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Leardi, K. Suzuki and D. Citterio, Analyst, 2018, DOI: 10.1039/C8AN00332G.

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11 Fabrication of paper-based analytical devices optimized by central
12 composite design
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14 Received 00th January 20xx, Vahid Hamedpoura, Riccardo Leardib, Koji Suzukia, Daniel Citterio*a
15 Accepted 00th January 20xx
In this work, an application of a design of experiments approach for the optimization of an isoniazid assay on a single-area
16
DOI: 10.1039/x0xx00000x inkjet-printed paper-based analytical device (PAD) is described. For this purpose, a central composite design was used for
17
evaluation of the effect of device geometry and amount of assay reagents on the efficiency of the proposed device. The
18

Analyst Accepted Manuscript

www.rsc.org/
factors of interest were printed length, width, and sampling volume as factors related to device geometry, and amounts of
19
the assay reagents polyvinyl alcohol (PVA), NH4OH, and AgNO3. Deposition of the assay reagents was performed by a thermal
20
inkjet printer. The colorimetric assay mechanism of this device is based on the chemical interaction of isoniazid, ammonium
21
hydroxide, and PVA with silver ions to induce the formation of yellow silver nanoparticles (AgNPs). The in-situ-formed AgNPs
22
can be easily detected by the naked eye or with a simple flat-bed scanner. Under optimal conditions, the calibration curve
23
was linear in the isoniazid concentration range 0.03–10 mmol L−1 with a relative standard deviation of 3.4% (n = 5 for
24
determination of 1.0 mmol L−1). Finally, the application of the proposed device for isoniazid determination in pharmaceutical
25
preparations produced satisfactory results.
26
27
In the current study, a DOE approach is demonstrated on a
28
Introduction model PAD targeting the detection of isoniazid, which is one of
29
the common first-line therapies for drug-susceptible
30 Paper-based analytical devices (PADs) can become inexpensive
tuberculosis. During recent decades, several analytical methods
31 portable sensing platforms for various applications like food
have been reported for the determination of isoniazid and some
32 safety control, environmental monitoring, disease diagnostics,
attempts to integrate proposed methods with sensors have
33 and water testing1–5. These devices, owing to features such as
gained attention11. To the best of our knowledge, this is the first
34 simple fabrication, low sample consumption, easy usage, and
report on the application of PADs for isoniazid determination
35 low cost, are mostly utilized for clinical, biological, or chemical
and the proposed device can be used as a portable platform for
36 analyses in developing countries or remote locations as well as
online dosage control for process quality control by
37 in emergency situations6,7. Making use of modern technologies,
pharmaceutical companies.
38 such as wax printing technology for substrate patterning and
The detection method of this device is based on the
39 inkjet technology for printing the assay reagents in small
plasmon resonance of in-situ-formed silver nanoparticles
40 amounts, as well as the combination of PADs with highly
(AgNPs)12. Since the colour intensity observed in the detection
41 sensitive detection methods, can improve the sensitivity of the
zone is highly dependent on several features of device design,
42 devices8,9. However, investigation into the parameters that may
attention should be given to them. For this purpose, an
43 affect the device performance, such as shape and size of device
experimental design, and more specifically a central composite
44 and assay reagent amounts, is still one of the challenges that
design (CCD), was employed for evaluating the impact of device
45 needs to be overcome.
geometry and amounts of assay reagents on the proposed
46 One of the aims of design of experiments (DOE) is to select
device’s sensitivity. CCD is an experimental design which
47 the optimum experimental conditions with limited
provides information on linear, interaction, and quadratic terms
48 experimental effort10. This approach is therefore superior to
and is widely used for formulation or process optimization by
49 traditional one-factor-at-time (OFAT) optimization strategies.
pharmaceutical companies13. The matrix of a CCD consists of an
50 Furthermore, DOE also allows to detect and to study
embedded full factorial or fractional factorial design, a star
51 interactions among experimental factors, which is not possible
design, and n replicates of the central point14. Star points have
52 by changing just one factor at a time.
a distance α from the centre point, and their values determine
53
the type of design, make it flexible, and allow estimation of
54
aDepartment of Applied Chemistry, Keio University, 3-14-1 Hiyoshi, Kohoku-ku, curvature15. Since the star points (α) represent new extreme
55 Yokohama 223-8522, Japan. E-mail: citterio@applc.keio.ac.jp; FAX: +81 45 566 values (low and high) for each factor in the design, all factors
56 1568; Tel: +81 45 566 1568.
are studied in five levels (−α, −1, 0, +1, +α). For illustration
57 bDepartment of Pharmacy, University of Genoa, Genoa, Italy.

