Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 245 (2021) 118900

Contents lists available at ScienceDirect

Spectrochimica Acta Part A: Molecular and Biomolecular

Spectroscopy
journal homepage: www.elsevier.com/locate/saa

Raman spectroscopy for the qualitative and quantitative analysis of solid

dosage forms of Sitagliptin
Muhammad Abu Bakkar a, Haq Nawaz a,⁎, Muhammad Irfan Majeed a,⁎, Ammara Naseem a, Allah Ditta a,
Nosheen Rashid b, Saqib Ali a, Jawad Bajwa c, Saba Bashir a, Shamsheer Ahmad a, Hamza Hyat a,
Kareem Shah Bukhari a, Franck Bonnier d
a
Department of Chemistry, University of Agriculture, Faisalabad, Pakistan
b
Department of Chemistry, University of Central Punjab, Faisalabad Campus, Faisalabad, Pakistan
c
Department of Chemistry, Government College University, Faisalabad, Pakistan
d
EA 6295 Nano-médicaments and Nano-sondes, Université de Tours, Tours, France

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

Article history: To demonstrate the potential of Raman spectroscopy for the qualitative and quantitative analysis of solid dosage
Received 10 July 2020 pharmacological formulations, different concentrations of Sitagliptin, an Active Pharmaceutical Ingredient (API)
Received in revised form 21 August 2020 currently prescribed as an anti-diabetic drug, are characterised. Increase of the API concentrations induces
Accepted 26 August 2020
changes in the Raman spectral features specifically associated with the drug and excipients. Principal Component
Available online 2 September 2020
Analysis (PCA) and Partial Least Squares Regression (PLSR), were used for the qualitative and quantitative anal-
Keywords:
ysis of the spectral responses. A PLSR model is constructed which enables the prediction of different concentra-
Sitagliptin tions of drug in the complex excipient matrices. During the development of the prediction model, the Root Mean
Solid dosage forms Square Error of Cross Validation (RMSECV) was found to be 0.36 mg and the variability explained by the model,
Raman spectroscopy according to the (R2) value, was found to be 0.99. Moreover, the concentration of the API in the unknown sample
Partial Least Squares Regression was determined. This concentration was predicted to be 64.28/180 mg (w/w), compared to the 65/180 mg (w/
w). These findings demonstrate Raman spectroscopy coupled to PLSR analysis to be a reliable tool to verify
Sitagliptin contents in the pharmaceutical samples based on calibration models prepared under laboratory
conditions.
© 2020 Elsevier B.V. All rights reserved.

1. Introduction drug formulation process. A number of studies report characterisation

Sitagliptin belongs to the gliptin group and is an orally active, potent,
anti-hyperglycemic drug of dipeptidyl peptidase-4 (DPP-4) inhibitors
that was discovered by optimisation of class β-amino-acid [1].
Sitagliptin phosphate monohydrate (C16H15F6N5O·H3PO4·H2O),
marketed under the brand name of Januvia®, was accepted by the
United States Food and Drug Administration (FDA) in October 2006
[2]. It inhibits the enzyme DPP-4, in order to prevent the inactivation
of GIP and GLP-1, thereby increasing insulin secretion from β-cells and
decreasing glucagon release from the pancreas to improve the glycemic
control in blood [3]. This drug is either used alone or in combination
with other hypoglycemic drugs for the treatment of type 2 diabetes.
In the pharmaceutical industry, qualitative and quantitative moni-
toring of the drugs and their precursors is required, especially in the
⁎ Corresponding authors.
E-mail addresses: haqchemist@yahoo.com (H. Nawaz), irfan.majeed@uaf.edu.pk
(M.I. Majeed).
of diabetic drugs like Metformin, pioglitazone, and glipizide by using an-
alytical methods including Reverse-Phase High performance liquid
chromatography (RP-HPLC), used and validated for direct estimation
of Metformin, Pioglitazone, and glipizide in solid forms for different
commercial brands [9]. Although the method is sensitive and accurate,
it requires considerable sample pre-treatment, remains time consum-
ing, and uses large volumes of solvents.
Specifically for the case of Sitagliptin phosphate, a detailed literature
survey has revealed that various spectroscopic as well as chromato-
graphic techniques, including High performance liquid chromatography
(HPLC), High performance thin layer chromatography (HPTLC), capil-
lary zone electrophoresis (CZE), liquid chromatography-mass spectros-
copy (LC-MS), gas chromatography- mass spectroscopy (GC–MS) and
ultraviolet (UV) visible spectroscopy methods have been reported for
determination in tablet formulations, along with different drugs in
pharmaceuticals formulations [4]. Although, these methods are relevant
analytical tools and provide accurate quantification, Process Analysis
Technology (PAT) is currently gaining interest for online, continuous

