Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 238 (2020) 118446

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Spectrochimica Acta Part A: Molecular and Biomolecular

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

Quantitative analysis of solid dosage forms of cefixime using

Raman spectroscopy
Jawad Bajwa a, Haq Nawaz b,⁎, Muhammad Irfan Majeed b,⁎, Abdullah Ijaz Hussain a, Sidra Farooq b,
Nosheen Rashid c, Muhammad Abu Bakkar b, Shamsheer Ahmad b, Hamza Hyat b, Saba Bashir b,
Saqib Ali b, Muhammad Kashif b
a
Department of Chemistry, Government College University, Faisalabad, Pakistan
b
Department of Chemistry, University of Agriculture, Faisalabad, Pakistan
c
Department of Chemistry, University of Central Punjab, Faisalabad Campus, Faisalabad, Pakistan

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

Article history: Quantification of antibiotics is of significant importance because of their use in the prevention and treatment of
Received 26 March 2020 different diseases. Cefixime (CEF) is a cephalosporin antibiotic that is used against bacterial infections. In the
Received in revised form 27 April 2020 present study, Raman spectroscopy has been applied for the identification and quantification of Raman spectral
Accepted 3 May 2020
features of cefixime with different concentrations of Active Pharmaceutical Ingredient (API) and excipients in
Available online 06 May 2020
solid dosage forms. The changes in Raman spectral features of API and excipients in the solid dosage forms of
Keywords:
cefixime were studied and Raman peaks were assigned based on the literature. Multivariate data analysis tech-
Cefixime niques including the Principal Component Analysis (PCA) and Partial Least Squares Regression analysis (PLSR)
Solid dosage forms have been performed for the qualitative and quantitative analysis of solid dosage forms of cefixime. PCA was
Raman spectroscopy found helpful in differentiating all the Raman spectral data associated with the different solid dosage forms of
PCA, PLSR cefixime. The coefficient of determination (R2), mean absolute error (MAE), and mean relative error (MRE) for
the calibration data-set were 0.99, 0.72, and 0.01 respectively and for the validation data-set were 0.99, 3.15,
and 0.02 respectively, that shows the performance of the model. The root mean square error of calibration
(RMSEC) and root mean square error of prediction (RMSEP) were found to be 0.56 mg and 3.13 mg respectively.
© 2020 Elsevier B.V. All rights reserved.

1. Introduction
Cefixime having molecular formula C16H15N5O7S2 is an important
semi-synthetic, stable beta-lactamase, broad spectrum, and orally active
third generation cephalosporin antibiotic [1,2]. Since 1989, it was syn-
thesized and marketed for the treatment of bacterial infections, that
was approved by the U.S. Food and Drug Administration as an antibiotic
in 1997 [1,3]. It is available for oral administration as capsules, tablets
(100 mg, 200 mg, and 400 mg of API) and suspensions (100, 200, 500/
5 ml spoonful) [1,4]. This drug is considered a treatment of different dis-
eases consisting of otitis, gonorrhea, pharyngitis, lower respiratory tract,
and urinary tract infections [5]. It attaches with penicillin-binding pro-
teins (PBP) of the bacterial cell wall and leads to the destruction of bac-
teria by inhibiting transpeptidase enzyme [6,7]. This antibiotic is a kind
of antibacterial compound with a molecular structure consisting of a
⁎ Corresponding authors.
E-mail addresses: haqchemist@yahoo.com (H. Nawaz), irfanmajeed2003@gmail.com
(M.I. Majeed).
cephem nucleus, aminothiazole ring, and acetic acid oxy-imino group,
which are responsible for antibacterial activity [8].
According to World Health Organization (WHO), cefixime is an es-
sential medicine but its higher concentration in pharmaceuticals dosage
form, excessive consumption by human beings may lead to the gastro-
intestinal side effects including the appearance of headaches, dizziness,
rashes, jaundice, allergy reactions, liver damage and digestive disorders
[4,7,9]. Furthermore, cefixime decomposition under storage conditions
might produce impurities; and patients have to face undesirable side ef-
fects due to these impurities [3]. Therefore, an optimum analysis of
cefixime is necessary for its quantitative and qualitative assurance in
pharmaceutical formulations. The concentration of cefixime in raw ma-
terials, pharmaceutical dosage, and biological samples has been deter-
mined by various techniques such as liquid chromatographic, spectro-
photometric, spectroscopic, capillary zone electrophoresis (CZE), fluo-
rometric and anodic voltametric methods [6,7,10–20]. These traditional
methods have been proved as a standard for the determination of
cefixime in dosage forms, raw materials, and biological samples. How-
ever, these methods are annulled because they are expensive, laborious,
poorly stable, time-consuming, and require sample preparation for the