Electronic Supplementary Information (ESI) available: Schematic illustration of a purposes, a generalized graphical representation of a three-
58 CCD, design matrix and the response for central composite design, calibration curve
factor CCD and a corresponding experimental matrix are shown
59 for isoniazid. See DOI: 10.1039/x0xx00000x
60

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3 in Figure S1 of the Electronic Supporting Information (ESI). It First, filter paper (Whatman grade 1, GE View Healthcare,
Article Online

4 should be noted that a CCD with a larger number of factors (as DOI: 10.1039/C8AN00332G
Buckinghamshire, U.K.) was cut and adjusted to A4 size; then,
5 applied here) can no longer be graphically represented. the designated pattern was printed by a wax printer. The
6 Although the DOE approach may contribute to PADs pattern was designed with Microsoft PowerPoint and printed
7 becoming powerful devices, only a few studies have used this only on the top side of the filter paper. For complete diffusion
8 optimization method. Moreover, in these studies, simplified of the wax into the thickness of the paper, the paper was heated
9 optimization procedures lacking the investigation of all effective on a hot plate (NHS-450ND, NISSIN Co., Tokyo, Japan) for 90 s.
10 factors such as size and shape of inlet area and the amount of Afterwards, the back of the paper was pouched with an
11 required assay reagents, have been applied16,17. adhesive lamination film (thickness = 150 µm) using a hot
12 We have previously reported the application of a Box- laminator (QHE325, Meikoshokai Co., Ltd., Tokyo, Japan).
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13 Behnken Design (BBD) on a PAD, where, because of chemistry- Finally, 11 printing cycles of PVA 2.8 g L−1, 8 printing cycles of
14 related experimental constraints, carrying out the experiments NH4OH 0.5 mol L−1 and 12 printing cycles of AgNO3 0.01 mol L−1
15 including all factors simultaneously at their highest or lowest were deposited consecutively on the same area (Figure 1).
16 levels had to be avoided18. Thus, the investigation of all
17 mentioned conditions, corner points, and star points was not Real sample preparation
18 possible. According to the practical applications and compared

