Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 293 (2023) 122457

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Spectrochimica Acta Part A:

Molecular and Biomolecular Spectroscopy
journal homepage: www.journals.elsevier.com/spectrochimica-acta-part-a-
molecular-and-biomolecular-spectroscopy

Surface-enhanced Raman spectroscopy of centrifuged blood serum samples

of diabetic type II patients by using 50KDa filter devices
Usama Ehsan a, Haq Nawaz a, *, Muhammad Irfan Majeed a, *, Nosheen Rashid b, *, Iram c,
Zain Ali a, Anam Zulfiqar a, Ayesha Tariq a, Muhammad Shahbaz a, Lubna Meraj a, Iqra Naheed a,
Nimra Sadaf a
a
Department of Chemistry, University of Agriculture Faisalabad, Faisalabad 38000, Pakistan
b
Department of Chemistry, University of Education, Faisalabad Campus, Faisalabad 38000, Pakistan
c
Government Graduate College for Women, Faisalabad 38000, Pakistan

H I G H L I G H T S G R A P H I C A L A B S T R A C T

• Surface enhanced Raman spectroscopy

is employed to characterize the centri­
fuged serum samples of diabetic patients
and healthy ones.
• Silver nanoparticles are used as Surface
enhanced Raman spectroscopy
substrates.
• Some Surface enhanced Raman spec­
troscopy spectral features are associated
with diabetic samples in comparison
with healthy ones which can be
employed for diagnostic purpose.
• Principal components analysis (PCA) is
found helpful for the differentiation be­
tween SERS spectral data of diabetic and
healthy samples.
• Partial Least discriminant Analysis
(PLSDA) model is validated and can be
used for the classification of diabetic
samples and healthy samples.

A R T I C L E I N F O A B S T R A C T

Keywords: Blood serum contains essential biochemical information which are used for early disease diagnosis. Blood serum
Centrifugal filtration consisted of higher molecular weight fractions (HMWF) and lower molecular weight fractions (LMWF). The
Filtrate portions of blood serum samples disease biomarkers are lower molecular weight fraction proteins, and their contribution to disease diagnosis is
surface-enhanced Raman spectroscopy
suppressed due to higher molecular weight fraction proteins. To diagnose diabetes in early stages are difficult
Type II diabetes
Multivariate data analysis
because of the presence of huge amount of these HMWF. In the current study, surface-enhanced Raman spec­
troscopy (SERS) are employed to diagnose diabetes after centrifugation of serum samples using Amicon ultra
filter devices of 50 kDa which produced two fractions of whole blood serum of filtrate, low molecular weight
fraction, and residue, high molecular weight fraction. Furthermore SERS is employed to study the LMW fractions

* Corresponding authors.
E-mail addresses: haqchemist@yahoo.com (H. Nawaz), irfan.majeed@uaf.edu.pk (M. Irfan Majeed), nosheenrasheed@yahoo.com (N. Rashid).

