1. Development of RP-HPLC method for
Simultaneous estimation of Bendamustine
Hydrochrolide and Omeprazole in Injection
formulation
Under the esteemed guidance of
Sri B. Thangabalan M.pharm, (Ph.d)
Presented by
K. Sampath Kumar
Y11MPH18060
SIMS COLLEGE OF PHARMACY
1 GUNTUR.

2. Aim & Objective
The literature survey indicates that there are no methods
reported for the simultaneous estimation of Bendamustine
Hydrochrolide and Omeprazole by RP – HPLC.
The present work is undertaken with an objective to develop
& validate a simple, sensitive, economical , precise & accurate
chromatographic method by HPLC.

3. Bendamustine Hydrochloride:
Description: white microcrystalline powder
Mol. Formula:C16H21Cl2N3O2
Mol. Weight: 358.262gm/mol
Iupac Name: (4-{5-[bis-(2-chloroethyl) amino]-1-methyl-
1Hbenzimidazol-2-yl} butanoic acid)
Cl-H2C-H2C
N N
Cl-H2C-H2C
.
(CH2)3-COOH HCl
N
CH3

4. Omeprazole:
Mol. Formula:C17H19N3O3
Mol. Weight: 345.42 g/mol
Iupac Name: 5-methoxy-2-(((4-methoxy-3, 5-dimethyl-2-pyridinyl)
methyl) sulphinyl)-1H-benzimidazole

5. UV method:
1. M. Mathrusri Annapurna et.al Derivative Spectrophotometric Methods for the
Determination of Bendamustine Hydrochloride
2. M. Mathrusri Annapurna et.al New analytical methods for the determination of
Bendamustine hydrochloride: An anti-neoplastic drug
3. D.Kumaraswamy Statistical assurance of process validation by analytical method
development and validation for omeprazole capsules and blend.
4. Satish d. Bhuva, mehul m. Patel spectrophotometric simultaneous estimation of
omeprazole and cinitapride in bulk and formulation
5. Nayan M Jagani et.al Development and Validation of Dual Wavelength Method
For Simultaneous Estimation of Omeprazole and Cinitapride in Combined
Capsule Dosage Form.
6. Y.G. Makani et.al development and validation of first order derivative
Spectrophotometric method for simultaneous estimation of Omeprazole and
cinitapride in pharmaceutical dosage form

6. HPLC:
7. Mathrusri Annapurna M. et.al Stability-Indicating Liquid Chromatographic
Method for the Determination of Bendamustine Hydrochloride in
Parenterals
8. E. Sasi Kiran Goud et. al Development and validation of RP-HPLC
method for determination of related substances of bendamustine
hydrochloride in bulk drug
9. Kalakonda Sri Nataraj et.al Development and validation of RP-HPLC
method for the estimation of omeprazole in bulk and capsule dosage forms
10. S. S. Pujeri et.al Development and Validation of LC Method for the Assay
of Omeprazole Enantiomers in Pharmaceutical Formulations

7. Plan of Work
1. Literature Survey
2. Selection of multi-component formulation
3. Procurement of selected drug for study
4. Selection of mobile phase
5. Optimization of chromatographic conditions
A. selection of column
B. selection of column temperature
C. selection of flow rate
D. fixing of injection volume
E. fixing of detection wave length
6. Validation
i. Accuracy iv. Linearity
ii. Precision v. LOD/LOQ
iii. Specificity vi. Ruggedness
vii. Robustness

8. References:
1. http://en.wikipedia.org/wiki/
2. www.chem.ucla.edu~bacherUV-visuv