https://doi.org/10.1016/j.saa.2020.118900
1386-1425/© 2020 Elsevier B.V. All rights reserved.
2 M.A. Bakkar et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 245 (2021) 118900
analysis. Therefore, the development of rapid, reliable analytical
methods not requiring any sample preparation of drug solid dosage
forms is required [5,6].
Vibrational spectroscopy, infrared absorption (IR) and Raman
scattering, are label free, cost effective, and non-destructive techniques
delivering molecular fingerprints of samples. While IR is a well-
established technique in the pharmaceutical industry for characterisa-
tion of samples, the analysis of thick samples is not possible, due to sat-
uration in the signal and loss of spectral information [7]. Attenuated
Total Reflectance (ATR-IR), can overcome this limitation, but the re-
quirement for deposition onto an ATR crystal and the low depth of anal-
ysis (about 2–3 μm) are obstacles to implementation of the technique
for high throughput analysis of solid pharmaceutical forms [8]. As an al-
ternative, Raman spectroscopy is usually performed with confocal mi-
croscopic systems, enabling collection of spectra through focusing
lenses without contact with the samples. Depending on the objective
and wavelength used, micrometric lateral resolution can be reached,
to perform mapping of large sample areas or investigation of small par-
ticles while analysis under the sample surface can also be achieved
[9,10].
Raman spectroscopy has been explored for a wide range of applica-
tions within various fields of agricultural products and food, including
fruits and vegetables, crops, meat and dairy products [11]. Moreover,
it has the important feature of direct measurement on solids and this ca-
pability is studied in the current application [12].
Raman spectroscopy has previously been used for qualitative and
quantitative analysis of drugs including antibiotics as ciprofloxacin,
norfloxacin, levofloxacin ampicillin, and amoxicillin [13] and Cefixime
[14]. The quantification of antibiotics in solid dosage form has also
been carried out by using Fourier transform Raman (FT-Raman) spec-
troscopy [5,15].
Qualitative and quantitative determination of an anti-diabetic drug,
Sitagliptin, in solid dosage forms using Raman spectroscopy has not
been described to date. In the present work, solid dosage forms are pre-
pared in different concentration by mixing of Sitagliptin (API) and ex-
cipients to analyse the spectral changes by using Raman spectroscopy
along with the multivariate data analysis techniques, including PCA
and PLSR. It is demonstrated that the method can be calibrated using a
range of reference samples prepared by mixing the different ingredients
and with known concentrations of Sitagliptin. Finally, the combination
of Raman spectroscopic analysis and PLSR is used to quantify the con-
centration of Sitagliptin in the self-made unknown sample.
2. Material and methods
2.1. Sample preparation
Pure Sitagliptin, used as a reference standard, and excipients includ-
ing starch, lactose, sugar, magnesium powder, talcum powder, and tita-
nium powder were used to prepare a range of concentrations of solid
dosage forms, as described in Table A-1. In the total excipient mixture,
all these excipients are taken in equal ratio, by weighing 1 g of each ex-
cipient and mixing them using a homogeniser to form a homogeneous
mixture of excipients for further analysis. While, calibration samples
and unknown sample (containing 65 mg of Sitagliptin) are prepared
by weighing and mixing ingredients.
2.2. Raman spectral acquisition
Raman spectral acquisition from Sitagliptin samples was performed
in the powdered form. The spectral measurements were performed
using a Peak Seeker Pro-Agiltron Raman spectrometer, (USA) equipped
with a 785 nm laser as source delivering 50 mW laser. About 50 mg of
each sample was deposited on aluminum slides at room temperature
for analysis. Samples were illuminated under a 10× objective for 30 s.
The instrument utilizes an air-cooled charge coupled (CCD) detector
to record Raman scattering. For each Sitagliptin sample, each Raman
spectrum was acquired from different position of the sample by chang-
ing the focus of the laser each time hence acquiring 15 Raman spectra in
total, in the 200 to 1800 cm−1 wavenumber range with a spectral reso-
lution of ~9 cm−1.
2.3. Data preprocessing
Data were pre-processed using MatLab 7.8 under established proto-
cols [16]. The data preprocessing include smoothing, subtraction of the
aluminum substrate signal (Savitzky-Golay smoothing order 3, 13 point
window), baseline correction (rubber band method) and followed by
vector normalization, [17].
2.4. Data analysis
PCA is a mathematical unsupervised procedure involving the trans-
formation of possibly correlated variables into a smaller number of un-
correlated variables, known as principal component (PC), basically to
reduce the dimensionality of the data while maintaining their variability
[18]. The dominant source of variability in the data is explained by the
1st principal component (PC-1) and each succeeding principal compo-
nent accounts for next highest source of remaining variability. The PC
loading can be understood as orthogonal dimensions of variability
which facilitate the separation of different groups of Raman spectral
data along their coefficient as each spectrum of Sitagliptin scores
along these dimensions [19].
PLSR is a chemometric tool which is used to develop a prediction
model for the concentration of the analyte (concentration of the API),
considered dependent variables, against the observed variables
(Raman spectral data), known as independent variables. The indepen-
dent variables (the Raman spectra) are regressed against the dependent
variables according to the regression parameter, leaving residuals (dif-
ferences between measured and predicted variables) which are suitable
to access the quality of the predictive model and the parameters for
model optimization. In this work, a PLSR model root mean squared
error of cross validation (RMSECV) as the fitness value. Leave-K-out
cross validation (LKOCV) was used to select the minimum number of la-
tent variables to retain in order to create a robust model but still avoid
over fitting. A LKOCV approach has been preferred to ensure all spectra
collected from a sample are either included in the calibration or the test
sets, but cannot be represented in both, to avoid bias during the analysis.
Ultimately, 2/3 of samples are randomly selected and identified as cali-
bration while the remaining 1/3 is used as test.
3. Results and discussion
3.1. Spectral characterisation
Sitagliptin is a crystalline, non-hygroscopic white powder [20–22]. It
is a biguanide containing a NH2 group and presenting a chiral center.
The two enantiomeric forms are (R)-enantiomer and (S)-enantiomer,
the former being used for medical purposes (Fig. 1), while the latter is
considered an impurity [23,24].
Fig. 2 shows mean Raman spectra of solid dosage forms for se-
lected concentrations of Sitagliptin. For clarity of presentation, the
Raman spectral features of pure Sitagliptin (S12) in (Fig. 2) are com-
pared to only S1, S4, and S8. The major differences in the Raman
spectral features of Sitagliptin are labeled with vertical lines, and
corresponding peak assignments, taken from literature [25–28], are
summarized in the supplementary material, Table A-2. A strong
Raman peak of Sitagliptin found at 307 cm−1, which represents the
C\\C bending vibrations of aliphatic the carbon chain, and medium
to weak intensity peaks at 409 cm−1 and 511 cm−1, are assigned to
the C\\N\\C deformation. Sitagliptin is a biguanide [29], therefore,
strong Raman peaks at 728 cm−1 , 760 cm −1 and 883 cm−1
 M.A. Bakkar et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 245 (2021) 118900 3