https://doi.org/10.1016/j.saa.2020.118446
1386-1425/© 2020 Elsevier B.V. All rights reserved.
2 J. Bajwa et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 238 (2020) 118446

determination of active constituents in pharmaceutical formulations each sample while 3 spectra were recorded for the unknown sample
[21]. Therefore, the development of a fast, selective, nondestructive, (L11) with an acquisition time of 20 s to ensure the stability and repro-
sensitive, and reliable method is required to quantify the cefixime in ducibility of the quantitative results. There were total 168 Raman spec-
solid dosage forms with or without sample preparation. tra acquired for 12 samples in the spectral range of 200–1800 cm−1
Recently, Raman Spectroscopy has become the most popular tech- because this spectral region accounts for most of the useful spectral fea-
nique in pharmaceuticals because of its great potential for solid dosage tures of cefixime. The Raman spectral data was imported into the
form analysis [22,23] and has been employed for the analysis of poly- MatLab 7.8 after extracting it in the form of binary data (.dat files).
morphic forms of the API, phase transformations, Quality Control oper-
ations, excipients' identification, API's quantitation, and homogeneity 2.2. Data preprocessing
determination in pharmaceutical formulations [24–27]. Raman Spec-
troscopy probes physical and chemical forms non-destructively, non- For the pre-processing of Raman spectral data of all the cefixime
invasively on a level that cannot be completed with traditional tech- samples, MatLab 7.8 (The MathWorks Inc., Natwick, MA, USA) and its
niques [28,29]. established protocols were employed [30]. This pre-processing of spec-
The proposed study is undertaken to develop a rapid, nondestruc- tral data consists of substrate removal, baseline correction, smoothing,
tive, and cost-effective spectroscopic method to analyze the solid dos- and vector normalization (VN). The Savitzky-Golay filtering (S-G) was
age forms of cefixime pharmaceutical formulations. There is no used for smoothing with 1st polynomial order and 11 point window
published literature found about the analysis of the solid dosage forms width [31]. Moreover, the substrate spectra were subtracted from
of cefixime with Raman spectroscopy, therefore this study would lead each spectrum and rubber band correction was carried out to remove
to establish the method, based on Raman spectroscopy, for the qualita- any residual baseline.
tive and quantitative analysis of this drug.
2.3. Data analysis
2. Materials and methods
The Raman spectral data of cefixime was analysed by comparing the
Cefixime API was obtained from CITI pharma, Limited, Lahore mean spectra of each sample including different concentrations of API
Pakistan, and the commercial cefixime capsule was purchased from a as well as excipients. Raman spectral features used for the interpreta-
pharmacy in Faisalabad, Pakistan. The homogeneous mixture of excipi- tion of cefixime results were taken from the literature [31–42] and are
ents including the talcum powder, magnesium powder, lactulose pow- assigned in Table 2. For the qualitative analysis of solid dosage forms
der, starch powder, titanium powder, and cefixime API were used for of cefixime, PCA was performed [43]. The PCA transforms a possibly
the preparation of solid dosage forms of cefixime as explained in large number of correlated variables into a small number of uncorre-
Table 1. Notably, for all these solid dosage forms/samples, these excipi- lated variables to reduce the dimensionality of data while maintaining
ents and API were mixed and homogenized and kept in the powder its variability [44]. The first principal component (PC-1) explains maxi-
form instead of compact tablets. About 50 mg of each sample (powder mum variability in the spectral data and the remaining variability is ex-
form) was put on the aluminium slide/substrate to acquire 5 Raman plained by the next principal components [45]. In order to obtain the
spectra and this process was repeated three times to get 15 spectra in maximum information about covariance of spectral data (x-variables)
total for each sample. During the acquisition of each spectrum, the ob- and known concentrations (y-variables), partial least squares regres-
jective lens was focused at different positions at the sample to consider sion (PLSR), a multivariate data analysis technique, was performed.
the heterogeneity of the sample. The quantitative analysis of the changes in the % age weight of API
was performed by building a regression model based on the changes
2.1. Raman spectral acquisition
Table 2
Raman spectrometer (Peak Seeker Pro-785; Agiltron, USA) was used Raman spectral peaks assignments observed in the spectra of cefixime samples.
for the acquisition of spectra from all samples by placing the sample on Peak cm−1 Assignment Reference
an aluminium slide at room temperature and employing 785 nm laser
308 NH2 tor [36,47]
as an excitation source delivering a laser power of ~40 mW through 398 Ti (anatase) [39,40]
40× objective. The charged coupled detector (CCD) was used in this in- 427 C-C str [31]
strument for recording the signal that helps to minimize the electrical 478 C-C sci, N-C-S sci [34]
noise. For samples L0–L10 (11 sample), 15 spectra were recorded for 515 Ti str (anatase) [40]
571 NH2 wag, C-S-C ip bend [36]
638 Ti bend (anatase) [40]
674 C-S sy str [36]
Table 1 737 (CH)w, sci C-N-C (lact + dihyd) [34]
Details of different concentrations of cefixime API and excipients for the preparation of dif- 785 O-C=O sci, β lactam ring def [33,34,38]
ferent solid dosage samples. 825 C-N-C (tert aliphatic) sy str [37]
889 p(CH2), C\ \C str [34]
Serial Sample name Excipients API Total Net
993 O-C-Me str [38]
