Trends in Analytical Chemistry 93 (2017) 212e227

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Trends in Analytical Chemistry

journal homepage: www.elsevier.com/locate/trac

Paper-based microfluidic analytical devices for colorimetric detection

of toxic ions: A review
Ganesan Sriram a, Mahesh P. Bhat a, Pravin Patil a, Uluvangada T. Uthappa a,
Ho-Young Jung b, Tariq Altalhi c, Tushar Kumeria d, Tejraj M. Aminabhavi e,
Ranjith Krishna Pai f, Madhuprasad a, **, Mahaveer D. Kurkuri a, *
a
Centre for Nano and Material Sciences, Jain University, Jain Global Campus, Bengaluru, 562112, Karnataka, India
b
Department of Environmental Energy Engineering, Chonnam National University, Gwangju, 500-757, Republic of Korea
c
Department of Chemistry, Faculty of Science, Taif University, Taif, Saudi Arabia
d
Department of Chemistry and Biochemistry, University of California, La Jolla, San Diego, CA, 92093, USA
e
Soniya College of Pharmacy, Dharwad, 580002, Karnataka, India
f
Technology Mission Division, Department of Science and Technology (DST), Ministry of Science and Technology, Government of India, Technology Bhavan,
New Mehrauli Road, New Delhi, 110016, India

a r t i c l e i n f o a b s t r a c t

Article history: Paper-based microfluidic analytical devices (mPADs) are becoming valuable tools in analytical area due to
Available online 19 June 2017 their attractive passive movements of analyte without any external forces due to capillary phenomenon.
Techniques used to design microfluidic patterns are cost-effective, less sample requirement, environ-
Keywords: mentally benign and rapid analysis. The application of mPADs with intricate fabrication methods have
Paper-based microfluidic devices (mPADs) emerged over a decade in several fields such as diagnostics, biological, food safety, environemental
Hydrophilic channel
analysis, electrochemical and colorimetric detections. The present review addresses colorimetric
Hydrophobic boundary
detection of metal ions and nitrites from various sources using a simple filter paper (Whatman®) as
Capillary action
Metal ions
mPADs, since filter paper is white in color, possibility of liquid transport through capillary action and high
Nitrite surface area offers high detection efficiency. Due to its importance as an analytical tool, an attempt has
Colorimetric detection been made here to critically discuss the present scenario and future prospects of this technique.
© 2017 Elsevier B.V. All rights reserved.

1. Introduction
Paper-based microfluidic analytical devices (mPADs) are gaining
enormous attention due to their applications in many interdisci-
plinary areas. The mPADs, also known as point-of-care devices or
health care devices have relevance in environment, diagnosis and
food assay [1e3]. Lab-on-a-chip devices are basically fabricated
using polydimethylsiloxane (PDMS), but mPADs or lab-on-a-paper
devices are fabricated on a filter paper [4,5]. These have gener-
ated a great deal of interest due to their hydrophilic nature, rapid
fabrication, easy to transfer complex patterns on a paper, ease of
handling, cost effectiveness, high wicking properties, high porosity,
disposability, non-toxicity, biodegradability and biocompatibility.
* Corresponding author.
** Corresponding author.
E-mail addresses: madhuprasad@jainuniversity.ac.in (Madhuprasad), mahaveer.
kurkuri@jainuniversity.ac.in (M.D. Kurkuri).
The white color of filter paper and its hydrophilic nature facilitates
its usage in rapid colorimetric assay without any requirement of
external forces to drive the analyte into the detection zone. Various
fabrication methods of mPADs and overall schematic of colorimetric
detection process using mPADs is shown in Fig. 1.
The working principle behind mPADs is that hydrophobic
boundary offers a physical barrier for sample dispersion pathway in
hydrophilic zones. Each mPADs are fabricated with different
detection zones that are coated with the respective organic re-
ceptors for detecting the target toxic ions. The solutions travel by
passive capillary action through a patterned path of filter papers to
the zone of detection for simultaneous multiple analyses. When the
analyte comes in contact with an organic receptor at the detection
zone there will be a rapid color change, which makes it a versatile
device (Fig. 2). This color change is noticeable by naked eye and can
be easily captured by electronic devices such as high resolution
camera or photo-sensors [6]. The fabrication of lab-on-a-chip de-
vices involves a tedious process [7] compared to mPADs fabrication

http://dx.doi.org/10.1016/j.trac.2017.06.005
0165-9936/© 2017 Elsevier B.V. All rights reserved.
 G. Sriram et al. / Trends in Analytical Chemistry 93 (2017) 212e227 213

Fig. 1. Various techniques for the fabrication of mPADs for colorimetric detection of toxic ions.

Fig. 2. Schematic representation of A) front and B) back side of mPADs: Capillary action of yellow color dye in mPADs and rapid color change in detection zone due to the reaction
with preloaded reagents [Reproduced with permission from Ref. [32], Copy right 2016, The Royal Society of Chemistry].