Fig. 1. Three dimensional chemical structure of Sitagliptin.

associated with the symmetric stretching vibrations of C\\F bonds
are observed. The prominent feature at 904 cm−1 is associated
with the C\\O\\C asymmetric vibrations. Another strong Raman
peak at 1019 cm −1 indicates the breathing of aromatic benzene
ring and the weak intensity peaks at 1156 cm−1 and 1450 cm−1 rep-
resents the stretching vibrations of C\\H bond and asymmetric de-
formation of CH3 respectively. In addition, the strong Raman peaks
of the API at 1637 cm−1 and 1673 cm−1respectively correspond to
stretching vibrations of C_O and C_C bonds.
Raman spectra encompass the spectral information from all ingredi-
ents including the API but also excipients found in the solid dosage. Ex-
cipients such as starch, lactose, sugar, magnesium powder, talcum
powder, and titanium powder are Raman active with specific features
observed as seen in Fig. 2. However, comparison of the mean spectrum
of S8 in (Fig. 2), corresponding to 70 mg Sitagliptin, with the mean spec-
trum of the pure API itself highlights that the excipients have limited
contribution to the spectra collected for higher concentrations of the
tested range. It can be observed that the two spectra are practically
identical, suggesting that the API features dominate. In the spectrum
of S1 (pure excipients), the major strong peaks include at 478 cm−1
and 665 cm−1 are observed for titanium stretching and bending vibra-
tions, respectively. An apparent change in intensities can be seen clearly
in the spectrum of S4 sample (30 mg of API) in Fig. 2 at 760 cm−1, as a
result of the increase in relative Sitagliptin concentration and a reduc-
tion of the feature of 478 cm−1, consistent with a decrease in relative
concentration of excipients.
As mentioned above that the excipients have limited contribution to
the spectra at higher concentrations and the API features dominate, In
order to further understand about the linearity of the Raman spectral
data as a function of changes in the ratios of the API and excipients,
PCA is employed. This would help to determine the suitability of the
spectral data for the application of PLSR analysis as it has been thought
to perform PLSR in the linear range where band intensities follow API
concentrations.

