Sensors for Drugs Screening and

Clinical Applications

Bambang Kuswandi
Chemo and Biosensors Group, Faculty of Pharmacy, University of Jember
Jl. Kalimantan 37 Jember, 68121 Indonesia

Presented at
Seminar Nasional Farmasi Unej 2015
28 November 2015
Hotel Aston, Jember , Indonesia
University of Jember
www.unej.ac.id
 Chemo & Biosensors Group
Food & agriculture
Low-cost & iFood, Halal sensor
disposable Environmental
sensing system Industry

Smart material & Pharmacy

Polymers
CBG (Natural products)
Clinical
Health

Wearable sensing
Microfluidics
system
Molecular recognition Lab on a Chip
Host-guest chemistry Lab on paper
Google Scholar
ResearchGate
 Out-line
• Introduction
• Sensors:
- Chemical sensors
- Biosensors
- Optical sensors
• Drugs sreening
- Anti-Alzemier
- Antioxidants
• Clinical applications:
- Lab on paper
- Smart pad
- Smart pact/koyo
- Wearable sensors
• Conclusions
 Sensor
- A small device used for direct measurement of a
physical quantity of an analyte in a sample matrix

- Response is continuous and reversible

- Sample is not perturbed

- Does not require sample collection and preparation

 Sensor
- Consists of a transduction element covered by a
recognition layer

- Recognition layer may be chemical or biological

- Recognition layer interacts with target analyte

- Transduction element translates the chemical

changes into electrical signals
 Definition of a Chemical Sensor
A Chemical sensor is an analytical device
incorporating
 A chemical material/reagent that can recognize or
react with chemical analytes in solution or in the
atmosphere with
 A physicochemical transducer that produces either
discrete or continuous electronic signals
proportional to the analyte

Chemical Sensor = Reagent + Transducer

Chemical Sensor Structure
Reactant

Analyte
R Transducer Electronics

Product
Recognitio
n
Element
 Classification

• Recognition Element Transducer

• Selective • Optical
membranes/films • Electrochemical
• Electromagnetic • Calorimetric
energy • Gravimetric
• Chemical reaction Resonance
 Definition of Biosensor
A biosensor is also an analytical device
incorporating
 A biological material that can recognize biological
or chemical analytes in any type of sample with
 A physicochemical transducer that produces either
discrete or continuous electronic signals
proportional to the analyte

Biosensor = Bioreceptor + Transducer

 Biosensor Principle

SIGNAL

RECOGNITION

TRANSDUCER
LAYER
TARGET
ANALYTE
 Principle of Biosensors

Electroactive  Electrode
Antibody
Substance
Cell
pH change  Semiconductor
Enzyme
pH electrode
Nucleic Acid
Heat  Thermistor
Tissue
Light  Photon counter
Mass change  Piezoelectric device

Solution or Recognizing Signal

Atmosphere Materials Transducers
 State of the art: BIOSENSOR

Recognition layer Transducer

 enzyme  electrochemical
 DNA  piezoelectrical
 antibody  optical
 cell  calorimetry
 tissue
B. Kuswandi et al. , Chimia. Journal, 59, 236 (2005).
 Type of Biosensor
• Enzyme/metabolic
biosensor
– Enzyme & Cell
– etc
• Affinity Biosensor
– Antibody
– DNA
– Lectine
– etc

B. Kuswandi et al. , Chimia. Journal, 59, 236 (2005).

 Schematic of Enzyme Based
Biosensor

Transducer
Biocatalytic Layer

Enzyme
Substrate(s) Product(s)

Substrate(s) Product(s)

B. Kuswandi, M. Mascini, Current Enzyme Inhibition, 1, 11 (2005).

Example of enzyme based Biosensor

Enzyme Substrate -
 kJ/mol)
Catalase Hydrogen peroxide 100.4
Cholesterol oxidase Cholesterol 52.9
Glucose oxidase Glucose 80.0
Hexokinase Glucose 27.6
Lactic dehydrogenase Pyruvate 62.1
 - Lactamase Penicillin G 67.0
Urease Urea 6.6
Uricase Uric acid 49.1
 Molecular Sensors

Techniques used for biomarker identification and detection.

