This document provides summaries of 5 research articles and 1 literature survey: 1. The first article compares 4 new spectrophotometric methods for the simultaneous determination of tamsulosin and solifenacin that have spectral overlap. 2. The second validates a new spectrofluorometric method for determining two drugs used to treat diabetes without prior separation. 3. The third characterizes impurities in the drug Enzalutamide, develops an HPLC method for quantification, and studies an oxidation impurity. 4. The fourth validates a sensitive LC-MS/MS method for measuring a novel anti-cancer drug in mice plasma and applies it to a pharmacokinetic study.

1. LITREATURE SURVEY
1

2. 2
1. Smart UV spectrophotometric methods based on simple mathematical filtration and
classical methods for the simultaneous determination of tamsulosin and solifenacin: A
comparative study of efficacy and spectral resolution
2. Validated spectrofluorometric determination of hypoglycemic combination, in pure form
and pharmaceutical formulation using 9,10-phenanthraquinone reagent
3. Structural elucidation of the impurities in Enzalutamide bulk drug and the
development, validation of corresponding HPLC method
4. Validation of an LC-ESI-MS/MS method for the determination of apalutamide, a novel
non-steroidal anti-androgen in mice plasma and its application to a pharmacokinetic study in
mice
5. Apalutamide-induced severe interstitial lung disease: A report of two cases from Japan

3. 3
Smart UV spectrophotometric methods based on simple
mathematical filtration and classical methods for the
simultaneous determination of tamsulosin and
solifenacin: A comparative study of efficacy and
spectral resolution
ABSTRACT
Treatment protocols combining tamsulosin and solifenacin proved better management of the complicated
urinary tract symptoms. The pharmaceutical preparations of tamsulosin and solifenacin suffered from the high
difference in their ratio, 0.4 mg tamsulosin and 6 mg solifenacin, and strong spectral overlap. Here, we
developed four simple, accurate and selective spectrophotometric methods based on simple mathematical
manipulations. These methods require the simplest mathematical filtration using short steps performed using
built-in functions of the spectrophotometer operating software utilizing zero-order or derivative spectra. These
methods are namely absorption correction method (ACM), induced dual-wavelength (IDW), absorptivity
factor method (AFM) and first derivative method (D1). The linear ranges were 15-70 μg/ml and 100-1200
μg/ml for TAM and SFN, respectively. The limits of quantitation were in the range of 3.8-4.05 μg/ml and
23.34-59.05 μg/ml, while the limits of detection were in the range of 1.25-1.34 μg/ml and 7.7-24.6 μg/ml for
TAM and SFN, respectively. All validation parameters investigated as per ICH guidelines. A statistical
comparison executed for the proposed methods with each other and with the reported methods showed no
significant difference between the proposed and the reported methods.

4. 4
Validated spectrofluorometric determination of hypoglycemic
combination, in pure form and pharmaceutical formulation using
9,10-phenanthraquinone reagent
ABSTRACT
The aim of this work is to develop and validate a simple, rapid, highly sensitive and selective spectrofluorometric
method for determination of Metformin HCl (MFH) and Glibenclamide (GLB) in their binary mixture without
prior separation. The proposed method based on measuring the native fluorescence intensity of GLB at λ emission=
348 nm, after excitation at λ excitation=226 nm and measuring the fluorescence intensity of the fluorescent product
produced from the derivatization of MFH using 9,10-phenanthraquinone in alkaline media at λ emission =
416 nm after excitation at λ excitation=240 nm. The proposed spectrofluorometric method allowed sensitive
determination
of the studied drugs with a limit of quantization of 0.04 and 0.01 μgmL−1 for MFH and GLB, respectively,
providing greater sensitivity than the reported one. Validation of the proposed method was carried out
according to ICH guidelines with respect to linearity, accuracy, precision, and selectivity. The developed method
was successfully applied for the determination of MFH and GLB in laboratory prepared mixtures and pharmaceutical
formulation and the results obtained were statistically compared to those of the reported HPLC method with
no significant difference between them.

5. 5
STRUCTURAL ELUCIDATION OF THE IMPURITIES IN
ENZALUTAMIDE BULK DRUG AND THE DEVELOPMENT,
VALIDATION OF CORRESPONDING HPLC METHOD
ABSTRACT
As the first approved androgen receptor(AR) signalling inhibitor, Enzalutamide was approved by the US Food and
Drug Administration as an anticancer drug used to treat castration-resistant prostate cancer in 2012. In this manuscript,
six potential impurities of Enzalutamide including process impurities and degradation products were studied. The
structures of six impurities obtained by synthesis were characterized and confirmed by IR, NMR and MS techniques. In
addition, an efficient chromatographic method to separate and quantify these impurities was developed, which
achieved on Inertsil ODS-3 column (250mm×4.6mm,5μm) in gradient mode with a mixture of acetonitrile and the
ammonium acetate buffer (10mM, pH adjusted to 4.0 with glacial acetic acid). The method was validated with respect to
specificity, precision, accuracy, and sensitivity and satisfactory result was achieved. The method was demonstrated to
be applicable in routine quality control and stability evaluation of Enzalutamide.
Highlights
1.Identification and characterization of process-related impurities and forced
degration impurities.
2.The pathway for the formation of the potential impurities have
3.A RP-HPLC method is developed for quantitatively determining impurities.
4.An important by-product and degradation product (oxidation
impurity) was studied in enzalutamide.

6. 6
Validation of an LC-ESI-MS/MS method for the
determination of apalutamide, a novel non-steroidal
anti-androgen in mice plasma and its application to a
pharmacokinetic study in mice
ABSTRACT
A sensitive, specific, selective and rapid LC-ESI-MS/MS method has been developed and validated for the
quantification of apalutamide in mice plasma using apalutamide-d3 as an internal standard (I.S.). Sample preparation
was accomplished through a simple protein precipitation process. Chromatography of apalutamide and the I.S. was
achieved on an
Atlantis dC18 column using an isocratic mobile phase comprising 0.2% formic acid in water and acetonitrile (20:80,
v/v) delivered at a flow rate of 0.8 mL/min. LC-MS/MS was operated under the multiple reaction-monitoring mode
(MRM) using the electrospray ionization technique in positive ion mode and the transitions of m/z 478 , 450 and m/z
481 , 453 were used to measure the derivative of apalutamide and the I.S, respectively.
The total chromatographic run time was 2.5 min and the elution of apalutamide and I.S. occurred at 1.10 and 1.09
min, respectively. Method validation was performed as per regulatory guidelines and the results met the acceptance
criteria. Linearity was established in the concentration range of 1.02-2030 ng/mL (r>0.995) for apalutamide. The intra-
and inter-day accuracy and precision for apalutamide in mice plasma were in the range of2.11-8.44 and 2.51-6.09%,
respectively. Apalutamide was found to be stable under various stability conditions. This novel method has been
applied to a pharmacokinetic study in mice.