Fig. 2. Representative mean Raman spectra of all Sitagliptin data; S12 pure API, S8: sample (70 mg), S4: sample (30 mg), and S1: sample corresponding to pure excipients (blank sample).
4 M.A. Bakkar et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 245 (2021) 118900

Fig. 3. PCA scatter plot of Raman spectra of all the samples of Sitagliptin.

3.2. Principal Component Analysis (PCA)
Fig. 3 depicts the PCA scatter plot of Raman spectra collected from
samples S1-S12. The spectra are clustered separately and clearly differ-
entiated from each other according to concentration. There is a reason-
ably good separation of spectral data as indicated according to the 1st
principal component (PC-1, 81.2%).
In the PCA loadings of whole Sitagliptin data shown in Fig. 4 labeled
as solid lines, negative and positive loadings can be associated with the
API and excipients. These loadings indicate and confirm the spectral fea-
tures of mean Raman spectra Fig. 2 as differences observed. PCA proves
the significance of the spectral features observed as positive loadings at
307 (C\\C bending vibration of aliphatic chain), 511 (C\\N\\C
deformation), 760 (stretching vibrations of C\\F bonds), 904 (C\\O\\C
asymmetric vibrations), 1019 (aromatic benzene ring breathing), 1450
(CH3 asymmetric deformation), 1522 (C_C stretching vibrations of ar-
omatic ring), 1637 (stretching vibrations of C_O) and 1673 cm−1 (C_C
stretching). Whereas, a spectral feature observed in PC-1 as negative
loading at 478 cm−1 which represent the stretching vibration of tita-
nium powder [30,31].
It can be seen that PCA scatter plot shows that the spectral data is not
linear around or above S8 sample (70/185 mg ratio of the API to excip-
ients). This indicates to select the range of the spectral data up to this
range for the PLSR analysis as it is the linear range of the data which is
in the range of interest matching to the dosages commonly used in
commercialised forms (between 0 and 70 mg) in drugs and tablets.

Fig. 4. PCA loadings associated with First Principal Component (PC-1) of Raman spectral data of Sitagliptin from S1 to S11 and pure API.
 M.A. Bakkar et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 245 (2021) 118900 5

Fig. 5. PLSR model for Raman spectral data of Sitagliptin for prediction of optimal number of latent variables.

PCA is found helpful to highlight the response of the Raman spectral
data to the ingredients and the linear range of this data is identified. No-
tably, a contribution from the excipients is required to construct a quan-
titative model as, it is the Raman peak ratios that help PLSR model to
perform and if there is only API features in spectra for high concentra-
tions it wouldn't be as relevant. This can be explained on the basis of
the limit of detection for excipients.
3.3. Partial Least Square Regression (PLSR) analysis
PLSR analysis is used for prediction of API concentrations in different
solid dosage forms based on the Raman spectroscopic response. For
performing PLSR prediction model, normalization of the spectral data
should be avoided in order to conserve the intensity to real concentra-
tion ratios in the data. For this purpose, the data in this study is proc-
essed without applying normalization for the PLSR analysis as this
method was not considered previously. However, for other spectral pre-
sentation (mean spectral analysis) and PCA, normalization of the spec-
tral data is done [32]. The first step is to construct the predictive
models using the standard samples containing known concentrations
of Sitagliptin (Table A-1). Two sets of samples, corresponding to calibra-
tion and test sets, are used for this study, a randomization of data col-
lected from samples S1 to S8 being applied to split the samples into
the 2 sets. Fig. 5 presents the RMSECV for the 20 first latent variables.
It is observed that the value tends to initially decrease before slightly in-
creasing above 10 latent variables. The optimal number of latent vari-
ables is usually selected to be where minimum RMSECV is reached.
However, the risk of over fitting should also be avoided. For this reason,
only 10 latent variables are selected, providing a RMSECV of 0.36. It can
be seen that, above 5 there is no significant improvement to the model.
A PLSR model of Raman spectral data acquired from the different
concentrations of Sitagliptin was constructed to evaluate the ability of
Raman spectral data to predict the level of concentrations of this API.
3.3.1. Root Mean Square Error of Cross Validation
The development of PLSR model with optimal numbers of latent var-
iables (LVs), found to be eight, shows the clear prediction of different
concentrations of drug along with unknown concentration and root