Abbreviations: FISH, fluorescence in situ hybridization; IHC,
immunohistochemistry
Why Chemical sensor & biosensor?

• Miniaturization
• Specificity
• Rapid Response
• Simplicity
• Sensitivity
• Regenerability
ETIOLOGI alzheimer
The Scope of Research...

Inhibition of
Acetylcholinesterase
AChE
Normal: Acetylcholine + H2O Acetate
+ Thiocholine+ H +

S
R’
ME
H EI ESS
Z R
AL OG
PR
 Curcuminoids
• Dari Ahmed dan Gilani (2008)
dalam penelititan yang
dilakukan secara in vitro dan
ex vivo pada senyawa
kurkuminoid, menghasilkan
data bahwa kurkuminoid
memberi aktivitas sebagai
inhibitor asetilkolinesterase.
• Hasil penelitian Abbasi et al.
(2012) terhadap kurkumin
bahwa kurkumin memiliki
aktivitas penghambatan
asetilkolinesterase (AChE),
butirilkolinesterase (BChE), Struktur kurkuminoid
(Ritmaleni dan Simbara, 2010)
lipoksigenase (LOX), dan
enzim tripsin
 Curcuma longa L. &
Curcuma xanthorrhiza R.
• Berdasarkan efek terapeutik,
senyawa kimia yang terkandung
dalam kunyit (Curcuma longa L.)
dan temulawak (Curcuma Curcuma longa L.
xanthorrhiza R.) berkhasiat untuk (Anonim 2, 2012)
mencegah berbagai penyakit
degeneratif seperti
kardiovaskular, sebagai anti
oksidan yang mengikat radikal
bebas, penurun kadar lipid darah,
meluruhkan plak pada otak
penderita penyakit Alzheimer,
mampu memerangi sel kanker
dan infeksi virus maupun bakteri
(Barmawie,et.al., 2006).

Curcuma xanthorriza Roxb.

(Ajizair,2012)
Biosensor Principles
Screen-printed electrodes
“screen-printed” or “silk-screen” Technology
 the possibility of mass production
 Extremely low cost
 Simplicity
 Complete electrochemical system

Reference Counter Work

Struture of screen printed electrodes

AChE Based Biosensor

SINYAL
TERUKUR

AChE +
ekstrak/RVT

Next...
Rivastigmin

Kurva kalibrasi penentuan linieritas RVT berbagai

konsentrasi secara DPV

Kurva kalibrasi penentuan linieritas RVT berbagai

konsentrasi secara CV
 Rivastignin Inhibition

Kurva kalibrasi pengukuran % inhibisi rivastigmin Kurva kalibrasi pengukuran % inhibisi rivastigmin dengan
dengan metode DPV metode CV
 Inhibition of drug candidate

Perbandingan % inhibisi AChE

Curcuma longa Linn. dengan Curcuma
xanthorrhiza Roxb. dengan metode
DPV

Perbandingan % Inhibisi
IC50 Curcuma longa Linn.
dan Curcuma xanthorrhiza
Roxb.
Perbandingan % inhibisi AChE Curcuma
longa Linn. dengan Curcuma xanthorrhiza
Roxb. dengan metode CV
 Comparison study
CLE = Ekstrak Curcuma longa L.
CXE = Eksrak Curcuma
xanthorrhiza R.
RVT = Rivastigmin

Gambaran perbandingan nilai IC50 pada herbal dan kontrol positif (RVT) (CLE=ekstrak Curcuma longa L.;
CXE=ekstrak Curcuma xanthorrhiza R.; RVT=Rivastigmin)
Internatiobal Seminar on Natural Products, 2013, Unair, Surabaya
 Antioxidant Sensors

