Pharmaceutica Analytica Acta

Quantitative Estimation of Dapagliflozin in Blood Plasma by Using UV Spectroscopy

Bhagwat J Bodade, Dhiraj A Kanade and Sandip S Chaudhari^* Department of Pharmaceutical Chemistry, KYDSCT’s College of Pharmacy, Sakegaon Bhusawal, Maharashtra, India
^*Correspondence: Sandip S Chaudhari, Department of Pharmaceutical Chemistry, KYDSCT’s College of Pharmacy, Sakegaon Bhusawal, Maharashtra, India, Tel: 9922246400, Email:

Received: 05-Apr-2019 Published: 29-Apr-2019, DOI: 10.35248/2153-2435.19.10.608

Introduction

Dapagliflozin is a drug of the Gliflozin class, used for the treat type 2 diabetes. Dapagliflozin tablets are approved as a once-daily oral medication in adult patients with Type 2 diabetes mellitus.

Dapagliflozin inhibits subtype 2 of the Sodium-Glucose Transport Proteins (SGLT2) which are responsible for at least 90% of the glucose reabsorption in the kidney. Blocking this transporter mechanism causes blood glucose to be eliminated through the urine. It was developed by Bristol-Myers Squibb in partnership with AstraZeneca Company [1].

Dapagliflozin is an inhibitor of the Sodium Glucose Co- Transporter-2 (SGLT-2), which is found almost exclusively in the proximal tubules of nephronic components in the kidneys (Figure 1).

Figure 1. Chemical structure of Dapagliflozin.

Materials and Methods

Chemicals and reagents

Dapagliflozin Tablet (Forxiga) 10 mg was obtained from local pharmacy and gives to patient and after 6 hours collect blood sample for analysis. All chemicals and reagents of analytical grade and were purchased from Merck Chemicals, Mumbai, India

Instrumentation

Samples were analysed on Shimadzu 1800 UV-visible spectrophotometer, equipped with UV spectroscopic detector, with 1 cm quartz cell and slit width 1.0 nm. The solvent was prepared freshly filtered through 0.45 μm membrane filter (Millipore, USA) and Sonicated for 30 min [2].

Preparation of standard stock solutions

A) Collection of blood plasma: Blood sample was transferred into purple top EDTA tubes and centrifuged (2000 rpm) at 4°C for 20 minutes. After centrifugation using clean pipette technique place 1.0 ml of plasma into 1.5 ml eppendorf tube labelled with tracking number and plasma and freeze immediately at –80º freezer [3]. After collecting blood plasma it can considered as working stock solution.

B) Preparation of working standard solution: By using the working standard solution it is further diluted by normal patient’s blood plasma and prepares concentration up to 20, 40, 60, 80 and 100 μg/m for linearity study.

Determination of λ_max of tablet

The sample of blood plasma containing dapagliflozin drug solution was scanned separately between 200 nm to 400 nm.

Study of beer-lambert’s law [linearity study]

A) Linearity Study for DAPA: An accurately measured aliquot portion of working standard solution blood plasma in 10 ml volumetric flask [4]. The volume was made up to the mark using blood plasma to obtain concentrations (20-100 μg/ml). Absorbance of this solution was measured at 224 nm, Calibration curve was plotted, absorbance vs. concentration (Table 1 and Figures 2-7).

Table 1: Linearity study of DAPA at 224 nm.

Figure 2. Calibration curve of Dapagliflozin.

Figure 3. Calibration curve of DAPA (20 μg/ml).

Figure 4. Calibration curve of DAPA (40 μg/ml).

Figure 5. Calibration curve of DAPA (60 μg/ml).

Figure 6. Calibration curve of DAPA (80 μg/ml).

Figure 7. Calibration curve of DAPA (100 μg/ml).

Results

Application of the proposed method for estimation of drug

Absorbance of drug in stock solution can study by amount of drug estimated. The results are reported in the Table 2.

Table 2: Results of estimation of dapagliflozin in sample.

Validation of proposed method

Validation is normally done to assure the reliability of the proposed method and was performed as per the ICH guide lines for the following criteria [5].

1) Accuracy: Accuracy of method is ascertained by recovery studies performed at different levels of concentrations (80%, 100% and 120%). Mean % recovery were found to be 98.33% for absorption method with % RSD less than 2.

2) Precision: Precision studies were calculated as SD and % RSD. The results within prescribed limit showed that the method was found to be precise and % RSD less than 2.

3) Ruggedness: The method was found to be rugged with no significant changes on test result upon change of analytical conditions like different time (Intraday), different day (Interday) and different analyst [6-8].

4) Linearity and range: The study of linearity and range was performed as per the USP/ICH recommendation. DAPA marketed formulation was found to be linear in the range of 20 to 100 μg/ml, with r²=0.994 for all drugs at selected wavelength for the methods [9,10].

5) LOD: Limit of detection of Dapagliflozin was found to be 1.09743 μg/ml.

6) LOQ: Limit of Quantitation of Dapagliflozin was found to be 0.01233 μg/ml.

Discussion

The management of various diseases and disorders are done by the use of multiple therapeutic agents which acts at different sites. It is necessary to measurement of amount of drug estimate in blood. So that blood plasma of patient given by dapagliflozin tablet is used as sample.

On extensive literature survey it was found that very few quantitative estimation of dapagliflozin by using UV spectroscopy method were used and it seems that this method is accurate, precise and linear.

Conclusions

It was made to develop a quantitative estimation of Dapagliflozin tablet (Forxiga) by using UV Spectroscopy. The developed method was validated for linearity, Accuracy, precision, ruggedness and results were within the limits according to ICH guidelines. The proposed method was cost effective, simple, rapid, economic, cheap, Precise and robust.

Acknowledgments

The author’s were thankful to Principle of KYDSCT’s College of Pharmacy, Sakegaon Bhusawal (MS) INDIA for providing necessary help for work.

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