where low cost and simple filter paper [8] been used. Thus, mPADs
fabrication includes development of hydrophobic boundaries and
hydrophilic channels by various techniques as discussed in recent
reviews [9e13].
The mPADs are first developed in 2007 by Martinez et al. using a
photolithographic technique for bioassay application [14].
Currently, researchers are moving towards designing novel mPADs
with new technological platforms for different applications
[15e17]. Over the past few years, development and functionality of
mPADs platform are embedded with nanoparticles for bioassay [18],
sensor probe for detection of cancer biomarkers [19] and integrated
electronics for diagnostic applications [20]. However, mass spec-
trometry (MS) and electrochemistry (EC) are playing crucial roles in
paper devices for biological applications. For instance, Wang et al.
214 G. Sriram et al. / Trends in Analytical Chemistry 93 (2017) 212e227
[21] have used paper spray method for the robust quantification of
complex mixtures using MS. A triangle shaped small paper was
used in which imatinib, a 2-phenyl-aminopyrimidine derivative
was identified using specific mass spectrum peak corresponding to
imatinib among several other compositions found in blood sample.
Further, the study was extended to separate two different dyes viz.,
methylene blue and methyl violet which was confirmed by MS. This
method has potential clinical applications such as therapeutic drug
monitoring, neonatal screening and personalized medicine. Simi-
larly, the paper based bio-chips were fabricated for cell culture
purpose, which has potential applications in bioassays and drug
screening process. The cell compatible paper chips for cell culture
were monitored using MS [22]. Furthermore, Liu et al. [23] used
wax printing to fabricate origami-paper based microfluidic devices
to make self-powered device for the detection of adenosine using
aptamer by electrochemical method. This method has advantages
of low cost, rapid detection and portability for biological assay. In
device fabrication, simple capacitor and digital multi-meter was
circuited with carbon electrode on the paper followed by lami-
nating a paper for the protection from external contaminants and
evaporation of fluids. Detection of adenosine was based on changes
observed in electrical signal via digital multi-meter due to the
presence of biomolecules on the device. In addition, commercially
available normal A4 size paper have been used for the analysis of
the enzyme inhibition assay and similar paper is modified by
plasma etching to fabricate mPADs [84,85]. For normal papers,
mainly ink jet printing technique has been used to fabricate mPADs.
On the other hand variety of filter papers available in the market,
which can be used for the fabrication of mPADs, filter papers with
less pore size, ash free, high purity and low thickness have been
used where proper capillary action drives the analyst molecules.
Since normal papers lacks in above mentioned qualities, the
application of such papers in colorimetric detection of toxic ions is
not possible. Another advantage with filter paper over normal pa-
per is that the filter paper is chemically resistant and has high liquid
holding capacity. Hence the filter papers have extensively been
used in the fabrication of mPADs for various applications.
Water is essential for all living beings, and water contamination
by toxic ions may cause severe damage to human health and hy-
giene if absorbed in the human body beyond a threshold level. The
World Health Organization (WHO) and United States-
Environmental Protection Agency (US-EPA) have set permissible
limits for toxic ions like Hg2þ (WHO: 1.0  103 and EPA:
2.0  103 ppm), Pb2þ (WHO: 1.0  102 and EPA: 1.0  102 ppm),
As3þ (WHO: 1.0  102 and EPA: 1.0  102 ppm), Cu2þ (WHO: 2.0
and EPA: 1.3 ppm), Ni2þ (WHO: 2.0  102 ppm), Cr ions (WHO:
5.0  102 and EPA: 1.0  101 ppm), Zn2þ (WHO: 3.0 ppm), Fe2þ
(WHO: 5.0  101 ppm) and nitrite (WHO: 50.0 and EPA: 1.0 ppm)
for potable water [86,87]. Thus, passive capillary action of filter
paper can be utilized for colorimetric detection of toxic metal ions
and nitrite in water. The objective of this review is to focus on
recent fabrication techniques of mPADs and their key roles in
colorimetric detection of toxic ions. Also it discusses on the
mechanism involved in colorimetric detection of toxic metal ions
and nitrites from environmental samples.
2. Fabrication of paper-based microfluidic devices (mPADs)
In 2007, Martinez et al. [14] first developed mPADs by photoli-
thography, but the method was complicated due to the usage of
photoresist, clean-room facility, UV light, mask and mask aligner. In
order to avoid such sophisticated techniques, new methodologies
were developed to fabricate these devices, giving rise to the
patterning of hydrophobic barrier and hydrophilic channel on filter
paper as discussed in recent reviews [9e13,24e26].
2.1. Lithography
Patterns designed through photolithography offers high reso-
lution features, and is well adopted method in the field of semi-
conductors to develop very small electronic devices. Same
technique was applied to create mPADs, but cost of photoresist and
clean room facilities are high, making it an expensive approach.
Kakoti et al. [27] adopted similar photoresist approach to fabricate
mPADs by Fast Lithographic Activation Sheets (FLASH), which re-
quires less than 30 min for device fabrication. The FLASH method
did not require sophisticated facilities. Sun light was used in
fabrication process instead of UV light. The patterns were devel-
oped on filter paper with a thickness of 180 mm using SU-8 and
photomask, followed by coating of SU-8 photoresist at the rear side
and cross linked by exposure to UV light, which would provide the
leak proof mPADs (Fig. 3A). This technique has limitations such as
high cost of photoresist and also involves multistep fabrication
process. Songok et al. developed rapid fluid flow in narrow con-
structed hydrophilic channel to eliminate the evaporation of sam-
ples via coating photocatalytic TiO2 nanoparticles because of which
liquid moves rapidly to the detection zone [28]. The TiO2 nano-
particles were coated on whole filter paper of 180 mm thickness by
liquid flame spray technique followed by UV irradiation through
photomask to develop a narrow hydrophilic channel for rapid de-
livery of samples into detection zone to enhance the colorimetric
detection. Asano et al. [29] have developed a different approach
using photolithographic method. A filter paper with thickness of
170 mm was soaked in octadecyltrichlorosilane (OTS)-n-hexane
mixture to make hydrophobic surface, which was dried under ni-
trogen atmosphere. Further the paper was exposed to UV light
through a photomask to create a hydrophilic region. The photo-
mask was custom-designed by 3D printer by using a mixture of
acrylic monomers. The advantage of this type of technique is easy
to fabricate with multiple channels. The dimensions of designed
hydrophilic region and hydrophobic barrier were 500 and 100 mm,
respectively. Again this technique is time consuming and non-eco-
friendly as OTS was used.
2.2. Three dimensional (3D) printer
In order to overcome the drawbacks of lithographic method,
desktop 3D printer was used. It is an inexpensive fabrication
technique for the development of 3D hydrophilic channels on
hollow structured polymeric substrate. The technique has advan-
tages as it can be used for patterning complicated 3D structure,
rapid fabrication and cost-effective. He et al. [30] followed this
technique using the filter paper in the form of cellulose powder
having the size of 74e125 mm, which acts as a sample transport
medium via capillary action such that analyte solution could move
very fast to the detection zone (Fig. 3B). This technique could offer
high resolution features depending on the 3D printer's printing
resolution and depth of the channel could also be controlled very
precisely.
Apart from using 3D printer for making channels, the printer
could also be used to make complicated masks. Salentijn et al. [31]
developed 3D printed mask to fabricate mPADs by alkyl ketene
dimer (AKD), which was used as a hydrophobic barrier for colori-
metric applications. Here, the area coated with low concentration
of AKD will define hydrophilic channels for permeability of sample
liquid.
2.3. Wax jetting
Li et al. [32] developed a wax jetting method that was useful in
bio-assay applications. This technique consists of a wax droplet
 G. Sriram et al. / Trends in Analytical Chemistry 93 (2017) 212e227
Fig. 3. Schematic fabrication of mPADs by (A) FLASH lithography [Reproduced with the permission from Ref. [27], Copy right 2015, American Institute of Physics Publishing], (B)
desktop 3D printing process [Reproduced from Ref. [30]] and (C) wax jetting process [Reproduced with the permission from Ref. [32], Copy right 2016, The Royal Society of
Chemistry].
generator, which has a piezo electric actuator (PZT), glass nozzle,
wax container, and one aluminum connector placed on a 3D
printing table. The cylindrical wax container was then filled with
paraffin and heated to melt. The molten wax was jetted by applying
voltage through glass nozzle, which was forged with micro-nozzle
of 150 mm diameter, creating a patterned hydrophilic channel of
around 600 mm (Fig. 3C). This method can be applied for colori-
metric detection of toxic ions as well. However, the technique in-
volves expensive set up due to the integration of electronics and
mechanical parts, which makes it expensive and also limits the
remote area applications.
2.4. Wax smearing
Wax has widely been used to create hydrophobic barriers in
mPADs technology, Chen et al. [33] for example have developed
mPADs by wax smearing method to detect urinary infection in
adult's diapers. The mold used was designed by AutoCAD software
and printed on a 3 mm thick poly(methyl methacrylate) (PMMA)
sheet and CO2 laser was used to cut printed pattern on PMMA
substrate. Solid wax was smeared on the surface of PMMA, fol-
lowed by sticking filter paper of thickness 180 mm and heating it on
a hot plate at 90 C for few min. The molten wax penetrated into the
filter paper to form hydrophilic and hydrophobic zones, which was
cooled at ambient temperature. PMMA mold and paper were then
removed to obtain patterned mPADs with an average width of 3 mm,
but the technique involved multistep process requiring sophisti-
cated facilities.
2.5. PDMS screen printing
Screen printing was also used to develop hydrophobic barriers
and hydrophilic channels on filter paper using hydrophobic PDMS
215
sheet, which was better than SU-8 photoresist because of the
flexibility nature of PDMS materials. Mohammadi et al. [34] intro-
duced PDMS screen printing for colorimetric detection in which the
fabrication process involved developing a mask using Adobe Illus-
trator software that was printed on screen stencil using a printer.
The mask printed as screen stencil was placed on filter paper of
thickness 180 mm followed by pouring PDMS solution, which was
rubbed on filter paper through stencil using squeegee followed by
gentle removal of stencil. The paper was kept in an oven for thermal
curing followed by drying in an ambient temperature. Thus
developed hydrophilic channels were 800 mm in width surrounded
by a hydrophobic barrier (Fig. 4A). Recently, Hu et al. [35] developed
mPADs for colorimetric detection of Fe3þ ions by fluorescent carbon
dots (FCD) and detection of ferritin by gold (Au) nanoparticles
coated on none-fluorescent carbon dots (nFCD) spiked in whole
blood samples. In briefly, the filter paper was placed between
plasma modified PDMS mask and bare PDMS sheet. Then, through
hydrothermal process the deposition of nFCD on exposed region of
filter paper and FCD on non-exposed region on PDMS was per-
formed. The biocompatible Au nanoparticles were coated on nFCD
regions, here the FCD regions were utilized for the analysis of Fe3þ
ions and Au nanoparticles coated nFCD regions for ferritin analysis
from whole blood samples. This fabrication process can be used for
the colorimetric detection of toxic ions as well.
2.6. Permanent marker pen
Permanent markers are usually water resistant, but soluble in
alcohol. Permanent markers are very cheap, have high contrast for
colors, different designs can be drawn manually, and can create
hydrophobic barrier on a filter paper due to the hydrophobic nature
of dyes. Hence these permanent markers can be used to fabricate
mPADs. Gallibu et al. [36] developed mPADs using permanent
216 G. Sriram et al. / Trends in Analytical Chemistry 93 (2017) 212e227
Fig. 4. The fabrication process of mPADs by A) PDMS screen printing [Reproduced from Ref. [34]] and B) ink-jet printing [Reproduced with the permission from Ref. [46], Copy right
2016, Elsevier].
marker for colorimetric detection of glucose. The patterns were
designed by Inkscape software and printed on filter paper by XY
plotter using a permanent marker. Further, the paper was dried to
evaporate the solvent present in the ink, which developed hydro-
phobic barrier and hydrophilic channels.
Nuchtavorn et al. [37] fabricated mPADs using technical drawing
pens by a software controlled desktop electronic plotter/cutter.
Filter paper with thickness of 180 mm was laminated with micro
level thick transparent film to obtain the desired patterns. The
laminated paper was placed on a cutting mat followed by technical
drawing pen filled with permanent ink, which was inserted into the
holder of electronic plotter for patterning. Similarly, Jiang et al. [38]
used permanent markers to develop mPADs, where pattering was
performed with the help of plastic template placed on filter paper,
followed by curing at room temperature to evaporate the solvents
and the remaining resins to create hydrophilic and hydrophobic
regions. The developed mPADs can be used for biosensing and
colorimetric applications. This type of techniques are cheaper than
other techniques (Table 1), and offers mass production of mPADs.
2.7. Wax printing
In this fabrication technique, wax is used as a hydrophobic
material that is patterned on paper as a barrier for analytes. Wax
has advantages of being cheap, water insoluble, low viscosity after
melting, can rapidly penetrate into the filter paper, is eco-friendly
for easy disposable in comparison with other hydrophobic sub-
stances such as SU-8 photoresist, PDMS and hydrophobic chem-
icals. Tenda et al. [39] developed mPADs by hot lamination method,
where wax was printed on 180 mm thick filter paper using wax
printer followed by hot lamination using transparent polymeric
sheet. During this process wax was melted on the surface of a filter
paper to create hydrophilic and hydrophobic patterns. Recently,
Kaewarsa et al. [40] developed 24 analytical circular zones on filter
paper by wax printing to diagnose the Glucose-6-phosphate de-
hydrogenase (G6PD), deficiency of which results in genetic hemo-
lytic disorder. Each circular zone have a diameter of 0.52 cm and
1 cm gap between the each other. The circular zones were designed
using Microsoft PowerPoint followed by wax printing. The wax
 G. Sriram et al. / Trends in Analytical Chemistry 93 (2017) 212e227 217

Table 1
Details of fabrication techniques of mPADs.

Methods Hydrophobic Filter Paper Time (min) Cost Advantage Disadvantage Reference
materials (Grade)

