This document presents the development and validation of an RP-HPLC method for the simultaneous estimation of a muscle relaxant drug and analgesic drug in pure form and pharmaceutical dosage forms. It discusses conducting a literature review on existing methods, determining the physicochemical properties of the drugs, optimizing the analytical method, and validating the developed method as per ICH guidelines. The expected outcomes are an accurate, precise, simple, cost-effective and fast method for simultaneously analyzing the two drugs.

1. PRESENTED BY
MR.VILAS H.KAMBLE
GIUDED BY
DR.MRS SONALI MAHAPARALE
DR.D.Y.PATIL COLLEGE OF PHARMACY AKURDI,PUNE.
DEVLOPMENT AND VALIDATION OF RP-HPLC
METHOD FOR SIMULTANEOUS ESTIMATION OF
MUSCLE RELAXANT AND ANALGESIC DRUG IN
PURE AND PHARMACEUTICAL DOSAGE FORM
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2. CONTENT
 Literature survey
 Principle
 Need of work
 Aim and objective
 Plan of work
 Drug profile
 Experiment
 Outcome of study expected
 References
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3. LITRATURE SURVEY
1 2
JOURNAL
NAME
International journal of pharmacy and
pharmaceutical science
International journal of
pharmacy and
pharmaceutical science
AOUTHER
NAME
Nimila Carolin,Balan P. et. al. Patel Madhavi Bhawani
Singh et. al.
METHOD NAME RP HPLC of Tolperisone and
Paracetamol
UPLC of Tolperisone and
Paracetamol
COLUMN NAME Symmetry C18 250×4.6mm UPLC BEH C18
50mm×2.1mm
MOBILE PHASE Water:ACN (60:40) Water:ACN (30:70)
WAVE
LENGTH
258 nm 254 nm
FLOW RATE 0.7 per min 0.20ml per min
RT Tolperisone=2.25,Paracetamol=3.29 Tolperisone=1.396,Paraceta12/7/2016 3

4. LITRATURE SURVEY
3 4
JOURNAL NAME Pelagia Research Library
International journal of
pharmacy and pharmaceutical
science
AOUTHER NAME
Satyanarayana M. V
Anuradha. V. et al
Thippeswamy M. Somanna P.
at al
METHOD NAME
RP HPLC of Tolperisone and
Paracetamol
RP HPLC of Paracetamol
COLUMN NAME
Hypersil BDS, 150 x
4.6mm C18
C18 colum (250 mm X 4.6 mm
i. d, 5 μm particle size)
MOBILE PHASE
Buffer:ACN (70:30) pH 3 with
TEA
ACN;Phospahte Buffer(15:85)
WAVE
LENGTH
267 nm. 210 nm
FLOW RATE
1.0 Per Min 1.0 ml/min
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5. CHROMATOGRAPHY
Principle-
 When a mixture of components are introduced into a
HPLC column, they travel according to their relative
affinities towards the stationary phase.
 The component which has more affinity towards
stationary phase travel slower.
 The component which has less affinity towards
stationary phase travel faster.
 Since no two component have the same affinity
towards the stationary phase, the component are
separated.
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6. CHROMATOGRAPHY
FIG.SEPARATION OF COMPONENT BY HPLC
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7. NEED OF WORK
 Simultaneous estimation of muscle relaxant and
analgesic drug combination
 Reduce time of analysis
 Increase accuracy of analysis
 Reduce cost of analysis
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8. AIM AND OBJECTIVE
 To develop method for simultaneous estimation of
muscle relaxant and analgesic drug by using RP-
HPLC method.
 To validate the developed method as per ICH
guidelines.
12/7/2016 8

9. PLAN OF WORK
Literature
survey
Selection of
drug
Study of
physicochemica
l properties of
drugs
Optimisation
of analytical
method
Validation of
developed
method
Force
degradation
study
Result and
discussion
Summary and
conclusion
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10. DRUG PROFILE
SR
NO
PARAMETER MASCLE RELAXANT
DRUG
ANALGESIC DRUG
1 Chemical
Structure
2 Chemical name 2-methyl-1-(4-methyl
phenyl)-3-(1-piperidinyl)-
1-propsnone
N-(4-hydroxyphenyl)-acetamine
3 Molecular
formula
C16H23NO.HCl C8H12 NO2
4 Category Muscle relaxant Antipyretic analgesic
5 Melting range Litrature-176-177 Litrature-169-170
6 Dissociation
constant /pKa
value
9.4 9.5
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11. EXPERIMENT
 Solubility studySR
NO
SOLVENT /BUFFER
MUSCLE RELAXANT
DRUG
ANALGESIC DRUG
1 Water Very soluble Sparingly soluble
2 Acetonitrile Very soluble Freely soluble
3 Methanol Very soluble Very soluble
4 Acetone Soluble Freely soluble
5 Chloroform Very soluble Practically insoluble
6 Tetra hydro furan Soluble Soluble
7 Ethyl acetate Very slightly
soluble
Soluble
8 Isopropyle alcohol Freely soluble Freely soluble
Solubility in solvent
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12. EXPERIMENT
 Solubility in buffer
SR
NO
NAME OF BUFFER pH SOLUBILITY OF DRUG
MUSCLE RELAXANT
DRUG
ANALGESIC DRUG
1 0.1M Ammonium acetate buffer 6.50 Soluble Sparingly
soluble
2 0.1M Potassium dihydrogen
phosphate buffer
4.75 Freely soluble Sparingly
soluble
3 0.1M Sodium dihydrogen
phosphate buffer
4.66 Freely soluble Freely soluble
4 0.1M Dipottasium hydrogen
phosphate
9.76 Sparingly soluble Soluble
5 0.1 M Disodium hydrogen
phosphate
9.82 Sparingly soluble Soluble
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13. EXPERIMENT
 Melting point determination
SR NO NAME OF DRUG OBSERVED
MELTING POINT
1 Muscle Relaxant
drug
176.5-177.7
2 Analgesic Drug 170.3-173.5
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14. EXPERIMENT
 λmax determination
By preparing 10 ppm solution of muscle relaxant drug
and analgesic drug individually and scanning in UV at
200nm-400nm
• Preparation of diluents for λmax determination
 Selected diluents ratio-70:30
SR
NO
DILUENT
RATIO
(WATER:ACN)
SOLUBILITY OF
MUSLE RELAXANT
DRUG
SOLUBILITY OF
ANALGESIC DRUG
1 90:10 Freely Soluble Very Slightly Soluble
2 80:20 Freely Soluble Soluble
3 70:30 Freely Soluble Freely Soluble
12/7/2016 14