Analyst Accepted Manuscript

For the evaluation of the proposed method’s accuracy, the
19 to BBD, CCD is more efficient and able to estimate new data concentration of isoniazid in both tablets (100 and 300 mg) and
20 points more reliably15,19. In the present work, due to the injection (100 mg mL−1) were determined using the fabricated PAD
21 features of isoniazid, the experimental restrictions mentioned and compared with results obtained using the standard method 20.
22 above are no longer encountered. Therefore, a CCD with the The samples were assayed by the international pharmacopeia
23 ability to study extreme conditions and better prediction quality method, which describes the titration of the isoniazid sample with
24 was applied. Finally, studies on the determination of isoniazid in potassium bromate in the presence of potassium bromide using
25 tablet and injectable formulations were carried out to illustrate methyl red as an indicator. For this purpose, five tablets were
26 the feasibility of the proposed device. weighed accurately, and the average weight of each tablet was found.
27 Then, the tablets were finely powdered and carefully mixed. A
28 portion of the mixture containing about 13.78 mg of isoniazid, was
29 Experimental
weighed accurately and put in a glass vessel; about 70 mL of ultra-
30 Reagents and materials pure water was added and stirred for 15 min. Then, after filtering,
31 All chemicals were used as obtained without further purification. the solution was transferred to a 100-mL volumetric flask using ultra-
32 A stock standard solution of isoniazid (0.1 mol L−1) was prepared pure water for dilution up to the mark, resulting in a nominal 1.0
33 by dissolving the appropriate amount of isoniazid (Sigma Aldrich, mmol L-1 solution. In the case of the injection type, solutions of five
34 Germany) in ultra-pure water. A stock solution of polyvinyl samples were mixed. Then, the appropriate volume to result in a
35 alcohol (PVA, 2.8 g L−1) was prepared by dissolving 0.14 g of PVA nominal 1.0 mmol L−1 solution of isoniazid was transferred to a 20-
36 (Kanto, Tokyo, Japan) in ultra-pure water and diluting to 50 mL. mL volumetric flask and diluted to the mark by ultra-pure water. It
37 A solution containing 0.5 mol L−1 NH4OH (Wako) was used. A should be mentioned that in all cases 9.2 µL of the samples were
38 stock solution of AgNO3 (0.01 mol L−1) was prepared by deposited on the paper device (Figure 2).
39 dissolving the proper amount of AgNO3 (Wako) in ultra-pure
40 water.
41 Software
42 Instrumentation The results of the experimental design were elaborated with an R-
43 based chemometric software developed by the Group of
44 Ultra-pure water was prepared by a PURELAB flex set (Veolia,
Chemometrics of the Italian Chemical Society, freely downloadable
45 Paris, France). A ColorQube 8570 wax printer (Xerox, Norwalk,
from http://gruppochemiometria.it/index.php/software.
46 CT, USA) was used for printing the wax barrier. A thermal Canon
47 ip2700 inkjet printer (Canon, Tokyo, Japan) was used for the
48 deposition of the assay reagents (PVA, NH4OH, and AgNO3). For
49 this purpose, the standard Canon printer’s black cartridge was
50 cut open and the inside sponge removed, and then the cartridge
51 was washed completely with ultrapure water and dried. Finally,
52 each cartridge was fed with about 2 mL of different assay
53 solutions, separately. Colour intensity was analysed by ImageJ
54 version 1.50f.
55
56 Device fabrication Figure 1. Schematic of PAD fabrication for the determination of isoniazid.
57
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3 reduce the number of experiments.28 Accordingly,View a CCD 6−1 +
(2Online
Article

4 DOI: 10.1039/C8AN00332G
12 star points + 2 centre points) with 46 experiments, which
5 enabled the estimation of the linear effects, the interactions
6 between pairs of factors and the quadratic terms, was applied
7 in this work. Table S1 of the ESI shows the full experimental
8 matrix for the CCD, in which the values of the factors are coded
9 according to Table 1.
10 The value of α was set to 2.4 for factors A, B and C and to 2.33
11 for factors D, E and F. The factors and their ranges were selected
12 according to preliminary experiments. The examined factors
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13 and their levels, both coded values and actual values, are shown
14 Figure 2. Schematic procedure for the determination of isoniazid using the PADs and in Table 1. In all experiments, the calculated response (∆B) is the
15 representative color scans of devices after exposure to different concentrations of blue colour intensity of the sample, which is corrected for the
16 isoniazid. corresponding blank.
17
18 Table 1. Factors and their investigated levels in CCD.