Fig. 6. Performance of PLSR model with (latent variable = 5), (A) calibration and (B) prediction for Raman spectral data for the direct quantification of solid dosage forms of Sitagliptin.
6 M.A. Bakkar et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 245 (2021) 118900
Fig. 7. Regression co-efficient for spectral data of solid dosage forms of different concentrations of Sitagliptin.
mean square error of cross validation of (0.36 mg). Fig. 6 shows the re-
gression plot resulting for 10 LV, indicating the predicted concentration
as a function of the true concentration. For PLSR model, the value of the
goodness of the model (R2) was found to be 0.983.
3.3.2. Prediction of unknown concentrations
The unknown concentration was predicted as 64.28% (w/w) by PLSR
model which is indicated by red bar in Fig. 6. The Raman spectra are not
averaged before employing the PLSR model. The spectra are used indi-
vidually and projected on the predictive model to estimate a concentra-
tion for each hence the prediction results appear as a mean ± standard
deviation.
3.3.3. Regression coefficients
Fig. 7 shows the regression coefficients obtained resulting from the
PLSR analysis. The positive features observed here including 307, 724,
760, 884, 903, 1019, 1518, and 1667 cm−1 are associated with the API
as observed in the Fig. 2 and have been discussed in detail earlier. It
can be seen that the negative features observed here can be associated
with excipients. Notably, the intensities of these negative features in-
cluding 359, 434, 478 and 665 cm−1 are found increasing in the negative
side indicating a decrease in their concentrations in the tested samples/
formulations.
4. Conclusions
Raman spectroscopy can be exploited for quantitative analysis of
solid dosage forms. This work shows that the proposed method is
rapid as it takes only 30 s to record one spectrum, simple, accurate,
and non-invasive to quantify drugs in different formulations. In this
study quantification of Sitagliptin (API) was carried out by forming dos-
age forms including different excipients which gradually decrease by in-
creasing concentration of API. Raman spectroscopy along PCA and PLSR
has been employed to identify the spectral features of the Sitagliptin
drug. PCA is found helpful to identify and select the linear range of the
Raman spectral data for the PLSR analysis, a basic requirement of the
PLSR model to work. The specific Raman spectral features of drug are
observed successfully in mean Raman spectra, and as well as PLSR
model. Raman spectral features of API appear prominently with an in-
crease in its concentrations and decrease of the concentrations of excip-
ients. This method has potential to replace the tedious and time-
consuming methods because of its non-invasiveness and non-
destructive analysis. This method is also helpful to determine the con-
centration kept as unknown for quantification by PLSR.
Funding
This research did not receive any specific grant from funding agen-
cies in the public, commercial, or not-for-profit sectors.
CRediT authorship contribution statement
Muhammad Abu Bakkar: Writing - original draft. Haq Nawaz: Su-
pervision, Project administration, Validation. Muhammad Irfan
Majeed: Conceptualization, Writing - review & editing, Validation.
Ammara Naseem: Software. Allah Ditta: Software. Nosheen Rashid:
Conceptualization, Resources. Saqib Ali: Data curation. Jawad Bajwa:
Software. Saba Bashir: Software. Shamsheer Ahmad: Formal analysis.