Relevant chain reactions in the formation of ROS (reactive oxygen species)and RNS (reactive nitrogen species) and the cellular
antioxidant scavenging mechanism. Once the lipid peroxyl radicals (ROO•/RO•) are generated, they are scavenged by
tocopherol (TocH) to produce the tocopheroxyl radical (Toc•), which is regenerated by ascorbate (AscH-). Ascorbate can
neutralize hydroxyl and water soluble peroxyl radicals and superoxides. The resulting ascorbate radical (DHA) is regenerated by
glutathione (GSH), which is oxidized to GSSG and converted back to GSH by the GSH reductase using NADPH.
Antioxidant Sensors
 Antioxidant Sensors

The schematic procedure for the preparation of dry reagent array of

DPPH (Patent Pending No. PI 2013000404).
Antioxidant Sensors
 Antioxidant Sensors
DPPH Based dry reagent array

Konsentrasi DPPH 50 ppm DPPH 100 DPPH 125 DPPH 150

asam galat ppm ppm ppm

1 ppm

5 ppm

8 ppm

10 ppm

15 ppm

23 ppm Calibration curve

 Antioxidant Sensors
Classification of antioxidant standards at 1 mmol/l
concentration based on their scavenging activity
towards the dry array reagent of DPPH, absorbance
was recorded at 517 nm.

The radical scavenging activity of food samples

as a results of reaction with fresh DPPH solution
(grey) and dry reagent array of DPPH (blue) at
517 nm. (For interpretation of the references to
colour in this figure legend, the reader is
referred to the web version of this article.)
 Integrated Optical Sensor
For Screening of Whitening Active Agent
Research collaboration with LIPI 2016
Near-Infrared Spectroscopy (NIRS)
Possibility of using it in a wide range of applications (physical and chemical),
and viewing relationships difficult to observe by other means.

Milled sugar Granular sugar

 Optical Sensor (NIRS)

http://www.arcoptix.com/ARCoptixSpectrometerNIRspectraExamples.pdf
NIRS Identification Procedure
 Quercetin:
The future of whitening active agent

• Lebih aktif daripada

OH standar (asam kojat)
OH

HO O
• Berfungsi ganda
(sebagai tabir surya juga)
OH
OH • Aman secara in vivo

• Pasokannya stabil
Quercetin Screening
in Plant Samples

KELEMAHAN:
• Sampel terbatas
• Perlu pelarut
• Menghasilkan
limbah
• Lama
Quercetin Screening
in Plant Samples

KEUNGGULAN:
• Sampel lebih
beragam
• Tidak perlu pelarut
• Tidak
menghasilkan
limbah
• Cepat
Integrated Optical Sensors
Based on NIRS

KEUNGGULAN:

• Praktis

• Lebih informatif
daripada spektroskopi
UV-Vis

• Dapat diterapkan
secara in situ
 NIR Spectra
Plant samples that contain Quercetin

Apel Malang

Johar

Kumis kucing
Jamblang
Salam
 Pocket-sized NIRS: Tells the Chemical
Makeup of Foods
• Tel Aviv-based startup Consumer Physics pulled the curtain back today on its first product: a tiny hand-held molecular sensor called Scio. The device, which
Consumer Physics has launched a Kickstarter project for, would allow users to scan practically anything –foods, drinks, pills, plants, and more– and get detailed
information on the object’s chemical makeup in just a few seconds.

It might sound like science fiction, but it’s actually

built around an age-old method of materials
analysis known as near-infrared spectroscopy.
Basically, this process involves shining a near-
infrared light onto the surface of a given material,
which causes the molecules to vibrate and bounce
back light in their own unique way. This reflected
light is then collected and passed through a
spectrometer (think of it like a prism) that
separates the light out into all the different
wavelengths it contains. By analyzing the unique
optical signature of the scanned material, it’s
possible to determine what it’s made out of.

http://www.digitaltrends.com/cool-tech/pocket-sized-molecular-spectrometer-tells-chemical-makeup-foods/#ixzz3n5bQ2eDh
 Clinical Application

