Manuscript 11032013-final (1)

1
Diagnosis of Skin Health Conditions Using Gold Nanoparticles#
Sudipa Panigrahi, Kavitha Murugesan, Samares C Biswas*
ITC R&D Centre, Personal Care Products Business Division, Peenya Industrial Area 1st Phase,
Bangalore – 560 058, India,
E-mail: samares.biswas@itc.in, Tel: +91 80427807335/9844500167, Fax: +918028394352
ABSTRACT
This study reports a novel non-invasive technique for
the detection of skin health conditions using gold
nanoparticles (GNP). The feasibility of the use of these
GNP in the diagnosis of skin health conditions was non-
invasively studied using skin samples collected through
tape-stripping method. The skin health conditions were
diagnosed based on the visual color change of GNP and the
same has been established spectroscopically (UV-Vis) by
monitoring the surface plasmon band (SPR) of GNP. This
novel non-invasive in-vitro method determines the skin
health conditions precisely and can guide the consumer to
select the suitable products fit for their skin conditions. It
diagnostic can be extended to demonstrate the performance
of personal care products. The proposed diagnostic kit can
also be used to demonstrate the dermatological conditions
to predict the onset of skin diseases. This tool would also be
applicable to predict any disturbances in the skin’s acid
mantle.
Keywords: nanoparticles, diagnosis, health, skin, skin pH,
skin moisture, acid mantle
1 INTRODUCTION
The skin is the outermost layer of vertebrates that
provides a vital barrier for protecting them from both
routine and extreme environmental assaults including
exposure to antigens, solvents, ultraviolet light, detergents,
microorganisms, toxins, nanoparticles and a variety of
physical and chemical insults [1-6]. All these insults alter
the skin barrier function and that in turn leads to change in
skin-health conditions. Therefore there is an utmost need to
diagnose and monitor the skin health conditions regularly
for maintaining skin homeostasis. There exist different
instruments that can facilitate the skin analysis through
measurement of intrinsic factors such as moisture content,
transepidermal water loss, skin elasticity, skin color, micro
circulation, skin thickness etc. [7-19]. In addition to this
there also exist a need of analysing skin conditions with
instruments to develop and recommend skin care products
for the consumers. The main limitation of using all these
instrumental methods in day to day use by the consumer
are (i) cost, (ii) portability, (iii) complexicity in operation
(iv) space etc.
Metal nanoparticles specifically gold nanoparticles have
occupied important position in the fields of chemistry,
physics, and biology because of their unique optical,
electrical, and photothermal properties. Nanoparticles have
found potential applications in analytical chemistry and the
same have been used as probes in mass spectroscopy, as
well as in the colorimetric detection for proteins and DNA
molecules [20-21]. Nanoparticles have high potential for
fabricating biological labels, biological sensors, bioanalysis
and biodiagnosis technologies, diagnosis and monitoring of
diseases, drug discovery, environmental detection of
biological reagents, and even medical and clinical diagnosis
and therapy [22-29].
The present work provides a in-vitro method for both
qualitatively and quantatively assessing skin health
conditions using the size dependent optical properties of
GNP in the presence of skin samples.
2 MATERIALS & METHODS
D(+) Glucose, sodium borohydride (NaBH4) and
chloroauric acid (HAuCl4, 3H2O) were purchased from
Sigma Aldrich (St. Louis, MO, USA). KOH was obtained
from SD Fine Chemicals, India Ltd. The adhesive tape of
2x2cm2
was used for collecting skin samples. Milipore
water was used throughout the experimental works. All
spectroscopic measurements were done using a Shimadzu
UV-vis spectrophotometer.
2.1 Preparation of Gold Nanoparticles (GNP)
The GNP was synthesise following two synthetic
methods, where gold salts such as was chemically reduced
usingreducing agents such as D(+) Glucose, sodium
borohydride (NaBH4) [30-34].
Method 1. In a 100 mL glass beaker 5 gm D(+) glucose
was dissolved in 48 mL water and mixed properly. Then 2
mL of 10-2
M HAuCl4 solution was mixed to it. Finally 100
µL of 1M KOH solution was added to it at a time under
vigorous stirring. The solution turned wine red indication
the formation of gold nanoparticles. The solution was
stirred for 10 minutes to ensure the complete formation of

