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Development and Validation of UV-Spectrophotometric Method for Estimation

of Metformin in Bulk and Tablet Dosage Form

Article in Indian Journal of Pharmaceutical Education and Research · December 2017

DOI: 10.5530/ijper.51.4s.109

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 Original Article

Development and Validation of

UV-Spectrophotometric Method for Estimation of
Metformin in Bulk and Tablet Dosage Form
Yuvraj Dilip Dange1, Sandip Mohan Honmane1*, Somnath Devidas Bhinge2, Vijay Rajaram Salunkhe2,
Dhanraj Raghunath Jadge1
1
Department of Pharmaceutics, Annasaheb Dange College of Pharmacy, Ashta, Shivaji University, Kolhapur, Maharashtra, INDIA.
2
Department of Pharmaceutical Chemistry, Rajarambapu College of Pharmacy, Kasegaon, Shivaji University, Kolhapur, Maharashtra,
INDIA.

ABSTRACT
Introduction: Diabetes mellitus, a metabolic disorder characterized by increased blood
sugar level. Metformin hydrochloride is used to treat type I Diabetes mellitus. Metformin
hydrochloride chemically 1, 1-dimethylbiguanide hydrochloride, is white crystalline
powder, hygroscopic and freely soluble in water, Officially UV spectrophotometric
method used for estimation of Metformin Hydrochloride from the bulk and tablets
formulations. Objective: Develop and validate a simple, rapid, accurate, economic and
precise UV/VIS method for Metformin Hydrochloride in bulk and tablets formulation.
Methodology: Choices of a common solvent were essential so various solvent ranges
including methanol, ethanol, acetonitrile and phosphate buffer and various concentrations
ranges of various buffers were analyzed. Conclusion: Among different solvents water
has showed better results, hence water was selected as a solvent for the proposed
method. Metformin Hydrochloride showed maximum absorbance at 234 nm. The
percentage recoveries for Metformin Hydrochloride were found in the range of 99-
101 %. Method was quantitatively evaluated in terms of linearity, accuracy, precision,
ruggedness, robustness and recovery. The method was simple, convenient and suitable
for the determination of Metformin Hydrochloride from bulk and tablet dosage forms.
Key Words: Metformin HCl, UV-Spectrophotometry, Tablet.

INTRODUCTION
Chemically Metformin Hydrochloride (HCl) HCl.3-18 Moreover reported methods were Submission Date: 29-08-2017;
Revision Date: 14-09-2017;
is a (N, N-dimethyl imidodicarbonimidic not much cost-effective in terms of solvent Accepted Date: 04-10-2017.
diamide monohydrochloride as shown in consumption. The present investigation DOI: 10.5530/ijper.51.4s.109
Figure 1. Metformin HCl is used in the was carried out in the view of establishing Correspondence:
treatment of diabetic’s mellitus–II, which Mr. Sandip Mohan Honmane,
a simple, rapid, accurate, economic, precise Annasaheb Dange College
works to decrease the glucose absorption of Pharmacy, Ashta, Shivaji
and robust UV method for estimation
in the small intestine, increase of glucose University, Kolhapur-416301.

transport into cells, decrease the plasma Metformin HCl in bulk and tablet dosage Maharashtra, INDIA.
Phone no: 918600392878
frees fatty acid concentrations and inhibition form using water as the solvent. E-mail: sandiphonmane@
gmail.com
of gluconeogenesis. Activation of AMPK
plays a vital role in these processes.1,2
Literature survey revealed that reports on MATERIALS AND METHODS19,20,21
analytical methods such as UV-Visible, Instrument
HPLC, LC-MS, LC-MS/MS and HPTLC
for the determination of Metformin HCl A Shimadzu UV-1800 240V UV/VIS
from the bulk and dosage form, very few spectrophotometer was used having two
www.ijper.org
analytical methods reported for Metformin matched 1 cm matches quartz cell.
S754 Indian Journal of Pharmaceutical Education and Research | Vol 51 | Issue 4S | Oct-Dec (Suppl), 2017
 Dange et al.: UV-Spectrometric validation of Metformin HCl

