Contemporary Trends in Analytical Method Development for Dapagliflozin and Pioglitazone: A Critical Review of Spectroscopic and Chromatographic Approaches

 

Prushti P. Bhakhar¹, Hiral S. Popaniya²*, Khushi V. Rathod³, Payal N. Vaja⁴

^1,3Research Scholar, School of Pharmacy, Dr. Subhash University, Junagadh (362001), Gujarat, India.

^2,4Assistant Professor, School of Pharmacy, Dr. Subhash University, Junagadh (362001), Gujarat, India.

 

ABSTRACT:

This review gives an overall overview of the different analytical methods available to determine dapagliflozin and pioglitazone, which are used to treat people suffering from type 2 Diabetes Mellitus. Accurate measurement of these two drugs is essential for testing their effect on humans in many ways; they can there for help researchers understand their safety profiles and help ensure that all drugs sold as medicine meet certain criteria (Quality Control). There are many different analytical chemistries available to measure dapagliflozin and pioglitazone in bulk and pharmaceutical preparations, as well as in human biological materials. Evaluating the various analytical methods available, we are specifically referring to methods of validation: For example, when evaluating an absolute measure of dapagliflozin and pioglitazone, we would evaluate the validity of the method by determining its precision, and Accuracy; therefore, this method would also be called Relative Precision and Relative Accuracy, using the validated dose concentration. Analytical methods are generally considered to be highly specific, therefore creating strong, reliable and reproducible analytical method should be selected based on the needs regarding accuracy, reliability, and cost. Some examples of analytical methods that could be used would include RP-HPLC, HPTLC, UV Spectroscopy, and Stability Testing, along with the new developments being made in analytical chemistry techniques.

 

 

 

 

 

INTRODUCTION:

Two oral antidiabetic medications, the combination of Dapagliflozin and Pioglitazone, are frequently taken together to good choice for managing Type 2 Diabetes. It is a smart idea to use Dapagliflozin and Pioglitazone because they are a dynamic duo that attacks high blood sugar from two completely different directions^1,2.

 

In addition to reducing blood glucose, dapagliflozin has several other beneficial effects, including modest weight loss (1 kg-5 kg), lowering of systolic blood pressure, and improved cardiovascular and renal outcomes. The combination of these Effects will allow dapagliflozin to be used not only as Monotherapy, but also in Combination with others such as metformin, DPP4 Inhibitors, and thiazolidinediones. As the Clinical Importance of Dapagliflozin (figure 1) Continues to Rise; it will be necessary to establish through validated analytical methods, the levels of dapagliflozin in bulk materials, drug products/formulations and biological fluids/samples^3,4,5.

 

 

Figure 1: Structure of Dapagliflozin⁸

 

Pioglitazone (figure 2) is an antihypergilycaemic agent that, in the presence of insulin resistance, increases hepatic and peripheral insulin sensitivity. Thereby inhibiting hepatic gluconeogenesis and increasing peripheral and splanchnic glucose uptake. It is an insulin-sensitizing thiazolidinedione that activates a particular nuclear receptor (peroxisome-proliferator activated receptor-ℽ [PPAR-ℽ]) found in adipose tissue, pancreatic B-cells, vascular endothelium, and macrophages. This evaluation focuses on its approved usages in the EU and the United States in the management of type 2 diabetes^6,7.

 

 

Figure 2: Structure of Pioglitazone⁹

 

This extensive research, we have collated the documented analytical methods available for the examination of formulations and biological samples of Dapagliflozin and Pioglitazone, both separately and in combination with other medications. These approaches comprise a number of techniques, including spectrophotometry, high performance chromatography (HPLC), liquid chromatography-mass spectrometry (LC-MS), and high-perfomance thin layer chromatography (HPTLC). Among the procedures, UV stands out as the most commonly utilized technology for analytical estimate. Table 1 lists the physicochemical characteristics and mode of action of the medications.