Analyst Accepted Manuscript

19 Results and discussion Levels
20 In-situ formation of silver nanoparticles Factor symbols
21 -ɑ -1 0 +1 +ɑ
According to previous studies12,21, [Ag(NH3)2]+ complex ions can
22
be reduced to silver nanoparticles at relatively high Area length (mm) A 4.6 6.0 7.0 8.0 9.4
23
concentrations of a reducing agent such as isoniazid. The
24 Area width (mm) B 4.6 6.0 7.0 8.0 9.4
proposed method is fundamentally similar to the Tollens
25 Sampling volume (µL) C 1.2 4.0 6.0 8.0 10.8
process, which was already reported for the synthesis of
26 Printing cycles of PVA D 2.0 6.0 9.0 12.0 16.0
AgNPs21. In this process, by having [Ag(NH3)2]+ as an oxidant and
27
isoniazid as a reductant, AgNPs giving rise to a yellowish colour Printing cycles of NH4OH E 2.0 6.0 9.0 12.0 16.0
28
can be obtained 22,23 (Figure 3).
29 Printing cycles of AgNO3 F 2.0 6.0 9.0 12.0 16.0
Compared to a uric acid study18, basic conditions are not
30
required because isoniazid possesses higher reducing strength.
31 Study of independent factors impact on the colour intensity
It is worth mentioning that in this method, PVA acts as a capping
32
agent and effectively stabilizes formed AgNPs 24,25. Due to the As mentioned before, with a CCD it is possible to estimate the
33
formation of AgNPs, a change from colourless to yellow occurs. coefficients of a quadratic model with interactions 29. The
34
The in-situ formation of AgNPs was verified by scanning relationship between the colour intensity (CI) and the factors
35
electron microscopy (SEM)26 as can be confirmed by comparing (as coded values) is explained by the model reported in Eq.1, in
36
the SEM images recorded of the paper surface before and after which the factors are coded according to Table 1. The
37
application of an isoniazid sample (Figure 4). The observed coefficients are also shown in Figure 5.
38
changes can be ascribed to a redox reaction between isoniazid
39
and [Ag(NH3)2]+. CI = 112.7 – 3.5 A – 1.4 B + 10.6 C (***) + 4.5 D (*) + 6.6 E
40
(**) + 20.2 F (***) + 9.7 AB (***) + 2.2 AC – 1.9 AD + 0.5 AE – 2.7
41
Structure of the experimental design AF – 0.8 BC – 1.0 BD + 2.9 BE – 0.2 BF + 3.6 CD – 4.2 CE + 6.3 CF
42
(**) + 2.4 DE + 3.8 DF – 2.6 EF – 0.9 A2 – 2.4 B2 + 3.8 C2 – 2.8 D2
43 The factors influencing the device efficiency, including area
– 4.6 E2 (*) – 6.3 F2 (**) (Eq.2)
44 length, area width, sampling amount, and amount of inkjet-
45 deposited assay reagents like PVA, NH4OH, and AgNO3, were
A is area length, B is area width, C is sampling volume, D, E
46 evaluated by performing an adequate DOE. In CCD, the total
and F are the amounts of deposited assay reagents consisting of
47 number of required experiments is determined by the following
PVA, NH4OH and AgNO3, respectively. It should be noted that
48 equation:27
these amounts are expressed as number of printing cycles used
49
for the inkjet-based deposition onto the PADs. The adjusted R 2
50 N = 2f + 2f + N0 (Eq. 1)
is 0.83 and the standard deviation of the residuals is 12.2. Stars
51
in this equation indicate the significance of the coefficients (* =
52 where, f and N0 represent the number of factors and centre
p < 0.05, ** = p < 0.01; *** = p < 0.001).
53 points (f= 6, N0 =2), respectively. In this equation, the factorial
54 part, star points and centre points are demonstrated
55 respectively. In this work, instead of the full factorial (2f), a
56 fractional factorial (2f-1) design has been utilized in order to
57
58
59
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16 Figure 3. The overall reaction between isoniazid and silver ions leading to formation of AgNPs.

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Analyst Accepted Manuscript

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33 Figure 4. Representative SEM images (500 × magnification) of a) the PAD after exposure to ultra-pure water b) the PAD after exposure to isoniazid.

