Electronic Supplementary Material (ESI) for RSC Advances.

This journal is © The Royal Society of Chemistry 2018

1. The construction of standard curve for the detection of enzyme activity.

Experiment: 0.15 mol L-1 Tris buffer (pH 8.0) was used to prepare 1.8 × 10-4 mol L-1
DTNB solution. Then 0.025, 0.050, 0.075, 0.100 and 0.125 mmol L-1 L-cysteine
hydrochloride was respectively dissolved in the DTNB solution. After that, the AChE-
functionalized 2D-PC was respectively placed into a series of L-cysteine
hydrochloride solutions and UV absorbance at 412 nm was monitored.

Table S1 Data of Standard Curve

Concentration of L-
cysteine Absorbance Absorbance Absorbance Absorbance
hydrochloride 1 2 3 average
(mmol.L-1)
0. 025 0.32253 0.3224 0.3225 0.322477
0. 050 0.65044 0.65027 0.65001 0.65024
0. 075 0.972 0.97182 0.97154 0.971787
0. 100 1.26311 1.26312 1.26234 1.262857
0. 125 1.59422 1.59169 1.59354 1.59315
0. 150 1.89241 1.89133 1.89098 1.891573
0. 175 2.21499 2.21065 2.21872 2.214787

Figure S1 Standard Curve of L-cysteine hydrochloride

2. Measurement of binding amounts of AChE

2.1. Standard curve
 Experiment: 2 mL obtained G-250 solution was mixed with 50 μL AChE solution
varied from 0.5 to 3.0 mg/mL (adjusted with 0.15 mol L-1 Tris-HCl buffer, pH=7.4)
and UV absorbance at 595 nm was measured.

Figure S2 Standard Curve of AChE solution

2.2. Detection of AChE-functionalized 2D-PC

The amount of AChE on film (mg cm-2) meets following equation:
𝑚 = 4(𝐴0 ‒ 𝐴1)/(0.25 × 12𝐾) (S1)
where 𝐴0 and 𝐴1 are AChE solution before and after modification respectively, K is
the slope of standard curve, 4 is the volume of AChE solution, 0.25 is the area of
every film, 12 represents the amount of films.
The change of absorbance (𝐴0 ‒ 𝐴1) and the amounts of AChE on film were depicted in
table S2.

Table S2 data of AChE-functionalized 2D-PC

sample
sample 1 sample 2 sample 3
average
Change of absorbance 0.1046 0.1517 0.1131 0.1231
The amounts of AChE on film (mg
0.36 0.53 0.39 0.43
cm-2)

3. water-solubility of organophosphates
 Table S3 Water-solubility of some organophosphates a.

glufosinate-
organophosphates dipterex dichlorvos malathion methidathion acephate
ammonium
4.39 × 10-4 7.94 × 10-4
mol/L mol/L
water-solubility soluble soluble soluble soluble
(slightly (slightly
soluble) soluble)
a Data sources: https://www.baidu.com/

4. Calculations of limit of detection (LOD)

We surmised that there was a linear correlation between particle spacing of our
sensor and dipterex concentration at the range of 0~10-14 mol L-1 before calculations.
We measured the responsivity of our AChE-PC sensor to a 50 mL of dipterex at a
concentration of 10-14 mol L-1 and a 38 nm decrease in particle spacing was observed.
The resulting responsivity was calculated as S=38 × 1014 nm/(mol L-1). At the blank
solution, the average standard deviation is σ=8.9 nm. As a consequence, the LOD=
3.3 × σ/S=7.7 × 10-15 mol L-1.