Construction of a biosensor: from materials, to devices, to systems

 Application
Clinical Diagnosis
Pregnancy, cancer, infectious disease, cardiovascular
problems, health monitoring etc

Food safety test

Allergens, food toxins, GMO, halal food
Ecological detection
Endocrine disturbing chemicals etc

Advantage ： User friendly format, easy to use

Rapid to obtain test result
Less interface due to chromatographic separation
Relative low cost
Problem ： Relative low sensitivity around 1 ng/mL
Semi-quantitative

B. Kuswandi, et al. Anal. Chim. Acta 601, 141 (2007).

Clinical Applications
Glucose monitoring
Glucose monitoring
Spectra may be obtained in non-invasive
manner.

Totally non-invasive analysis of blood glucose by NIR

 Example of Optical Sensor
Non-invasive blood glucose monitor

Schematic overview of
operation of noninvasive Absorbance Spectrum of Glucose
blood glucose monitor
 Miniaturization
• Lab-on-a-chip (LOC) the miniaturization of fluidic
networks (microfluidics) for chemical and biochemical
processing and analysis—offers a means for mass-
produced, low-cost, single-use (disposable) devices for
screening and diagnostics, providing easily interpreted
test results in a time frame of 10 to 60 min.
Microfluidics as key performance

• B. Kuswandi, et al. Anal. Chim. Acta 601, 141 (2007).

Microfluidics vs Macrofludics

• Analytical Lab • Lab on a Chip

• Instrument • Instrument
dimension ~ 10 cm dimension ~ 100 um
• Sample volume • Sample volume
~ 10 mL ~ 1 nL
LOC in the Future
 Microfluidics in Analysis
Sample

Cell

Molecule
• Analytical Lab • Lab on a Chip
• considerable • small sample volumes.
Data
• rapid response & real-
volumes of time.
biological samples • reduced reagent used.
• large, expensive, Time • minimal risk of sample
and require contamination.
Cost • convenient disposal.
trained personnel
• low cost.
Various Lab on a Chips
LOC based on chemical sensor and biosensor
Optical Sensing in Microfluidics
System
 LOC Fabrication
• Usually LOC has been fabricated with silicone,
glass and polymers (PDMS & PMMA) in specific
lab (clean room) with photolithography
technology and wet chemical etching.
• Therefore, the fabrication of LOC is difficult to be
perform in general lab out side clean room.
• In this direction, there is need for new approach in
LOC fabrication out-side a clean room.

http://www.nanoterra.com/i/soft_lithography_diagram.jpg
 Pattern Fabrication
• The crucial step in LOC development is
fabrication of pattern/circuit using generic
technology available out side clean room,
with rapid, simple process & low-cost.
 Our approach:
Screen Printing Technology
• Simple
• Low cost
• Rapid fabrication

Printed LOC – The Dream

 Paper Based Microfluidics
• We believe that paper based microfluidics may be one of
the low-cost sensor platforms available for developing
chemical & biochemical analysis.
• We develop sensing devices based on paper by
patterning paper to form defined areas of hydrophilic
paper separated by hydrophobic lines or “walls” using
screen printing technology at both sides.
• These patterns provide spatial control of biological fluids
and enable fluid transport, without pumping, owing to
capillary action in the millimeter-sized channels
produced.
 Strip Tests

Patterns in paper make it feasible to run analyses on one strip of paper

while still using only small volumes of a single sample.
Furthermore, in a less developed technology lab setting, patterning paper
with screen printing technology is convenient and useful for prototyping
these devices in new micro-patterning technology.

