ChemistrySelect

Supporting Information

Electrochemical Sensor for the Anti-tuberculosis Drug

Rifampicin on CuO@rGO-Nanocomposite-Modified GCE by
Voltammetry Techniques
Pinky Sagar, Monika Srivastava, and Sanjay K. Srivastava*
EXPERIMENTAL SECTION

Reagents and Chemicals

CuO, graphite powder and RFP were procured from Merck, India. KMnO4 and Hydrazine

hydrate were of analytical grade and used without further any purification. Conc. H2SO4, Conc.

H3PO4, Conc. HCl, KOH, Na2HPO4, NaH2PO4 and hydrogen peroxide were purchased from

Merck, India. R-CIN 300 capsule was purchased from local medical shop in Varanasi, India.

Instruments

CuO, rGO and CuO@rGO were investigated by X-RD diffraction instrument (Rigaku, Mini-

Flex 600, Japan) from 2θ = 10˚ to 90˚. Raman spectroscopic measurement were taken on

instrument WITec alpha 300 RAS spectrophotometer, Germany ranging from wavenumber 0 –

4000 cm-1. Further, for the elemental analysis, X-ray photoelectron spectroscopy was

performed on XPS K-Alpha, Thermo Fisher Scientific, UK. All the measurement was taken in

triplicates. After that, to check the chemical bonds and functionality of the samples, FT-IR

spectroscopy was conducted on instrument Nicolet iS5 (Thermo Fisher Scientific Instrument).

Transmission electron microscopy was carried out to check the structural properties of the

samples on instrument TEM Technai G20 (FEI, USA).

All the electrochemical measurements were taken on Autolab (PGSTAT, 101, The

Netherlands). At the time of experimentation, three electrode assembly was considered

equipped with glassy carbon electrode as working electrode (d = 3mm), Pt-foil as counter

electrode and Ag/AgCl as reference electrode.

Synthesis of Materials

Synthesis of rGO

rGO was synthesized by one of the most popular method ‘Modified Hummer’s method’ with

slight changes as cited earlier by our group.[58] To synthesize rGO, we first synthesized GO

through exfoliation of graphite powder. Here, we have taken 1.5 g of graphite powder in a flask

followed by addition of K2SO4 and P2O5 (1:1) and then conc. H2SO4 (40 mL). After the 4 h

reaction at 80 ˚C, the reaction mixture was collected and washed thoroughly multiple times,

(until pH ~ 7) and then vacuum dried at 50 ˚C. Next, conc. H2SO4 (180 mL) and conc. H3PO4

(13 mL) were added to pre-oxidized graphite powder and stirred well followed by gradual

addition of KMnO4 at 50 ˚C for 15h. The obtained mixture was left to cool at room temperature

(RT) and then ice cubes (~ 200 mL) were added. The colour of the mixture was changed to

grey. Further, H2O2 (30%, 1.5 mL) was dripped to the mixture until grey colour changed to

pale yellowish colour. This colour change is the signal of GO formation. Finally, the mixture

was washed with DI water several times and dried in ambient conditions to collect GO powder.

Hydrazine hydrate was used to reduce rGO from GO at 100 ˚C. Hydrazine hydrate was added

to the suspended GO heated at 100 ˚C for 1h. Thereafter, the suspension was filtered with
cellulose filter and washed via DI water and 0.1 M HCl until pH~7. Hence obtained precipitate

was dried and stored for experimentation.

Synthesis of CuO@rGO composite

The as-synthesized rGO was dispersed as 1 mg/ ml of DI water followed by rigorous

sonication. CuO powder as 1 mg/ ml in DI was also prepared separately and followed by several

times sonication until it appeared well dispersed. Afterward, both the samples were mixed in

equal volumetric ratio (1:1 ratio) and then again allowed to sonicate for 1.5 h at R.T. Hence

prepared sample was kept safe in a vial and was used for experimental purposes without any

further treatment.

Modification of Electrode

Before using GCE for experiment, the bare electrode was polished well on the wet pad with

alumina slurry. Subsequently, GCE was rinsed with DI water and treated for ~2 minutes in

ethanol and DI water. For the modification, 5 µL of homogeneous CuO@rGO composite was

drop casted on the naked and cleaned surface of GCE and left to dry for 1h at R. T. After drying

process, the GCE was used for further experimental procedures.

Preparation of real sample

Prior to analyse the effect of commercially available antibiotic drug on the sensing of RFP by

CuO@rGO/GCE, we have procured RFP from local therapeutic drug store. Next, to make the
solution, a complete capsule was uncovered and powder (~705 mg) was collected in a separate

vial. Following the procedure, the 705 mg powder was then dissolved in 10 ml DI and

centrifuged at 3000 RPM for ~15 minutes. The precipitate was removed and supernatant was

further filtered with the help of syringe filter (pore size = 450 µm). When calculated, the

concentration of RFP in above mentioned prepared solution (later named as ‘real sample’) was

found to be 72.90 mM. Hence prepared solution was kept as stock and used to make solutions

of desired concentrations by further diluting it.