5. Performance comparison of different materials

Table S4 Comparison of various analytical methods for sensing Sarin analogs
Method Strengths/
Analyte Sensitivity References
Names Weakness
S: detecting
organophosphate with high
sensitivity; miniaturized;
AChE- LOD: 0.77 x 10-
Diptere simple detection device;
functionali 14 mol L-1 This study
x simple preparation;
zed 2D-PC
W: can’t detect dipterex in
complex environment until
now.
S: detecting a real LOD: 10-15 mol
3D-PC
Sarin chemical warfare with high L-1 [1]
biosensor
sensitivity;
 W: can’t detect Sarin in
complex environment until
now.
S: high sensitivity; using a
portable device; LOD: ~13 fmol
VX;
SERS W: relatively high cost; (VX); ~670 fmol [2]
tabun
require extensive sample (tabun)
pre-treatment;
S: enormous electro-
DMMP
magnetic enhancement;
; PMP; LOD: lower than
SERS W: relatively high cost; [3]
DEPA; 1 ppm;
require extensive sample
CEES
pre-treatment;
VX and S: could distinguish the
its nerve agent VX and its
hydroly hydrolysis products;
SERS 50-100ug L-1 [4]
sis W: relatively high cost;
product require extensive sample
s pre-treatment;
S: high sensitivity; large
emission shift;
W: time-consuming; LOD: 17 nmol L-
CIEE/FS DCP [5]
require extensive sample 1

pre-treatment; relying on
sophisticated instruments
S: sufficient separation of
these four compounds in
environmental forensic LOD: 0.015-
DMT;
HPLC- analysis of samples with 0.025mg L-1
DET;
MS/CE- minimum sample pre- (HPLC-MS); [6]
DPT;
UV treatment; 1.5-2mg L-1 (CE-
DIT
W: time-consuming; UV)
require extensive sample
pre-treatment; relying on
 sophisticated instruments
S: could detect real
samples; good alternative
extraction method; LOD: 0.2 ng L-1
SPME/GC TnBP;
W: time-consuming; (TnBP); 1.5ng L- [7]
-MS TEHP
require extensive sample 1 (TEHP)
pre-treatment; relying on
sophisticated instruments
S: realized qualitative and
quantitative determination
14
of pesticide residues in
pesticid
SPME/GC mangoes; LOD: 1.0-3.3
e [8]
-MS W: time-consuming; μg kg-1
residue
require extensive sample
s
pre-treatment; relying on
sophisticated instruments
S: gas detection; short
Tabun, detection time (2.8s)
Sarin, W: time-consuming;
IMS/DMS LOD: 20 μg m-3 [9]
Soman, require extensive sample
VX pre-treatment; relying on
sophisticated instruments
S: could detect sample in a
variety of sample matrixes
(water, kerosene, gasoline,
IM(tof)M diesel); LOD: lower than
DMMP [10]
S W: time-consuming; 1000 ppm;
require extensive sample
pre-treatment; relying on
sophisticated instruments
Abbreviation: surface-enhanced Raman scattering (SERS); dimethyl methylphonate
(DMMP); pinacolyl methylphosphonate (PMP); diethyl phosphoramidate (DEPA);
chloroethyl ethylsulfide (CEES); cyclization-induced emission enhancement (CIEE);
fluorescence spectrum (FS); diethyl chlorophosphate (DCP); high-performance liquid
chromatography-mass spectrometry (HPLC-MS); capillary electrophoresis with direct
ultraviolet detection (CE-UV); N, N-(dialkyl)aminoethanesulfonicacids, where alkyl
= methyl, ethyl, n-propyl or iso-propyl (DMT, DET, DPT, and DIT, re-spectively);
solid-phase microextraction (SPME); gas chromatography–mass spectrometry (GC–
MS); tri-n-butyl phosphate (TnBP); tris (2-ethylhexyl) phosphate (TEHP); ion
mobility spectrometry (IMS); differential mobility spectrometry (DMS); ion
mobility orthogonal reflector time-of-flight mass spectrometer (IM-(tof)MS);
References:
[1] C. X. Yan, F. L. Qi, S. G. Li, J. Y. Xu, C. Liu, Z. H. Meng, L. L. Qiu, M. Xue, W.
Lu and Z. Q. Yan, Talanta, 2016, 159, 412-417.
[2] A. Hakonen, T. Rindzevicius, M.S. Schmidt, P.O. Andersson, L. Juhlin, M.
Svedendahl, A. Boisen, M. Kall, Detection of nerve gases using surface-enhanced
Raman scattering substrates with high droplet adhesion, Nanoscale 8 (2016) 1305-
1308.
[3] F. Yan, T. Vo-Dinh, Surface-enhanced Raman scattering detection of chemical
and biological agents using a portable Raman integrated tunable sensor, Sensor.
Actuat. B-Chem. 121 (2007) 61-66.
[4] S. Farquharson, A. Gift, P. Maksymiuk, F. Inscore, Surface-enhanced Raman
spectra of VX and its hydrolysis products, Appl. Spectrosc. 59 (2005) 654-660.
[5] A.K. Mahapatra, K. Maiti, S.K. Manna, R. Maji, S. Mondal, C. Das
Mukhopadhyay, P. Sahoo, D. Mandal, A cyclization-induced emission enhancement
(CIEE)-based ratiometric fluorogenic and chromogenic probe for the facile detection
of a nerve agent simulant DCP, Chem. Commun. 51 (2015) 9729-9732.
[6] I. Rodin, A. Stavrianidi, R. Smirnov, A. Braun, O. Shpigun, I. Rybalchenko, New
Techniques for Nerve Agent Oxidation Products Determination in Environmental
Water by High-Performance Liquid Chromatography-Mass Spectrometry (HPLC-MS)
and Capillary Electrophoresis (CE) with Direct Ultraviolet (UV) Detection, Environ.
Forensics 14 (2013) 87-96.
[7] Y.C. Tsao, Y.C. Wang, S.F. Wu, W.H. Ding, Microwave-assisted headspace solid-
phase microextraction for the rapid determination of organophosphate esters in
aqueous samples by gas chromatography-mass spectrometry, Talanta 84 (2011) 406-
410.
[8] A. Menezes Filho, F.N. dos Santos, P.A. de Paula Pereira, Development,
validation and application of a methodology based on solid-phase micro extraction
followed by gas chromatography coupled to mass spectrometry (SPME/GC-MS) for
the determination of pesticide residues in mangoes, Talanta 81 (2010) 346-354.
[9] M. Maziejuk, M. Ceremuga, M. Szyposzynska, T. Sikora, A. Zalewska,
Identification of organophosphate nerve agents by the DMS detector, Sensor. Actuat.
B-Chem. 213 (2015) 368-374.
[10] W.E. Steiner, S.J. Klopsch, W.A. English, B.H. Clowers, H.H. Hill, Detection of
a chemical warfare agent simulant in various aerosol matrixes by ion mobility time-
of-flight mass spectrometry, Anal. Chem. 77 (2005) 4792-4799.