34
35
36 strong interaction between the two factors can be easily
37 The significant coefficients are the linear terms for C, D, E interpreted. The fact that the best response is obtained on the
38 and F, plus the interactions AB and CF and the quadratic terms diagonal of the plot means that the ratio of the factors is crucial.
39 for E and F. This means that all factors have a significant effect It means that by increasing the value of A, the value of B must
40 on the response. Two factors—sampling volume (coded as C) also be increased. Since the values of the responses on that
41 and printing cycles of AgNO3 (F)—are by far the most relevant, direction are very similar, it can be concluded that the relevant
42 with the largest linear effects and a significant interaction (CF). parameter is the shape, and not the size of the area. In the case
43 Factor F also shows a large quadratic effect (FF). Other factors, of unequal area length and width, the observed “coffee ring”
44 which are related to assay reagents, such as printing cycles of phenomenon, a pattern caused by solute left on a substrate
45 NH4OH (E) and PVA (D) have a significant linear effect, and after evaporation of the solvent droplet, was more pronounced
46 factor E also has a significant quadratic effect (EE). Device than in the case of equal length and width, corresponding to a
47 geometry factors, including area length (A) and area width (B), square reaction area. Hence, a smaller device with a square
48 do not have a significant linear effect; however, their shape, which also requires lower consumption of the assay
49 interaction is highly significant (AB). reagents, is preferred.
50 Except for D, having only a linear significant effect (positive, The response surface on the plane C – F (Figure 7) shows
51 meaning that the higher, the better), all the other factors are that an increase of the sampling volume only produces an effect
52 involved in significantly higher terms (interactions or quadratic while the number of printing cycles of AgNO 3 is high. From a
53 terms). Therefore, to obtain a better understanding of the chemical viewpoint, by increasing the amount of AgNO 3 and
54 phenomenon, response surfaces must be drawn. In each plot, sample volume, the amount of available silver ions that can
55 the effect of two factors can be studied while the remaining four participate in the redox reaction and available isoniazid to act
56 factors were set at the centre levels. as reducing agents will be increased. Therefore, the obtained
57 The impact of area length and area width (AB) on the colour intensity will be higher.
58 obtained colour intensity is shown in Figure 6. In this figure, the A similar explanation can be given for the amounts of
59 circle defines the experimental domain. From this plot, the ammonium hydroxide (E) and PVA (D). From the plot, it can be
60

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Analyst Accepted Manuscript

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30 Figure 5. Plot of the coefficients (the green bracket indicates the confidence interval at p = 0.05; * = p < 0.05, ** = p < 0.01; *** = p < 0.001); the y-axis (dimensionless) shows the
31 magnitude of the coefficients, which is identical to the coefficient values reported in equation 2.
32
33 Response Surface: Contour Plot
NH4OH 8 printing cycles (−0.33), and AgNO3 12 printing cycles
34 (+1).
35 Performing replicated experiments (n=8) at these conditions
36 produced an average response of 179 ± 1.5 in good agreement with
60 13
2

37 70 0 the predicted value.

38 It has to be noticed that this result is significantly higher than the
80
39 90 1 20
best result obtained by the actually performed experiments within
1

40 1 00 the design matrix (165; Table S1). This clearly shows that the
41 1 10
possibility of building a predictive model allows the detection of the
110
42
B

120 1 00 real optimum inside the experimental domain. Owing to the

43 presence of several significant interactions and quadratic terms, the
44 1 30 90
OFAT approach would not have been able to find an acceptable
45
-1

14 solution.
46 0
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48 50 Study of interferences
0

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49 For this purpose, the determination of 1.0 mmol L−1 isoniazid in

50 -2 -1 0 1 2 the presence of potentially interfering substances was
51 examined. The tolerance limits of various ions and organic
52 A
substances were selected as the concentrations of the
53 Figure 6. Response surface on the plane A – B (area length vs. area width).
interfering species that cause less than ±5% relative error. The
54 tolerance limits for analyte ratios were over 500 for NO3−, Na+,
55 When trying to keep A and B as small as possible, the
K+, fructose, glucose, and sucrose; 100 for SO42− and urea; 75 for
56 conditions inside the experimental domain corresponding to
Mg2+, Ca2+; 50 for NH4+; 35 for Cl−; 10 for PO43−; 5 for citric acid
57 the highest predicted response (184) are the following: area
and 1 for ascorbic acid.
58 length 6.1 mm (coded value −0.9), area width 6.1 mm (−0.9),
59 sampling volume 9.2 µL (+1.6), PVA 11 printing cycles (+0.67),
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Figure 8. Response surface on the plane D – E (printing cycles of PVA vs. printing cycles
3 Analytical figures of merit
of ammonium hyroxide).
View Article Online

