6.

EXPERIMENTAL WORK
 6. EXPERIMENTAL WORK

6.1 Instruments and Materials:

6.1.1 Apparatus and Instruments:

Table 5.1Instruments
Instrument Manufacturer

UV Visible Spectrophotometer Shimadzu 1800

HPLC Shimadzu LC10AD

FTIR Shimadzu
Ultrasonicator Thermo

Analytical weight balance Thermo

pH meter Remi

Volumetric flask (graduated) 10, 25, 50, 100 ml

Pipettes 1, 2, 5,10 ml
Beakers 250, 500, 1000 ml
Measuring cylinders 50, 100, 1000 ml

6.1.2 Materials and Reagents:

Table 5.2 Chemicals

S.NO. NAME MANUFACTURER GRADE

1. Selumetinib Astra Zeneca

-
Potassium dihydrogen
2. phosphate Merck GR

3. NaOH Merck GR

4. HCL Merck GR

5. Ether Merck HPLC

6. Methanol Merck HPLC

7. Acetonitrile Merck HPLC

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8. Water Merck HPLC

9. 0.45µmNylonfilter Thermo P0023

10. 0.45µmPVDFfilter Thermo SA0078

6.2 Identification of APIS

6.2.1 Melting Point Determination of Selumetinib:
Melting point of Selumetinib has been determined by using micro controller based
melting point apparatus. The melting points of the drugs were also taken by open
capillary method.

Table 5.3Melting Point of Drugs

Melting point (˚C)

Sr.No. APIs
Reported Measured
1 Selumetinib 219 °C 218- 219 ˚C

6.2.2 Identification by FTIR Spectroscopy:

6.2.2.1 IR Spectra of Selumetinib:

Fig No. 6.1 IR Spectra of sample Selumetinib

Table 6.4 IR Spectral interpretation of Selumetinib

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Sr. No. Functional Group Frequency (cm-1)

1 C-H (stretch) 2915

2 H-O (stretch) 1232

3 N-H (stretch) 3156

4 C=O 1632
5 C-C 1509
6 C=C 3341
7 F 1117
8 Cl 1091
9 Br 662

6.3 Solubility Study

The solubility of Selumetinib practically determined separately by taking 100 mg of both

the drugs in 100 ml volumetric flasks, adding required quantity of solvent at room
temperature and shaken for few minutes. Solubility data for each study was observed
and recorded.

Table 6.6 Solubility table as per IP 2018 specification

6.3.1 Solubility data of Selumetinib:

Table 5.7 Solubility Data
Solvent Selumetinib
Water Soluble
0.1 N HCl sparingly soluble
0.1 N NaOH Slightly soluble
Methanol Slightly soluble
Eather Soluble

6.4. Development and Optimization of RP-HPLC method

6.4.1 Selection of Wavelength:
42µg/mL solution of Selumetinib was prepared using methanol as solvent. The above
mentioned solutions were scanned individually from 190 to 400 nm in UV-Visible

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spectrophotometer. The optimal response for the overlain spectrum of Selumetinib

was obtained at 243 nm. Spectrums are shown in Fig

Fig. 6.4 (A) UVspectrum of Selumetinib for detection of wavelength at 243nnm

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