no. net percentage
1063 CN str, CH def [34]
weight
1147 C-N-C asy str [37]
1. L0 200 mg 0 mg 200 mg 0% 1207 C-N str, C\\H, N\\H ip bend [36]
2. L1 180 mg 20 mg 200 mg 10% 1248 CH2 wag [34]
3. L2 160 mg 40 mg 200 mg 20% 1303 CH2 wag [38]
4. L3 140 mg 60 mg 200 mg 30% 1339 C-O str [38]
5. L4 120 mg 80 mg 200 mg 40% 1408 NH ip bend, C\ \N str [36]
6. L5 100 mg 100 mg 200 mg 50% 1430 CH3 sci [34]
7. L6 80 mg 120 mg 200 mg 60% 1573 C-C str [34]
8. L7 60 mg 140 mg 200 mg 70% 1614 C=O str [32–34,41]
9. L8 40 mg 160 mg 200 mg 80% 1670 C=O str [32–34,41,48]
10. L9 20 mg 180 mg 200 mg 90% 1785 C=O str [32–35,41]
11. L10 0 mg 200 mg 200 mg 100%
Abbreviations tor, torsion; str, stretching; sci, scissoring; p, rocking; ip, in plane; wag, wag-
12 L11 60 mg 140 mg 200 mg 70%
ging; sy, symmetric; asy, asymmetric; def, deformation; op, out of plane; lact, lactam; dihy,
(unknown)
dihydro-thiazine.
 J. Bajwa et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 238 (2020) 118446
in the Raman spectral peaks of the different concentrations of solid dos-
age forms of cefixime [46]. PLS regression analysis involves calibration
and validation steps. The pertinent information for prediction is ex-
tracted from the calibration step in a few PLS components and then
these newly extracted components are used in the validation step to
check the performance of the model. In the present study, leave one
out cross-validation has been performed with the validation step. The
performance of the built model was evaluated by calculating the root
mean square error (RMSE) by the following equation.
sffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
N 2
∑i¼1 ybi −yi
RMSE ¼
N stretching of tertiary aliphatic amine (C-N-C) bond and a medium in-
in this equation, N is the total number of samples used in the calibration
step, ybi is the predicted or calculated value and yi is the actual value.
3. Results and discussion
3.1. Mean Raman spectra
Fig. 1 shows the mean Raman spectra of all samples containing dif-
ferent concentrations of API and excipients, having a net weight of
50 mg for each sample. The major differences in the Raman spectral fea-
tures of cefixime are labelled as vertical lines and are assigned in Table 2.
The changes in Raman spectral features are associated with different
concentrations of cefixime by increasing concentration of API and de-
creasing concentration of excipients as shown in Table 1. The changes
in peak intensities associated with pure excipients (L0), different ratios
of the API and excipients (L1–L9) and pure API (L10) have appeared
more prominently in all samples in Fig. 1. The Raman spectral features
of high intensities for API appeared at 1785, 1614, 1573, 1430, 1339,
1303, 1248, 1207, 825, 785, 737, 571, 478, 427 and 308 cm−1. Their in-
tensities are found gradually increasing as the concentration of API in-
creases. Literature has revealed that a strong intensity Raman peak of
API at 1614 cm−1 represents C_O stretching vibrations of amide-I
and a medium intensity peak at 1573 cm−1 represents C\\C stretching
vibrations [34]. The cephem nucleus in the cefixime is characterized by
a weak intensity peak at 785 cm−1 which represents combined
Fig. 1. Mean Raman spectra of all cefixime samples (L0; pure excipient; L1–L9; different concentrations of API and excipients; L10; pure API).
3
scissoring and deformation of the carboxyl group and beta-lactam ring
[33]. The medium intensity peak at 1785 cm−1 represents the Raman
spectral feature of the lactam ring indicating stretching vibrations of
C_O [32]. The peak intensity of the Raman feature at 1339 cm−1 can
be associated with C\\O stretching vibrations [38]. The combined
stretching and in-plane bending vibrations of CN, CH, and NH are repre-
sented with a medium intensity peak at 1207 cm−1, and the scissoring
vibrations of the methyl group appear prominently at 1430 cm−1. The
Raman spectral feature of the wagging vibration of CH2 represents
two peaks in the mean Raman spectra, medium intensity peak at
1303 cm−1 and a weak intensity peak at 1248 cm−1 respectively. An-
other weak intensity peak at 825 cm−1 indicates the symmetric
tensity peak at 737 cm−1 represents CH wagging and scissoring vibra-
tions of the lactam ring containing C-N-C bond respectively. The
Raman spectral features of medium intensity peak at 571 cm−1 indicate
the wagging and in-plane bending vibrations of NH2 as well as C\\S
bond of 2-aminothiazole ring respectively [36]. In addition, Raman
spectral features of excipients found at 638 cm−1, 515 cm−1, and
398 cm−1 are representing stretching vibrations of titanium powder
[40]. However, some weak intensity features at 1147, 1063, 993 and
889 cm−1 can be associated with the asymmetric stretching vibrations
of tertiary aliphatic amines, stretching and deformation of CN and CH
bond, stretching vibrations of methoxy group, combined rocking and
stretching vibrations of CH2 and C\\C groups respectively. The Raman
spectral features of high intensity for API and excipients have been suc-
cessfully used for the identification of API in the solid dosage forms.
3.2. Principal component analysis
In order to explore the ability of PCA to differentiate solid dosage
forms of cefixime, PCA was performed by using all the spectral data
that could identify each spectral feature of cefixime and excipients
[49]. Fig. 2 presents the PCA scatter plot of Raman spectral data of eleven
different concentrations of cefixime, in which all samples have been dif-
ferentiated in the form of different clusters indicated by different colors.
In PCA scatter plot, each cluster represents Raman spectral data of one
concentration. Notably, the PCA scatter plot differentiates all the
4 J. Bajwa et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 238 (2020) 118446