4 The relation between the colour intensity and isoniazid

DOI: 10.1039/C8AN00332G

5 concentration, under the optimal conditions, was determined

6 by using different standard solutions. Under the optimum
7 conditions, the linearity of the calibration graph ranged Application to real samples
8 between 0.03 and 1 mmol L−1 with a correlation coefficient of The optimized method was employed for the quantification of
9 0.991 (Figure S2 of the ESI), which shows a good linear isoniazid in real samples. For evaluating the accuracy of the
10 regression between the colour intensity and the concentrations. proposed method, the obtained results were compared with
11 The calibration equation was Y=72.6 C + 347.4, where Y is the the reference method (Table 2). The obtained results show the
12 colour intensity, and C is the isoniazid concentration. The capability of the device in determination of isoniazid in
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13 relative standard deviation (n=5 for the determination of 1.0 pharmaceutical samples.
14 mmol L−1) was estimated to be 3.4%.
15 Table 2. Determination of isoniazid in real samples by PADs and reference method (n=3).
16 Sample Sample taken Found Reference method19
17 Response Surface: Contour Plot (mmol L−1) (mmol L−1) (mmol L−1)
18

Analyst Accepted Manuscript

19 Tablet (300 mg) 1.0 0.99 ± 0.06 1.01 ± 0.03
90
20 Tablet (100 mg) 1.0 0.98 ± 0.08 1.00 ± 0.03
2

2 00

21
1 80

1 00 Injection (100 mg ml−1) 1.0 1.02 ± 0.15 0.99 ± 0.05

22
19

23
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0
1

24
Conclusions
16

25
0

26
14 In most reports on the development of PADs, the focus is on the
0
F

0 15
0
27 100 12 implementation of a particular sensing mechanism on a paper
0 13
28 0 substrate, while the optimization of device geometry and
90 1 10
-1

29 amounts of deposited reagents remains a challenge. In other

80
30 70 words, although the mentioned factors are often optimized, this
31
60 is mostly done using the traditional OFAT approach, which
-2

50
32 40 cannot provide any information about interactions and
30
33 quadratic effects. Therefore, it often happens that an “optimal
34 -2 -1 0 1 2
condition” far from the ideal one is selected. In the current work,
35 after performing only 46 experiments, the colorimetric
C response obtained at the optimum conditions detected by the
36
Figure 7. Response surface on the plane C – F (sampling volume vs. printing cycles of model was significantly higher than any of the responses
37 AgNO3).
38 experimentally obtained and in good agreement with the
39 predicted value. According to these results, it can be concluded
Response Surface: Contour Plot
40 that the application of design of experiments on PADs can lead
41 to low cost but sensitive devices, which are the main goal of
42 fabrication of PADs.
85
70
2

115
43 The proposed device, due to its simplicity, provides a rapid and
44 user-friendly method for on-site isoniazid quantification in
80

45 pharmaceutical preparations.
1

46
47 Acknowledgements
48 We acknowledge professor Lutgarde M.C. Buydens and her
E

49 1 10 group (Radboud University, Nijmegen, The Netherlands) for

50 105 help and advice. The authors thank Dr. Med. Ursula Nagel for
-1

51 100 her help with isoniazid tablets.

95
52 90 85
53 References
-2

80
1 A. W. Martinez, S. T. Phillips, M. J. Butte and G. M. Whitesides,
54 65 70
75
70 65 60
Angew. Chem. Int. Ed., 2007, 46, 1318–1320.
55 2 J. R. Kraly, R. E. Holcomb, Q. Guan and C. S. Henry, Anal. Chim.
56 -2 -1 0 1 2
Acta, 2009, 653, 23–35.
57 D 3 E. W. Nery and L. T. Kubota, Anal. Bioanal. Chem., 2013, 405,
58 7573–7595.
59
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3 4 N. Sharma, T. Barstis and B. Giri, Eur. J. Pharm. Sci., 2018, 111, View Article Online

4 46–56. DOI: 10.1039/C8AN00332G

5 M. Sher, R. Zhuang, U. Demirci and W. Asghar, Expert Rev. Mol.
5
Diagn., 2017, 17, 351–366.
6 6 R. A. G. de Oliveira, F. Camargo, N. C. Pesquero and R. C. Faria,
7 Anal. Chim. Acta, 2017, 957, 40–46.
8 7 G. G. Morbioli, T. Mazzu-Nascimento, A. M. Stockton and E.
9 Carrilho, Anal. Chim. Acta, 2017, 970, 1–22.
10 8 K. Yamada, T. G. Henares, K. Suzuki and D. Citterio, Angew. Chem.
11 Int. Ed., 2015, 54, 5294–5310.
12 9 K. Yamada, H. Shibata, K. Suzuki and D. Citterio, Lab Chip, 2017,
Published on 31 March 2018. Downloaded by UNIVERSIDAD DE BUENOS AIRES on 31/03/2018 08:32:46.