https://doi.org/10.1016/j.saa.2023.122457
Received 2 January 2023; Accepted 3 February 2023
Available online 6 February 2023
1386-1425/© 2023 Elsevier B.V. All rights reserved.
U. Ehsan et al.
of healthy and diseased samples. Some prominent SERS bands are observed at 725 cm− 1, 842 cm− 1, 1025 cm− 1,
959 cm− 1, and 1447 cm− 1 due to small molecular weight proteins, and these biomarkers helped to diagnose the
disease early stage. Moreover, chemometric techniques such as principal component analysis (PCA) and partial
least square discriminant analysis (PLS-DA) are employed to check the potential of surface-enhanced Raman
spectroscopy for the differentiation and classifications of the blood serum samples. SERS can be employed for the
early diagnosis and screening of biochemical changes during type II diabetes.
1. Introduction
Diabetes type II is non-insulin-dependent diabetes and is a major
health problem. It has recently emerged as one of the most common and
serious chronic disease, which causes life-threatening, devastating,
costly complications and reduction in life expectancy [1]. According to
the International Diabetes Federation (IDF), an estimated 537 million
individuals suffer diabetes around the world and it is expected that this
value will rise to 783 million in 2045. Among different types of diabetes,
diabetes type II is lethal and accounts for the majority of cases [2]. Major
risk factors for diabetes type II are overweight, a lack of physical activity
and a family history of diabetes [3]. The long-term impact causes many
complications including loss of vision; kidney failure, amputations,
gastrointestinal, genitourinary, cardiovascular symptoms and sexual
dysfunction [4]. Furthermore, it is usually diagnosed at later stages of
the disease, causing many problems in treatment. The diagnosis of this
disease in early stages is critical for extending the life, reducing com­
plications and for its diagnosis diabetes screening and blood glucose
testing is frequently advised [5].
Raman spectroscopy is used for the biochemical analysis and provide
information about the molecular structure and conformation as a unique
fingerprint [6–12]. Both Raman Spectroscopy and Surface Enhanced
Raman spectroscopy can be used for the disease diagnosis as explained
earlier in different studies. These studies include applications of Raman
Spectroscopy for diagnosis of Hepatitis B [13], breast Cancer [14] and
diabetes [15].Moreover, in a recent study Raman spectroscopy is used
for quantification of glucose by employing centrifugally filtered serum
samples [16]. Surface enhanced Raman spectroscopy (SERS) has been
used for the diagnosis of different diseases such as hepatitis C [17,18],
hepatitis B [19], dengue infection [20], typhoid infection [21] and
breast cancer [22]. Although, FTIR, NIR can also be used for the bio­
logical studies but have the problem of interference by water if present
in the samples [23]. SERS is widely used in disease diagnosis because it
clearly differentiates between healthy and disease spectral data sets
[24–26]. Moreover, this technique is low cost, requires little or no
sample preparation and enables easy sample identification [27]. Raman
Spectroscopy and Surface Enhanced Raman spectroscopy were also
employed for the detection of diabetes by using serum [28,29] plasma
[30] and erythrocytes [31] samples, after carefully reviewing no study
on the centrifugally filleted serum of diabetes type II samples.
A variety of disease-related biomarkers can be found in body fluids
such as urine, blood and serum [32]. In this regard, blood serum is
significant for early monitoring or screening of diabetes patients in order
to identify disease-related biomarkers which are lower molecular weight
fraction biomolecules such as DNA [33–35], aliphatic hydrocarbon of
lipid molecules [36–39] and cholesterols [40,41]. Blood serum contains
an extensive collection of essential disease related information deter­
mination of which can aid in early disease detection and its biochemical
characterization. Blood serum contains high molecular weight protein
fractions (HMW) and some low molecular weight protein fractions
(LMW). These LMW protein fractions are considered as disease bio­
markers but the investigation of human whole serum for disease specific
biomarkers seems to be quite difficult. This is due to the reason that in
blood serum, albumin and globulin are high molecular weight proteins
suppress the low molecular weight proteins making their detection
difficult [42].
Approximately 90 % of the serum’s total protein composition is
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Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 293 (2023) 122457
made up of six proteins. Each one weighs than 50 kDa.. These proteins
are Albumin (66 kDa), immunoglobulin G (IgG) with 150 kDa, trans­
ferrin (80 kDa), α1-Antitrypsin (52 kDa), immunoglobulin A (IgA)
180–500 kDa and immunoglobulin M (IgM) (950 kDa) [43,44].In this
study 50 kDa filter devices used because a majority of diabetes type II
related disease proteins range in size less than 50 kDa[45,46].In diabetes
type II, more than fifteen proteins with molecular weights ranging from
9 to 100 kDa are present, of which six major proteins have molecular
weights of 45, 37, 25, 24.7, 20, 13.9, and 9.4 kDa which are Hapto­
globin, Immunoglobulin heavy constant alpha, Serum amyloid P-
component, Glucagon-like peptide-1 amide, Transthyretin and apoli­
poprotein respectively [47]. These proteins could be potential targets in
the current study following centrifugal filtration with a filter size of 50
kDa. Therefore, serum samples performed centrifugal filtration to
separate the LMWF and HMWF. The SERS spectral characteristics
related to the diabetes type II disease biomarkers were subsequently
carried out to analyse the LMWF. After centrifugation two fractions are
obtained including high molecular weight protein fractions (HMW) and
some low molecular weight protein fractions (LMW). The later contains
possible disease related proteins. The LMW are then analyzed with
surface enhance Raman spectroscopy (SERS) to identify the biochemical
changes based on lower molecular weight protein fractions in diabetic
patients in comparison with healthy ones. Moreover, different chemo­
metric techniques such as principal component analysis (PCA) and
partial least square regression discriminant analysis (PLS-DA) are