Chemical and reagents Method validation

All the reagents and solvents were of analytical grade The method was developed and validated according to
high purity deionized water was obtained from Millipore, the analytical procedure as per the ICH guidelines for
Milli-Q (Bedford, MA, USA) purification system. validation of analytical procedures in order to determine
Metformin HCl was purchased from Pawar supplier linearity, accuracy precision, ruggedness, and robustness
Karad, Maharashtra, India. All other chemicals used for the analyte.19,20,21
were of analytical grade. Linearity
Preparation of Standard Stock Solutions The linearity was evaluated by analyzing the different
100 mg of Metformin HCL was weighed separately and concentration of the standard solution of Metformin
transferred in 100 mL volumetric flasks. The drugs were HCl. The Beer-Lambert’s concentration range was found
dissolved in 50 mL of distilled water by sonication and to be 10-50 µg mL-1 for Metformin HCl respectively.
then the volume was made up to the mark with the same The linearity of the relationship between absorbance’s
solvent to obtain final concentration 1000 µg mL-1 of and concentration was determined by plotting the
the component. calibration curves for Metformin HCl are shown in
Figure 3 and Table 1.
Sample Solution
Accuracy (% Recovery)
Powder of twenty tablets (Marketed tablets of metformin;
Bigomet), containing 500 mg Metformin HCl, was The accuracy study was performed using the standard
weighed. A quantity of powder equivalent to 10 mg addition method.22 The pre-quantified 2 µg mL-1 sample
of Metformin HCl was taken in different 10 mL solution of Metformin HCl were spiked with an extra
volumetric flasks containing about 5 mL distilled water 80, 120, and 100 % of the standard Metformin HCl.
Absorbances were measured at 234 nm and the concen-
for analysis and sonicated for 15 min. After sonication,
tration of drug was determined. These mixtures were
the volume was made up to the mark with the same
analyzed by the developed method. The experiment was
solution to obtain sample stock solution of Metformin
performed in six times. The percentage recovery of the
HCl (1000 mg mL-1). Further, 0.010 mL solution was
samples, % RSD and the percentage were calculated at
quantitatively transferred to a 10 mL volumetric flask to
each concentration level shown in Table 2.
get final concentration 10 mg mL-1 Metformin HCl. The
resulting solution was used for absorbance and results Precision
were recorded. Repeatability measurement was carried out by analyzing
six different solutions containing same concentration
Preparation of Working Standard Solution
20 ppm Metformin HCl and % RSD was calculated.
Suitable aliquots of 1000 mg mL-1 solution were diluted Repeatability of the method was established by ana-
up to the mark with water to get the concentration range lyzing various replicates samples of metformin HCl.
of 10, 20, 30, 40 and 50 mg mL-1 for Metformin HCL. Precision was carried out by performing interday and
The absorbance was measured at 234 nm. intraday variation. In Inter day variation the sample was
analyzed on three consecutive days. In an intraday varia-
Selection of wavelength
tion in the absorbance was measured three times in a
The wavelength for the analysis of Metformin HCl day. Inter and intraday precision was determined using
(20 ppm) was selected from the UV spectrum. The 20 ppm concentration.
standard solution of Metformin HCl was scanned in
Intraday Precision
the range of 200-400 nm and the λmax was found to be
234 nm against water shown in Figure 2. In the intraday variation study was determined for a
solution (20 ppm) and was analyzed three times for
Amount of Metformin in each tablet was calculated the consecutive days (i.e. morning, afternoon, evening).
by using following formula Mean, standard deviation and % RSD was calculated
and shown in Table 3.
Sample Absorbance Standard Dilution
× × Average Weight (1)
Standard Absorbance Sample Dilution Interday Precision
The interday precision was determined for a solution
Amount Present (2) (20 ppm) and was analyzed for the three times on a
% Content = × 100
Label Claim different day. % RSD was calculated shown in Table 4.
Indian Journal of Pharmaceutical Education and Research | Vol 51 | Issue 4S | Oct-Dec (Suppl), 2017 S755
 Dange et al.: UV-Spectrometric validation of Metformin HCl

Ruggedness of correlation coefficient for Metformin HCl was found

The ruggedness of the method was determined by to be 0.9998 shown in Table 1 and Figure 3.
carrying out the analysis by the different analyst in The standard addition method was employed for accuracy
different laboratories using different UV spectropho- measurement. The percentage recoveries for Metformin
tometer and the respective absorbance of 20 mg mL-1
was noted. % RSD was calculated. The Ruggedness data HCl were found in the range of 99-101 %. The values
and analytical performance parameters of Metformin of the recovery (%) and %RSD were shown in Table 2,
HCl were shown in Table 5. which indicates the accuracy of the proposed method.
Robustness
The robustness was carried out to evaluate the influence
of a small but deliberate variation in the spectrometric
condition for determination of Metformin HCl bulk
and tablet dosage form. The Robustness data for varia-
tions in wavelength of detections (±5nm) and the
absorbance and its analytical performance parameters
of Metformin HCl were shown in Table 6.

