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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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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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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)

WAVELENGTH

258 nm 254 nm

FLOW RATE 0.7 per min 0.20ml per min RT Tolperisone=2.25,Paracetamol=3.

29Tolperisone=1.396,Parac

etamol=2.625

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LITRATURE SURVEY 3 4

JOURNAL NAME

Pelagia Research LibraryInternational 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 C18C18 colum (250 mm X 4.6 mm i. d, 5 μm particle size)

MOBILE PHASE Buffer:ACN (70:30) pH 3

with TEAACN;Phospahte Buffer(15:85)

WAVELENGTH

267 nm. 210 nm

FLOW RATE1.0 Per Min 1.0 ml/min

RT Tolperisone=4.6,Paracetamol=2.3

5.7 minutes

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CHROMATOGRAPHYPrinciple- 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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CHROMATOGRAPHY

FIG.SEPARATION OF COMPONENT BY HPLC

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NEED OF WORK Simultaneous estimation of muscle relaxant

and analgesic drug combinationReduce time of analysis Increase accuracy of analysis Reduce cost of analysis

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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.

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PLAN OF WORK Literature survey

Selection of drug Study of

physicochemical

properties of

drugs

Optimisation of

analytical method Validatio

n of developed method

Force degradation study

Result and

discussion

Summary and

conclusion

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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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EXPERIMENT

Solubility study SR NO

SOLVENT /BUFFER SOLUBILITY

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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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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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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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

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EXPERIMENT Muscle relaxant drug

λmax of Muscle relaxant drug was found to be 260nm

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EXPERIMENT Analgesic Drug

λmax of Analgesic Drug was found to be 246nm

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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 6.353 Water 6.50

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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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EXPERIMENT Blank

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EXPERIMENT Muscle relaxant drug standard

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EXPERIMENT Analgesic drug standard

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WORK DONE Observation

Sr No

Drug Category

Retention Time

Area Theoretical Plates

Asymmetry

1 Blank 1.60 1725 2844 1.342 Muscle

Relaxant Drug Std

- - - -

3 Analgesic Drug Std

0.95 34933940

784 1.35

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OUTCOME OF STUDY EXPECTED

The accurate and precise method for simultaneous estimation of both the drug

Simple methodCost effective method Less time consuming method

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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, fifth edition, 2.624-2.639

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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.

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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.

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