Hamza Hyat: Formal analysis. Kareem Shah Bukhari: Formal analysis.
Franck Bonnier: Writing - review & editing.
Declaration of competing interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influ-
ence the work reported in this paper.
Appendix A. Supplementary data
Supplementary data to this article can be found online at https://doi.
org/10.1016/j.saa.2020.118900.
 M.A. Bakkar et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 245 (2021) 118900
References
[1] A. Makdissi, H. Ghanim, M. Vora, K. Green, S. Abuaysheh, A. Chaudhuri, S. Dhindsa, P.
Dandona, Sitagliptin exerts an antinflammatory action, J. Clin. Endocrinol. Metab. 97
(2012) 3333–3341.
[2] N.C. Stofella, A. Veiga, L.J. Oliveira, E.F. Montin, I.F. Andreazza, M.A. Carvalho Filho,
L.S. Bernardi, P.R. Oliveira, F.S. Murakami, Solid-state characterization of different
crystalline forms of Sitagliptin, Materials 12 (2019) 2351.
[3] G. Herman, P. Stein, N. Thornberry, J. Wagner, Dipeptidyl peptidase-4 inhibitors for
the treatment of type 2 diabetes: focus on sitagliptin, Clin. Pharmacol. Ther. 81
(2007) 761–767.
[4] Y. Ozaki, R. Cho, K. Ikegaya, S. Muraishi, K. Kawauchi, Potential of near-infrared Fou-
rier transform Raman spectroscopy in food analysis, Appl. Spectrosc. 46 (1992)
1503–1507.
[5] W. Ji, L. Wang, H. Qian, W. Yao, Quantitative analysis of amoxicillin residues in foods
by surface-enhanced Raman spectroscopy, Spectrosc. Lett. 47 (2014) 451–457.
[6] E.L. Izake, Forensic and homeland security applications of modern portable Raman
spectroscopy, Forensic Sci. Int. 202 (2010) 1–8.
[7] S. Wartewig, R.H. Neubert, Pharmaceutical applications of Mid-IR and Raman spec-
troscopy, Adv. Drug Deliv. Rev. 57 (2005) 1144–1170.
[8] Y. Yamamoto, T. Fukami, T. Koide, T. Suzuki, Y. Hiyama, K. Tomono, Pharmaceutical
evaluation of steroidal ointments by ATR-IR chemical imaging: distribution of active
and inactive pharmaceutical ingredients, Int. J. Pharm. 426 (2012) 54–60.
[9] T. Vankeirsbilck, A. Vercauteren, W. Baeyens, G. Van der Weken, F. Verpoort, G.
Vergote, J.P. Remon, Applications of Raman spectroscopy in pharmaceutical analysis,
TrAC Trends Anal. Chem. 21 (2002) 869–877.
[10] J. Breitenbach, W. Schrof, J. Neumann, Confocal Raman-spectroscopy: analytical ap-
proach to solid dispersions and mapping of drugs, Pharm. Res. 16 (1999)
1109–1113.
[11] Y. Hu, S. Feng, F. Gao, E.C. Li-Chan, E. Grant, X. Lu, Detection of melamine in milk
using molecularly imprinted polymers–surface enhanced Raman spectroscopy,
Food Chem. 176 (2015) 123–129.
[12] S. Sahoo, C.K. Chakraborti, S.C. Mishra, Qualitative analysis of controlled release cip-
rofloxacin/carbopol 934 mucoadhesive suspension, J. Adv. Pharm. Technol. Res. 2
(2011) 195.
[13] V. Desai, O.E. Afieroho, B. Dagunduro, T. Okonkwo, C. Ndu, A simple UV spectropho-
tometric method for the determination of levofloxacin in dosage formulations,
Tropic. J. Pharm. Res. 10 (2011).
[14] J. Bajwa, H. Nawaz, M.I. Majeed, A.I. Hussain, S. Farooq, N. Rashid, M.A. Bakkar, S.
Ahmad, H. Hyat, S. Bashir, Quantitative analysis of solid dosage forms of cefixime
using Raman spectroscopy, Spectrochim. Acta A Mol. Biomol. Spectrosc. 118446
(2020).
[15] S.G. Skoulika, C.A. Georgiou, Rapid quantitative determination of ciprofloxacin in
pharmaceuticals by use of solid-state FT-Raman spectroscopy, Appl. Spectrosc. 55
(2001) 1259–1265.
[16] H. Nawaz, F. Bonnier, P. Knief, O. Howe, F.M. Lyng, A.D. Meade, H.J. Byrne, Evaluation
of the potential of Raman microspectroscopy for prediction of chemotherapeutic re-