FLASH SU-8 photoresist 1 60 High Low wicking property, leak Multistep process, high cost [27]
proof, sun light can be used of photoresist
to cure photoresist and low
sample volume
Photolithography TiO2 1 60 High Higher resolution and rapid Liquid flame spray and spin [28]
fluid flow coater used for fabrication
process and Multistep
process
Photolithography OTS-n-hexane 51B 1440 Low Higher resolution and good Multistep process, time [29]
device stability consuming, non
environment friendly
3D printing Polylactide Cellulose powder 90 Moderate Higher resolution, complex Large quantity of PDMS is [30]
and PDMS patterning and multi required
analysis
3D printing AKD 1 30 Moderate Rapid process, high Oxygen plasma and masks [31]
resolution, moderate are required
hydrophobic barriers
Wax jetting Wax Filter paper e Moderate Multi assay on a single Electronic devices are [32]
paper, no mask, rapid required
fabrication, high resolution
Wax smearing Wax 1 40 Low High resolution Laser cutting and mold are [33]
required
PDMS screen printing PDMS 1 30 Moderate Multi-analysis, rapid Handling viscous PDMS, [34]
fabrication, ‘n’ number of thermal curing is required
designs through screen
stencil on a single paper,
good stability
PDMS mask PDMS Filter paper 1450 Moderate Rapid fabrication, easy Multi-step fabrication [35]
handling and multi- process and large volume of
detection in a mPAD PDMS used in fabrication
process
Permanent marker pen Hydrophobic ink 1 30 Very low Raid process, ease of Low resolution and ink may [36,38]
fabrication, complex not penetrate into the
channels and possibility of paper due to rapid
mass production evaporation of solvent
Technical drawing pen Hydrophobic ink 1 Short time Very low Simple process, can Low resolution [37]
produce ‘n’ number of
designs on a single paper
and no need of hot plate
Wax printing Wax 1, 4, 1, 1, 1, 1, 1, 1 5 Low High resolution, very low Limited only up to 0.9 mL of [39e45]
sample volume (0.5 mL) sample in channel, hot
required lamination/hot plate
method used to melt the
wax into the paper
Ink-jet printing HMDS and TEOS 4 10 Low Rapid process, one step Toxicity of TEOS, low [46,47]
printing and possibility of resolution, multiple
mass production printing could damage the
barrier wall
Eyeliner pencil Wax 41 5 High Simple process, easy to CO2 laser used for cutting [48,49]
draw complex channels the template makes the
through template, multi- process high cost and
detection and non-toxic difficult to control the wax
barrier due to high
temperature
Cutter printer e 1 Less than 5 Low Mass production, Used polymer sheets for [50]
fabrication of 2D and 3D lamination on filter paper
mPADs, no sample leakage which makes it non-
due to plastic adhesive to ecofriendly
seal the paper and no
tearing problem
Solegel method Fluoroalkyl silica sol, Filter paper 240 Low Produces superior The prepared pattern of sol [51]
methyl silica sol hydrophobic barrier, simple need one month of aging
and wax process, no need of
template or any other mask
to fabricate the pattern
Scholar glue spray Scholar glue Filter paper 130 Low Instrument free fabrication Usage of UV light, usage of [52]
process, non-toxic, lamination makes the
hydrophobic barrier highly process non eco-friendly
resistant for organic
solvents
218 G. Sriram et al. / Trends in Analytical Chemistry 93 (2017) 212e227
printed filter paper was kept on a hot plate at 150 C for few minutes
to melt the wax into the filter paper in order to develop hydrophilic
and hydrophobic regions for diagnosing the G6PD. Further, Kong
et al. [41] developed mPADs by wax screen printing technique. The
screen stencil was developed by Adobe Illustrator software and the
stencil was used as a mask on chromatograph filter paper to make a
wax pattern on paper by rubbing the wax through stencil. The wax
screen printed paper was placed in an oven for few seconds to melt
the wax and penetrate into the paper which creates hydrophilic
and hydrophobic barrier. Raman spectroscopy has been used to
analyze the structure of a molecule, however, drawbacks of Raman
spectroscopy is low sensitivity during the interaction of molecules.
Hence, Lee et al. [42] have addressed this problem to enhance the
sensitivity of Raman spectroscopy by coating metallic (Au) nano-
structure on mPADs: the hydrophobic barrier was patterned on filter
paper by wax printing followed by curing it in an oven at 130 C for
few seconds. Then, Au nanoparticles were coated on hydrophilic
path by successive ionic layer absorption and reaction method
(SILAR) to enhance the Raman signal for wastewater assay.
mPADs are low-cost analytical devices for fluid transport and
colorimetric analysis. However, mPADs have few drawbacks such
as maintaining flow of fluid in particular direction into the
detection zones, controlling the hydrophobic barrier formation
during heating and reproducibility. To address this problem,
Akyazi et al. [43] have optimized the temperature to control the
wax barrier formation followed by applying ionogel based mate-
rial by drop casting technique to maintain controlled fluid flow
direction of analytes. The desired pattern was developed by
AutoCAD software and the pattern was printed on filter paper by
wax printer. Later, wax printed filter paper was placed on a hot
plate to form controlled wax barrier by controlling temperature at
125 C with 6 min. The optimized temperature gave the uniform
wax barrier on front and back side of the filter paper. The drop
casted ionogel material on the detection zone was photo-
polymerized by irradiation of light, the shape of ionogel on the
detection zone was controlled by gasket. The ionogel material on
the detection zone offers quick absorption of sample liquids from
the inlet zone.
Recently, Kudo et al. [44] embedded cellulose material and
mPADs in between 3D holder to process large amount of sample
liquids for high sensitivity in colorimetric detection applications.
Adobe Illustrator CC software was used to develop the hydrophobic
pattern followed by printing on filter paper using wax printer. The
hot lamination was performed to melt the wax into the filter paper
to create barrier. Then, different sizes of glass fiber were stacked on
mPADs. Further, highly water absorbing cellulose porous materials
were place under mPADs. The above materials were embedded
between 3D holders for further application. In this process,
cationically charged polymeric nanoparticles were coated on the
detection zone by ink-jet printer. Chromogenic indicators were
electrostatically attached to the surface of cellulose surface for
colorimetric detection.
mPADs play very important role in the environmental and bio-
logical applications in resource limited areas. The enhancement of
flexibility, strength, evaporation of sample liquid and external
contamination can be avoided by laminating mPADs. Thus, Wang
et al. [45] addressed the mPADs fabrication by covering polymer
composite film through lamination. In this technique poly-
propylene film was used as a base material and surface was treated
by oxygen plasma to enhance the hydrophilic property. Further,
polyvinyl butyral solution was coated on modified film by spin
coater. The composite film was drilled with pinhole of diameter of
1 mm for liquid inlet, the wax printer was used to print the wax on
filter paper followed by curing on hot plate to form hydrophobic
barrier, and finally the device is sealed through lamination.
2.8. Ink-jet printing
Ink-jet printing technology has advantages such as cost effec-
tive, time-saving, mass production, direct patterning of hydrophilic
and hydrophobic barrier on paper, multi analyte detection without
having the necessity of specific mask and clean room facilities.
Malekghasemi et al. [46] used 250 mm thick filter paper to develop
mPADs using ink-jet printer followed by microwave irradiation.
Hexamethyldisilazane (HMDS) and tetraethylorthosilicate (TEOS)
were used to produce hydrophobic paper by three different
methods such as heating, plasma treatment and microwave irra-
diation. Ink-jet printer was used to develop hydrophilic channel on
modified hydrophobic paper. However, the entire fabrication pro-
cess of ink-jet printing followed by microwave irradiation requires
more than 10 min (Fig. 4B).
A small volume of liquid sample would move through hydro-
philic path by capillary action to the detection zones for colori-
metric application. The usage of small volume might restrict the
rapid flow of analyte. Hence, Henares et al. [47] have used bulk
volume of liquid sample in hydrophilic regions to enhance liquid
transport to the detection zone by ink-jet printing for colorimetric
detection of metal ions. The pattern was designed using Microsoft
PowerPoint and printed on filter paper by ink jet printer, thus
developed channel widths range from 0.3 to 3.5 mm. The UV
curable ink was used to develop hydrophobic barrier. The solution
of cationically charged polystyrene nanoparticles were printed on
the detection zones in mPADs to enable for the chromogenic indi-
cator to bind electrostatically onto the substrate which will further
interact electrostatically with metal ions. The bulk liquid sample
used in mPADs will enhance the liquid flow much faster than the
small volume used with higher sensitivity and lower evaporation
during the capillary flow.
2.9. Eyeliner pencil
Recently, wax containing eyeliner pencils have been used to
develop the mPADs for colorimetric detection application. The
pencil is inexpensive, could be used to design many paper devices,
easy to use and eco-friendly. Hence, Ostad et al. [48] used simple
eyeliner pencil to develop mPADs for colorimetric detection of
metal ions. AutoCAD software was used to design the patterns,
CO2 laser was used to cut the designs on white sticker. The
patterned sticker was pasted on filter paper, followed by applying
the wax through eyeliner pencil. The filter paper was cured on the
hot plate at 150 C for few minutes to melt the wax and to form
hydrophobic and hydrophilic areas. Finally, the white sticker was
peeled off and used for colorimetric applications. Recently, Aba-
dian et al. [49] developed digital mPADs by eyeliner pencil. The
mask was developed by CorelDraw followed by lithography pro-
cess to develop the polyester screen mesh. The filter paper was
placed under polyester screen mesh and rubbed with eyeliner
pencil to develop patterns on the filter paper. Later, it was heated
on a hot plate at 100 C for few seconds to allow the wax to
penetrate into the filter paper. Further, the developed mPADs were
combined with electrode patterns designed by polyester screen
mesh using conductive ink. These hybrid digital mPADs were used
for colorimetric detection of biological assay and could be used for
metal ion detection as well.
2.10. Cutter printer
Though filter paper is inexpensive material, techniques such as
lithography and wax jetting used to develop mPADs makes the
whole process expensive. So, Oliveira et al. [50] develop mPADs by
cutter printer which is low-cost technique and easy to cut soft filter
 G. Sriram et al. / Trends in Analytical Chemistry 93 (2017) 212e227
papers with desired pattern. The adhesive tape was used to cover
one side of filter paper and cutter printer was used to cut the exact
design on the filter paper. Then, filter paper was heated on hot plate
at 120 C to melt the adhesive tape followed by sealing the back side
of filter paper by heat press machine to prevent solvent leakage, to
avoid contamination and the evaporation. Cutter printer provides
minimum zone diameter of 0.5 and channel width of 0.3 mm. The
cutter printer has been used to fabricate 2D and 3D mPADs for
colorimetric detection applications.
2.11. Hydrophobic solegel fabrication
The solegel process was used to fabricate hydrophobic/lip-
ophobic boundaries on filter paper, which has gained much
attention of researchers in analytical fields. Solegel process has
advantages such as it is a simple process, low-cost, no need of clean
room facilities, no need of sophisticated instruments and simple
synthetic methods. Zhang et al. [51] used solegel process to fabri-
cate hydrophobic/lipophobic barrier on filter paper for biological
and environmental applications. Three different hydrophobic ma-
terials such as fluoroalkyl-silica, methyl-silica and wax were used
to develop the patterns on filter paper. But, fluoroalkyl-silica gave
high hydrophobic boundaries compared to methyl-silica and wax.
The fluoroalkyl-silica and methyl-silica were applied on filter pa-
pers by writing brush. The colored wax presents in crayon pencil
was also used to pattern on the filter paper. Where, fluoroalkyl-
silica hydrophobic boundaries showed more hydrophobicity than
other two materials.
2.12. Scholar glue spray method
The fabrication methods such as wax printing, ink-jet printing,
polymer based screen printing etc. gives hydrophobic barriers in
defined boundaries. However, in some cases the organic solvents
may destroy or cross the barrier resulting in reliability of the
mPADs. Hence, Cardoso et al. [52] addressed this problem by
developing the mPADs with strong hydrophobic barrier by using
scholar glue spray method. The scholar glue is low-cost, non-toxic
and is white colored poly(vinyl acetate) solution which is highly
hydrophobic in nature. In fabrication process, the magnetic mask
was used to pattern the hydrophobic design on filter paper. Then,
polymeric magnetic sheet and neodymium magnet was placed
under the filter paper for holding the whole set up. The scholar
glue was sprayed on filter paper over magnetic mask. The sprayed
filter paper was heated at 130 C for 1 h followed by UV light
irradiation to form strong hydrophobic barriers for the colori-
metric detection.
In this section various methods to fabricate mPADs were dis-
cussed. Among all the fabrication techniques published to date,
wax printing offers cost effective, simple, stable, high resolution
and eco-friendly and simple fabrication process. This technique
provides relatively high resolution patterns without sophisticated
instruments. On the other hand, FLASH, wax jetting, 3D printing are
expensive due to the sophisticated instruments involved in the
fabrication process. Few fabrication techniques like ink-jet printing
using hydrophobic chemical and photolithography involve the use
of hazardous chemicals, solegel method gives good hydrophobic
barriers but the method requires a month time for aging. Eyeliner
pencil method needs expensive instrument to develop template/
mask for hydrophobic patterns, cutter printer and scholar glue
method using hot lamination technique requires polymer films
where the production cost would be moderate, but are not eco-
friendly. The whole summary of recent developments in fabrica-
tion techniques of mPADs for colorimetric detection is shown in
Table 1.
219
3. Colorimetric detection using mPADs
In recent years, colorimetric detection is gaining much impor-
tance due to its rapid detection efficiency, naked eye detection, low
cost, remote area applications, easy to operate, and stability after
detection. These mPADs have been used for colorimetric detection
of various target analytes including biomolecules and toxic ions. It
is defined by the passive movement of the analyte solution to test
zones by capillary action and thereby reacting with precisely
loaded reagents followed by noticeable color change. The recent
reviews on the applications of mPADs are mostly focused on bio-
logical [3,11,12,53e55], electrochemical [56,57] and environmental
applications [58,59]. Though there has been review published
recently on colorimetric detection of heavy metals from water
samples using mPADs, the major focus was on the fabrication and
detection of metal ions using colorimetric, fluorescent and elec-
trochemical methods, however in this review all the fabrications
techniques were not summarized with the specific application of
metal ion and nitrite ions detection [60]. Thus, the current review
emphasizes on the most recent advances in fabrication techniques
and colorimetric detection of toxic metal ions and nitrites.
3.1. Detection of metal ions
Recently, Rattanarat et al. [61] used a filter paper of thickness
180 mm to develop mPADs by wax printing for colorimetric detec-
tion of total chromium (Cr3þ ions) from baghouse samples. The
Cr3þ ions are toxic to humans and hence, detection of Cr3þ ions
colorimetrically using mPADs is important. Initially, baghouse dust
sample containing metal was suspended in a 10 mm air filter punch
of mixed cellulose ester (MCE) and the filter was placed on a
detection zone of mPADs. PDMS lid with four detection holes and
one sample reservoir hole of diameter 5 and 2 mm respectively
were placed on filter sample along with mPADs. Then, acetate buffer
solution was added through reservoir hole such that metal ions
were eluted from the filter sample to the detection zones followed
by complete color change within 10 min. Tetravalent cerium (Ce4þ)
was treated with chromium in pretreatment zone to oxidize
chromium to Cr6þ which reacts with 1,5-diphenylcarbazide (1,5-
DPC) to form 1,5-diphenylcarbazone and Cr3þ. The appearance of
purple color confirms the presence of Cr3þ ions and the limit of
detection is 1.2  104 ppm, which is very sensitive (Table 2).
Furthermore, Feng et al. [62] fabricated enrichment-based
mPADs using a filter paper of thickness 215 mm by wax printing
for colorimetric detection of heavy-metal ions using pyridylazo
indicators. The sensitivity of mPADs was enhanced by pasting water-
adsorbent resin at the end of each channel using a hydrophilic gum.
Enrichment based mPADs shows excellent sensitivity for the heavy
metal ions in comparison to conventional mPADs. The fabricated
enrichment-mPADs could reserve up to 800 mL of sample solution,
wherein the closed end type of mPADs have the maximum limit of
20 mL. The Hg2þ ion is one of the heavy-metal ions, which was
tested in the range of 50 mM in enrichment approach as well as
conventional mPADs having eight types of pyridylazo indicator
compounds. The indicators are immobilized on mPADs by coating
polymer matrix on all zones in order to enhance the sensitivity of
colorimetric detection. Also, the same concentration of eight
different heavy-metal ions such as Agþ, Cd2þ, Cu2þ, Hg2þ, Ni2þ,
Pb2þ, Zn2þ, and Co2þ with pyridylazo indicator were used to test
with the enrichment-based mPADs, which showed good sensitivity.
Gold nanoparticles were also employed in mPADs fabricated
using 340 mm thick filter paper developed by wax printing for rapid
colorimetric detection of Hg2þ ions from pond and river water [63].
Au nanoparticles were used for rapid detection of ions due to its
high affinity with analyte. The Au nanoparticles were
Table 2

