1 Supporting Information for

2 Point-of-Care Photoresistive and Colorimetric Sensor for

3 Iron Detection in Environmental and Biological Samples
4 with Spectroscopic Validation
5 Shrikant Kashyapa, Harendra Singha, Purushotam Shivshakti Mahana, Sibnath Kayalb, Tapas K
6 Mandala*
a
7 Department of Chemical Engineering, Indian Institute of Technology, Guwahati, Assam,
8 India 781039

b
9 Sharjah Maritime Academy, Khor Fakkan, Sharjah, United Arab Emirates

*
10 Corresponding author tapasche@[Link]

11

12

13 Supporting Information (SI)

14

15 Content of SI
16 Total pages: 6
17 Number of figures: 06 (Figure S1-S6)
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S1
26 S1. Pretreatment of Samples

27 We employed a two-step protocol for the samples collected from environmental sources
28 (river, borewell, tap) to remove particulates and suspended solids that could interfere with the
29 analysis. In the first step, samples were passed through a Whatman filter paper grade 1 (20–
30 25 μm pore size) to remove debris, sediment, and finer particles. This was followed by
31 acidification of the samples using nitric acid (69%) in the ratio of 1% v/v. This process
32 helped us to suppress the organic interference and reduce the colloidal Fe to Fe 2+ (if Fe is
33 present inherently or contaminated). The treated sample was then collected in a pre-labelled
34 container for immediate analysis or stored at 4°C if the analysis was delayed beyond 24
35 hours. The protocol ensured the removal of interfering particulates before preserving the
36 samples for quantification. Also, the urine sample of a healthy person was collected and
37 filtered with Whatman filter paper grade-1 of pore-size 20-25 µm. This filtration is essential
[1]
38 to remove cellular extracts and any particulate matter that could lead to turbidity of urine.
39 The filtered urine was centrifuged at 10000 rpm for 30 minutes and then digested with nitric
[2]
40 acid (65%) in a volume ratio of 1:19 to suppress any organic interference. The digested
41 urine was then diluted 10-fold with mili-Q water before investigation.

42 Some of the samples, namely urine, urea, creatinine, and microbial enzymes, pose challenges
43 to the stability of the samples for delayed sample handling. To mitigate these effects, sodium
44 fluoride (NaF) (0.05% w/v) was added to the treated urine samples to inhibit urease activity
45 from potential microbial growth, and samples were stored at 4 ℃ until further use.

46 S2. Reagent Optimization

47 The reagent concentrations were optimized to stabilize the Fe(II) phenanthroline complex at
48 pH 2, balancing ligand availability and redox control (Figure S1). A saturated 1,10-
49 phenanthroline solution (0.3% w/v ) ensures a substantial molar excess over Fe 2+, overcoming
[53]
50 protonation effects (pKa ≈4.86) at this pH while preventing ligand depletion or
51 precipitation.

S2
52

53 Figure S1 Phen optimization

54 S3. AAS Calibration

55 Iron solutions in the concentration ranging from 0.5 mg L -1 to 4 mg L-1 were prepared and
56 studied under AAS. A standard calibration plot was prepared to validate the developed
57 prototype of the iron sensor, as shown in Figure S2.

58

59
60 Figure S2 AAS Calibration

61 S4. Structure of ferroin

62 Ferroin, ([Fe(C12H8N2)3]2+ [3], is a coordination complex featuring a central low-spin Fe 2+ ion
63 octahedrally coordinated to three bidentate 1,10-phenanthroline ligands, each donating two
64 nitrogen atoms to form a chelate structure (Figure S3). This arrangement confers D 3
65 symmetry, with Fe–N bond lengths around 197.3 pm. [3]

S3
66 S5. Pourbiax diagram
67 The Pourbaix diagram (Figure S4) for iron illustrates the thermodynamic stability of different
[4]
68 iron species as a function of pH and electrode potential (E) in aqueous environments. It
69 identifies three main regions: immunity, where metallic iron (Fe0) is stable; corrosion,
70 where soluble ions like Fe2+ and Fe3+ dominate; and passivation, where insoluble compounds
71 such as Fe(OH)2 and Fe(OH)₃ form protective layers. Active corrosion occurs at low pH and
72 low potential, while passivation prevents further degradation at higher pH and moderate
73 potential. The diagram also incorporates water stability boundaries, highlighting regions
74 where hydrogen or oxygen evolution occurs. Though based on thermodynamics, it does not
75 predict corrosion rates, making it a valuable tool for understanding iron's behaviour in
76 various aqueous environments.

77

78 Figure S3 Structure of Ferroin (Adapted from Wikipedia) [3]

S4
79
80 Figure S4 Pourbiax diagram for iron (Adapted from Libretext) [4]

81 S6. Selectivity Study with Potential Interferent

82 Fresh experiments at 0.2 mgL-1 and 200 mgL-1 of Cu2+ are one-tenth lower and 100 times
83 higher than the WHO's permissible limits. We specifically targeted Cu 2+ as it may interfere if
84 the reaction's pH is not controlled. The results are shown in Figure S5. These results
85 demonstrate the sensor's robustness and ability to maintain accuracy even under extreme
86 conditions, which are unlikely to be encountered in real-world samples.

87
88 Figure S5 Selectivity study with potential interferent

89 S7. Calibration of the Real samples

90 All the collected real field samples were spiked with iron, and its calibration plot was
91 prepared to estimate the LOD of each type of sample. Figure S6 (a-d) shows the calibration
92 plot for (a) Brahmaputra water, (b) Tap water, (c) Borewell, and (d) Urine. R 2 values
93 indicated in each figure prove the good linearity of the prototype.

S5
 94

95 Figure S6 Calibration plot: (a) Brahmaputra water, (b) Tap water, (c) Borewell, and (d) Urine

96

97 References
98 [1] S. Mitra, M. Basak, S. Biswas and P. S. G. Pattader, "Digital electronic based portable device
99 for colorimetric quantification of ketones and glucose level in human urine" Measurement. 2023,
100 214, 112848. [Link] 112848
101 10.1016/[Link].2023.112848
102 [2] M. R. Patel, M. D. Upadhyay, S. Ghosh, H. Basu, R. K. Singhal, T. J. Park and S. K. Kailasa,
103 "Synthesis of multicolor silver nanostructures for colorimetric sensing of metal ions (Cr(3+), Hg(2+)
104 and K(+)) in industrial water and urine samples with different spectral characteristics" Environ Res.
105 2023, 232, 116318. [Link]
106 [3] Ferroin, [Link] 5th April, 2025)
107 [4] Libretext in INTRODUCTION TO INORGANIC CHEMISTRY, (Eds.: Penn State University, 2025,
108

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