13 17, 1206–1249.
14 10 R. Leardi, Anal. Chim. Acta, 2009, 652, 161–172.
11 K. C. M. S. Lima, A. C. F. Santos, R. N. Fernandes, F. S. Damos and
15
R. de Cássia Silva Luz, Microchem. J., 2016, 128, 226–234.
16
12 A. Amirjani, M. Bagheri, M. Heydari and S. Hesaraki, Sensor.
17 Actuat. B Chem., 2016, 227, 373–382.
18

Analyst Accepted Manuscript

13 G. Singh, R. Pai and K. Devi, J. Adv. Pharm. Technol. Res., 2012, 3,
19 30–40.
20 14 M. A. Bezerra, R. E. Santelli, E. P. Oliveira, L. S. Villar and L. A.
21 Escaleira, Talanta, 2008, 76, 965–977.
22 15 J. Zolgharnein, A. Shahmoradi and J. B. Ghasemi, J. Chemometr.,
23 2013, 27, 12–20.
24 16 M. Shariati-Rad, M. Irandoust and S. Mohammadi, Chemometr.
25 Intell. Lab. Syst., 2016, 158, 48–53.
17 A. Avoundjian, M. Jalali-Heravi and F. A. Gomez, Anal. Bioanal.
26
Chem., 2017, 409, 2697–2703.
27
18 V. Hamedpour, G. J. Postma, E. van den Heuvel, J. J. Jansen, K.
28 Suzuki and D. Citterio, Anal. Bioanal. Chem., 2018, 410, 2305–
29 2313.
30 19 T. Rakić, I. Kasagić-Vujanović, M. Jovanović, B. Jančić-Stojanović
31 and D. Ivanović, Anal. Lett., 2014, 47, 1334–1347.
32 20 The International Pharmacopoeia (Ph. Int.), Seventh Edition,
33 2017, Isoniazid tablets,
34 http://apps.who.int/phint/pdf/b/6.1.194.Isoniazid-
35 (Isoniazidum).pdf or
36 http://apps.who.int/phint/pdf/b/6.2.2.65.Isoniazid-tablets-
(Isoniazidi-compressi).pdf, accessed on March 22, 2018.
37
21 M. Amjadi and E. Rahimpour, Microchim. Acta, 2012, 178, 373–
38
379.
39 22 L. P. Wu, Y. F. Li, C. Z. Huang and Q. Zhang, Anal. Chem., 2006, 78,
40 5570–5577.
41 23 J. He, T. Kunitake and A. Nakao, Chem. Mater., 2003, 15, 4401–
42 4406.
43 24 V. K. Sharma, R. A. Yngard and Y. Lin, Adv. Colloid Interfac. Sci.,
44 2009, 145, 83–96.
45 25 Y. Yin, Z.-Y. Li, Z. Zhong, B. Gates, Y. Xia and S. Venkateswaran, J.
46 Mater. Chem., 2002, 12, 522–527.
47 26 N. Pourreza, H. Golmohammadi, T. Naghdi and H. Yousefi,
Biosens. Bioelectron., 2015, 74, 353–359.
48
27 F. N. Arslan and H. Azak, Food Anal. Method., 2018, 11, 1163–
49 1179.
50 28 S. M. Sanchez and P. J. Sanchez, ACM T. Model. Comput. S., 2005,
51 15, 362–377.
52 29 D. Baş and İ. H. Boyacı, J. Food Eng., 2007, 78, 836–845.
53
54
55
56
57
58
59
60

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