http://img.alibaba.com/photo/11325478/Hcg_Pregnancy_Test_Urine_And_Serum.jpg
 Lab on Paper
• Here, we develop simple paper based
microfluidics for detection of single analyte
(e.g. uric acid, urea & protein) in clinical
samples, such as urine, blood serum & saliva.
kontrol
kontrol
Immobilisasi
reagen

imobilisasi
reagen

1cm 1 cm

Design 1 Design 2
B. Kuswandi and A. A. Gani, Asiasense 2009, p. 41.
 Experimental
• The filter paper has been patterned with rubber
base ink in both side using screen printing
technology.
• This screen printing technology, offers rapid,
simple and low cost in developing pattern on the
paper as microfulidics.
 Experimental
• After printing, the patterned paper was dried for 3 min using hair dryer.
• The paper was more hydrophobic after it was patterned, while the un-
patterned side was still hydrophilicity region of the paper.
• After this process, the patterned paper can be used as LOC for clinical
analysis by adding appropriate reagents to the test region.
 Lab on Paper
• The lab on paper based could be used for
dual and multiple analytes detection
simultaneously.

Single analyte Dual analytes Multiple analytes

B. Kuswandi and A. A. Gani, Asiasense 2009, p. 41.

 Image Analysis

The image Data

1000 ppm

• Data can be send to

the doctor or physician

B. Kuswandi, Asianalysis X, p. 24-25.

 Color Analysis
• The color of the image
could be further
analyzing by photo UA conc. (ppm) 1000 ppm Δ Mean Red

software for quantitative 100 4,69

assays.
• The paper chip has 200 5,16

been tested for UA,

300 8,22
using the value of Δ
mean of Red using 400 13,22
“Adobe Photoshop 7.0”.
500 16,15
Tele-Health Care
 Wearable Sensors

Woman i-pad, which is hoped will

be used to monitor:
- women healths
- anemia
- to detect the initiation of cancer-
forming cells prior to any
substantial proliferation (cervix
cancers)
- help to detect the end of blood
menstruation, which is very
important for women according
to Islamic law (syari’ah)
Smart Pad
 Smart Pad (1)

Urease in
Buffer pH 7 & buffer pH 7,5
Chlorophenol & Bromothymol
red 5000 ppm Blue 3000 ppm

Before After
Smart Pad (1)
 Smart Pad (2)

Bromothymol blue

1,10 – Fenantrolin

Before After
Smart Pad (2)
Smart Pact (Koyo Pintar)
Batik Technology Based LOC
Smart Koyo with LOC
Smart Koyo with LOC
 Wearable Sensors
iBra: Sensors for monitoring breast health & cancers

The iBra will be integrated with sensors that can be used to monitor breast health & to detect of
minor temperature changes that occur to the breast tissue. As tumors begin to grow, they
demand a higher blood flow to develop. This increased blood flow will result in an elevated
temperature, signifying the initial stages of cancerous growth.
The cancer-preventing bra will be worn instead of a "normal" undergarment, and will incorporate
a discrete alarm feature to alert the wearer to seek medical attention if necessary.
Conclusions
Sensors (chemical sensors and biosensors) are rapid
analytical tools that can be used as drugs screening.
Microfluidics are versatile platform for chemical sensors &
biosensors as analytical tools in clinical applications
Sensors are at the forefront of multi-disciplinary science
involving the marriage of the chemical & biological world
and the physical electronic world
Smart function could be build in conjunction with
microprocessors and telecommunications technology,
including Image analysis, Data acquisition and processing
Broad Application in various field ranging from drugs
screening, clinical, environmental, industry, health etc
 Acknowledgment
• Thanks to all funded institutions:
- KNAW the Netherlands via SPIN &
Postdoctoral Program
- DP2M Higher Education via Hibah Kompetensi ,
& HKLN
• The author also thanks:
• CBG Staffs & students members for assisting and
performing this work.
 Reference
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sensors, Sensors, 6, 978 (2006).
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microfluidic devices, Anal. Chim. Acta 601, 141 (2007).
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samples using a microfluidic device containing a selective tripodal chromoionophore-PVC film,
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Platform, Asiasense 2009, The 4th International Conference on Sensors, July, 29-31, 2009, The
Imperial Queen’s Park Hotel, Bangkok, Thailand, p. 41.
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Putra World Trade Centre, Kuala Lumpur, Malaysia, p. 24-25.
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Thank You
For time & attention !!!