Spectrophotometric Method Development For The Estimation

of Metformin in its Bulk and Tablet Dosage Form

R .SAIKIRAN GOUD

IV year B.PHARMACY CMR COLLEGE OF PHARMACY

CONTENTS Abstract Introduction Review of Literature Drug Profile Need for Study Aim & Objective Plan of Work Materials & Methods Results & Discussion Conclusion Bibliography

ABSTRACTMetformin is a biguanide antihyperglycemic agent used for treating non-insulin dependent diabetes mellitus (NIDDM). . The present work is designed to

develop a new validated, simple, rapid, efficient, reliable and economic UV spectrophotometric method for the estimation of metformin from

pharmaceutical dosage forms. The absorption maxima was measured at 646 nm and is linear over concentration range 8-16µg/ml. The limits of detection and quantification were calculated to be 0.0218 µg/ml& 0.066 µg/ml respectively, percentage drug recovery is 99.2%.

Key words: NIDDM , Metformin, biguanide, antihyperglycemic,.

INTRODUCTIONPharmaceutical AnalysisPharmaceutical analysis is a branch of analytical chemistry that involves a series of process for identification, determination, quantification and purification of a substance, separation of the components of a solution or mixture, or determination of structure of chemical compounds.

There are main two types of chemical analysis.1. Qualitative (identification)2. Quantitative (estimation)

UV VISIBLE SPECTROSCOPYPrinciple : Beer-Lambert law Beer’s Law :intensity of incident radiation decreases exponentially with the

increase in the concentration of the absorbing medium I=I0e-kc

Lambert’s law

Rate of decrease in the intensity of the incident radiation(I) with the thickness ofthe medium (t) is directly proportional to the intensity of the incident light. dI/dt= kI I/I0 =e-kct

A=εctINSTRUMENTATION OF UV

INSTRUMENTATIONRadiation sources (UV)Filter or MonochromatorSample cellsDetectorMETHOD DEVELOPMENT OF UV SPECTROSCOPY1.Selection of Equipment2.Equipment Validation 3.Nature Of Sample4.Selection of solvent5.Selection of wavelength 6.Sample Handling and measurementOPTIMIZATION OF UV-VISIBLE SPECTROSCOPY• Wavelength• Solvent• Concentration• pH• Temperature

Method validation Specificity AccuracyPrecision Detection of LimitQuantitation LimitLinearityRangeRobustness

Review of Literature

Development and Validation of Analytical Method for Determination of Metformin Hydrochloride in Bulk and Tablet Dosage Form by Using UV-Visible Spectrophotometer.

AUTHORS :Bhadresh A. Koli*, Pravin N. Sable.METHOD:UV-VIS METHOD ƛmax :233nm Development of Fixed Time Kinetic Spectrophotometric Method for Selective Determination ofMetformin in Pharmaceutical Formulations.AUTHORS : SHIVAJI R. LABHADE1*, KAILAS R. LABHADE2 and VISHWAS B.GAIKWAD3METHOD :UV-VIS METHOD ƛmax:685nm

Spectrophotometric determination of metformin hydrochloride via oxidative coupling reaction with 1-naphthol in pharmaceutical and environmental water sample.AUTHOR: Farha Khalaf Omar METHOD:UV-VIS METHOD ƛmax:580nm DEVELOPMENT AND VALIDATION OF UV SPECTROSCOPIC METHOD FOR THE DETERMINATION OF METFORMIN HYDROCHLORIDE IN TABLET DOSAGE FORMAUTHORS: Reatul Karim*, Nurunnahar Poly and Rebecca Banoo METHOD :UV-VIS METHOD ƛmax:233nm

SECOND DERIVATIVE SPECTROPHOTOMETRIC METHOD FOR THE ESTIMATION OF

METFORMIN HYDROCHLORIDE IN BULK AND IN TABLET DOSAGE FORMAUTHORS:NARENDER SHARMA*, ANURAG MISHRA, RAVINDER KUMAR et.al METHOD:UV-VIS METHOD ƛmax :233.8nm

Quantitative Determination of Metformin Hydrochloride in Tablet Formulation ContainingCroscarmellose Sodium as Disintegrant by HPLC and UV Spectrophotometry AUTHORS:P Umapathi , J.Ayyappan and S Darlin Quine METHOD: HPLC

Spectrophotometric Method for Estimation of Metformin Hydrochloride AUTHORS: *Mubeen .G, Khalikha Noor and Vimala M NMETHOD : UV-VIS METHOD ƛmax :400nm

Spectrophotometric method for analysis of metformin hydrochlorideAUTHORS: Mubeen .G, Khalikha Noor METHOD: UV-VIS METHOD ƛmax :570nm

Drug Profile

 

Pka:12.33 Official status : Indian pharmacopoeia

Molecular weight: 129.163g/mol

Chemical Formula: C4

H11

N5

CAS number : 657-24-9

Metformin

Solubility studies

soluble

Methanol

Ethanol

Distilled wate

r

insoluble

Acetone

EtherDichlorometh

ane

Chloroform

Mechanism of action

Pharmacokinetices Adverse drug reactions

Contraindications

It decreases hepatic and renal glucose uptake

Absorption: Absorbed over 6 hours ,. Peak action occurs 3 hours after oral administration.Distribution: steady state plasma concentrations of metformin are reached within 24-48 hours.Metabolism: Metformin is not metabolized.Excretion: metformin is excreted unchanged in the urine , T1/2 -1.5 to 3 hrsDuration of action is 8-12 hours

More commonNausea, vomitingAbdominal discomfort,Less commonBlurred visionChest discomfortRareBehavior change similar to being drunkDrowsiness

I. Renal disease or renal dysfunction2. Congestive heart failure requiring pharmacologic treatment.3. Known hypersensitivity to metformin hydrochloride.4. Acute or chronic metabolic acidosis

Need for Study

Metformin is a biguanide antihyperglycemic agent used for treating non-insulindependent diabetes mellitus (NIDDM). It improves glycemic control by decreasinghepatic glucose production, decreasing glucose absorption and increasing insulinmediated glucose uptake.