15. EXPERIMENT
 Muscle relaxant drug
 λmax of Muscle relaxant drug was found to be
260nm
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16. EXPERIMENT
 Analgesic Drug
 λmax of Analgesic Drug was found to be 246nm
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17. EXPERIMENT
• pH determination
By preparing 10% solution of each drug in water
and observed pH on pH meter
SR NO DRUG CATEGORY OBSERVED pH
Of 10% solution
1 Muscle relaxant
drug
4.70
2 Analgesic drug 6.35
3 Water 6.50
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18. EXPERIMENT
Selection of mobile phase
• Selected mobile phase-Water:ACN
• Ratio-50:50
• Chromatographic condition
SR NO PARAMETER DISCRIBTION
1 Mode Isocratic
2 Column Hypersil BDS,
C18,150×4.6,5Micrion
3 Wavelength 254nm
4 Flow Rate 1ml Per Min
3 Column Oven Temperature 30 Degree Celsius
4 Injection Volume 20 Micro Litter
5 Run Time 60 Mins
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19. EXPERIMENT
 Blank
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20. EXPERIMENT
 Muscle relaxant drug standard
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21. EXPERIMENT
 Analgesic drug standard
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22. WORK DONE
 Observation
Sr
No
Drug
Category
Retention
Time
Area Theoretical
Plates
Asymmetry
1 Blank 1.60 1725 2844 1.34
2 Muscle
Relaxant
Drug Std
- - - -
3 Analgesic
Drug Std
0.95 3493394
0
784 1.35
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23. OUTCOME OF STUDY
EXPECTED
 The accurate and precise method for simultaneous
estimation of both the drug
 Simple method
 Cost effective method
 Less time consuming method
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24. REFRENCES
 Carolin N, Balan P. Development and Validation of A
Reverse Phase HPLC Method of Simultaneous
Estimation of Tolperisone Hydrochloride and
Paracetamol in tablet dosage form, International
Journal of Pharmacy and Pharmaceutical
Science,2012,4(5),84-88.
 Patel M. Parmar R. The Simultaneous Estimation of
Paracetamol and Tolperisone Hydrochloride in Tablet
by UV Spectrophotometric Methods, Jouranl of
Pharmaceutical Science and Bioscientific Research,
2012, 2(2),63-67.
 Chatwal G. Anand S. Instrumental Method of Chemical
Analysis, Himalaya Publishing House, 2012, fifth12/7/2016 24

25. REFRENCES
 Thippeswamy M. Somanna P. A New Method
Development and Validation for Estimation of
Paracetamol In Pharmaceutical dosage form by RP-
HPLC, International Journal of Pharmacy and
Pharmaceutical Sciences, 2015, 7(8), 190-194.
 Beckett A . Stenlake J . Practice Pharmaceutical
Chemistry. CBS Publisher, 4th Edition, Part II. New
Delhi,285-288.
 Sharma K. Patel P. First Derivative Spectrophotometric
Method for the Simultaneous Estimation of Tolperisone
and Paracetamol in their Combined Dosage Form,
Journal of Pharmaceutical Sciences and Bioscientific
Research, 2012,2(2),92-96.12/7/2016 25

26. REFRENCES
 Mathew M. Puthusseri S. Validated HPTLC Method
for Simultaneous Estimation of Tolperisone
Hydrochloride And Paracetamol In Tablet Dosage
Form, World Journal of Pharmaceutical Research,
2012,3(10), 513-522.
 Patel M. Singh B. Stability Indicating UPLC Method for
Quantification of Tolperisone HCl and Paracetamol
from muscle relaxant combination tablet, International
Journal of Pharmacy and Pharmaceutical Sciences,
2015,7(8),300-304.
 www.pelagiaresearchlibrary.com
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