 

Table 1: Physicochemical properties of Dapagliflozin and Pioglitazone^10-13

Parameters	Description
Drug Name	Dapagliflozin	Pioglitazone
CAS Number	461432-26-8	111025-46-8
Category	Antidiabetic agent SGL2 inhibitor	Thiazolidinedione
Chemical Formula	C21H25ClO6	C19H20N2O3S
Molecular Weight	408.9 gm/mole	356.44g/mol
Physical State and Appearance	White to Pale Yellow Solid	White of White
Melting Point	74-78℃	183℃-184℃
Solubility	Methanol, Ethanol, Dimethyl formamide	Soluble in organic solvent
Mechanism of Action	In high-risk individuals with type 2 diabetes, SGLT2 inhibitors lower heart failure and cardiovascular events, but DPP-4 inhibitors have no such benefit and may raise the risk of heart failure. In individuals with type 2 diabetes and established coronary artery disease, this study compared the cardiometabolic effects of the SGLT2 inhibitirs dapagliflozin and the DPP-4 inhibitors vildagliptin.	Pioglitazone, a thiazolodinedione, stimulates the PPARℽ nuclear receptor. activation of PPARℽ-RXR heterodimers leads to improved insulin sensitivity in adipose tissue, muscle, and liver. as a result, glucose absorption increase while hepatic glucose production decreases, and lowering lipotoxicity. it increases adiponectin levels and inhibits NF-Kb, providing anti-inflammatory actions.
Drug Approved by CDSCO	29 November 2022	CDSCO (16 May 2023)

 

 

Table 2: Analytical Methods for Estimation of Dapagliflozin

Sr. No.	Drug Name	Analytical Methods	Description	Ref. No.
1.	Dapagliflozin	UV-Spectrophotometry	Linearity: 5-40 μg/ml Solvent: Methanol: water Wavelength: Method I (Zero order): 224 nm Method II (Area Under Curve): 218-230 nm Method III (1º derivative): 220 nm Method IV (2ºderivative): 224 nm, 235.5 nm	14
2.	Dapagliflozin	UV-Spectrophotometry	Linearity: 10-35 μg/ml Solvent: Ethanol: Phosphate buffer (1:1) (pH 7.2) Wavelength: 233.65 nm	15
3	Dapagliflozin	RP-HPLC	Stationary phase: Water S C18 5 μm particle size, 25 cm×4.6 mm i.d. Mobile phase: Phosphate buffer: acetonitrile (60:40 % v/v) Flow rate: 1.0 ml/min Detection: 237 nm Concentration range: 10-60 μg/ml	16
4	Dapagliflozin	RP-HPLC	Stationary phase: Princeton C18 column Mobile phase: Acetonitrile: 0.1% Triethylamine pH-5.0 (50:50 % v/v) Flow rate: 1.0 ml/min Detection: 224 nm Concentration range: 10-70 μg/ml	17
5	Dapagliflozin	Stability indicating RP-HPLC	Stationary phase: BDS column Mobile phase: Acetonitrile: Orthophosphoric acid (55:45 % v/v) Flow rate: 1.0 ml/min Detection: 245 nm Concentration range: 25-150 μg/ml	18
6	Dapagliflozin	RP-HPLC	Stationary phase: hypersil BDS (250 mm × 4.6 mm, 5 μm) Mobile phase: Ortho phosphoric acid buffer: Acetonitrile (60:40 % v/v) Flow rate: 1.0 ml/min Detection: 245 nm Concentration range: 25-150 μg/ml	19
7	Dapagliflozin	RP-HPLC and UV-Spectrophotometry	Stationary phase: BDS column Mobile phase: Ortho phosphoric acid buffer: Acetonitrile (45:55 % v/v) Flow rate: 1.0 ml/min Concentration range: 25-150 μg/ml Concentration range: 1-5 μg/ml Wavelength: 203 nm Solvent: Methanol	20
8	Dapagliflozin	RP-HPLC	Stationary phase: Symmetry C18, 25 cm x 4.6 mm i.d.5 µm, Mobile phase: Methanol: Acetonitrile: OPA (75:25:05 % v/v) Flow rate: 1.0 ml/min Detection: 246 nm Concentration range: 20-70 μg/ml	21
9	Dapagliflozin	Stability indicating HPLC	Stationary phase: C18 (4.6 mm* 150,5 μm) Mobile phase: Acetonitrile: di-potassium hydrogen phosphate with pH - 6.5 adjusted with OPA (40:60 %v/v) Flow rate: 1.0 ml/min Detection: 222 nm Concentration range: 50-150 μg/ml	22
10	Dapagliflozin	HPTLC	Stationary phase: Merck precoated silicagel aluminum plate 60F254 Mobile phase: Chloroform: Methanol (9:1 % v/v) Detection: 223 nm using Camag TLC Scanner. Concentration range: 400 ng/band to 1200 ng/band	23