Fig. 2. PCA scatter plot of Raman spectra of different solid dosage form samples of cefixime.

clusters of different concentrations of cefixime in sequence from L0-L10
explaining 83.3% variability in the data by PC-1 and 10.24% by PC-2.
Fig. 3 shows pairwise PCA separation of pure API and excipients in
which these samples show clear differentiation as positive and negative
loadings in Fig. 4. In the PCA loading of API and excipients as shown in
Fig. 4 labelled with solid lines, positive and negative loadings are associ-
ated with pure API and excipients. These loadings indicate the clear sep-
aration as observed in the mean Raman spectra Fig. 1. The spectral
features observed in PC-1 as positive loadings at 1782 (C_O str) of
lactam ring, 1673 (C_O str) of the carboxyl group, 1614 (C_O str) of
amide-I, 1574 (C\\C str), 1430 (CH3 sci), 1408 (NH ip bend & C\\N
str), 1338 (C\\O str), 1303 (CH2 wag), 1245 (CH2 wag), 1207 (C\\N
str, CH & NH ip bend), 1152 (C-N-C asy str), 1064 (CN str & CH def),
992 (O-C-Me str), 892 (CH2 rock, CC str), 825 (C-N-C sy str), 786 (O-
C=O & deforming of lactam ring), 736 (CH wag, C-N-C sci), 568 (NH2
wag, C-S-C ip bend), 478 (sci of C\\C and N-C-S), 431 (C\\C str) and
308 (NH2 tor) respectively appeared more prominently and have been
assigned to the API. However, the Raman spectral features of excipients