220
Summary of colorimetric detection of different metal ions and nitrite from various samples in mPADs.

Methods Receptor Nano particle Detection Mechanism Limit of detection Time (min) Sample Reference

Wax printing 1,5-diphenylcarbazide (1,5-DPC) e Cr3þ Tetravalent Ce(IV) was oxidized Cr to Cr(VI). It 1.2  104 ppm 10 Baghouse [61]
reacts with 1,5 e diphenylcarbazide to form
1,5-diphenylcarbazone and Cr(III), the
formation of purple color confirmed the
presence of Cr(III) ions
Wax printing Pyridylazo indicators e Hg2þ The enrichment based mPADs with eight e 5 Sewage water [62]
detection zones was coated by eight different
indicators in each zone for high sensitivity
towards metal ions. The identification of metal
ion by using chemometric approach
Wax printing e Au Hg2þ The detection of Hg2þ is based on Au 0.005 mM e Pond and river water [63]
nanoparticles combined with oligonucleotide
by Thymine-Hg2þeThymine coordination
chemistry, where upon addition of NaCl Au
particles form aggregation which leads to
change in color from purple to blue.

G. Sriram et al. / Trends in Analytical Chemistry 93 (2017) 212e227

Cutting machine 3,3,5,5-tetramethylbenzidine Pt Hg2þ Pt nanoparticles oxidizes TMB in the detection 0.01 mM 3 Pond and tap water [64]
(TMB) zone, the color formed is blue and if Hg2þ ions
present the blue color is reduced.
Chemical vapor HOTT e Hg2þ The yellow colored chromogenic reagent HOTT 5.0  104 ppm 10 Tap and municipal [65]
deposition loaded in the detection zones reacts with the water
Hg2þ ion resulted in the color change from
yellow into brick-red.
Ink-jet printing e Ag Hg2þ The Ag nanoparticles were oxidized by Hg2þ 1.0  103 ppm e Drinking and tap [66]
ions resulted in the disintegration of Ag water
nanoparticles and Hg2þ ion to Hg0. Due to this
mechanism there was a color change from deep
yellow color to bright yellow color that
confirms the presence of the Hg2þ ions
Ink-jet printing Zincon, sodium e Hg2þ, Cu2þ, The chlorophenol red b-galactopyranoside were 1.0  103, 2  102, 1.5  101 10 Distilled, tap, lake [67]
diethyldithiocarbamate, Cr6þ and Ni2þ hydrolyzed by adding B-GAL to get red- and 2.3  101 ppm water and waterfalls
1,5-diphenyl carbazide and magenta. The respective chromogenic reagents
dimethyl glyoxime were added into bioactive coated detection
zones. Color was changed when respective
metal ions comes in contact with reagents.
Wax printing e Ag Cu2þ Ag nanoparticles were catalytically etched by 1.0  103 ppm [16], 2 Mineral and ground [16,68]
the addition of thiosulfate in the detection zone. 4.12  103 ppm [68] water [16], Stream
Cu2þ ions oxidizes Ag nanoparticles thus and ground
particles were disintegrated. Resulted in color water [68]
change from pinkish-violet to colorless
Knife cutting e Ag Cu2þ Modified Ag nanoparticles with homocysteine 5.0  104 ppm 5 Tap and pond water [69]
and dithiothreitol binds with Cu2þ ions due to
their higher affinity. In the presence of Cu2þ
ions, oxidation of homocysteine and
aggregation of Ag nanoparticles takes place,
hence the color changed to orange. Excess of
Cu2þ ions changes color to greenish brown
Drop casting Chrysoidine-G e Sr2þ The chysoidine-G in the detection zone reacts 2.0  101ppm Less than 1 Real stream [70]
with Sr2þ ions resulting in color change from
orange to dark orange
Ink-jet printing Sodium rhodizonate (NaR) Zirconium Pb2þ The zirconium silicate was used as adsorbent 1.0  102 ppm 15 Tap, drinking and [71]
silicate and chromogenic reagent NaR forms the metal surface water
complex with Pb2þ ions, due to which color
changes to pink.
Y-shaped filter e Au-TA-TG As3þ The Au-TA-TG solution reacts with As3þ ions 1.0  103 ppm 5 e [72]
paper resulted in the aggregation of Au nanoparticles,
due to which bluish-black color appears
3D printing 1, 10-phenanthroline e Fe2þ The hydroxylammonium chloride is used as 3.96 mM e River and tap water [29]
reducing agent for Fe3þ to Fe2þ and
chromogenic reagent phenanthroline reacts
with Fe2þ ions resulting in color change
Wax printing 1,10-phenanthroline e Fe3þ The hydroxylamine and chromogenic reagent e e Natural water [73]
1,10-Phenanthroline are used as a reducing
agent and color indicator respectively in the
detection zone: Fe3þ ions reduced to Fe2þ ions
resulted in color change
Wax printing 1,10-phenanthroline, e Fe, Cu and Hydroxylamine was used for reducing Fe3þ to 1.0  103 ppm 2 Airborne [2]
bathocuproine and Ni ions Fe2þ upon reacting with 1,10-phenanthroline a
dimethylglyoxime (DMG) reddish orange color appears. Same molecule
reduced Cu2þ to Cuþ upon reacting with
bathocuproine the orange color formed. For
Ni2þ, DMG was used to reduce Ni2þ to Niþ upon
reacting with sodium fluoride produced bright

