Derivative spectrophotometric methods for the assay of Glibenclamide (An anti-diabetic agent)

 

Mukthinuthalapati Mathrusri Annapurna*, Palyam Bhanu, Kandula Venkat Navya

Department of Pharmaceutical Analysis & Quality Assurance, Gandhi Institute of Technology and Management, GITAM Institute of Pharmacy, GITAM (Deemed to be University),

Visakhapatnam, Andhra Pradesh-530045, India.

 

ABSTRACT:

Glibenclamide is an anti-diabetic agent used for the treatment of diabetes mellitus type 2. First derivative spectrophotometric techniques were developed for the assay of Glibenclamide tablet dosage forms in three different reagents and the methods were validated. Glibenclamide has shown linearity over the concentration range 1-80, 1-100 and 1-80 µg/ml respectively in phosphate buffer pH 2.0 (Method A), phosphate buffer pH 8.0 (Method B) and borate buffer pH 9.0 (Method C) respectively. The methods were validated by precision and accuracy studies and are suitable for the determining the assay of Glibenclamide tablets.

 

 

 

INTRODUCTION:

Glibenclamide (Figure 1) also known as Glyburide is an oral hypoglycemic drug and it acts by stimulating the pancreatic beta cells and secretes insulin. Glibenclamide is chemically 5-chloro-N-(4-[N-(cyclo hexyl carbamoyl) sulphamoyl] phenethyl)-2-methoxy-benzamide. Glibenclamide is a sulphonylurea derivative consisting of two moieties such as a sulphonylurea moiety and a benzamide moiety^1-2. In literature various analytical methods were found for the estimation of Glibenclamide in dosage forms. Spectrofluorimetry³, Spectrophotometry^4-9, HPLC^10-14 and LC-MS¹⁵ methods were developed for the assay of Glibenclamide as well as biological fluids^16-18 and nano emulsions^19-20. The present study summarizes three new validated²¹ first order derivative UV spectrophotometric methods for the determination of Glibenclamide in tablet dosage forms.

 

Figure 1: Chemical structure of Glibenclamide

 

MATERIALS AND METHODS:

Glibenclamide is available as tables with brand names Afidex (Label claim: 2.5 mg) (Cadila pharmaceuticals Ltd) and Giclamide (Label claim: 5.0 mg) (Leben laboratories Pvt. Ltd). Model No. UV-1800 Shimadzu double beam UV-VIS spectrophotometer with quartz cells is used for the study and all the solutions were scanned (200-400 nm). 

 

Preparation of solutions

Solutions such as phosphate pH 2.0, phosphate pH 8.0 and borate buffer were prepared as per IP 2010. Stock solution of Glibenclamide was prepared by dissolving 25 mg of Glibenclamide in methanol (1000 µg/ml) in 25 ml volumetric flask and further dilutions were prepared using phosphate buffer pH 2.0 (Method A), phosphate buffer pH 8.0 (Method B) and borate buffer pH 9.0 (Method C) respectively.

 

Method validation

Linearity

A series of Glibenclamide solutions were prepared in phosphate buffer pH 2.0 (Method A), phosphate buffer pH 8.0 (Method B) and borate buffer pH 9.0 (Method C) (10-80 µg/ml for Method A and Method C; 10-100 µg/ml for Method B) from the stock solution and scanned (200-400 nm) against their reagent blanks. The absorption spectra obtained in Method A, Method B and Method C were transformed in to first order derivative spectra with the help of inbuilt software of the instrument. The resultant first derivative spectra has shown maxima and minima and therefore the amplitude was chosen for Method A (238.60-285.80) and Method B (238.80-286.0) for the calculation purpose whereas for Method C minima (238.20) was chosen for the spectral calculations.

 

Precision and accuracy studies

The intra-day and inter-day precision studies were performed at three different concentration levels (40, 60 and 80 µg/ml) and accuracy studies were carried out by standard addition method (50%, 100%, and 150%). The % recovery was calculated for all techniques Method A, B and C.

