6, respectively. Several studies used a


Fig 6. Calibration curve for (RhB)
2-Zn(CNS)4
Table 1. Recovery percentages of TCS3200 sensor and UV-vis spectrophotometer
Concentration (mg/L)
Actual concentration
(mg/L)
Recovery (%)
TCS3200 UV-Vis TCS3200 UV-Vis
1 1.030 0.818 103.5 81.80
10 11.03 10.06 110.3 100.6
20 19.25 19.03 96.25 95.15

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Table 2. VC values of TCS3200 sensor and UV-vis spectrophotometer obtained from intra-day and inter-day repetition
[Rhodamine B] (mg/L)
Intra-day variation coefficient (%) Inter-day variation coefficient (%)
TCS3200 UV-Vis TCS3200 UV-Vis
1 0.291 0.721 7.966 8.563
10 0.268 0.521 1.294 1.664
20 0.253 0.357 0.509 0.851

non-linear calibration curve because the sensor system
formed an exponential response [33]. However, in the
present study, the quantitative analysis was conducted
based on linear regression.
Sensitivity
The sensitivity value is shown from the slope of the
complex standard curve of (RhB)
2-Zn(CNS)4 for each
method. Based on the linear regression standard curve
equation, the slope value for the TCS3200 color sensor
measurement method was obtained from the regression
equation y = 0.0028x + 0.3411, which was 0.0028. While
the slope value for the UV-Vis spectrophotometer
measurement method was obtained from the regression
equation y = 0.0253x + 0.0773 is 0.0253. Based on the
constructed standard curve, we calculated the limit of
detection (LOD) by multiplying the standard deviation of
response by 3.3 and dividing with the slope. Meanwhile,
the limit of quantification (LOQ) could be obtained by
multiplying the standard deviation of response by 10 and
dividing it with the slope. The LOD obtained for the
TCS3200 color sensor and UV-Vis spectrophotometer
was 2.766 and 1.715 mg/L, respectively. These values
explain why the inter-day precision for the 1 mg/L
rhodamine B sample obtained for both methods exceeded
the acceptable maximum variation coefficient (< 2%). As
for the LOQ, the values reached 8.383 and 5.196 mg/L for
the TCS3200 sensor and UV-Vis spectrophotometer,
respectively. Lower LOD and LOQ in UV-Vis
spectrophotometer suggest its superiority in comparison
to the TCS3200 color sensor, in terms of sensitivity.
Quantitative Analysis Using the TCS3200 Color
Sensor
Samples were measured using a series of tools that had
been readily connected to the TCS3200 color sensor. The
measurement was carried out by dipping the filter paper
into the sample solution to which 3 mL of Zn(CNS)
2
reagent had been added, then dried and measured using
the TCS3200 color sensor in dark conditions.
Measurements were carried out three times on each
sample with 3 cm-long distance between the sensor and
the sample. Such distance was given to allow even
distribution of the emitted light from four Light
Emitting Diodes (LEDs) to the sample and the
photodiode, in which the sample could emit a current
proportional to the basic color of received light.
Table 3 shows that the RGB value obtained from
each sample is a code to indicate a specific color. The
HIS value in the table was obtained using Eq. (1-3). The
I
R value was used to determine the concentration of
rhodamine B in the sample because it had the best R
2

(0.9792) among the others (Fig. 6). The total
concentration of rhodamine B obtained from the
measurement using the TCS3200 color sensor based on
the I
R value can be observed in Table 4, showing the
concentration of each sample with five repetitions. The
red index value obtained from Eq. (1) has the same
function as the absorbance value, the dependent variable
in determining the concentration. Therefore, the
concentration of rhodamine B in the sample was
calculated by substituting the red color index value of the
sample into the standard curve regression equation
(RhB)
2-Zn(CNS)4 R index.
Following the analysis, we found that samples A, B,
and C contained rhodamine B with an average of 1.74,
16.74, and 5.10 mg/L, respectively. However, sample A
had a rhodamine B concentration lower than the LOD
of both the TCS3200 and UV-Vis spectrophotometer
(2.766 and 1.715 mg/L, respectively). In this case, the
response generated from sample A could not be
differentiated from that of the blank standard. Hence,
the presence of rhodamine B in sample A could not be

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Table 3. RGB value samples
Repetition
RGB Measurement HIS Value Measurement
Color
R G B I R I G I B
Sample A
1 232 213 224 0.346 0.317 0.334
2 233 213 224 0.347 0.317 0.334
3 233 212 224 0.348 0.317 0.334
4 231 211 222 0.343 0.312 0.330
5 230 210 223 0.346 0.316 0.336
A 231.8 211.8 223.4 0.345 0.316 0.334
Sample B
1 206 108 215 0.389 0.203 0.406
2 206 107 216 0.388 0.202 0.407
3 206 108 216 0.386 0.203 0.407
4 205 106 215 0.389 0.201 0.408
5 206 107 215 0.388 0.203 0.407
D 205.2 107.2 215.4 0.388 0.203 0.407
Sample C
1 222 173 228 0.356 0.278 0.366
2 221 173 227 0.355 0.278 0.365
3 221 172 226 0.357 0.276 0.366
4 220 171 227 0.355 0.276 0.367
5 220 172 228 0.354 0.277 0.367
F 220.8 172.2 227.2 0.355 0.276 0.366
Table 4. Sample concentration value of TCS3200 color sensor
Sample (X)
Repetition (mg/L)

X (mg/L)
1 2 3 4 5
A 1.75 2.10 2.46 0.67 1.75 1.74
B 17.10 16.75 16.03 17.10 16.75 16.74
C 5.32 4.96 5.67 4.96 4.61 5.10

confirmed by either method. As for sample C, the
calculated concentration was lower than the LOQ of the
TCS3200. Although its presence was confirmed, its
quantitative concentration value was not reliable.
Therefore, for the following analysis of comparing
TCS3200 with UV-Vis spectrophotometer, samples A and
C were excluded.
Comparing Methods Betw een the TCS3200 Color
Sensor with UV-Vis Spectrophotometry Using the
Two-Way t-Test
Method comparisons were carried out to see whether
the TCS3200 color sensor had similar results to a UV-Vis
spectrophotometer. The prepared samples were measured
for five repetitions with UV-Vis at a wavelength of 590 nm
and TCS 3200. Concentrations of rhodamine B in sample
B were 16.74 and 17.26 mg/L for measurements using
TCS3200 and UV-Vis spectrophotometer, respectively.
T-test (a = 8.95%) performed on the obtained data
revealed that the t
experimental and ttheoretical values were 1.21
and 2.31, respectively. Therefore, H
0 is accepted because
the value of t
experimental < ttheoretical. H0 states that differences
of data obtained from TCS3200 and the UV-Vis
spectrophotometer are not meaningful or significant.
This analysis validates the high concentration of
rhodamine B in sample B, calculated using TCS3200. It
is worth mentioning that high concentrations of
rhodamine B exposed to the human body could cause
adverse health effects [34].
■ CONCLUSION
The analytical performance of the newly developed
TCS3200 color sensor was satisfactory, considering that

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638
the analysis could be run in situ and available at an
affordable cost, and the components were free to access.
The results showed that the color gradation only occurred
in the R (Red) component, while the other components,
G (Green) and B (Blue), were not concentration-
depended. The t-test results showed that t
experimental <
t
theoretical suggesting the absence of statistical significance
between the results obtained from the TCS3200 color
sensor and the UV-Vis spectrophotometric method. The
syrup samples procured from the local stores in Banda
Aceh were tested qualitatively and quantitatively and was
found to contain rhodamine B with high concentrations.
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