G. Sriram et al. / Trends in Analytical Chemistry 93 (2017) 212e227

pink color.
Wax printing DMG, 1,10-phenanthroline, e Ni, Fe, Cu, and For Ni, Fe and Cu used the same mechanism [2]. 7.5  104 ppm for Ni, e Baghouse [74]
bathocuproine Cr ions For Cr detection, Cr3þ was oxidized to Cr6þ Fe, Cu ions and 1.2  104 ppm
and 1,5-DPC using Ce4þ in the detection zone and then for Cr ions
reacted with 1,5-DPC for a color change.
Wax printing 1,10-phenanthroline, DMG, e Fe, Ni, Cr and Above mechanism was used for this method 1.1  103, 1.1  103, 3.7  103 e Welding fumes [75]
1,5-DPC and bathocuproine Cu ions [74] and 1.5  103 ppm respectively
Ink-jet printing Sulfanilamide, Zn Nitrate and Zn microparticles react with nitrate resulting in 1.0 and 19 mM respectively 5 Pond and [76]
N-(1-naphthyl)-ethylenediamine Nitrite reduction to nitrite. Through this reduction mineral water
dihydrochloride (Greiss reagent) mechanism nitrate ion was detected. For nitrite
detection, Greiss reagent was used.
Laser printer Greiss reagent e Nitrite Above mechanism was used for nitrite 10 mM 1 Artificial Saliva [79]
detection [76]
Permanent N-(1-naphthyl)-ethylenediamine e Nitrite Nitrite ion reacts with sulfonamide 1000 mM e Pond water [80]
marker pen and p-amino benzene bydiazotization mechanism, followed by the
sulfonamide reaction between the chromogenic indicators
which resulted in the color change from
colorless to red.
PDMS stamping Greiss reagent e Nitrite Above mechanism was used for nitrite 5.2  101 ppm e Drinking water [81]
detection [80]
Metal stamping Greiss reagent e Nitrite Above mechanism was used for nitrite 5.6 mM e River water [82]
detection [80]
Craft cutting Tetrazine reagent e Nitrite The method involves the detection of nitrite in 1.30 mM 5 Mineral and [83]
an acid medium, the citric acid was used in the tap water
detection zone then added with tetrazine
reagent to generate nitrous acid, the nitrite ions
oxidises tetrazine reagent. Thus there was color
change from colorless to pink, which confirms
the presence of nitrite ions.

221
222 G. Sriram et al. / Trends in Analytical Chemistry 93 (2017) 212e227
functionalized with oligonucleotide sequence by using thymine-
Hg2þethymine co-ordination chemistry to enhance the detection
capability. The limit of detection of Hg2þ ion was found to be
0.005 mM. Recently, Chen et al. [64] developed mPADs using glass
fiber paper by using cutting machine for the detection of Hg2þ ions.
The detection zone was preloaded with receptor 3,3,5,5-
tetramethylbenzidine (TMB) and hydrogen peroxide (H2O2) for
catalytic activity. Platinum (Pt) nanoparticles were mixed with
water containing Hg2þ ions followed by 2 min incubation. The so-
lution mixture was allowed to the detection zone which resulted in
color change from dark blue to light blue due to oxidation of TMB.
The change in color confirms the presence of Hg2þ ions with the
limit of detection of 0.01 mM. Mujawar et al. [65] reported hex-
amethyldisilazane (HMDS) modified mPADs fabricated using
210 mm thick filter paper, which was developed by chemical vapor
deposition for colorimetric detection of Hg2þ, Co2þ and Zn2þ ions
using chromogenic reagents. They found trace amount of Hg2þ ions
in tap and municipal water sample with the limit of detection of
5.0  104 ppm. The detection zone was preloaded with yellow
colored 6-hydroxy-3-(2-oxoindolin-3-ylideneamino-2-thio-2H-
1,3-thiazin-4(3H)-one (HOTT) molecule, when this molecule comes
in contact with water containing Hg2þ ions there was instant color
change from yellow to brick-red. In another study, Meelapsom et al.
[66] developed silver (Ag) nanoparticles modified double layered
mPADs using 180 mm thick filter paper by ink-jet printing method
for the detection of Hg2þ ions from water samples, which showed
the detection limit of 1  103 ppm. Here, Ag nanoparticles were
oxidized by Hg2þ ions resulting in the disintegration of Ag nano-
particles and reduction of Hg2þ to Hg0. The deep yellow colored Ag
nanoparticles were added to the detection zone followed by Hg2þ
ions, resulting in color change from deep yellow to bright yellow
which confirms the presence of Hg2þ ions. Hossain et al. [67] used
180 mm thick filter paper and developed multiplexed b-galactosi-
dase (B-GAL) modified mPADs substrate by ink-jet printing for
colorimetric detection of mixture of heavy metals. Chromogenic
reagents such as zincon, sodium diethyldithiocarbamate, 1,5-
diphenylcarbazide and dimethylglyoxime were used to detect of
Hg2þ, Cu2þ, Cr6þ, and Ni2þ from water sample (Fig. 5A). The limits of
detection for these metal ions were found to be 1.0  103,
2.0  102, 1.5  101 and 2.3  101 ppm respectively. The
chlorophenol red b-galactopyranoside was coated on each detec-
tion zones of mPADs and it was hydrolyzed by adding B-GAL
resulting in the formation of red-magenta. Further, each detection
zones were loaded with respective chromogenic reagent for the
detection of desired metal ions. As the metal ion samples Hg2þ,
Cu2þ, Cr6þ, and Ni2þ come in contact with the respective detection
zones, there was an immediate color changes blue, brown, red-
magenta and pink colors respectively.
The metal nanoparticles such as Pt, Au and Ag nanoparticles
plays important role in colorimetric detection of toxic ions. How-
ever, Ag nanoparticles is the best choice for colorimetric detection
application compared to Au and Pt nanoparticles due to low-cost
and higher extinction coefficient. Chaiyo et al. [16] fabricated
mPADs from 180 mm thick filter paper by wax printing for the
detection of Cu2þ ions using Ag nanoplates. The synthesized Ag
nanoplates were modified by hexadecyltrimethylammonium bro-
mide (CTAB) and coated on test zone of paper followed by adding
thiosulfate which is used as catalytic agent. In the presence Cu2þ
ions there was an instantaneous color change from violet-red to
colorless. The device could detect up to 1.0  103 ppm of Cu2þ ions
(Fig. 5B). The color change is due to Cu2þ ions which can oxidize the
modified Ag nanoparticles/thiosulfate resulting in the reduction of
particle size.
In another study, Chaiyo et al. [68] used filter paper of same
thickness to fabricate mPADs by wax printing for electrochemical
and colorimetric detection of Pb2þ, Cd2þ and Cu2þ ions. In colori-
metric detection, CTAB modified Ag nanoplates with thiosulfate
were added at the detection zone followed by the addition of Cu2þ
ions. The color was changed from pinkish-violet to colorless with
the limit of detection of 4.12  103 ppm for Cu2þ ions. The
mechanism of colorimetric detection of Cu2þ ion is as follows: In
presence of Cu2þ, preloaded Ag nanoparticles were etched by the
presence of thiosulphate which were disintegrated into smaller
particles due to oxidation. Thus, there were color changes from
pinkish-violet to colorless.
In addition to the above observations, Ratnarathorn et al. [69]
used filter paper with thickness of 180 mm to develop mPADs by
knife cutting for the colorimetric detection of Cu2þ ions using Ag
nanoparticles. Ag nanoparticles were adopted due to their low cost
and higher extinction coefficient than that of Au nanoparticles. The
higher extinction coefficient of Ag nanoparticles resulted in higher
color visibility. The synthesized Ag nanoparticles were modified by
homocysteine and dithiothreitol, which were loaded at the detec-
tion zone. In the presence of Cu2þ ions the color change was from
yellow to orange due to oxidation of homocysteine and aggregation
of Ag nanoparticles. The increase in the concentration of Cu2þ ions
resulted in a sudden color change from orange to greenish brown
due to the Ag particle aggregation. The aggregation of Ag nano-
particles due to the presence of Cu2þ ions, reduces the surface
Plasmon resonance coupling resulting in the color change. The
visibility of color depends on the concentration of metal ions in the
analyte solution. In addition, Cu2þ ions were detected from tap and
pond water using modified Ag nanoparticles using mPADs with a
detection limit of 5.0  104 ppm for Cu2þ ions (Fig. 5C). Recently,
Kang et al. [70] have developed chrysoidine-G (CG) modified mPADs
by drop casting method for the colorimetric detection of strontium
(Sr2þ) ions from water samples with a detection limit of
2.0  101 ppm. The detection zone was previously loaded with
orange colored CG solutions. The paper devices consist of hydroxyl
and carboxylic acid groups within fiber networks. These groups
offered space for the immobilization of CG on paper devices. As the
Sr2þ ions were added to the detection zone, the color changes from
orange to dark orange, which confirms the presence of Sr2þ ions.
Recently, Satarpai et al. [71] developed mPADs by ink-jet printing for
colorimetric detection of Pb2þ ions from water samples, which of-
fers limit of detection of 1.0  102 ppm. Here, zirconium silicate
was used as an adsorbent material and sodium rhodizonate (NaR)
as a chromogenic reagent for Pb2þ ions. The adsorbent material was
coated on to the detection zone of mPADs followed by NaR. As the
detection zone comes in contact with Pb2þ ions, the color changed
from orange-brown to pink, which confirms the presence of Pb2þ
ions.
In another study, Nath et al. [72] developed Y-shaped mPAD for
colorimetric detection of arsenic (As3þ) using Au nanoparticles
with thioctic acid (TA) and thioguanine (TG) (Au-TA-TG). The mPAD
could detect As3þ ions up to 1.0  103 ppm level in the presence of
Au-TA-TG. There was a formation of dark bluish-black precipitate
due to aggregation of Au nanoparticles by interaction of TG with
As3þ ions.
Asano et al. [29] used filter paper with a thickness of 170 mm to
fabricate mPADs by 3D printer for the detection of iron from water
sample using phenanthroline indicator. The detection zone was
preloaded by adding 20 mL of hydroxylammonium chloride, phe-
nanthroline and acetate buffer solutions. After drying, river and tap
water were analyzed for the detection of iron. The visible color
change was captured in high resolution camera and was found that
iron concentration in tap water was lower than that in river water.
The limit of detection and limit of quantitation were found to be
3.96 and 13.8 mM, respectively. Ogawa et al. [73] fabricated mPADs
using 180 mm thick filter paper by wax printing for colorimetric
 G. Sriram et al. / Trends in Analytical Chemistry 93 (2017) 212e227 223

Fig. 5. Colorimetric detection of (A) individual metals and mixture of metals using a multiplexed paper substrate [Reproduced with permission from Ref. [67], Copy right 2011,
American Chemical Society], (B) detection of Cu2þ ions in different concentration [Reproduced with the permission from Ref. [16], Copy right 2015, Elsevier], (C) detection of
different concentrations of Cu2þios using Ag nanoparticles modified mPADs from water samples [Reproduced with the permission from Ref. [69], Copyright 2012, Elsevier], (D) Fe, Ni
and Cu metals from airborne samples [Reproduced with permission from Ref. [2], Copyright 2012, American Chemical Society] and (E) total Cr metal ion from welding fumes
[Reproduced from Ref. [75]].