RESULT AND DISCUSSION

To optimize the UV parameters, several conditions were
tried to achieve a good absorption and peak shape for
Metformin HCl. Several solvents of different composi-
tions were tried to provide sufficient selectivity towards
the drugs. Distilled water components resulted in better
Figure 2: UV Spectrum of Metformin
sensitivity.
The methods discussed in the present work provide a
convenient and accurate way for the analysis of Metformin
HCl from bulk and tablet dosage form by UV Spectro-
photometry method. The 234 nm wavelength was
selected for analysis of Metformin HCl (Figure 2). The
absorbance of Metformin HCl was found to be 1.102 to
5.710 (Table 1). Selected methods linearity was observed
in the concentration range of 10-50 mg mL-1. In this
method, the concentration of the drug was determined
at 234 nm using the respective absorptivity value shown
in Figure 3. Figure 3: Calibration curve of Metformin at 234 nm
A Linear correlation was obtained between absorbance
Vs concentration. Calibration curve for Metformin HCl Table 1: Linearity study for Metformin.
showed linearity in the concentration range 10-50 mg Sr.No. Concentration (ppm) Absorbance
mL−1. The linearity of the calibration curve was validated 1 10 1.102
by the value of correlation coefficients (r2). The value 2 20 2.280
3 30 3.215
4 40 4.541
5 50 5.710
Mean 3.3696
SD 1.8167
RSD 0.5390
%RSD 53.90
Correlation coefficient 0.9998
Figure 1: Structure of Metformin Slope 0.07877

S756 Indian Journal of Pharmaceutical Education and Research | Vol 51 | Issue 4S | Oct-Dec (Suppl), 2017
 Dange et al.: UV-Spectrometric validation of Metformin HCl

Table 2: Statistical Validation for recovery studies of Metformin (n = 6).

Sr. Concentration Absorbance Mean SD RSD %RSD
No. (ppm)
3.810
1 80% 3.790 3.795 0.01322 0.00348 0.34

3.785
4.250
2 120% 4.285 4.269 0.01778 0.00416 0.41

4.273
3.913
3 100% 4.000 3.947 0.04650 0.01170 1.17

3.928

Table 3: Intraday Precision for Metformin (n = 6).

Sr. No Conc.(ppm) Abs-I Abs-II Abs-III
1 20 2.232 2.231 2.199
2 20 2.222 2.260 2.251
3 20 2.251 2.260 2.251
4 20 2.260 2.197 2.257
5 20 2.257 2.251 2.260
6 20 2.255 2.195 2.226
Average 2.2451 2.2253 2.2421
SD 0.0147 0.0336 0.0247
RSD
%RSD 0.0065 0.0151 0.0110
0.65 1.50 1.10
Average %RSD 1.08

Table 4: Interday Precision for Metformin (n = 6).

Sr. No Conc.(ppm) Day I Day II Day III
1 20 2.232 2.270 2.280
2 20 2.222 2.260 2.275
3 20 2.251 2.265 2.277
4 20 2.260 2.281 2.280
5 20 2.251 2.280 2.261
6 20 2.255 2.278 2.271
Average 2.245 2.251 2.272
SD 0.0150 0.0086 0.0066
RSD 0.0066 0.0038 0.029
%RSD 0.66 0.38 0.29

The precision of the method was determined by analyzing of precision. The low values of the % RSD indicate the
the drug formulation by replicate injections and preci- repeatability of the proposed method.23
sion of the system was determined by mixed standard Ruggedness was determined by performing the assay
with the same condition on different days, by different
solutions. % RSD of the analyte was found to be within
analysts, different instrument and different time.24 The
the limit of 2 %, shown in Table 3 and Table 4 thus the test results were found within limit 99–101% shown in
developed method was found to be in the high degree Table 5.
Indian Journal of Pharmaceutical Education and Research | Vol 51 | Issue 4S | Oct-Dec (Suppl), 2017 S757
 Dange et al.: UV-Spectrometric validation of Metformin HCl

Table 5: Statistical Validation for Ruggedness studies of Metformin.

