Electronic Supplementary Material

Amidosulfonic acid-capped silver nanoparticles for the spectrophotometric determination of lamotrigine in exhaled

breath condensate

Abolghasem Jouyban1,2, Azam Samadi1,*, Maryam Khoubnasabjafari3, Vahid Jouyban-Gharamaleki4, Fatemeh Ranjbar5

1Pharmaceutical Analysis Research Center and Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran

2Kimia Idea Pardaz Azarbayjan (KIPA) Science Based Company, Tabriz University of Medical Sciences, Tabriz 51664, Iran

3Tuberculosis and Lung Disease Research Center, Tabriz University of Medical Sciences, Tabriz, Iran

4Liver and Gastrointestinal Diseases Research Center, Tabriz University of Medical Sciences, Tabriz, Iran

5Research Center of Psychiatry and Behavioral Sciences, Tabriz University of Medical Sciences, Tabriz, Iran

* Corresponding author

Email: Samadi_azam@yahoo.com; Tel: +984133379323

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1. Optimization of experimental conditions

1.1. Effect of the ASA concentration

The color change and detection capability of AgNPs are highly sensitive to the concentration of their stabilizing agent. Thus, ASA-AgNPs were synthesized based on four different molar ratios (nAgNO3:nASA). According to Fig. S1A, the UV-Vis spectra of AgNPs in the presence of different concentration of ASA did not change sharply. In addition, their sensitivity to the concentration of LTG was not differ seriously (Fig. S1B). It shows that the size and concentration of the resulting ASA-AgNPs is almost the same. In fact, the different sizes and concentrations of nanoparticles that have particular SPR wavelengths leads to varying sensitivity to analyte concentration [1]. However, an excess or deficit of stabilizing agent leads to quick aggregation of silver nanoparticles and the stability of ASA-AgNPs declines. The molar ratio of 1:1 was selected as the optimized conditions, which remained

stable at least for two months (Fig. S1C).

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Fig.S1. (A) Absorption spectra of ASA-AgNPs prepared with various molar ratios

(nAgNO3:nASA). (B) The absorption intensity ratio (A450nm/A390nm) of these different types of

ASA-AgNPs as a function of LTG concentrations. (C) The UV-Vis spectra of ASA-AgNPs

stored at 4 ºC for different times

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1.2. Effect of the ASA-AgNPs concentration

The concentration of the testing AgNPs solution was estimated to be 3.5 nM according to the extinction coefficient based on particle diameter (ε=Adγ, for AgNPs with diameter less than or equal to 38 nm, A=2.3×105

M-1cm-1,γ=3.48, [2]). In our experiment, the average AgNPs particle size (30 particles) is 10.2 ± 2.3 nm and the absorbance of the testing solution is 2.64. The effect of the concentration of the AgNPs solution on the signal intensity was also investigated. The results showed that the sensitivity is higher for the lower concentration AgNPs solution, but because of the quick aggregation of nanoparticles the linearity range is narrower. Analytical parameters including slopes of the calibration graphs, linear ranges, and correlation coefficients (r) for the three amounts of AgNPs, are listed in Table S1. The concentration of 17.5 × 10-10 M (corresponding to 0.5 mL of prepared silver nanoparticle solution in 1.0 mL total volume) was chosen as optimum for subsequent work.

Table S1. Analytical parameters for the determination of LTG with different amounts of

AgNPs

LOQ

(µg·mL-1)

LOD

(µg·mL-1)

Correlation

coefficient (r)

Sensitivity

(slope of

calibration graph)

Linear

range

(µg·mL-1)

Concentration of

AgNPs (×10-10 M)

0.0090.0030.9909.120.01–0.17

0.0160.0050.9982.060.02–0.417.5

0.0370.0120.9911.100.04–0.824.5

Experimental conditions: pH=8.5; incubation time=30 min.

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1.3. Effect of pH and reaction timeThe influence of the media pH (4.0–11.0) was also investigated. According to Fig. S2A, pH 8.5 was found to be the optimal value for further experiments. LTG is a weak base with a dissociation constant (pKa) of 5.5 [3] and at basic pH it can act as a better electron pair donor toward the ASA acceptor. However, at pH higher than 9.5, competition of hydroxyl ions in solution against LTG probably leads to decreased signal intensity. Among the three buffers tested (i.e. phosphate, Britton robinson and bicarbonate buffer ), bicarbonate buffer gave the best results in terms of sensitivity and linear range, so100 µL of 0.01 M bicarbonate solution was used for adjustment of pH whenever it was required. To obtain the maximum response, the influence of incubation time on signal intensity was studied. According to the obtained results, the signal intensity is enhanced rapidly following the addition of LTG and reaches a plateau in about 30 min (Fig.S2B).

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Fig.S2. Effect of (A) pH and (B) time on the absorption intensity ratio (A450nm/A390nm); ASA-AgNPs concentration = 17.5 × 10-10 mol·L-1, [LTG] = 0 .1 µg·mL-1

Fig. S3. The UV-vis spectra of LTG (20 µg·mL-1) in the presence of ASA (80 µg·mL-1)

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Fig.S4. (A) Absorbance spectra of the ASA-AgNPs in the presence of LTG in EBC samples with various concentrations (0, 0.2, 0.5, 1, 1.5, 2, 4, 5 μg·mL -

1). Inset is the selected photograph of colorimetric response of ASA-AgNPs to the different concentration of LTG. (B) The plot of intensity ratio (A450nm/A390nm) of ASA-AgNPs versus the concentration of LTG in EBC. Concentration of ASA-AgNPs (1:1) = 17.5×10-10 M; pH=8.5; incubation time=30 min.

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Table S2. List of some reported compounds and their detection range in EBC media

Material Concentration range(ngmL-1)

Ref.

Pepsin 0.45–1.91 [4]

Leukotriene B4 0.000006–0.0000180.0053–0.00750.0023–0.00630.003–0.044

[5][6][7][8]

Interleukin 5 – [9]Interleukin 8 – [9]Matrix metallopeptidase 9 – [9]Lipoxin A4 0.0013–0.004 [7]Tumor necrosis factor alpha (TNF-α ) 0.0038–0.005 [6]NH4

+ 0–55.43 [10]K+ 0.026–3.84 [10]Na+ 0.39–8.69 [10]Ca2+ 0.5–7.5 [10]Mg2+ 0–0.42 [10]Cl- 0–5.64 [10]NO2

- 0–0.22 [10]NO3

- 0–0.16 [10]SO4

2- 0–0.1 [10]Iron 0.007–0.053 [11]Acetate 0.085–3.38 [10]Lactate 0.055–2.77 [10]Phosphate 0.01–1.58 [10]pH 8.05–8.16

6.17–7.48[8][11]

H2O2 0.011–0.025 [8]8-Isoprostane 0.0036–0.0093 [8]4-Hydroxynonenal 0.000013–0.000016 [8]Malondealdehyde 0.000075–0.00012 [8]Urea 39–4190 [12]Ammonia – [13]Acetic acid – [13]Formic acid – [13]Protein ¿1000 [14]Amino acids – [15]Ferritin 0–0.9 [11]Fractional concentration of exhaled nitric oxide 3.0–14.3 [11]Acetone – [16]Ethanol – [16]Methanol – [16]Propanol – [16]Isoprene – [16]Hydrogen cyanide – [16]Formaldehyde – [16]Acetaldehyde – [16]

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