 

Table 3: Analytical Methods for estimation Dapagliflozin with other drug combination

Sr. No	Drug Name	Analytical Methods	Description	Ref. No.
1	Dapagliflozin and Metformin HCL	RP-HPLC	Stationary phase: Phenomenex C18 250 mm x 4.6 mm Mobile phase: Water: Methanol (50:50 % v/v) Flow rate: 1.0 ml/min Detection: 230 nm Concentration range: 2-7 μg/ml (Metformin HCl), 60–210 μg/ml (Dapagliflozin)	24
2	Dapagliflozin and Metformin HCL	UV-Spectrophotometry including force degradation	Simultaneous equation method: Concentration range: 2-32 μg/ml (Dapagliflozin) and 1– 20 μg/ml (Metformin). Detection Wavelength: 222 nm (Dapagliflozin) and 232nm (Metformin). Solvent: Water	25
3	Dapagliflozin propanediol monohydrate and Sitagliptin	UV-Spectrophotometry	1storder derivative spectroscopic Method: Sitagliptin at zero cross over- 275 nm and Dapagliflozin zero cross over point- 232 nm Concentration range:25-125 μg/ml (Dapagliflozin) 2.5-12.5 μg/ml and (Sitagliptin) Solvent: Methyl alcohol	26
4	Dapagliflozin and Saxagliptin	Stability indicating RP-HPLC	Stationary phase: BDS C18 (150 x 4.6 mm, 5.0 μ) Mobile phase: Ammonium acetate buffer: CAN (40:60% v/v) Flow rate: 1.0 ml/min Detection: 220 nm Concentration range: 0-15 μg/ml (Dapagliflozin) and 0-8 μg/ml (Saxagliptin)	27
5	Dapagliflozin propanediol monohydrate and Sitagliptin	RP-HPLC	Stationary phase: Inertsil ODS C18 Mobile phase: Methyl Nitrile (25 parts) and 0.02 M KH2PO4 buffer 0.02 M having 1 ml triethylamine with neutral pH adjusted by orthophosphoric acid (75parts) isocratic mode Flow rate: 1.0 ml/min Detection: 210 nm Concentration range: 5–15 μg/ml (Dapagliflozin) and 50-150 μg/ml (Sitagliptin)	28
6	Dapagliflozin and Saxagliptin	RP-HPLC	Stationary phase: Symmetry C8 (4.6×150 mm, 3.5 μm, Make: XTerra) Mobile phase: Buffer: acetonitrile 70:30% v/v (pH 3) Flow rate: 1.0 ml/min Detection: 221 nm Concentration range: 25-125 μg/ml (Dapagliflozin) and 12.5-62.5 μg/ml (Saxagliptin)	29
7	Dapagliflozin and Saxagliptin	RP-HPLC	Stationary phase: XT erra C18 column (150 mm x 4.6 mm x 5 μm particle size) Mobile phase: Phoaphate buffer: Acetonitrile (50:50v/v) Flow rate: 1.0 ml/min Detection: 225 nm Concentration range: Dapagliflozin and Saxagliptin 10-20μg/ml (Dapagliflozin), 20-60 μg/ml respectively	30