Sr. No Parameter Set I Set II
1 System Shimadzu-1700 Systronics -119
2 Sample Batch No-X Batch No –Y
3 Day Monday Tuesday
4 Date 30/01/2017 31/01/2017
5 Time 11.45am 3.45 pm
6 Lab Analysis Chemistry
7 Analyst Dange Y. D. Honmane S. M.
8 Sample 20 ppm 20 ppm
9 Absorbance 2.286 2.280
10 Assay 99.65% 99.85%

Table 6: Statistical validation for Robustness studies of metformin.

Sr.No Set. No Wavelength (nm) Conc. Absorbance Average STDEV RSD %RSD % Assay
( µg/ml) Abs
1 239 20 1.927
2 l 239 20 1.931 1.929 0.002 0.0010 1.03 97.98

3 239 20 1.929
4 234 20 2.199
5 ll 234 20 2.185 1.888 0.019 0.0100 1.00 98.10

6 234 20 2.170
7 2029 20 2.150
8 lll 229 20 2.157 2.154 0.014 0.0069 0.60 98.02

9 229 20 2.154

Robustness was determined by carrying out the assay ABBREVIATIONS USED

during change wavelength. The % RSD was found to HCl: Hydrochloride; UV: Ultraviolet; RSD: Relative
be not more than 2 % which was within the limit shown standard deviation; HPLC: High-performance liquid
in Table 6. chromatography; HPTLC: High-performance Thin
layer liquid chromatography; LC-MS; liquid chroma-
CONCLUSION tography-mass spectroscopy; AMPK: AMP-activated
protein kinase.
The proposed method quantitatively evaluated in terms
of linearity, accuracy, precision, ruggedness, robustness
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PICTORIAL ABSTRACT SUMMARY

• UV/VIS method has been developed and validated
for Metformin Hydrochloride. The Linearity was
observed in the concentration range of 10-50 mg
mL-1 for Metformin Hydrochloride. The correlation
coefficients were found to be 0.9998. The
percentage recoveries for Metformin Hydrochloride
were found in the range of 99-101 %. Method
was quantitatively evaluated in terms of linearity,
accuracy, precision, ruggedness, robustness and
recovery. The method was simple, convenient
and suitable for the determination of Metformin
Hydrochloride from bulk and tablet dosage forms.

About Authors

Mr. Sandip M. Honmane: M. Pharm. Asst. Professor at Annasaheb Dange College of B. Pharmacy,
Ashta, Sangli (MS). He had total 5 years of experience amongs that 3 years industrial experience
in Multinational pharmaceutical companies. He has > 7 International and national Publications and
one book on his credits.

Mr. Yuvraj D Dange: M. Pharm. Asst. Professor at Annasaheb Dange College of Pharmacy, Ashta,
Sangli (MS). He has > 5 International and national Publications.

Dr. Somnath D. Bhinge: M. Pharm Ph.D. Professor & HOD at Rajarambapu college of Pharmacy
Kasegaon, Sangli (MS). He had 9 years in teaching experience. He is Approved PG Teacher. He
has > 20 International and national Publications. He is a Life member of various professional
associations like IPA, APTI & IPS.

Indian Journal of Pharmaceutical Education and Research | Vol 51 | Issue 4S | Oct-Dec (Suppl), 2017 S759
 Dange et al.: UV-Spectrometric validation of Metformin HCl

Dr. Vijay R. Salunkhe: M. Pharm. Ph.D. Professor & HOD at Rajarambapu college of Pharmacy
Kasegaon, Sangli (MS). He had 22 years in teaching experience. He is Approved PG Teacher and
Ph.D Guide. He has > 32 International and national Publications. He is a Life member of various
professional associations like IPA, APTI & IPS.

Dr. Dhanraj R. Jadge: M. Pharm. Ph.D. Professor and Principal at Annasaheb Dange College of
B. Pharmacy, Ashta, Sangli (MS). He had 24 years in teaching experience. He is Approved PG
Teacher and Ph.D Guide. He has > 20 International and national Publications. He is a Life member
of various professional associations like IPA, APTI & IPS.

Cite this article: Dange YD, Honmane SM, Bhinge SD, Salunkhe VR, Jadge DR. Development and Validation
of UV-Spectrophotometric Method for Estimation of Metformin in Bulk and Tablet Dosage Form. Indian J of
Pharmaceutical Education and Research. 2017;51(4S):S754-S60.

S760 Indian Journal of Pharmaceutical Education and Research | Vol 51 | Issue 4S | Oct-Dec (Suppl), 2017

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