gold nanoparticles in water. Finally the solution
overnight at room temperature under dark.
Method 2. 30 mL solution of 2.5×10-4
M HAuCl4 in water
was prepared in a 100 mL round bottle conical flask and an
ice cold solution of NaBH4was addedinto it under constant
stirring until its concentration reached to 2.5×10
appearance of immedite wine red colour indicating the
formation of GNP. The synthesized nanoparticl
stored overnight at 4˚C for further use.
2.2 Diagnosis of Skin Health using GNP
Collection of skin samples using tape-stripping method
2×2 cm2
area were marked on volar forearm and elbow
Apply adhesive tape of dimension 2×2 cm2
on the marked
region and pressed the adhesive tape gently
and then remove the adhesive tape containing skin samples.
Treatment of skin samples with GNP
In a 2.0 mL ependorf tube 1.0 mL of nanoparticle
dispersion (wine redcolor) were taken. The skin samples
collected through tape stripping technique was added to it
(skin samples from forearm and elbow). The solution was
mixed for 30 seconds using a vortex mixture. The
color of the GNP solution was followed visually as well as
spectrophotometrically by recording the change in the
pattern of SPR band of the GNP.
3 RESULTS& DISCUSSION
The UV-Vis spectra for GNP were recorded. The
characteristic SPR of the GNP was observed at 520 nm
(Figure 1) indicating the presence of spherical GNP
of 10-20 nm. [35-36]. The color of GNP as well the SPR
band remains unafected in the presence of the only
tape without any skin samples..
From Figure 2, one can note that the wine red
the GNP solution turns blue, when treated with skin
samples collected from elbow through tape stripping while
the colour changed to violet while traeted with the sk
samples collected from volar forearm. The color of GNP
gradually changes from wine red to blue, when treated with
skin samples collected from panelists having different skin
moisture level. The change of color of GNP in the presence
of skin samples indicating the change in the aggregation
state of GNP under different conditions offered by the skin
samples from different regions.[37- 42].
water. Finally the solution was stored
HAuCl4 in water
was prepared in a 100 mL round bottle conical flask and an
was addedinto it under constant
2.5×10-3
M.An
indicating the
The synthesized nanoparticles were
Diagnosis of Skin Health using GNP
stripping method
on volar forearm and elbow.
on the marked
gently using finger
skin samples.
In a 2.0 mL ependorf tube 1.0 mL of nanoparticle
The skin samples
ng technique was added to it
he solution was
The change in
visually as well as
change in the
& DISCUSSION
were recorded. The
observed at 520 nm
indicating the presence of spherical GNP of size
color of GNP as well the SPR
only adhesive
wine red color of
the GNP solution turns blue, when treated with skin
through tape stripping while
the colour changed to violet while traeted with the skin
The color of GNP
en treated with
lected from panelists having different skin
The change of color of GNP in the presence
the change in the aggregation
ffered by the skin
Figure 1.Plasmon resonance absorbance Spectra of GNP
the absence and presence of tape
The surface properties of skin depends on the
anatomical location in human body.
surface of elbow differ from forearm with
moisture content, skin integrity, pH etc
different skin barrier function [43]. Skin of volar forearm
more moisturized when compared with elbow skin.
addition to this, pH of the skin surface of elbow
be higher than forearm. Therefore the change in color
well as the SPR of GNP in the presence of skin samples
(forearm and elbow) could be attributed
difference in the biochemical properties (skin mo
level, pH, water soluble protein content
between the two skin surfaces.
Figure 2. Change in color of the GNP in the presence of
skin samples
2
lasmon resonance absorbance Spectra of GNP in
The surface properties of skin depends on the
anatomical location in human body. It is known that skin
ffer from forearm with respect to
, skin integrity, pH etc and they have
. Skin of volar forearm is
more moisturized when compared with elbow skin. In
of the skin surface of elbow reported to
Therefore the change in color as
of GNP in the presence of skin samples
attributed due to the
biochemical properties (skin moisture
level, pH, water soluble protein content, lipid content etc)
Change in color of the GNP in the presence of

3
Figure 3. SPR of GNP in the presence of skin samples.
Colorimetric responses of GNP towards skin samples of
different types (dryness level). The GNP was treated with
skin samples for 30-45 sec.
The SPR of GNP was recorded in the presence of
different skin samples as described in meterials and
methods section. The SPR of GNP undergone a red shift,
when treated with dry skin sample. A gradual red shift on
the characteristic SPR was observed, when GNP was
treated with skin samples of increasing dryness level.
It is reported in the literature that the biochemical
properties and barrier function of skin surface differ from
normal to dry skin [43]. The major differences between
normal and dry skin are moisture level, skin surface pH.
The red-shift of the SPR band can be attributed to changes
in the shape, size and different state of aggregation of GNP.
Therefore, the observed change in spectral profile of GNP
in the presence of skin samples might be due to the
differences in the biochemical properties of skin samples
having different dryness level.The effect of different bulk
parameters on the morphological changes of nanoparticles
has been described elsewhere [35-42].
Therefore from Figure 3, one can noted that the change in
SPR of GNP can be correlated with the skin type having
different dryness level.
The effect of pH on the SPR of GNP has been studied and
the data has been presented in Figure 4. The change in of
SPR of GNP with pH is of great interest. The SPR of GNP
was found to be slightly blue shifted with increase in pH.
From Figure it can be seen that change in spectral pattern of
GNP with buffer solution of different pH is not similar to
change in spectral pattern of SPR of GNP treated with skin
samples collected from different region of skin. Hence it
can be concluded that the change in SPR pattern along with
the change in colour of GNP is due to the different skin
conditions which is a combination of factors discussed
earlier. F The study was conduceted over 30 panelists and
similar change in colour and spectral pattern was observed.
Figure 4. Plasmon resonance absorbance spectra of GNP
under varying bulk solution pH
4 CONCLUSIONS
In this work it has been demonstrated that GNP can be
used for the detection of skin health conditions both
visually and spectroscopically. This non-invasive in-vitro
method determines the skin health conditions precisely and
can guide the consumer to select the suitable products. This
diagnostic method can be extended to demonstrate the
performance of personal care products. This diagnostic kit
can also be used to demonstrate the dermatological
condition to predict the onset of skin diseases. This tool
would also be applicable to predict any disturbances in the
skin acid mantle. Finally this technique would be useful to
evaluate the performnace of different skin care products and
recommend the correct product to the consumer depending
on their skin conditions.]’
5 ACKNOWLEDGEMENT
The authors are thankful to Dr. V. Krishnan for his
valuable advice and helpful stimulating discussions. The
authors are also thankful to ITC Ltd for the financial
support and providing the necessary laboratory facilitiesfor
carrying out this work.

4
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#Patent filed, * All correspondence should be addressed to
Samares C Biswas, ITC R&D Centre, Peenya Industrial
Area, 1st
Phase, Bangalore – 560 068, samares.biswas@itc.in

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