8	Dapagliflozin and Saxagliptin	RP-HPLC	Stationary phase: Discovery C18 column (250 mm, 4.6 mm, and 5μm). Mobile phase: acetonitrile: orthophosphoric acid (0.1%) 50:50 Flow rate: 0.98 ml/min Detection: 210 nm Concentration range: Dapagliflozin and Saxagliptin25-150 μg/mland12.5-75 μg/mL respectively	31
9	Metformin, Dapagliflozin, and Saxagliptin	Stability indicating RP-HPLC	Stationary phase: Kromasil C18 column (150 mm× 4.6 mm, 5 μm) Mobile phase: phosphate buffer: acetonitrile (60:40%) Flow rate: 1 ml/min Detection: 230 nm Concentration range: Metformin, Dapagliflozin, and Saxagliptin 125–750 μg/mL, 1.25–7.5 μg/ml, and 0.625–3.75 μg/mL respectively	32
10	Dapagliflozin and Saxagliptin	RP-HPLC	Stationary phase: XT era C18 column (150 mm x 4.6 mm x 5 μm) Mobile phase: phosphate buffer: Acetonitrile (50:50 % v/v) Flow rate: 1 ml/min Detection: 225 nm Concentration range: Dapagliflozin, and Saxagliptin 100-1500 μg/ml, 20–300 μg/ml respectively	33
11	Dapagliflozin and Saxagliptin	RP-HPLC	Stationary phase: Phenomene x Hyperclone C18 column (250×4.6 mm,5 μm) Mobile phase: methanol: phosphate buffer (pH 3.0) (70:30, v/v) Flow rate: 1 ml/min Detection: 220nm Concentration range: Dapagliflozin, and Saxagliptin 4-24 μg/mL, 2-12 μg/mL respectively	34
12	Dapagliflozin, Saxagliptin, and Metformin	UV Spectrophotometric method (Simultaneous estimation)	Simultaneous equation method: Concentration range: 5-25 μg/ml (Dapagliflozin) and 10-50 μg/ml (Metformin) and 1-5 μg/ml (Saxagliptin) Detection Wavelength: 272 nm (Dapagliflozin), 232nm (Metformin) and 212 nm (Saxagliptin) Solvent: methanol: water 80:20 % v/v	35
13	Dapagliflozin and Linagliptin	LC-MS/MS	Stationary phase: Phenomenex Gemini C18 column (50 mm× 4.6 mm, 5 µm) Mobile Phase: Methanol: water 80:20 % v/v Flow rate: 0.5 ml/min Concentration range: 5 µL	36
14	Dapagliflozin and Vildagliptin	RP-HPLC	Stationary phase: BDS C18 column (4.6 mm × 150 mm, 5 µm) Mobile Phase: Acetonitrile: Na2HPO4 70:30 % v/v Flow rate: 0.8 ml/min Concentration range: 1.25 -7.5 μg/ml (Dapagliflozin), 12.5 -75 μg/ml (Vildagliptin)	37
15	Dapagliflozin and Bisoprolol fumarate	RP-HPLC	Stationary phase: Agilent zorbax ODS C18 column (4.6 mm× 150 mm, 5 µm) Mobile Phase: Triethyl amine: Acetonitrile (66:33, v/v) (pH 3.0) Flow rate: 1.5 ml/min Concentration range: 5 – 15 μg/ml	38
16	Dapagliflozin and Linagliptin	RP-HPLC	Stationary phase: Nova C18 column (250 mm × 4.6 mm, 5 µm) Mobile phase: Methanol: Phosphate buffer 65:35 % v/v Flow rate: 1 ml/min Concentration range: 4-40 μg/ml (Linagliptin) 8-80 μg/ml (Dapagliflozin),	39