employed to check the potential of surface-enhanced Raman spectros­
copy for this purpose. Furthermore, PLS-DA approach is employed for
differentiating and classification of SERS spectral data sets of filtrate
portions of centrifuged serum samples of diabetic patients and healthy
ones. Various studies are being conducted to investigate the use of
Raman spectroscopy [15,28,48] and surface enhanced Raman spec­
troscopy (SERS) [29,49] for the analysis and diagnosis of diabetes while
using blood components including serum and plasma. After careful re­
view of these paper, it is observed that analysis of diabetes of centrifuged
serum samples by using surface enhanced Raman (SERS), has not been
explored and no such research article has been published yet.
2. Material and methods
2.1. Sample preparation
The blood samples of healthy and diabetic patients were obtained
from the Rehman Clinical Lab, Faisalabad, Pakistan. In total were 60
samples, including 15 healthy samples and 45 diabetic samples were
used. For this study, we have used a 3 ml venous blood sample drawn
into an EDTA/Sodium fluoride tube. To separate blood serum, centri­
fugation at 2000 rpm, 4℃ for 10 min [50]. For further centrifugation of
serum samples to get low molecular weight protein fraction called
filtrate parts and high molecular weight proteins called as residue,
Amicon Ultra-2 ml centrifugal devices (Merck, Millipore) were used
which has cutoff value of 50 kDa. For the centrifugation of the serum,
250 µl of serum was taken in a centrifugation tube and centrifuged at
6500 rpm for 30 min. After that two fractions were found, one was
residue which have a high molecular weight (>50KDa) and the other
was filtrate which have low molecular weight (less than50KDa) [51].
U. Ehsan et al.
Fig. 1. Mean SERS spectra of centrifugally filtered serum (filtrate) and uncentrifuged serum (whole Serum) and Residue of healthy samples.
2.2. Synthesis of silver nanoparticles
The chemical reduction method was used to synthesize the silver
nanoparticles (Ag-NPs). As a precursor silver nitrate (AgNO3) was used
and reduced by trisodium citrate (Na3C6H5O7). For this, 0.0085 g of
AgNO3 was added to distilled water followed by heating the solution to
100 ◦ C and then added one percent of trisodium citrate (Na3C6H5O7).
The solution was continuously stirred with the magnetic stirrer over the
hot plate for one and half hours to produce grey coloured silver (Ag)
nanoparticles [17].
2.3. SERS spectral acquisition
A Peak Seeker Pro-785; Agiltron, USA Raman spectrometer having a
785 nm diode laser delivering a power of 50 mW through a 40X
objective was used to get SERS spectra of the filtrate portions of the
blood serum samples of the healthy volunteers and diabetic patients. For
SERS spectral acquisition, a 10 µl fraction of each filtrate sample was
placed inside the groove on the aluminum slide and SERS spectra were
obtained. SERS spectra were recorded from all the samples in the range
of 400–1800 nm, and 15 SERS spectra were taken for each sample with
an integration time of 15 s for each SERS spectrum.
2.4. Data preprocessing
By Using Matlab 7.8, the SERS spectral data was preprocessed ac­
cording to the established protocols [52]. The preprocessing of SERS
spectra included smoothing, baseline correction, substrate removal and
normalization,. Savitzky-Golay smoothing method is used for smoothing
of spectra [18]. The smoothing methods of Savitzky-Golay (order 1, 17-
point window) were applied to smooth the data. Moreover, vector
normalization method for normalizing the spectra, and subtraction
method for th removal of background/substrate. The baseline correction
was applied to all of the spectra using the Rubberband algorithm [53].
2.5. SERS data analysis
To compare the mean SERS spectra of centrifuged serum samples of
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Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 293 (2023) 122457
healthy/control with that of centrifuged serum samples of diabetes
positive patients, Raman spectral data was analyzed by mean SERS plot.
Moreover, PCA and PLS-DA are also employed for the multivariate data
analysis. PCA is a statistical technique that transforms correlated vari­
ables into uncorrelated ones. It maintains the variability of the data by
reducing its dimension. The first principal component (PC) -explains the
highest variances in data, while each subsequent PC explains the
remaining variance in the data. The PC loadings are illustrating the
orthogonal dimensions of variability that can be used to separate SERS
spectral data into groups based on their variability. While partial least
square discriminant analysis (PLS-DA) was further employed for the
differentiation and classification of the SERS spectral data sets of the
filtrate portions of the centrifuged serum samples of healthy and dia­
betes positive samples.
3. Results and discussion
Blood serum contains LMW and HMW fractions.Notably, as the LMW
fractions are suppressed by HMW fractions in the uncentrifuged serum
samples so not identifies as SERS spectral features. On centrifugal
filtration, the HMW fraction is removed (go in residue) so LMW fraction
related SERS features are now easily identified in centrifugally filtered
serum samples.
Fig. 1 shows the SERS spectral features of healthy whole serum
(blue), filtrate (red), and residue (black). Which clearly indicates that in
the filtrate, SERS spectral features are more prominent than in the serum
due to the removal of HMW fractions of the serum, and also, some SERS
spectral features that the absent in the filtrate and present in the residue
and serum are due to the HMW fractions of the Serum and these SERS
spectral features are 497 cm− 1 (Glycogen),592 cm− 1(Amide-VI),630
cm− 1(Glycerol),725 cm− 1 (Glycosidic ring vibrational mode (adenine,
polyadenine, DNA),850 cm− 1(tyrosine),890 cm− 1,970 cm− 1,1003
cm− 1(phenylalanine),1102 cm− 1(v(CC)and v(CN)), 1133 cm− 1,1205
cm− 1 (Collagen),1280 cm− 1(C–H bending),1450 cm− 1(deformation
mode of CH4),1512 cm− 1.Some SERS spectral features 850 cm− 1 (tyro­
sine), 1003 cm− 1 (phenylalanine),1102 cm− 1 (v(CC)and v(CN)) 1450
cm− 1 (deformation mode of CH4) are present only in uncentrifuged
serum samples which are due to higher molecular weight proteins
U. Ehsan et al. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 293 (2023) 122457