(a) (b)

rGO-CuO C

(c) (d)

O Cu

Figure S1. EDS mapping of (a) CuO@rGO comprises of, (b) C, (c) O and (d) Cu elements
 (b)

Transmittance (%)
(a)

Epoxide
(c)

-OH
C-O-H
Cu-O

C-OH

C=O
1000 2000 3000 4000

Wavenumber (cm-1)

Figure S2. FTIR spectra of (a) rGO (b) CuO and (c) CuO@rGO

(a) Measured
Calculated
(b) Measured
Calculated
2000 2000

1500 1500
Z"

Z"

1000 1000

500 500

0 0
0 500 1000 1500 2000 0 500 1000 1500 2000
Z' Z'

(c) Measured
Calculated
(d) Measured
Calculated
2000 2000

1500 1500
Z''

Z''

1000 1000

500 500

0 0
0 500 1000 1500 2000 0 500 1000 1500 2000
Z' Z'

Figure S3. Electrochemical impedance fitting plots for (a) bare GCE, (b) bare GCE + RFP, (c)
CuO@rGO/GCE and (d) CuO@rGO/GCE + RFP
 0.51

0.50

Ea (V)
0.49 y = 0.277x +0.44
R2 = 0.98

0.48

0.47

0.8 1.0 1.2 1.4 1.6 1.8 2.0 2.2 2.4 2.6
log [n/mVs-1]

Figure S4. Peak potential vs. logarithm of scan rate plot

(a) (b)0.65
5

Potential, V vs. Ag/AgCl

0.60
0
Current (mA)

0.55
-5
0.50
8.0
-10 7.4 y = -0.053pH + 0.82
6.5 0.45 R2 = 0.99
5.5
-15
4.5 0.40
4.0
0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 4 5 6 7 8
Potential, V vs. Ag/AgCl pH

(c)
5.0

4.5
Current (mA)

4.0

3.5

3.0

2.5

2.0

4 5 6 7 8
pH

Figure S5. (a) CV response of CuO@rGO/GCE at different pH (values are shown in inset) of
0.1 M PBS, (b) corresponding potential calibration plot with respect to pH and (c)
corresponding current calibration plot with respect to pH
 6.0 6.0
6
(a) 5.5
(b) 5.5
(c)
4
2 5.0 5.0

Current (mA)

Current (mA)
Current (mA)

0
4.5 4.5
-2
4.0 4.0
-4
-6 3.5 3.5

-8 3.0 3.0
Y = (0.07±0.001) X + (2.74±0.02) Y = (0.08±0.004) X + (2.70±0.06)
-10
2.5 R2 = 0.99 2.5 R2 = 0.98
-12
0.2 0.4 0.6 0.8 0 10 20 30 40 50 0 10 20 30 40 50
Potential (V) vs. Ag/AgCl Concentration (mM) Concentration (mM)

Figure S6. (a) CV response of CuO@rGO/GCE upon addition of various concentration of

RFP, (b) corresponding calibration plot for oxidation current and (c) reduction current, in 0.1
M PBS (pH = 7.4)

6.0
6 (a) (b) 6.0 (c)
4 5.5
5.5
2 5.0
Current (mA)

Current (mA)
Current (mA)

5.0
0
4.5
-2 4.5
-4 4.0 4.0
-6 3.5 3.5
-8
3.0 Y = (0.10±0.003) X + (2.74±0.04) 3.0 Y = (0.09±0.003) X + (2.98±0.05)
-10 R2 = 0.99
R2 = 0.99
-12 2.5 2.5
0.2 0.4 0.6 0.8 0 10 20 30 40 50 0 10 20 30 40 50
Potential (V) vs. Ag/AgCl Concentration (mM) Concentration (mM)

Figure S7. (a) CV response of CuO@rGO/GCE upon addition of various concentration of

RFP-capsule, (b) corresponding calibration plot for oxidation current and (c) reduction current,
in 0.1 M PBS (pH = 7.4)

Table S1. Extracted data from the impedance curves for bare GC, Bare GCE+RFP,
CuO@rGO/GCE and CuO@rGO/GCE +RFP.

Parameter Bare GCE Bare GCE + CuO@rGO/GCE CuO@rGO/GCE

RFP + RFP

Rs (Ohm) 8.78 9.1 8.9 9.5

CPE (γ0) 8.13×10-7 6.618×10-7 0.0009567 0.001182

(S-sec.n )
Freq. power, 0.8 0.8 0.6866 0.8

R1 (Ohm) 667.7 600.3 560.3 538.4

Warburg (γ0) 0.001684 0.001747 0.001627 0.001846

(S-sec.5)

C (F) 1.32×10-7 3.319×10-7 0.02847 0.02639

R2 (Ohm) 198.4 496 158 251.6

χ2 2.546×10-5 1.367×10-4 1.896 ×10-5 7.803×10-5