 

 

Table 4: Analytical methods for estimation of Pioglitazone

Sr. No.	Drug Name	Analytical Methods	Description	Ref. No.
1	Pioglitazone	UV Spectrophotometer	Concentration range: 2–14 µg /ml Solvent: 0.1 M HCl Drug + Phthalate buffer pH 2.4 And Extracted Into chloroform Wavelength: 419 nm	40
2	Pioglitazone	UV Spectrophotometer	Concentration range: 2.5 - 20 µg /ml Solvent: 0.1 M HCl Wavelength: 269 nm	41
3	Pioglitazone	UV Spectrophotometer	Concentration range: 5 – 30 µg /ml Solvent: Ethanol Wavelength: 224.4 nm	42
4	Pioglitazone	UV Spectrophotometer	Concentration range: 10-50 µg /ml Solvent: Phosphate Buffer (pH: 7.4) Wavelength: 238 nm	43
5	Pioglitazone	UV Spectrophotometer	Concentration range: 10-50 µg /ml Solvent: Phosphate Buffer (pH:7.4) Wavelength: 238 nm	44
6	Pioglitazone	HPLC	Mobile Phase: Toluene: Ethylacetate: Formic Acid 10:3:1 % v/v Wavelength: 254 nm Concentration Range: 100-3000 µg /ml	45
7	Pioglitazone	HPLC	Mobile Phase: Water: Tri fluoro acetate Acid (100:0.05 v/v) Wavelength: 225nm Flow Rate: 1.0 ml/Min Concentration range: 10-100 µg /ml	46
8	Pioglitazone	HPLC	Mobile Phase: Potassium Dihydrogen Phosphate Buffer: Methanol (pH=3) 55:45 v/v Wavelength: 241 nm Flow Rate: 1.5 ml/Min Concentration range: 0.06 - 250 µg /ml	47
9	Pioglitazone	RP-HPLC	Stationary Phase: Hypersilc-8 (250) Mobile Phase: Acetonitrile: Water 60:40 %v/v Wavelength: 266 nm Flow Rate: 1 ml/Min Concentration range: 5- 20 µg/ml	48
10	Pioglitazone	LC-MS	Mobile Phase: Water: Trifluoroacetate Acid 100:0.05 %v/v Wavelength: 225nm Flow Rate: 1.0 ml/Min Concentration range: 10-1000 µg /ml	49
11	Pioglitazone	HPLC	Stationary Phase: RP C18 (25 cm× 0.46 mm) Mobile Phase: Methanol: Water 65:35 %v/v Wavelength: 230 nm Flow Rate: 1.0 ml/Min Concentration range: 10-50 µg /ml	50
12	Pioglitazone	RP-HPLC	Stationary Phase: XBP C18 column, 250×4.6 mm Mobile Phase: Acetonitrile: Phosphate buffer 50:50 % v/v Flow Rate: 1.0 ml/min Concentration Range: 25-75 µg/ml	51

 

 

 

Table 5: Analytical Methods for estimation Pioglitazone with other drug combination

Sr. No.	Drug Name	Analytical Methods	Description	Ref. No.
1	Pioglitazone, Metformin and Glimepiride	UV Spectrophotometry	Concentration range: Pioglitazone (6 -36 µg/ml), Metformin (10 – 60 µg/ml), Glimepiride (1 -6 µg/ml) Solvent: Methanol Wavelength: 265.4 nm	52
2	Pioglitazone and Glimepiride	UV Spectrophotometry	Concentration range: Pioglitazone (10 - 60 µg/ml), Glimepiride (1 - 10 µg/ml) Solvent: Methanol Wavelength: 250 nm	53
3	Pioglitazone and Glimepiride	HPLC	Stationary Phase: C18 reversed-phase column, 250×4.6 mm Mobile Phase: Acetonitrile: Ammonium Acetate (pH=4.5) 60:40 v/v Flow Rate: 1.0 ml/min Detection: 230nm Concentration Range: Pioglitazone (2.0 To 200 µg/ml) Glimepiride (0.5 To 50 µg/ml)	54
4	Pioglitazone and Metformin	UV Spectrophotometry	Concentration range: Pioglitazone (5 -30 µg/ml) Metformin (10 - 70 µg/ml) Solvent: Sodium hydroxide Wavelength: 233 nm	55
5	Pioglitazone and Metformin	RP-HPLC	Stationary Phase: BDS Hypersil C18column,250×4.6 mm Mobile Phase: Acetonitrile: Potassium dihydrogen ortho phosphate buffer (pH=3) 50:50 % v/v Flow Rate: 1.0 ml/min Detection: 230 nm Concentration Range: Pioglitazone (12-7 µg/ml) Metformin (40-240 µg/ml)	56
6	Pioglitazone and Glimepiride	RP-HPLC	Stationary Phase: C18 reversed-phase column,250×4.6 mm Mobile Phase: Acetonitrile: Phosphate buffer (pH=7) 60:40 % v/v Flow Rate: 0.8 ml/min Detection: 230 nm Concentration Range: Pioglitazone (10 -50 µg/ml) Glimepiride (3- 15 µg/ml)	57
7	Pioglitazone and Telmisartan	RP-HPLC	Stationary Phase: Phenomenex C18 reversed-phase column, 250×4.6 mm Mobile Phase: Acetonitrile: Triethyleamine (pH=4.5) 65: 35 % v/v Flow Rate: 1.3 ml/min Concentration Range: Pioglitazone (7.5 - 37.5 µg/ml) Telmisartan (10 - 50 µg/ml)	58
8	Pioglitazone, Metformin and Nateglinide	HPLC	Stationary Phase: C18 reversed-phase column,250×4.6 mm Mobile Phase: Methanol: Potassium dihydrogen ortho phosphate 85 :15 % v/v Flow Rate: 1.2 ml/min Concentration Range: Pioglitazone (2-10µg/ml) Metformin (50 - 250 µg/ml) Nateglinide (3-15 µg/ml)	59
9	Pioglitazone, Aspirin and Atrovastatin	RP-HPLC	Stationary Phase: Zorbax SBCN C18 reversed-phase column, 250×4.6 mm Mobile Phase: Acetonitrile: Phosphate buffer (pH=3.5) 40:60 % v/v Flow Rate: 1 ml/min Detection: 261 nm Concentration range: Pioglitazone (30 µg/ml) Aspirine (150 µg/ml) Atrovastatin (20 µg/ml)	60