Fig. 2. Mean SERS spectra of centrifuged serum samples of diabetic patients and healthy/control.

from that healthy ones. A solid line represents peaks that are present in
Table 1
one spectrum while absent in others, and a dotted line represents peaks
SERS spectral peaks observed in the SERS spectra of Centrifuged serum samples
that are present in both spectrums but vary in peak intensities. Some
of healthy and diabetic patients.
higher intensities SERS spectral features in diabetic samples include 725
Peak Peak Assignment References
cm− 1 (Glycosidic ring vibrational mode adenine, polyadenine, DNA)),
position
(cm¡1)
959 cm− 1 (Cholesterol), 1025 cm− 1 (C–H deformation/adenine ring
stretching), 1095 cm− 1 (Lipids), 1332 cm− 1 (C–H stretching vibration,
356 δ(C–C–C) ring vibration of glucose [69]
Nucleic acid bases), 1447 cm− 1 (A Phospholipid), 1528 cm− 1 (carot­
362 C–C stretching [70]
392 C–COO − stretching mode [71] enoid (absent in normal tissues)), 1587 cm− 1 (Protein, Tyr) and 1703
418 Cholesterol [72] cm− 1 (ν(C–– O)OH).
429 Cholesterol, cholesterol ester [72] Similarly, some SERS spectral features including 356 cm− 1 (C–C–C
497 Glycogen [73]
ring vibration in glucose), 548 cm− 1 (Cholesterol), 1290 cm− 1 (Cyto­
526 S–S disulfide stretching in proteins [74]
548 Cholesterol [72] sine) and 1703 cm− 1 (ν(C– – O)OH) are present only in diabetic type- II
566 Tryptophan [75,76] but absent in filtrate portion of healthy centrifuged serum samples.
592 Amide-VI [58] However, some SERS spectral features are observed both in healthy and
630 Glycerol [72] diabetic type- II samples, including 362 cm− 1 (C–C stretching), 418
643 C–C twisting mode of tyrosine [73,77]
cm− 1 (Cholesterol), 429 cm− 1 (Cholesterol), 566 cm− 1 (Tryptophan),
659 C–S stretching mode of cysteine [78]
725 Glycosidic ring vibrational mode (adenine, [79,80] 959 cm− 1 (Cholesterol), 1095 cm− 1 (Lipid) and 1572 cm− 1 (v(C– – C)
polyadenine, DNA) Retinal). However, SERS spectral peak due to 725 cm-1 (adenine) can be
802 Uracil-based ring breathing mode [81] considered as a biomarker that can differentiate diabetic type- II from
842 Glucose [73]
healthy ones [58]. Some other SERS spectral features are observed with
893 Backbone, C–C skeletal [82]
959 Cholesterol [83]
increased peak intensities including 418 cm− 1 (Cholesterol), 429 cm− 1
1025 C–H deformation/adenine ring stretching [84] (Cholesterol), 959 cm− 1 (Cholesterol), 1095 cm− 1 (Lipid) which shows
1095 Lipid [85] that cholesterol and lipids concentration is increased in the diabetic
1174 Tyrosine, phenylalanine, C H bend (protein) [82] patients as compared to the normal ones [59]. Although human serum
1204 Collagen [86]
has over 4000 metabolites [60], some of which have a strong affinity for
1290 C–H bending [87]
1332 C–H stretching vibration, Nucleic acid bases [88] metal surfaces, and produce sharp SERS signal when used with metal
1400 NH in-plane deformation [32] nanoparticles. When the ratio of nanoparticles to biofluid volume is
1447 Phospholipid [58,76] high, there will be enough surface available to adsorb a range of analytes
1528 Carotenoid (absent in normal tissues) [89]
from those with high binding affinity (adsorbing quickly) to those with
1572 v(C–
–C) Retinal [90]
1587 Protein, Tyr [58]
low binding affinity (adsorbing slowly). The analytes with the highest
1703 ν(C–
–O)OH [91] binding affinities will adsorb first, potentially saturating the available
surface. When this ratio is low, there will be less surface available for
adsorption. These two effects lead to adsorption of different ratios of
including immunoglobulins and albumin [54–57]. biofluids at the surface of nanoparticles and, consequently, different
Mean SERS spectra of filtrate portions of the centrifuged serum SERS spectra are produced, which has recently been reported for the
samples of diabetic patients and healthy individuals are shown in Fig. 2. spectra of serum and plasma [61]. SERS of biofluids predominantly
References for their peak assigments of SERS spectral features shown in measures the biochemical constituents which have an affinity to adhere
Fig. 1 and Fig. 2 are provided in Table 1. There are two types of lines to the metal nanoparticle surface [62–64].”.
solid lines and dotted lines, used for labelling the SERS features differ­ In the filtrate portions of centrifuged blood serum samples, an
entiating between filtrate portions of serum samples of diabetic patients