detection of Fe3þ ions in natural water using 1,10-phenanthroline
and poly(acrylic acid) as indicators in the detection zone followed
by drying. Natural water samples were added to the same reservoir
zone, which resulted in the reduction of Fe3þ to Fe2þ ions, while the
limit of quantification of Fe3þ ion was 40 ppm and color saturation
intensity occurred after 350 ppm. Further, Mentele et al. [2] also
used filter paper of same thickness to develop mPADs by wax
printing for the colorimetric detection of particulate metals such as
iron, nickel and copper ions in air filters. The sample punches were
placed on paper device, then water was flushed to eject the
digested metal ions from filter and transported to the detection
zone. Each detection zones were pre-loaded with chromogenic
reagents for the detection of particulate metal ions. Iron was
detected by the formation of ferroin complex resulted in an orange
color due to the reaction between Fe2þ and 1,10-phenanthroline
(Fig. 5D). The detection of Ni ions by using dimethylglyoxime
resulted in the formation of bright pink color for Ni2þcomplex. The
Cu ions detection was performed by bathocuproine, which resulted
in the formation of light orange color. The limit of detection of Fe2þ,
Cu2þ and Ni2þ ions were found 1  103, 1  103 and 1  103 ppm
respectively. In another study, the same mechanism was used for
the colorimetric detection of metal ions by Rattanarat et al. [74].
The filter paper with a thickness of 180 mm was used to develop
multilayer mPADs by wax printing for colorimetric and electro-
chemical detection of metal ions from re-suspended baghouse dust
samples The device could detect Fe2þ, Ni2þ, Cr2þ and Cu2þ ions
colorimetrically. For detecting Fe2þ ions, hydroxylamine,
poly(acrylic acid) and 1,10-phenanthroline were added to the
detection zone. For Ni2þ ion detection, sodium fluoride, acetic acid,
ammonium hydroxide and dimethylglyoxime were added to the
detection zone. For Cr2þ ion detection, ceric (IV) ammonium ni-
trate, poly(diallyldimethylammonium chloride), 1,5-
diphenylcarbazide and phthalic anhydride were added. For Cu2þ
ion detection, hydroxylamine, acetic acid, bathocuproine and poly
ethylene glycol were added. The limit of detection for Fe2þ, Ni2þ
and Cu2þ ions were 7.5  104 ppm, while for Cr2þ, it was
1.2  104 ppm (Table 2).
Further, Cate et al. [75] attempted to detect Fe2þ, Cu2þ, Ni2þ and
2þ
Cr ions present in welding fumes collected from air filter sample.
The 10 mm filter punch collected from air filter samples was
mounted on the detection zone of mPADs developed from 180 mm
thick filter paper by wax printing, followed by fixing the desired
size of PDMS layer on a punch containing paper device. The
required reagents were then added to the detection zone and dried
for few min. Water was used to elute the metal from air filter
sample to reach the detection zone. When the sample reached the
detection zone, there was a change in color with respect to the
presence of metal ions. The reagents used were ferroin complex
and 1,10-phenanthroline to detect Fe2þ ions in air filter sample. The
limit of detection for Fe2þ ions was 1.1  103 ppm and the satu-
ration was about 1.0  102 ppm. For Ni2þ ion detection, reagents
such as acetic acid and sodium fluoride were used with limit of
detection of 1.1  103 ppm. In case of Cr2þ ions, 1,5-
diphenylcarbazide was used, but its limit of detection was less
224 G. Sriram et al. / Trends in Analytical Chemistry 93 (2017) 212e227

than that of Fe2þ ions (Fig. 5E). The bathocuproine was used to
detect Cu2þ ions with a limit of detection of 1.5  103 ppm.
3.2. Nitrite detection
Innumerable methods have been developed for the detection of
nitrite. Jayawardane et al. [76] used a 180 mm thick filter paper to
fabricate mPADs by ink-jet printing method for the detection of
nitrite and nitrate from environmental samples. Nitrates are
poisonous to human body as they are easily reduced to nitrite,
which is more toxic than nitrate [77]. According to US-EPA,
permissible limit of nitrate and nitrite in drinking water are 1 and
10 ppm, respectively [78]. The nitrite ions are determined using
Griess reaction i.e. nitrite ions were react with Greiss reagents
called sulphanilamide to form diazonium salt which reacts with
amino group of N-(1-naphthyl)-ethylenediamine dihydrochloride
to produce pink color which confirms the presence of nitrite. For
nitrate ion detection, nitrate was reduced to nitrite in the detection
zone of mPADs using immobilized zinc microparticles. The limit of
detection and quantification for nitrite were 1.0 and 7.8 mM and for
nitrates 19 and 48 mM, respectively.
Bhakta et al. [79] also used filter paper of same thickness to
develop mPADs by laser printer for the detection of nitrite in arti-
ficial saliva by adding Griess reagent to the detection zone. As
sample raised to the detection zone through capillary action, there
was appearance of pink color. The color intensity depends on the
amount of nitrite present in the samples and the limit of detection
was found up to 10 mM L1. Colorimetric detection of nitrite [80]
was also done by adding standard sodium nitrite solutions to six
detection zones of mPADs which was developed by permanent
marker pen and color change was observed from colorless to red
using N-(1-naphthyl)-ethylenediamine and p-amino benzene-
sulfonamide as indicators. The color intensity depends on the
concentration of nitrite ions and the same device was checked for
the presence of nitrite ions in pond water (Fig. 6A).
Further, Ruiz et al. [81] used same sized filter paper to develop
mPADs by PDMS contact stamping method for the colorimetric
detection of nitrite as well as pH from drinking water sources.
Initially, pH determination was carried out by two indicators such
as phenol red and chlorophenol red. Water sample was added to
four detection zones followed by phenol red for the first two zones
and chlorophenol red to the other two zones with a response time
for both indicators set at 15 min. The color change was observed
from yellow to purple for chlorophenol red and yellow to pink for
phenol red due to change in pH. The colorimetric detection of ni-
trite was determined by Griess reaction. For nitrite ion detection,
three different indicators viz., (i) mixture of nafion, 2-propanol,
water containing polyethylene glycol, (ii) sulphanilamide with
citric acid and (iii) N-(1-napthyl)ethylenediamine dihydrochloride
were used. The three different solutions were added to the

