INTRODUCTION:

Canagliflozin, chemically (2S,3R,4R,5S,6R)-2-[3-[[5-(4-fluorophenyl)thiophen-2-yl]methyl]-4-methylphenyl]-6-(hydroxymethyl)oxane-3,4,5-triol, is an oral selective Sodium-Glucose co-transporter 2 (SGLT2) inhibitor used for the management of type 2 Diabetes Mellitus.

Metformin HCl is N, N-Dimethylimidodicarbonimidic diamide hydrochloride, is biguanide Derivative used for the management of type 2 Diabetes Mellitus.

Figure 1. Structure of Canagliflozin

Figure 2. Structure of Metformin

The literature reveals that there is various UV Spectrophotometric method and HPLC methods has been developed for Canagliflozin and metformin in its combined Dosage form^6-12. The purpose of this work was to develop and validate simple, specific, sensitive, accurate, precise and novel UV Spectroscopic method for the estimation of Canagliflozin and metformin in bulk and it’s Table Dosage Form by Q Absorbance Ratio Method.

Q-Absorbance ratio method:

Q Absorbance ratio method has been used in the present study where absorbance for a drug is measured at to wavelength.

i) Isoabsorptive point of the two drugs and

ii) At the absorption

Maximum of any one of the two drugs. The concentration of each drug was calculated using the equations, Where X and Y are the two selected drugs concentration . Cx and Cy are the concentration of drugs X and Y respectively.Qm represents the ratio of the absorbances.

Qx, Qy represents the ratio of the absorbptivity’s for the drugs X, Y at their respective isoabsorptive points. A1 and ax1 are the absorbance and absorptivity of drug X at isoabsorptive point.

MATERIALS AND METHODS:

Materials:

Canagliflozin API was procured as a gift sample from Alembic Pharmaceutical Ltd., Vadodara. Tablet Formulation of Canagliflozin and metformin (Invokanamet Canagliflozin 50mg and Metformin 500mg) manufactured by Jenseen Pharmaceuticals Ltd. was purchased from local market.

Chemicals and reagents:

All chemicals were of analytical grade

INSTRUMENTATION:

A Shimadzu 1800 visible Spectrophotometer was used for spectral measurements. A high precision analytical balance was used for weighing the reagents. Ultrasonication was used for solubilization of drug.

Procedure:

Procedure for preparation of solution:

· Preparation of Canagliflozin stock solution:

The standard stock solution of Canagliflozin was prepared by dissolving 100mg drug in 100ml methanol which gives 1000µg/ml concentration. From that 100 µg/ml solution of Canagliflozin was prepared by taking 10ml of above solution into 100ml volumetric flask and was diluted up to the mark with Methanol.

· Preparation of calibration curve:

Different aliquots were taken from the 100µg/ml of Canagliflozin stock solution in separate 10ml volumetric flask and finally diluted with Methanol to prepare a series of concentration ranging from 2-10µg/ml. The UV spectrum for Canagliflozin was recorded by placing drug solution in Sample cell and methanol in reference cell at 235(isoabsorptive point) nm and 260nm. Calibration curve was plotted by taking concentration of drug on X-axis and Absorbance in Y-axis

· Preparation of Metformin Hydrochloride stock solution:

The standard stock solution of Metformin Hydrochloride was prepared by dissolving 100mg drug in 100ml methanol which gives 1000µg/ml concentration. from that 100µg/ml solution of Metformin Hydrochloride was prepared by taking 1ml of above solution into 10ml volumetric flasks and were diluted up to the mark with Methanol.

· Preparation of calibration curve:

Different aliquots were taken from the 100µg/ml of Metformin Hydrochloride stock solution in separate 10ml volumetric flask and finally diluted with Methanol to prepare a series of concentration ranging from 2-10 µg/ml. The UV spectrum for Metformin Hydrochloride was recorded by placing drug solution in Sample cell and methanol in reference cell at 235nm and 260nm. Calibration curve was plotted by taking concentration of drug on X-axis and Absorbance in Y-axis.

Figure 3. Overlay Spectra of Canagliflozin and Metformin HCl.