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U. Ehsan et al. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 293 (2023) 122457

Fig. 3. PCA scatter plot of SERS spectra of filtrate portions of healthy (green dots) individuals and diabetic patients (red dots).

Fig. 4. PCA loadings of SERS spectral data SERS spectra of filtrate portions of healthy individuals and diabetic patients.

intense SERS feature was observed at 725 cm− 1, which corresponds to
the C–H bending mode of adenine. An increase in blood glucose seems
to be directly related to the intensity of this band, indicating changes in
adenine nucleotide A SERS contents in the centrifuged serum samples of
diabetic individuals [58,65]. A SERS feature at 842 cm− 1, is related to
carbohydrate (glucose), and its intensity is increased in diabetic spec­
trum as compared to normal spectrum.. As a result of this, it is critical to
explain that impaired glycoprotein component metabolism is expected
to play a key role in type- II diabetes diagnosis [66]. The other SERS
band at 1447 cm− 1 is assigned to the CH2 bending mode of the aliphatic
hydrocarbon of lipid molecules. This increases in intensity levels of this
band is an indication of diabetes. This may imply that an increase in lipid
content is closely related to diabetes [65,67]. Moreover, the prominent
SERS band at 1095 cm− 1 represent higher contents of lipids which shows
that diabetic patients have enhanced levels of lipids. The SERS bands at
418 cm− 1,429 cm− 1, 548 cm− 1, 959 cm− 1 are due to cholesterol which
differentiate that diabetic patients have more concentration of choles­
terol as compared to the normal ones [68]. Moreover, all of the peaks in
different SERS spectra of centrifuged serum samples of diabetes may also
be considered significant biomarkers for the diagnoses of diabetes type-

5
U. Ehsan et al. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 293 (2023) 122457

Fig. 5. PLS-DA scatter plot of SERS spectral data sets of filtrate portions of healthy individuals and diabetic patients.

II on the basis of relatively high blood glucose levels As a result, the SERS
spectra have been successfully used to analyze centrifuged blood serum
samples in order to identify people with high diabetic conditions
compared to healthy ones.
3.1. Principal component analysis (PCA)
Principal component analysis (PCA) is used to differentiate SERS
spectral data of different groups/classes. PCA was used on entire data­
sets to classify and verify SERS features related to diabetes. The SERS
spectra of different groups/classes are clustered and differentiated by
PC-1 along X-axis. Maximum variability is explained by PC-1 which can
be seen in the PCA scatter plot as shown in Fig. 3. The SERS spectra of
diabetes positive samples are clustered on the negative side of PC-1,
while those of healthy samples are clustered on the positive side of
PC-1, explaining the majority of the variance, 66.65 %, in the SERS
spectral data of disease and healthy samples, with PC-2 accounting for
the 20.41 % variance.
Fig. 4 Shows PCA loadings identifying SERS spectral features as
loadings of centrifuged serum of healthy samples and centrifuged serum
samples of diabetic patients. The SERS spectral features observed in PCA
loadings include 548 cm− 1 (Cholesterol), 725 cm− 1 (Glycosidic ring

Fig. 6. Receiver operating characteristic (ROC) curve showing area under the curve (AUC).