Fig. 6. mPADs representing the colorimetric detection of (A) detection of nitrite in different concentration from pond water [Reproduced with permission from Ref. [80], Copy right
2015, American Chemical Society], (B) detection of pH and nitrite from drinking water [Reproduced with permission from Ref. [81], Copy right 2014, American Chemical Society], (C)
detection of nitrite using Greiss reaction from river water [Reproduced with permission from Ref. [82], Copy right 2015, The Royal Society of Chemistry] and (D) detection of nitrite
from water samples [Reproduced with permission from Ref. [83], Copy right 2016, Elsevier] using mPADs.
 G. Sriram et al. / Trends in Analytical Chemistry 93 (2017) 212e227
detection zone followed by Griess reagents. As the sample water
was added to the detection zone, the color intensity was changed
due to the presence of nitrite ions and the limit of detection of
nitrite ion was found 5.2  101 ppm (Fig. 6B).
In another study, Cardoso et al. [82] have fabricated mPADs by
metal stamping for the detection of nitrites by Griess reaction from
various sources such as saliva, ham, sausage and river water for
clinical, food and environmental applications. The detection of ni-
trites was done colorimetrically with the help of Griess reaction
using mixtures of indicators containing sulfanilamide, hydrochloric
acid and N-(1-napthyl)ethylenediamine, which were added to the
detection zones followed by drying under ambient temperature.
Then, the analyte was allowed to flow to the detection zone for
nitrite assay (Fig. 6C). Recently, Ortiz-Gomez et al. [83] used filter
paper with thickness of 170 mm to develop mPADs for the detection
of nitrite ions present in water samples using tetrazine, which was
developed by craft cutting method. The tetrazine is a non-
carcinogenic reagent, and it can rapidly react with analytes over
Griess reagent. The formation of pink color with the limit of
detection of nitrite ions from water sample was found to be 1.30 mM
(Fig. 6D). Table 2 shows the summary of colorimetric detection of
different metal ions and nitrite from various samples.
It is worth mentioning here that fabrication of mPADs using
different types of filter papers by different techniques with various
designs may result in increased limit of detection, higher efficiency
with quick assay and requirement of very less analyte and receptor
solutions. The dimension of hydrophilic channel and hydrophobic
barrier on mPADs will result in rate of capillary action. Smaller the
dimension of hydrophilic channel faster will be the capillary rise of
analyte from sample zone to detection zone. The detection effi-
ciency also depends on the type of receptor solution preloaded on
the detection zone, i.e., higher the receptor affinity towards ions,
better will be the sensitivity. The filter paper with high thickness
may give low wicking, low color resolution, less detection limit and
requires more analyte solutions over thin filter papers.
4. Conclusions and future directions
In summary, the mPADs are attractive as they are inexpensive,
flexible, and white in color, which allow passive fluid transport
through capillary phenomenon and offers rapid detection. In market,
various types of filter papers with different thickness are available,
which can be utilized to fabricate mPADs for colorimetric applications.
The hydrophilic channels in mPADs are developed with different
structures and dimensions by various fabrication techniques. The
greatest advantages of mPADs are that they do not need any sophis-
ticated facilities including instrumentation, clean-room facilities etc.
The detection process involves simple color change which could be
observed through naked eye. Due to above advantages, this tech-
nique has created immense interest in analytical sciences as a
powerful detection tool especially for remote area applications.
Among the many materials including hazardous hydrophobic
chemicals, wax is used to create hydrophobic boundaries, but wax
printing is the cheapest and most widely used in toxic ion detec-
tion. The toxic ions can be detected from various environmental
samples such as water samples collected from tap, river, pond and
welding fumes, etc. The limit of detection and color intensity de-
pends on the thickness of the paper, detection mechanism and type
of receptors used, concentration of metal ions in samples, and the
dimensions of the channel. As a future prospectus, this technique
plays a major role for the detection of toxic ions and offers un-
precedented advantages of having miniature design, low cost pro-
duction, portability, eco-friendly, rapid detection without labeling
in a real-time format. There is a huge scope for commercialization
of mPADs for metal ion detection and quantifications especially for
225
remote area applications and in preliminary estimation of water
quality.
Acknowledgements
The authors acknowledge the financial support from DST
Nanomission, India (SR/NM/NS-20/2014), DST, India (DST-TM-WTI-
2K14-213) and SERB-DST, India (YSS/2015/000013) for financial
support. We also thank Jain University, India for providing facilities.
References
[1] J.C. Jokerst, J.A. Adkins, B. Bisha, M.M. Mentele, L.D. Goodridge, C.S. Henry,
Development of a paper-based analytical device for colorimetric detection of
select foodborne pathogens, Anal. Chem. 84 (2012) 2900e2907.
[2] M.M. Mentele, J. Cunningham, K. Koehler, J. Volckens, C.S. Henry, Microfluidic
paper-based analytical device for particulate metals, Anal. Chem. 84 (2012)
4474e4480.
[3] A.K. Yetisen, M.S. Akram, C.R. Lowe, Paper-based microfluidic point-of-care
diagnostic devices, Lab. Chip 13 (2013) 2210e2251.
[4] P. Patil, Madhuprasad, T. Kumeria, D. Losic, M. Kurkuri, Isolation of circulating
tumour cells by physical means in a microfluidic device: a review, RSC Adv. 5
(2015) 89745e89762.
[5] W. Zhao, A.V. der Berg, Lab on paper, Lab. Chip 8 (2008) 1988e1991.
[6] A.W. Martinez, S.T. Phillips, E. Carrilho, S.W. Thomas, H. Sindi,
G.M. Whitesides, Simple telemedicine for developing regions: camera phones
and paper-based microfluidic devices for real-time, off-Site diagnosis, Anal.
Chem. 80 (2008) 3699e3707.
[7] T. Fujii, PDMS-based microfluidic devices for biomedical applications, Micro-
electron. Eng. 61e62 (2002) 907e914.
[8] T. Songjaroen, W. Dungchai, O. Chailapakul, W. Laiwattanapaisal, Novel,
simple and low-cost alternative method for fabrication of paper-based
microfluidics by wax dipping, Talanta 85 (2011) 2587e2593.
[9] D.M. Cate, J.A. Adkins, J. Mettakoonpitak, C.S. Henry, Recent developments in
paper-based microfluidic devices, Anal. Chem. 87 (2015) 19e41.
[10] Y. He, Y. Wu, J.Z. Fu, W.B. Wu, Fabrication of paper-based microfluidic analysis
devices: a review, RSC Adv. 5 (2015) 78109e78127.
[11] Y. Xia, J. Si, Z. Li, Fabrication techniques for microfluidic paper-based analytical
devices and their applications for biological testing: a review, Biosens. Bio-
electron. 77 (2016) 774e789.
[12] Y. Xu, M. Liu, N. Kong, J. Liu, Lab-on-paper micro- and nano-analytical devices:
fabrication, modification, detection and emerging applications, Microchim.
Acta 183 (2016) 1521e1542.
[13] X. Jiang, Z.H. Fan, Fabrication and operation of paper-based analytical devices,
Annu.Rev. Anal. Chem. 9 (2016) 203e222.
[14] A.W. Martinez, S.T. Phillips, M.J. Butte, G.M. Whitesides, Patterned paper as a
platform for inexpensive, low-volume, portable bioassays, Angew. Chem. 46
(2007) 1318e1320.
[15] B.G.A. Murphy, K. de Guzman, N. Kelly, E.P. Nesterenko, A. Morrin, Micro-
fluidic paper analytical device for the chromatographic separation of ascorbic
acid and dopamine, RSC Adv. 5 (2015) 93162e93169.
[16] S. Chaiyo, W. Siangproh, A. Apilux, O. Chailapakul, Highly selective and sen-
sitive paper-based colorimetric sensor using thiosulfate catalytic etching of
silver nanoplates for trace determination of copper ions, Anal. Chim. Acta 866
(2015) 75e83.
[17] S. Kar, T.K. Maiti, S. Chakraborty, Capillarity-driven blood plasma separation
on paper-based devices, Analyst 140 (2015) 6473e6476.
[18] E. Evans, E.F. Moreira Gabriel, T.E. Benavidez, W.K. Tomazelli Coltro,
C.D. Garcia, Modification of microfluidic paper-based devices with silica
nanoparticles, Analyst 139 (2014) 5560e5567.
[19] Y. Wu, P. Xue, Y. Kang, K.M. Hui, Paper-based microfluidic electrochemical
immunodevice integrated with nanobioprobes onto graphene film for ultra-
sensitive multiplexed detection of cancer biomarkers, Anal. Chem. 85 (2013)
8661e8668.
[20] G. Jenkins, Y. Wang, Y.L. Xie, Q. Wu, W. Huang, L. Wang, X. Yang, Printed
electronics integrated with paper-based microfluidics: new methodologies for
next-generation health care, Microfluid. Nanofluid. 19 (2015) 251e261.
[21] H. Wang, J. Liu, R.G. Cooks, Z. Ouyang, Paper spray for direct analysis of
complex mixtures using mass spectrometry, Angew. Chem. 122 (2010)
889e892.
[22] Q. Chen, Z. He, W. Liu, X. Lin, J. Wu, H. Li, J.M. Lin, Engineering cell-compatible
paper chips for cell culturing, drug screening, and mass spectrometric sensing,
Adv. Healthc. Mater. 4 (2015) 2291e2296.
[23] H. Liu, Y. Xiang, Y. Lu, R.M. Crooks, Aptamer-based origami paper analytical
device for electrochemical detection of adenosine, Angew. Chem. 124 (2012)
7031e7034.
[24] X. Li, D.R. Ballerini, W. Shen, A perspective on paper-based microfluidics:
current status and future trends, Biomicrofluidics 6 (2012), 011301.
[25] D.D. Liana, B. Raguse, J.J. Gooding, E. Chow, Recent advances in paper-based
sensors, Sensors 12 (2012) 11505.
226 G. Sriram et al. / Trends in Analytical Chemistry 93 (2017) 212e227
[26] P. Lisowski, P.K. Zarzycki, Microfluidic paper-based analytical devices (mPADs)
and micro total analysis systems (mTAS): development, applications and
future trends, Chromatographia 76 (2013) 1201e1214.
[27] A. Kakoti, M.F. Siddiqui, P. Goswami, A low cost design and fabrication method
for developing a leak proof paper based microfluidic device with customized
test zone, Biomicrofluidics 9 (2015), 026502.
[28] J. Songok, M. Toivakka, Enhancing capillary-driven flow for paper-based
microfluidic channels, ACS Appl. Mater. Interfaces 8 (2016) 30523e30530.
[29] H. Asano, Y. Shiraishi, Development of paper-based microfluidic analytical
device for iron assay using photomask printed with 3D printer for fabrication
of hydrophilic and hydrophobic zones on paper by photolithography, Anal.
Chim. Acta 883 (2015) 55e60.
[30] Y. He, Q. Gao, W.B. Wu, J. Nie, J.Z. Fu, 3D Printed paper-based microfluidic
analytical devices, Micromachines 7 (2016) 108.
[31] G.I. Salentijn, N.N. Hamidon, E. Verpoorte, Solvent-dependent on/off valving
using selectively permeable barriers in paper microfluidics, Lab. Chip 16
(2016) 1013e1021.
[32] Z.A. Li, J. Yang, L. Zhu, W. Tang, Fabrication of paper micro-devices with wax
jetting, RSC Adv. 6 (2016) 17921e17928.
[33] C. Chen, T. Dong, Microfluidic paper-based analytical devices for colorimetric