 

Table 6: Reported Analytical Methods for Pioglitazone with Dapagliflozin drug Combination

Sr. No.	Drug Name	Analytical Methods	Description	Ref. No.
1.	Dapagliflozin and Pioglitazone	RP-HPLC	Mobile phase: ACN: Potassium dihydrogen phosphate (pH:4) 25:75 % v/v Flow rate: 1 ml/min Wavelength: 228 nm Concentration range: Dapagliflozin (2 - 10 µg/ml) Pioglitazone (3 - 15 µg/ml)	61
2.	Dapagliflozin and Pioglitazone	UPLC	Stationary Phase: C18 Acquity BEH column (2.1 mm × 100 mm, 1.7 μm) Mobile phase: Ethanol: Ammonium sulphate buffer 43:56 % v/v Flow rate: 0.246 ml/min Concentration range: Dapagliflozin (3.5- 50 μg/ml) and Pioglitazone (2.5 - 50 μg/ml)	62
3.	Dapagliflozin and Pioglitazone	RP-HPLC	Stationary phase: Kromstar Vertex C18 analytical column Mobile phase: Methanol: Potassium dihydrogen phosphate (pH:4) 25:75 % v/v Flow rate: 1 ml/min Wavelength: 228 nm Concentration range: Dapagliflozin (2-10 µg/ml) and Pioglitazone (3–15 µg/ml)	63

 

CONCLUSION:

The methods available in this review included both spectroscopy and chromatography methodologies to quantify dapagliflozin and pioglitazone. While various methods can individually be used to detect dapagliflozin and pioglitazone, there are no UV spectroscopy methods are present that allow for a combination technique that can determine both drugs at the same time. However, the currently available methods can be classified as being relatively simple, cost-effective, accurate, and reproducible. Most of the methods reported in the literature are based on RP-HPLC using UV absorbance detection, which provides an optimal balance of repeatability, speed, and sensitivity for the determination of these compounds. Because of the lack of available combination techniques for the simultaneous quantification of dapagliflozin and pioglitazone, future development of such methods stands to improve upon the quality control of those dosage forms that combine both drugs, and would provide much-needed improvements in sensitivity and selectivity of those drugs as they are analysed in.

 

CONFLICT OF INTEREST:

The authors have no conflicts of interest regarding this investigation.

 

ACKNOWLEDGEMENT:

We are thankful to Dr. Subhash University for guiding and supporting this review work.

 

REFERENCE:

1.      Dhillon S, Dapagliflozin: A Review in Type 2 Diabetes. Drugs. 2019; 79(10): 1135-1146. DOI: 10.1007/s40265-019-01148-3

2.      Jabbour S, Durability of response to dapagliflozin: a review oflong-term efficacy and safety. Current Medical Research and Opinion. 2017; 33(9): 1685-1696. DOI:10.1080/03007995.2017.1354822

3.      ode BW, Garg SK. The emerging role of adjunctive noninsulin antihyperglycemic therapy in the management of type 1 diabetes. Endocr Pract. 2016; 22(2): 220–30.

4.      Ahmed-Sarwar N, Nagel AK, Leistman S, et al. SGLT-2 inhibitors: is there a role in type 1 diabetes mellitus management? Ann Pharmacother. 2017; 51(9): 791–6.

5.      Chen J, Fan F, Wang JY, et al. The efficacy and safety of SGLT2 inhibitors for adjunctive treatment of type 1 diabetes: a systematic review and meta-analysis. Sci Rep. 2017; 7: 44128.