Fig. 3. PCA scatter plot of Raman spectral data of API versus excipients.
 J. Bajwa et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 238 (2020) 118446 5

Fig. 4. PCA loadings associated with the first Principal Component (PC-1) of Raman spectral data of API versus excipient.

are observed as negative loading in PC-1 at 398, 515 and 638 cm−1 as-
sociated with stretching vibrations of excipients. Hence PCA has been
used to successfully differentiate Raman spectral features of API and
excipients.
3.3. Partial least squares regression analysis
Raman spectroscopy can analyze changes in the spectral features as-
sociated with different samples as a function of the concentration of the
API as demonstrated by PCA. In order to construct a predictive model for
the quantification of different solid dosage forms of API, the PLSR model
was built for the Raman spectral data of nine different concentrations of
cefixime (L1–L9). The PLSR analysis was performed with the SIMPLS al-
gorithm by employing all the 1499 wavenumbers as predictors (x-var-
iables) with API concentrations as a response (y-variables). The
development of the regression model was based on spectral data of all
the samples of cefixime, except pure API (L0) and excipients (L10), as
these samples may cause biasedness of the analysis. All the spectra of
different concentrations of cefixime were compiled into a matrix and
were randomly selected for modelling to avoid the data biasedness
[50]. The total data (135 spectra) was divided into 60% (81 spectra) cal-
ibration dataset and 40% (54 spectra) test dataset. The method of leave
one out cross-validation (LOOCV) with the calibration dataset was used
to determine the optimal model complexity for validation of test dataset