Linearity of Canagliflozin:

Table 2. Linearity parameters of Cana at 235nm and 260nm 235nm and 260nm

Parameter	Value (260nm)	Value (235 nm)
Range	2-10µg/ml	2-10µg/ml
Correlation Coefficient(R²)	0.998	0.998
Slope of regression line	0.017	0.006
Y-intercept	0.012	0.006

Figure 4. Calibration Curve of Canagliflozin

Linearity of Metformin:

Table 3. Linearity parameters of Metformin HCl and 235 nm at 260nm and 235nm

Parameter	Value (260nm)	Value (235nm)
Range	2-10µg/ml	2-10µg/ml
Correlation Coefficient(R²)	0.998	0.997
Slope of regression line	0.003	0.097
Y-intercept	0.018	0.021

Figure 5. Calibration Curve of Metformin HCl

Analysis of Commercial Dosage form^6-9

Twenty tablets were weighed and powdered finely. A quantity of tablet powder equivalent to 10mg of Canagliflozin and metformin was accurately weighed and transferred to 10ml volumetric flask and sonicated for 10 min. The solution was further filtered by whattmann filter paper. The solution was further diluted with methanol to get required concentration. The amount of drug present in the sample solution was determined using the calibration curve of standard drug and applying Q Absorbance Ratio method.

Method Validation^{10 -25}

The developed method was validated for linearity, accuracy, precision, and repeatability, limit of detection and limit of quantification. To check the accuracy of the proposed method recovery studies were carried out by spiking 50%, 100% and 150% of test concentration. The interday and intraday precision of the method was ascertained by taking three concentrations, three replicates within beer’s range and finding out the % RSD.

Applicability of Developed method:

Applicability of the proposed method was tested by analyzing the commercially available Canagliflozin and Metformin tablet. The UV spectrum of is shown in Figure 6

Figure 6. UV Spectrum of Canagliflozin and Metformin HCl Tablet Dosage form

Table 4. Assay of Canagliflozin and Metformin HCl combined dosage form

Brand Name

Label claim

Amount estimated

Average

% RSD

% recovered

Invokamat

Canagliflozin 50mg

Metformin 500mg

49mg

498mg

99.01

99.61

0.102

0.012

99.75±0.92%

Table 5. Optical characteristic and validation parameter of Canagliflozin and metformin HCl

Parameters	Canagliflozin		Metformin HCl
Wavelength (nm)	260nm	235nm	260nm	235nm
Beer’s law limit ( µg /ml)	2-10µg/ml		2-10µg/ml
Régression équation(Y)	y = 0.017x + 0.014	y = 0.011x + 0.006	y = 0.003x + 0.018	y = 0.097x + 0.021
Correlation coefficient (r²)	0.998	0.998	0.998	0.996
LOD (µg/ml)	0.387134	0.196419	0.547723	0.055098
LOQ (µg /ml)	1.290445	0.654729	1.825742	0.183661
Repeatability (RSD), n=6)%	1.536224	1.6508	1.175391	0.997124
Precision (RSD)% Interday (n=3) Intraday (n=3)	1.2993- 1.7069 0.8206- 1.0243	1.1904-1.3546 0.5244-0.71157	0.6794-1.4526 0.5879-1.4526	1.1133-1.3595 1.0791-1.1994
Accuracy	99.75±0.92	99.84±101.36	99.44±100.83	98.36±100.92

RESULT AND DISCUSSION^26-28

A simple, precise and accurate spectrophotometric method has been developed for the estimation of Canagliflozin and metformin in formulation. In this method the standard deviation of the slope and intercept were low. The correlation coefficient exceeded 0.9990. The mean percent label claims estimated for the formulation was 98.86%. This value is very close to 100 indicating the accuracy of the proposed method. The mean percent recovery was within the range of 99.94% to 99.86%, thus it is concluded that the proposed method of analysis is new, simple, accurate and reproducible, successfully applied in the routine analysis of Canagliflozin and metformin in Tablet formulation.

CONCLUSION:

A new, simple and sensitive spectroscopic method was developed for the analysis of Canagliflozin and metformin in bulk and in its tablet dosage form. At 260 nm and 235nm Canagliflozin and metformin was analyzed and it was found that method was accurate, precise and reproducible and successfully applied to the pharmaceutical formulation as per ICH guideline.

AKNOWLEDGEMENT:

The authors are thankful to K.B.Raval College of Pharmacy for providing research facilities.

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Received on 17.05.2021 Modified on 04.09.2021

Research J. Pharm. and Tech 2022; 15(10):4637-4640.

DOI: 10.52711/0974-360X.2022.00778