detection of urinary tract infection biomarkers on adult diapers, Conf. Proc.
IEEE Eng. Med. Biol. Soc. (2015) 5892e5895, http://dx.doi.org/10.1109/
EMBC.2015.7319732.
[34] S. Mohammadi, M. Maeki, R.M. Mohamadi, A. Ishida, H. Tani, M. Tokeshi, An
instrument-free, screen-printed paper microfluidic device that enables bio
and chemical sensing, Analyst 140 (2015) 6493e6499.
[35] S.W. Hu, S. Qiao, B.Y. Xu, X. Peng, J.J. Xu, H.Y. Chen, Dual-functional carbon
dots pattern on paper chips for Fe3þ and ferritin analysis in whole blood, Anal.
Chem. 89 (2017) 2131e2137.
[36] C. Gallibu, C. Gallibu, A. Avoundjian, F. Gomez, Easily fabricated microfluidic
devices using permanent marker inks for enzyme assays, Micromachines 7
(2016) 6.
[37] N. Nuchtavorn, M. Macka, A novel highly flexible, simple, rapid and low-cost
fabrication tool for paper-based microfluidic devices (mPADs) using technical
drawing pens and in-house formulated aqueous inks, Anal. Chim. Acta 919
(2016) 70e77.
[38] P. Jiang, M. He, L. Shen, A. Shi, Z. Liu, A paper-supported aptasensor for total
IgE based on luminescence resonance energy transfer from upconversion
nanoparticles to carbon nanoparticles, Sens. Actuators B 239 (2017) 319e324.
[39] K. Tenda, R. Ota, K. Yamada, T.G. Henares, K. Suzuki, D. Citterio, High-reso-
lution microfluidic paper-based analytical devices for sub-microliter sample
analysis, Micromachines 7 (2016) 80.
[40] P. Kaewarsa, W. Laiwattanapaisal, A. Palasuwan, D. Palasuwan, A new paper-
based analytical device for detection of glucose-6-phosphate dehydrogenase
deficiency, Talanta 164 (2017) 534e539.
[41] Q. Kong, Y. Wang, L. Zhang, S. Ge, J. Yu, A novel microfluidic paper-based
colorimetric sensor based on molecularly imprinted polymer membranes
for highly selective and sensitive detection of bisphenol A, Sens. Actuators B
243 (2017) 130e136.
[42] J.C. Lee, W. Kim, S. Choi, Fabrication of a SERS-encoded microfluidic paper-
based analytical chip for the point-of-assay of wastewater, Int. J. Precis. Eng.
Manuf. Green Tech. 4 (2017) 221e226.
[43] T. Akyazi, N.G. Gonza
lez, L.B. Desmonts, E. Castan
~ o, M.C.M. Min
~ ana, F.B. Lopez,
Manipulation of fluid flow direction in microfluidic paper-based analytical devices
with an ionogel negative passive pump, Sens. Actuators B 247 (2017) 114e123.
[44] H. Kudo, K. Yamada, D. Watanabe, K. Suzuki, D. Citterio, Paper-based analyt-
ical device for zinc ion quantification in water samples with power-free an-
alyte concentration, Micromachines 8 (2017) 127.
[45] C.M. Wang, C.Y. Chen, W.S. Liao, Paper-polymer composite devices with
minimal fluorescence background, Anal. Chim. Acta 963 (2017) 93e98.
[46] S. Malekghasemi, E. Kahveci, M. Duman, Rapid and alternative fabrication
method for microfluidic paper based analytical devices, Talanta 159 (2016)
401e411.
[47] T.G. Henares, K. Yamada, S. Takaki, K. Suzuki, D. Citterio, “Drop-slip” bulk
sample flow on fully inkjet-printed microfluidic paper-based analytical de-
vice, Sens. Actuators B 244 (2017) 1129e1137.
[48] M.A. Ostad, A. Hajinia, T. Heidari, A novel direct and cost effective method for
fabricating paper-based microfluidic device by commercial eye pencil and its
application for determining simultaneous calcium and magnesium, Micro-
chem. J. 133 (2017) 545e550.
[49] A. Abadian, S.S. Manesh, S.J. Ashtiani, Hybrid paper-based microfluidics:
combination of paper-based analytical device (mPAD) and digital microfluidics
(DMF) on a single substrate, Microfluid. Nanofluid. 21 (2017) 65.
[50] R.A. de Oliveira, F. Camargo, N.C. Pesquero, R.C. Faria, A simple method to
produce 2D and 3D microfluidic paper-based analytical devices for clinical
analysis, Anal. Chim. Acta 957 (2017) 40e46.
[51] Y. Zhang, T. Li, T. Ren, D. Fang, J. He, Hydrophobic/lipophobic barrier capable of
confining aggressive liquids for paper-based assay, Colloids Surf. A 520 (2017)
544e549.
[52] T.M.G. Cardoso, F.R. de Souza, P.T. Garcia, D. Rabelo, C.S. Henry, W.K.T. Coltro,
Versatile fabrication of paper-based microfluidic devices with high chemical
resistance using scholar glue and magnetic masks, Anal. Chim. Acta 974
(2017) 63e68.
[53] M. Dou, S.T. Sanjay, M. Benhabib, F. Xu, X. Li, Low-cost bioanalysis on paper-
based and its hybrid microfluidic platforms, Talanta 145 (2015) 43e54.
[54] C. Rozand, Paper-based analytical devices for point-of-care infectious disease
testing, Eur. J. Clin. Microbiol. Infect. Dis. 33 (2014) 147e156.
[55] J.R. Choi, J. Hu, S. Wang, H. Yang, W.A. Wan Abas, B. Pingguan-Murphy, F. Xu,
Paper-based point-of-care testing for diagnosis of dengue infections, Crit. Rev.
Biotechnol. (2016) 1e12, http://dx.doi.org/10.3109/07388551.2016.1139541.
[56] J. Adkins, K. Boehle, C. Henry, Electrochemical paper-based microfluidic de-
vices, Electrophoresis 36 (2015) 1811e1824.
[57] B. Liu, D. Du, X. Hua, X.Y. Yu, Y. Lin, Paper-based electrochemical biosensors:
from test strips to paper-based microfluidics, Electroanalysis 26 (2014)
1214e1223.
[58] N.A. Meredith, C. Quinn, D.M. Cate, T.H. Reilly, J. Volckens, C.S. Henry, Paper-
based analytical devices for environmental analysis, Analyst 141 (2016)
1874e1887.
[59] J. Mettakoonpitak, K. Boehle, S. Nantaphol, P. Teengam, J.A. Adkins, M.S. Art,
C.S. Henry, Electrochemistry on paper-based analytical devices: a review,
Electroanalysis 28 (2016) 1420e1436.
[60] Y. Lin, D. Gritsenko, S. Feng, Y.C. Teh, X. Lu, J. Xu, Detection of heavy metal by
paper-based microfluidics, Biosens. Bioelectron. 83 (2016) 256e266.
[61] P. Rattanarat, W. Dungchai, D.M. Cate, W. Siangproh, J. Volckens,
O. Chailapakul, C.S. Henry, A microfluidic paper-based analytical device for
rapid quantification of particulate chromium, Anal. Chim. Acta 800 (2013)
50e55.
[62] L. Feng, X. Li, H. Li, W. Yang, L. Chen, Y. Guan, Enhancement of sensitivity of
paper-based sensor array for the identification of heavy-metal ions, Anal.
Chim. Acta 780 (2013) 74e80.
[63] G.H. Chen, W.Y. Chen, Y.C. Yen, C.W. Wang, H.T. Chang, C.F. Chen, Detection of
mercury (II) ions using colorimetric gold nanoparticles on paper-based
analytical devices, Anal. Chem. 86 (2014) 6843e6849.
[64] W. Chen, X. Fang, H. Li, H. Cao, J. Kong, A simple paper-based colorimetric
device for rapid mercury (II) assay, Sci. Rep. 6 (2016) 31948.
[65] L.H. Mujawar, A.A. Felemban, M.S. El-Shahawi, Hexamethyldisilazane modi-
fied paper as an ultra-sensitive platform for visual detection of Hg2þ, Co2þ,
Zn2þ and the application to semi-quantitative determination of Hg2þ in
wastewater, Anal. Sci. 32 (2016) 491e497.
[66] R. Meelapsom, P. Jarujamrus, M. Amatatongchai, S. Chairam, C. Kulsing,
W. Shen, Chromatic analysis by monitoring unmodified silver nanoparticles
reduction on double layer microfluidic paper-based analytical devices for
selective and sensitive determination of mercury (II), Talanta 155 (2016)
193e201.
[67] S.M.Z. Hossain, J.D. Brennan, b-Galactosidase-based colorimetric paper sensor
for determination of heavy metals, Anal. Chem. 83 (2011) 8772e8778.
[68] S. Chaiyo, A. Apiluk, W. Siangproh, O. Chailapakul, High sensitivity and
specificity simultaneous determination of lead, cadmium and copper using
mPAD with dual electrochemical and colorimetric detection, Sens. Actuators B
233 (2016) 540e549.
[69] N. Ratnarathorn, O. Chailapakul, C.S. Henry, W. Dungchai, Simple silver
nanoparticle colorimetric sensing for copper by paper-based devices, Talanta
99 (2012) 552e557.
[70] S.M. Kang, S.C. Jang, Y.S. Huh, C.S. Lee, C. Roh, A highly facile and selective
Chemo-Paper-Sensor (CPS) for detection of strontium, Chemosphere 152
(2016) 39e46.
[71] T. Satarpai, J. Shiowatana, A. Siripinyanond, Paper-based analytical device for
sampling, on-site preconcentration and detection of ppb lead in water,
Talanta 154 (2016) 504e510.
[72] P. Nath, R.K. Arun, N. Chanda, A paper based microfluidic device for the
detection of arsenic using a gold nanosensor, RSC Adv. 4 (2014)
59558e59561.
[73] K. Ogawa, T. Kaneta, Determination of iron ion in the water of a natural hot
spring using microfluidic paper-based analytical devices, Anal. Sci. 32 (2016)
31e34.
[74] P. Rattanarat, W. Dungchai, D. Cate, J. Volckens, O. Chailapakul, C.S. Henry,
Multilayer paper-based device for colorimetric and electrochemical quantifi-
cation of metals, Anal. Chem. 86 (2014) 3555e3562.
[75] D.M. Cate, P. Nanthasurasak, P. Riwkulkajorn, C. L'Orange, C.S. Henry,
J. Volckens, Rapid detection of transition metals in welding fumes using
paper-based analytical devices, Ann. Occup. Hyg. 58 (2014) 413e423.
[76] B.M. Jayawardane, S. Wei, I.D. McKelvie, S.D. Kolev, Microfluidic paper-based
analytical device for the determination of nitrite and nitrate, Anal. Chem. 86
(2014) 7274e7279.
[77] K.V.H. Sastry, R.P. Moudgal, J. Mohan, J.S. Tyagi, G.S. Rao, Spectrophotometric
determination of serum nitrite and nitrate by copperecadmium alloy, Anal.
Biochem. 306 (2002) 79e82.
[78] D. Tsikas, Analysis of nitrite and nitrate in biological fluids by assays based on
the Griess reaction: appraisal of the Griess reaction in the l-arginine/nitric
oxide area of research, J. Chromatogr. B 851 (2007) 51e70.
[79] S.A. Bhakta, R. Borba, M. Taba Jr., C.D. Garcia, E. Carrilho, Determination of
nitrite in saliva using microfluidic paper-based analytical devices, Anal. Chim.
Acta 809 (2014) 117e122.
[80] B. Wang, Z. Lin, M. Wang, Fabrication of a paper-based microfluidic device to
readily determine nitrite ion concentration by simple colorimetric assay,
J. Chem. Educ. 92 (2015) 733e736.
[81] N.L. Ruiz, V.F. Curto, M.M. Erenas, F.B. Lopez, D. Diamond, A.J. Palma,
L.F.C. Vallvey, Smartphone-based simultaneous pH and nitrite colorimetric
determination for paper microfluidic devices, Anal. Chem. 86 (2014)
9554e9562.
 G. Sriram et al. / Trends in Analytical Chemistry 93 (2017) 212e227
[82] T.M.G. Cardoso, P.T. Garcia, W.K.T. Coltro, Colorimetric determination of ni-
trite in clinical, food and environmental samples using microfluidic devices
stamped in paper platforms, Anal. Methods 7 (2015) 7311e7317.
[83] I.O. Gomez, M.O. Mun ~ oz, A.S. Castillo, J.A.A. Bermejo, M.A. Avidad, I. de
Orbe Pay
a, F.S. Gonzalez, L.F.C. Vallvey, Tetrazine-based chemistry for ni-
trite determination in a paper microfluidic device, Talanta 160 (2016)
721e728.
[84] J. Lee, A.A.S. Samson, J.M. Song, Inkjet-printing enzyme inhibitory assay based
on determination of ejection volume, Anal. Chem. 89 (2017) 2009e2016.
227
[85] P. Dimitrakellis, Travlos, V.P. Psycharis, E. Gogolides, Superhydrophobic paper
by facile and fast atmospheric pressure plasma etching, Plasma Process.
Polym. (2017), http://dx.doi.org/10.1002/ppap.201600069.
[86] Lenntech, Water treatment solutions, Netherland. http://www.lenntech.com/
applications/drinking/standards/who-s-drinking-water-standards.htm, 2017
(Accessed 12 June 2017).
[87] United States, Environmental protection agency, United States. https://www.
epa.gov/ground-water-and-drinking-water/national-primary-drinking-
water-regulations, 2017 (Accessed 12 June 2017).