The present work is designed to develop a new validated, simple, rapid, efficient,reliable and economic UV spectrophotometric method for the estimation ofmetformin from pharmaceutical dosage forms.

Aim & Objective

• To develop a simple, accurate, precise spectrophotometric method.

• To optimize and validate the spectrophotometric method as per ICH guidelines.

Plan of Work

Review of literature Selection of drug Solubility

Selection of reagents Determination of ƛmax

Color development Linearity

Assay

Analytical method validation

Materials & Methods

Solvents

• Distilled water, Methanol, Ethanol, Acetone, Chloroform, Ether, Concentrated Hydrochloric acid , Ferric chloride (1%,2%), Sodium Hydroxide (2%,4%,10%), Sodium Nitrite (10%), ß-Naphthol (1%).

Equipments

• UV spectrophotometer,• Sonicator• Weighing balance,

Preparation of metformin standard solution 100mg drug +100ml methanol=1000µg/ml concentration(primary stock)

8,10,12,14,16µg/ml are prepared 100µg/ml(secondary stock)

2ml from each conc. 2-3 drops of conc.Hcl , 10% sodium nitrite , was withdrawn & 10% sodium hydroxide with 1% of ß-naphthol

make upto final volume

Absorbance was measured maintained at 0°C @ 646nm

Preparation of metformin sample solution

20 tablets weighed & powdered weighed quantity equivalent to 100mg powder + 100ml methanol

100µg/ml(secondary stock) 1000µg/ml conc.(primary stock)

10µg/ml was prepared 2ml was withdrawn

maintained at 0°C 2-3 drops of conc.Hcl , 10% sodium nitrite

& 10% sodium hydroxide with 1% of ß-naphthol

make upto final volume Absorbance was measured @ 646nm

Results & Discussion

Calibration curve

Linearity

7 8 9 10 11 12 13 14 15 16 170

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1

f(x) = 0.0889500000000001 x − 0.5268R² = 0.99886032674631

Series1Linear (Series1)Series3

concentration µg/ml x

Abs

orba

nce

nm

y

Assay of MeforminIn Dosage Form

METHOD VALIDATION Accuracy

To study the accuracy of the proposed methods, recovery studies were carriedout by adding a known amount of drug to the pre analyzed tablet powder andpercentage recoveries are calculated.

Brand of

Drug

Label

Claim

Amount

Added

Amount of

Drug

Estimated

%Recovery

Glyciphage 250 mg 248 mg 244mg 99.2%

Precision

The reproducibility of the proposed method were determined by performing thetablet assay at different time intervals on the same day (intra-day precision) and onthree different days (inter-day precision).the results of intra-day and inter-dayprecisions were expressed in %RSD.

Brand of

Drug

Label

Claim

Amount

Added

Amount of

Drug

Estimated

%Recovery % RSD

Glyciphage 250 mg 248 mg 244mg 99.2% 0.654

INTRA-DAY PRECISION INTER-DAY PRECISION

Amount

µg/ml

Amount

found

%RSD

10 0.351

10 0.356

10 0.354 0.654

10 0.357

10 0.355

10 0.352

Amount

µg/ml

Amount

found

%RSD

10 0.352

10 0.357

10 0.354 0.646

10 0.357

10 0.356

10 0.351

LINEARITY : linear of the range of 8-16 µg/mL.

Limit Of Detection And Limit Of Quantification :LOD & LOQ for metformin were found to be 0.0218 µg/ml & 0.066 µg/ml

respectively .

7 8 9 10 11 12 13 14 15 16 170

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1

f(x) = 0.0889500000000001 x − 0.5268R² = 0.99886032674631

Series1Linear (Series1)Series3

concentration µg/ml x

Abs

orba

nce

nm

y

Conclusion

The absorption maxima was measured at 646 nm and is linear overconcentration range 8-16µg/ml. The limits of detection and quantification werecalculated to be 0.0218 µg/ml & 0.066 µg/ml respectively The method havebeen successfully applied for determination of drug in tablet and resultsobtained are good and percentage drug recovery is 99.2%.

Bibliography

• Review of literature (online)- http://connection.ebscohost.com/c/articles/72102256/• Review of literature (online)- https://www.academia.edu/9691171• Review of literature (online)- www.ajptr.co• Drug profile(online) – www.drugbank.com• Drug profile(online) –www.Rxlist.com• Drug profile(online) – www.pubmed.com• Drug profile(online)-www.chemwikki.com• Drug profile(online)- www.webmd.com• Different types of dosage forms of metformin(online)- www.mayoclinic.org• Different types of dosage forms of metformin(online)-

www.drugs.com/metformin.html• Different types of dosage forms of metformin(online)- reference.medscape.com• Different types of dosage forms of metformin(online)- www.medicinenet.com• pharmacokinetics of metformin(online)-https://www.pharmgkb.org• pharmacodynamics of metformin(online)- onlinelibrary.wiley.com• pharmacodynamics of metformin (online)-bmcpharmacoltoxicol.biomedcentral.com