Fig. 5. Performance (calibration vs prediction) of the PLSR model for Raman spectral data for the quantification of solid dosage forms of cefixime.
6 J. Bajwa et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 238 (2020) 118446
Table 3
Details of performance evaluation parameters of the PLSR model.
Evaluation parameters Calibration data-set Test data-set
RMSE 0.56 3.13
R2 0.99 0.99
MAE 0.72 3.15
MRE 0.01 0.02
[51]. The optimal number of latent variables (LVs) or PLS components
are very important for every PLSR model because too many or few LVs
will affect the prediction ability of the PLSR model. The optimal number
of LVs was chosen by performing cross-validation on the calibration
dataset [52]. After cross-validation, the 8 optimal LVs were selected be-
cause they provided the lowest root mean square error. The model was
trained on the calibration dataset and was employed on the validation
dataset that was retained independent to check the performance of
the established model for predicting unknown samples. The PLSR
model with 8 optimal LVs has been shown in Fig. 5 and it was found
helpful in predicting the levels of different concentrations of cefixime
in the solid dosage forms. Different parameters for the performance of
the prediction model are presented in the Table 3. The determination
coefficient of calibration (R2 cal) and prediction (R2 val) were found to
be 0.99 and 0.99 respectively that shows acceptable and excellent per-
formance of the regression model. The low values of the errors for the
calibration data-set (RMSEC = 0.56, MAE = 0.72, MRE = 0.01) and
for the test data-set (RMSEP = 3.13, MAE = 3.15, MRE = 0.02) also in-
dicated the excellent performance and robustness of the model.
To evaluate the performance of our established model, one sample
was kept blind. The PLSR was performed on that unknown sample
(L11) and it was predicted around 143.29 mg as shown in Fig. 5
(B) that is the concentration of API in it while its expected value was
140 mg. This shows the good predictive ability of the established model.
4. Conclusion
This study indicates the high efficiency of Raman spectroscopy com-
bined with multivariate data analysis techniques including PCA and
PLSR to differentiate and quantify the API contents in the solid dosage
forms of cefixime. Based on the multivariate data analysis, commercial
preparations were successfully quantified. Raman spectroscopy accu-
rately characterized the spectral signatures of the antibiotic cefixime
at 1785, 1614, 1573, 1430, 1248, 1209, 825, 785, 737, 515 and
478 cm−1. Based on these features, Raman spectroscopy can be used
for the successful evaluation of the cefixime contents as an active phar-
maceutical ingredient in the pharmaceutical formulations. In this re-
gard, the PLSR model was successfully developed and Root Mean
Square Error of Calibration (RMSEC) and Root Mean Square Error of Pre-
diction (RMSEP) were found to be 0.56 mg and 3.13 mg respectively.
CRediT authorship contribution statement
Jawad Bajwa: Writing - original draft. Haq Nawaz: Conceptualiza-
tion, Writing - review & editing, Validation. Muhammad Irfan Majeed:
Supervision, Project administration, Validation. Abdullah Ijaz Hussain:
Resources. Sidra Farooq: Conceptualization, Resources. Nosheen Ra-
shid: Writing - review & editing. Muhammad Abu Bakkar: Data
curation. Shamsheer Ahmad: Software. Hamza Hyat: Software. Saba
Bashir: Formal analysis. Saqib Ali: Formal analysis. Muhammad Kashif:
Formal analysis.
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.
References
[1] N.K. Tan, D.N. Nhuan, A Novel Synthesis of Cefixime From 7-Amino-3-vinyl
cephalosporanic acid (7-AVCA), 2018.
[2] M. Ramalingam, V. Sethuraman, N. Sundaraganesan, Molecular structure, vibra-
tional spectroscopic, first order hyperpolarizability and HOMO–LUMO studies of
7-amino-8-oxo-3-vinyl-5-thia-1-azabicyclo [4.2. 0] oct-2-ene-2-carboxylic acid,
Spectrochim. Acta A Mol. Biomol. Spectrosc. 78 (2011) 660–669.
[3] Z. Talebpour, H. Pourabdollahi, H. Rafati, A. Abdollahpour, Y. Bashour, H. ABOUL-
ENEIN, Determination of cefixime by a validated stability-indicating HPLC method
and identification of its related substances by LC-MS/MS studies, Sci. Pharm. 81
(2013) 493–504.
[4] S.N.H. Azmi, B. Iqbal, N.S.H. Al-Humaimi, I.R.S. Al-Salmani, N.A.S. Al-Ghafri, N.
Rahman, Quantitative analysis of cefixime via complexation with palladium (II) in
pharmaceutical formulations by spectrophotometry, J. Pharm. Anal. 3 (2013)
248–256.
[5] R.N. Brogden, D.M. Campoli-Richards, Cefixime, Drugs 38 (1989) 524–550.