6
U. Ehsan et al.
vibrational mode (adenine, polyadenine, DNA)), 842 cm− 1 (Glucose),
959 cm− 1 (Cholesterol), 1025 cm− 1 (C–H deformation/adenine ring
stretching), 1095 cm− 1 (Lipids), 1174 cm− 1 (Tyrosine, phenylalanine, C
H bend (protein)), 1290 cm− 1 (C–H bending), 1332 cm− 1 (C–H
stretching vibration, Nucleic acid bases), 1447 cm− 1 (Phospholipid),
1587 cm− 1 (Protein, Tyr), 1703 cm− 1 (ν(C– – O)OH) on positive side of
the loadings while the PCA loadings at 592 cm− 1 (Amide-VI), 802 cm− 1
(Uracil-based ring breathing mode), 893 cm− 1 (backbone, C–C skel­
etal), 1133 cm− 1 (d-mannose, γ (C–N)), 1203 cm− 1 (proteins), 1260
cm− 1 (CH2 in-plane deformation (lipids), 1572 cm− 1 (v(C–– C) Retinal),
are on negative side of the loadings. Mean plot SERS spectral features
correspond with these SERS spectral features as show in Fig. 3.
3.2. Partial least square discriminant analysis (PLS-DA)
Partial least square discriminant analysis (PLS-DA) seems to be a
more valuable algorithm that is used for predictive and descriptive
modeling of data into different groups to distinguish the different levels/
stages of SERS spectral datasets. PLS-DA is a method that has been
widely used in several fields, including forensic and medical sciences
[92]. This model was used to investigate the authenticity of a classifi­
cation system that included both healthy and disease classes/groups.
Fig. 5 shows a clear distinction between SERS spectral data sets of dia­
betes (red dots) samples and SERS spectral data from healthy/control
samples (green dots). The number of latent variables chosen are 4 and
the method of validation is Leave one sample (15 SERS spectra) out
Cross-validation method (LOOCV).”. The classification of SERS spectral
data is achieved with an accuracy of 96 %, sensitivity of 94 %, and
specificity of 99 %, indicating that the PLS-DA model is reliable and can
be used to classify diabetic samples and healthy samples.
The classification of diabetic and control/healthy centrifuged serum
samples has been explained by obtaining a ROC curve that illustrates
validity of this model. The value obtained from area under the curve
(AUC) for PLS-DA model for SERS spectral data is 0.85, as shown in
Fig. 6. The maximum value for AUC is 1, which indicates 100 % accu­
racy and validation of the proposed model, while a value of zero in­
dicates that the model is insufficiently valid.
4. Conclusions
The findings of the current study showed that surface enhanced
Raman spectroscopy (SERS) along with chemometric tools such as
principal component analysis (PCA) and Partial least discernment
analysis (PLS-DA) can be used to clearly differentiate between filtrate
portions of the centrifuged serum samples of diabetes patients from
those of healthy ones with high sensitivity and specificity. According to
the research results, the major SERS spectral features as molecular dif­
ferences between diabetes and healthy centrifuged serum samples were
identified at 497 cm− 1, 630 cm− 1, 725 cm− 1, 791 cm− 1,959 cm− 1,1095
cm− 1, 1143 cm− 1, 1447 cm− 1 and 1587 cm− 1.
These SERS markers of diseased may perform an important role in
the early detection of type- II diabetes. The findings of this study show
that surface enhanced Raman spectroscopy of centrifuged serum sample
has the potential to be used as non-invasive complementary tool used for
characterization of type- II diabetes. Surface enhanced Raman spec­
troscopy combined with statistical tools has the potential to provide
rapid screening for diabetic conditions in the community.
Funding Information
This research did not receive any specific grant from funding
agencies in the public, commercial, or not-for-profit sectors.
Declaration of Competing Interest
The authors declare that they have no known competing financial
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Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 293 (2023) 122457
interests or personal relationships that could have appeared to influence
the work reported in this paper.
Data availability
Data will be made available on request.
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