sponse to cisplatin in lung adenocarcinoma, Analyst 135 (2010) 3070–3076.
7
[17] T. Mahmood, H. Nawaz, A. Ditta, M. Majeed, M. Hanif, N. Rashid, H. Bhatti, H. Nargis,
M. Saleem, F. Bonnier, Raman spectral analysis for rapid screening of dengue infec-
tion, Spectrochim. Acta A Mol. Biomol. Spectrosc. 200 (2018) 136–142.
[18] J. Saade, M.T.T. Pacheco, M.R. Rodrigues, Identification of hepatitis C in human blood
serum by near-infrared Raman spectroscopy, J. Spectrosc. 22 (2008) 387–395.
[19] H. Nawaz, N. Rashid, M. Saleem, M. Asif Hanif, M. Irfan Majeed, I. Amin, M. Iqbal, M.
Rahman, O. Ibrahim, S. Baig, Prediction of viral loads for diagnosis of Hepatitis C in-
fection in human plasma samples using Raman spectroscopy coupled with partial
least squares regression analysis, J. Raman Spectrosc. 48 (2017) 697–704.
[20] G. Schernthaner, J.L. Gross, J. Rosenstock, M. Guarisco, M. Fu, J. Yee, M. Kawaguchi,
W. Canovatchel, G. Meininger, Canagliflozin compared with sitagliptin for patients
with type 2 diabetes who do not have adequate glycemic control with metformin
plus sulfonylurea: a 52-week randomized trial, Diabetes Care 36 (2013) 2508–2515.
[21] J.L. Atwood, Phosphoric acid salts of sitagliptin, in, Google Patents, 2014.
[22] J.G.D. Chávez, K.M. Vásquez, J.P.S. Peláez, New solid forms of sitagliptin, in, Google
Patents, 2018.
[23] M.K. Mone, P. Jain, S. Kurhade, D.P. Sonune, R.D. Kaduskar, Development and valida-
tion of a liquid chromatographic enantiomer separation method for the estimation
of (S)-enantiomer in sitagliptin, Int. J. Pharm. Sci. Res. 5 (2014) 2382.
[24] T. Zerilli, E.Y. Pyon, Sitagliptin phosphate: a DPP-4 inhibitor for the treatment of type
2 diabetes mellitus, Clin. Ther. 29 (2007) 2614–2634.
[25] S. Liu, M. Rong, H. Zhang, N. Chen, F. Pang, Z. Chen, T. Wang, J. Yan, In vivo Raman
measurement of levofloxacin lactate in blood using a nanoparticle-coated optical
fiber probe, Biomed. Opt. Exp. 7 (2016) 810–815.
[26] L. Yang, M. Gong, X. Jiang, Y. Chen, X. Han, K. Song, X. Sun, Y. Zhang, B. Zhao, SERS
investigation and detection of levofloxacin drug molecules on semiconductor
TiO2: charge transfer contribution, Colloids Surf. A Physicochem. Eng. Asp. 508
(2016) 142–149.
[27] I.J. Hidi, M. Jahn, K. Weber, D. Cialla-May, J. Popp, Droplet based microfluidics: spec-
troscopic characterization of levofloxacin and its SERS detection, Phys. Chem. Chem.
Phys. 17 (2015) 21236–21242.
[28] V. Renganayaki, S. Srinivasan, HF, DFT computations and spectroscopic study of the
vibrational and thermodynamic properties of metformin, Int. J. PharmTech. Res. 3
(2011) 1350–1358.
[29] B. Hernández, F. Pflüger, S.G. Kruglik, R. Cohen, M. Ghomi, Protonation–
deprotonation and structural dynamics of antidiabetic drug metformin, J. Pharm.
Biomed. Anal. 114 (2015) 42–48.
[30] M. De Veij, P. Vandenabeele, T. De Beer, J.P. Remon, L. Moens, Reference database of
Raman spectra of pharmaceutical excipients, J. Raman Spectrosc. 40 (2009)
297–307.
[31] A.O. Surov, N.A. Vasilev, A.V. Churakov, J. Stroh, F. Emmerling, G.L. Perlovich, Solid
forms of ciprofloxacin salicylate: polymorphism, formation pathways and thermo-
dynamic stability, Crystal Growth & Design, 2019.
[32] A.A. Makki, F. Bonnier, R. Respaud, F. Chtara, A. Tfayli, C. Tauber, D. Bertrand, H.J.
Byrne, E. Mohammed, I. Chourpa, Qualitative and quantitative analysis of therapeu-
tic solutions using Raman and infrared spectroscopy, Spectrochim. Acta A Mol.
Biomol. Spectrosc. 218 (2019) 97–108.