[6] S. Dey, P.K. Pradhan, U. Upadhayay, S. Shah, K. Goswami, UV spectrophotometric de-
termination of cefixime in bulk and its dosage form, J. Pharm. Res. 5 (2012)
5419–5422.
[7] M. Dehghani, N. Nasirizadeh, M.E. Yazdanshenas, Determination of cefixime using a
novel electrochemical sensor produced with gold nanowires/graphene oxide/
electropolymerized molecular imprinted polymer, Mater. Sci. Eng. C 96 (2019)
654–660.
[8] H.M. Arshad, S. Gauhar, R. Bano, I.N. Muhammad, Development of HPLC-UV method
for analysis of cefixime in raw materials and in capsule, Jordan J. Pharm. Sci. 2
(2009) 53–65.
[9] W.H. Organization, WHO Model List of Essential Medicines: 17th List, March 2011,
2011.
[10] A.A. Kandhro, A.H. Laghari, S.A. Mahesar, R. Saleem, A. Nelofar, S.T. Khan, S. Sherazi,
Application of attenuated total reflectance Fourier transform infrared spectroscopy
for determination of cefixime in oral pharmaceutical formulations, Spectrochim.
Acta A Mol. Biomol. Spectrosc. 115 (2013) 51–56.
[11] A.A. Ramadan, H. Mandil, M. Dahhan, UV–VIS spectrophotometric study for deter-
mination of cefixime in pure form and in pharmaceuticals through complexation
with Cu (II) using acetate-NaOH buffer in water: methanol, Int J Pharm Pharm Sci
5 (2013) 428–433.
[12] M.M. Deshpande, V.S. Kasture, S.A. Gosavi, Application of HPLC and HPTLC for the si-
multaneous determination of cefixime trihydrate and ambroxol hydrochloride in
pharmaceutical dosage form, Eurasian J. Anal. Chem. 5 (2010) 227–238.
[13] A. Solangi, S. Memon, M. Khuhawar, M. Bhanger, Quantitative analysis of eight ceph-
alosporin antibiotics in pharmaceutical products and urine by capillary zone electro-
phoresis, Acta Chromatogr. 19 (2007) 81.
[14] V. Pareek, S. Tambe, S. Bhalerao, Role of different hydrotropic agents in spectropho-
tometric and chromatographic estimation of cefixime, Int. J. Pharm. Bio Sci. 1 (2010)
1–10.
[15] Z. Masoudyfar, S. Elhami, Surface plasmon resonance of gold nanoparticles as a col-
orimetric sensor for indirect detection of cefixime, Spectrochim. Acta A Mol. Biomol.
Spectrosc. 211 (2019) 234–238.
[16] G. Shahnaz, H.M. Arshad, N.S.B. Shyum, R. Bano, M. Shoukat, M.I. Naeem, Pharma-
ceutical evaluation of commercial brands of cefixime 400MG capsules marketed in
Karachi (Pakistan), J. Pharm. Res. 8 (2009) 98–104.
[17] D.T. Gadhiya, H.L. Bagada, Simultaneous equation method for the estimation of
cefixime trihydarte and linezolid in their combined tablet dosage form by UV–
visible spectrophotometry, Int. Bull. Drug Res. 3 (2013) 29–38.
[18] M. Attimarad, B.E. Al-Dhubiab, I.A. Alhaider, A.B. Nair, Simultaneous determination
of moxifloxacin and cefixime by first and ratio first derivative ultraviolet spectro-
photometry, Chem. Central J. 6 (2012) 105.
[19] L. Manna, L. Valvo, Development and validation of a fast reversed-phase ion-pairing
liquid chromatographic method for simultaneous determination of eight cephalo-
sporin antibiotics in pharmaceutical formulations, Chromatographia 60 (2004)
645–649.
[20] Y.B. Wani, D.D. Patil, An experimental design approach for optimization of spectro-
photometric method for estimation of cefixime trihydrate using ninhydrin as
derivatizing reagent in bulk and pharmaceutical formulation, J. Saudi Chem. Soc.
21 (2017) S101–S111.
[21] G.-k. Liu, H. Zheng, J.-l. Lu, Recent progress and perspective of trace antibiotics de-
tection in aquatic environment by surface-enhanced Raman spectroscopy, Trends
Environ. Anal. Chem. 16 (2017) 16–23.
[22] A. Heinz, C.J. Strachan, K.C. Gordon, T. Rades, Analysis of solid-state transformations
of pharmaceutical compounds using vibrational spectroscopy, J. Pharm. Pharmacol.
61 (2009) 971–988.
[23] C.J. Strachan, T. Rades, K.C. Gordon, J. Rantanen, Raman spectroscopy for quantitative
analysis of pharmaceutical solids, J. Pharm. Pharmacol. 59 (2007) 179–192.
[24] J. Cailletaud, C. De Bleye, E. Dumont, Y. Sacré, L. Netchacovitch, Y. Gut, M. Boiret, M.
Ginot, P. Hubert, E. Ziemons, Critical review of surface-enhanced Raman spectros-
copy applications in the pharmaceutical field, J. Pharm. Biomed. Anal. 147 (2018)
458–472.
[25] A. Hédoux, Y. Guinet, M. Descamps, The contribution of Raman spectroscopy to the
analysis of phase transformations in pharmaceutical compounds, Int. J. Pharm. 417
(2011) 17–31.
[26] T. Vankeirsbilck, A. Vercauteren, W. Baeyens, G. Van der Weken, F. Verpoort, G.
Vergote, J.P. Remon, Applications of Raman spectroscopy in pharmaceutical analy-
sis, TrAC Trends Anal. Chem. 21 (2002) 869–877.