6. Calculated wavelength of 2D-PC

6.1 The 2D Bragg diffraction equation.
The diffraction process of 2D-PC was illustrated in figure S3, while white source (S1,
S2) illuminated below the sample at an incidence angle of 45° (𝜃), and the structural
color was recorded above the sample along the normal.

Figure S3 Illustration of the diffraction process of 2D-PC

The path difference ∆ was given by

3
∆ = 𝑛𝑎𝑖𝑟(𝐴𝐵) = 𝑛𝑎𝑖𝑟(𝑝sin 𝜃) = 𝑝sin 𝜃 = 𝑑sin 𝜃
2 (S2)
Where 𝑛𝑎𝑖𝑟 is the refractive index of air (𝑛𝑎𝑖𝑟=1), 𝑝 is the distance between adjacent
lattice rows. 𝜃 is the incidence angle. According to reference1, 𝑝 is relate to the lattice
spacing (𝑑) by
3
𝑝= 𝑑 (𝑆3)
2
According to Bragg’s law, while the path difference (∆) is equal to a whole number of
wavelengths, the diffraction will be strengthened. So the path difference meets
following equation:
3
𝑑sin 𝜃 = 𝑚𝜆 (𝑆4)
2
Where 𝑚 is the diffraction order, 𝜆 is the diffracted wavelength. While the incidence
angle was fixed, and we surmise the diffraction was the first-order diffraction.
According to the determination of particle spacing, we can also calculate the
diffracted wavelength of 2D-PC in different condition.

Figure S4 AChE-functionalized 2D-PC (◆) and unfunctionalized 2D-PC (▲) for

dipterex detection.