 

Assay of Glibenclamide tablets

Twenty tablets of Glibenclamide were weighed accurately, powdered and powder equivalent to 25 mg of Glibenclamide was extracted with methanol in a 25 ml volumetric flask and dilutions were made using phosphate buffer pH 2.0, phosphate buffer pH 8.0 and borate buffer pH 9.0 for Method A, Method B and Method C respectively. The assay was carried out using the above analytical procedures of Method A, Method B and Method C and the percentage recovery was calculated.

 

RESULTS AND DISCUSSION:

Three new first order derivative spectrophotometric methods have been developed for the assay of Glibenclamide. A review of published methods in the literature was given in Table 1.

 

Table 1: Review of spectrophotometric methods in the literature

 

The first order derivative absorption spectra obtained in phosphate buffer pH 2.0, phosphate buffer pH 8.0 and borate buffer pH 9.0 for Method A, B and C respectively was shown in Figure 2. The calibration curves for Method A, B and C were shown in Figure 3. Beer-Lambert’s law was obeyed over the concentration range 10-80 µg/ml in Method A and C and 10-100 µg/ml Method B (Table 2). The percentage RSD in precision and accuracy studies was found to be less than 2 indicating that the methods are precise (Table 3) and accurate (Table 4). The characteristics and the assay results of Glibenclamide for all the three methods were given in Table 5.

 

 

 

Method A (Phosphate buffer pH 2.0)

 

 

Method B (Phosphate buffer pH 8.0)

 

 

Method C (Borate buffer pH 9.0)

Figure 2: Overlay first derivative spectra of Glibenclamide

 

 

 

 

 

Method A  (Phosphate buffer pH 2.0)

 

 

 

Method B  (Phosphate buffer pH 8.0)

 

 

 

Method C  (Borate buffer pH 9.0)

Figure 3: Calibration curves of Glibenclamide

 

 

 

 

Table 2: Linearity of Glibenclamide (First order derivative spectroscopy)

 

 

Table 3: Precision study of Glibenclamide (First order derivative spectroscopy)

Interday precision study
Conc. (µg/mL)	A	B	C	Statistical parameters: Mean ± Standard deviation(%RSD)
	Amplitude	Amplitude	Minima	A	B	C
40	0.048	0.038	0.033	0.048  ± 0.00044 (0.93)	0.0318  ± 0.00031 (0.84)	0.033  ± 0.00023 (0.72)
60	0.07	0.055	0.049	0.075 ± 0.00082 (1.08)	0.059 ±0.00082 (1.38)	0.054 ±0.00082 (1.51)
80	0.095	0.072	0.065	0.098 ±0.00082 (0.83)	0.078 ±0.00082 (1.04)	0.071 ±0.00082 (1.14)
Intraday precision study
40	0.048	0.038	0.033	0.054  ± 0.00048 (0.89)	0.056  ± 0.00042 (0.75)	0.047  ± 0.00044 (0.94)
60	0.07	0.055	0.049	0.07±0.000816 (1.14)	0.055±0.000816 (1.48)	0.049±0.000816 (1.6)
80	0.095	0.072	0.065	0.095±0.000816 (0.85)	0.072±0.000816 (1.13)	0.065±0.000816 (1.2)

 

 

Table 3: Accuracy study of Glibenclamide (First order derivative spectroscopy)

                                              

Table 5: Characteristic parameters of Glibenclamide (First order spectroscopy)

 

CONCLUSION:

The three first order derivative spectrophotometric methods for the determination of Glibenclamide are found to be economical, simple, precise and accurate for the routine analysis of Glibenclamide tablets.

 

ACKNOWLEDGEMENT:

The authors are grateful to M/s GITAM (Deemed to be University), Visakhapatnam for providing the research facilities Cadila pharmaceuticals Ltd (India) for supplying gift samples of Glibenclamide. There is no conflict of interest.

 

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Accepted on 02.10.2019           © RJPT All right reserved

Research J. Pharm. and Tech 2019; 12(10):4827-4830.