[27] A.P. Ayala, Polymorphism in drugs investigated by low wavenumber Raman scatter-
ing, Vib. Spectrosc. 45 (2007) 112–116.
 J. Bajwa et al. / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 238 (2020) 118446
[28] B. D Patel, P. J Mehta, An overview: application of Raman spectroscopy in pharma-
ceutical field, Curr. Pharm. Anal. 6 (2010) 131–141.
[29] T. Lipiainen, S.J. Fraser-Miller, K.C. Gordon, C.J. Strachan, Direct comparison of low-
and mid-frequency Raman spectroscopy for quantitative solid-state pharmaceutical
analysis, J. Pharm. Biomed. Anal. 149 (2018) 343–350.
[30] 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.
[31] 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.
[32] P.K. Bhattacharyya, W.M. Cort, Amoxicillin, Analytical Profiles of Drug Substances,
Elsevier 1978, pp. 19–41.
[33] B.B. Koleva, T.M. Kolev, M. Spiteller, Determination of cephalosporins in solid binary
mixtures by polarized IR-and Raman spectroscopy, J. Pharm. Biomed. Anal. 48
(2008) 201–204.
[34] T. Iliescu, M. Baia, I. Pavel, Raman and SERS investigations of potassium
benzylpenicillin, J. Raman Spectrosc. 37 (2006) 318–325 (An International Journal
for Original Work in all Aspects of Raman Spectroscopy, Including Higher Order Pro-
cesses, and also Brillouin and Rayleigh Scattering).
[35] S.J. Clarke, R.E. Littleford, W.E. Smith, R. Goodacre, Rapid monitoring of antibiotics
using Raman and surface enhanced Raman spectroscopy, Analyst 130 (2005)
1019–1026.
[36] X. Chen, Y. Hu, J. Gao, Tautomers of 2-aminothiazole molecules in aqueous solutions
explored by Raman, SERS and DFT methods, J. Mol. Struct. 1049 (2013) 362–367.
[37] B. Kure, M.D. Morris, Raman spectra of phenothiazine and some pharmaceutical de-
rivatives, Talanta 23 (1976) 398–400.
[38] A. Raj, K. Raju, H.T. Varghese, C.M. Granadeiro, H.I. Nogueira, C.Y. Panicker, IR, Raman
and SERS spectra of 2-(methoxycarbonylmethylsulfanyl)-3,5-dinitrobenzene car-
boxylic acid, J. Braz. Chem. Soc. 20 (2009) 549–559.
[39] U. Balachandran, N. Eror, Raman spectra of titanium dioxide, J. Solid State Chem. 42
(1982) 276–282.
[40] 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 (An International Journal for Original Work in all Aspects of Raman
7
Spectroscopy, Including Higher Order Processes, and also Brillouin and Rayleigh
Scattering).
[41] A.L. Filgueiras, D. Paschoal, H.F. Dos Santos, A.C. Sant’Ana, Adsorption study of anti-
biotics on silver nanoparticle surfaces by surface-enhanced Raman scattering spec-
troscopy, Spectrochim. Acta A Mol. Biomol. Spectrosc. 136 (2015) 979–985.
[42] J.M. Chalmers, H.G. Edwards, M.D. Hargreaves, Infrared and Raman Spectroscopy in
Forensic Science, John Wiley & Sons, 2012.
[43] Y. Roggo, A. Edmond, P. Chalus, M. Ulmschneider, Infrared hyperspectral imaging for
qualitative analysis of pharmaceutical solid forms, Anal. Chim. Acta 535 (2005)
79–87.
[44] 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.
[45] E. Deconinck, A. Van Nederkassel, I. Stanimirova, M. Daszykowski, F. Bensaid, M.
Lees, G. Martin, J. Desmurs, J. Smeyers-Verbeke, Y. Vander Heyden, Isotopic ratios
to detect infringements of patents or proprietary processes of pharmaceuticals:
two case studies, J. Pharm. Biomed. Anal. 48 (2008) 27–41.
[46] J. Gabrielsson, N.O. Lindberg, T. Lundstedt, Multivariate methods in pharmaceutical
applications, J. Chemom. 16 (2002) 141–160.
[47] J.F. Arenas, J. Perez-Peña, M. Gonzalez-Davila, Vibrational spectra and thermody-
namic properties of thiazole, 2-aminothiazole, 2-amino-[2H]-thiazole and 2-
amino-[2H2]-thiazole, Collect. Czechoslov. Chem. Commun. 54 (1989) 28–41.
[48] J.T. Edsall, Raman spectra of amino acids and related compounds I. The ionization of
the carboxyl group, J. Chem. Phys. 4 (1936) 1–8.
[49] J.M. Amigo, Practical issues of hyperspectral imaging analysis of solid dosage forms,
Anal. Bioanal. Chem. 398 (2010) 93–109.
[50] Z. Bouhsain, S. Garrigues, M. de la Guardia, PLS-UV spectrophotometric method for
the simultaneous determination of paracetamol, acetylsalicylic acid and caffeine in
pharmaceutical formulations, Fresenius J. Anal. Chem. 357 (1997) 973–976.
[51] A. Meade, C. Clarke, H. Byrne, F. Lyng, Fourier transform infrared microspectroscopy
and multivariate methods for radiobiological dosimetry, Radiat. Res. 173 (2010)
225–237.
[52] H. Martens, E. Anderssen, A. Flatberg, L.H. Gidskehaug, M. Høy, F. Westad, A. Thybo,
M. Martens, Regression of a data matrix on descriptors of both its rows and of its
columns via latent variables: L-PLSR, Comput. Stat. Data Anal. 48 (2005) 103–123.