6.      European Medicines Agency. Actos: summary of product characteristics [online].

7.      Takeda Pharmaceuticals America Inc. Actos (pioglitazone hy-drochloride) tablets prescribing information [online].

8.      Nationallibraryofmedicine.Dapagliflozin[Imageoninternet]Availablefrom:https://pubchem.ncbi.nlm.nih.gov/compound/Dapagliflozin

9.      Tushar Raut, Dr. Deokar G.S, Rupali Kadam* and Pranali Patil - Scientific Figure on ResearchGate. Available from: https:// www.researchgate.net/figure/Chemical-structure-of-Pioglitazone-hydrochloride.

10.   TABLE 1 Obermeier M, Yao M, Khanna A, Koplowitz B, Zhu M, Li W, Komoroski B, Kasichayanula S, Discenza L, Washburn W, Meng W. In vitro characterization and pharmacokinetics of dapagliflozin (BMS-512148), a potent sodium-glucose cotransporter type II inhibitor, in animals and humans. Drug metabolism and disposition. 2010; 38(3): 405-14.

11.   Al-Majed A, Bakheit AH, Abdel Aziz HA, Alharbi H, Al-Jenoobi FI: Pioglitazone. Profiles Drug Subst Excip Relat Methodol. 2016; 41: 379-438. doi: 10.1016/bs.podrm.2015.11.002. Epub 2016 Feb 2.

12.   Yue Sun, Dong Yan, ZiruiHao, Lijuan Cui, GuipingLiMedical Science Monitor: International Medical Journal of Experimental and Clinical Research 26, e921891-1, 2020

13.   Smith U. Pioglitazone: mechanism of action. International journal of clinical practice. Supplement. 2001 Sep 1(121): 13-8

14.   Mante GV, Gupta KR, Hemke AT. Estimation of dapagliflozinfrom its tablet formulation by UV-spectrophotometry. Pharm Methods. 2017; 8(2): 102-7.

15.   Karuna PC, China E, Rao MB. Unique UV spectrophotometric method for reckoning of Dapagliflozin in bulk and pharmaceutical dosage forms. Journal of Chemical and Pharmaceutical Research.2015; 7(9): 45-9.

16.   Debata J, Kumar S, Jha SK, Khan A. A New RP-HPLC method development and validation of dapagliflozin in bulk and tablet dosage form International Journal of Drug Development and Research. 2017; 9(2): 48-51.

17.   Mante G, Hemke A, Umekar M .RP-HPLC Method for Estimation of Dapagliflozin from its Tablet. International Journal of Chemtech Research. 2018; 11(1): 242-248.

18.   Sanagapati M, Dhanalakshmi K, Reddy NG, Sreenivasa S. Development and Validation of stability- Indicating RP-HPLC method for determination of Dapagliflozin. Journal of Advance d Pharmacy Education & Research. 2014; 4(3): 350-353.

19.   Basha S, Sravanthi P. Development and validation of dapagliflozin by reversed-phase high-performance liquid chromatography method and it’s forced degradation studies. Asian Journal of Pharmaceutical and Clinical Research. 2017; 10(11):101-5.

20.   Sanagapati M, Dhanalakshmi K, Reddy NG, Sreenivasa S. Method development and validation of dapagliflozin in API by RP-HPLC and UV-spectroscopy. International Journal of Pharmaceutical Sciences and Drug Research. 2014; 6(3): 250-252

21.   Sarkar S, Patel V. Method development and validation of dapagliflozine drug in bulk and tablet dosage form by RP-HPLC International Journal of Pharma Research and Health Sciences. 2017; 5(4):1755-9.

22.   Verma MV, Patel CJ, Patel MM. Development and stability indicating HPLC method for dapagliflozin in API and pharmaceutical dosage form. International Journal of Applied Pharmaceutics. 2017; 9(5): 33-41.

23.   Suma BV, Deveswaran R, Premnath SH. Anewhigh-performancethin layer chromatographic method development and validation ofdapagliflozin in bulk and tablet dosage form. International Journal of Pharmacy and Pharmaceutical Sciences. 2019; 11: 58-63.

24.   Khagga B, Thaneeru S, Samreen B, Mogili S. RP-HPLC Methodfor Dapagliflozin and Metformin HCL in Bulk and Combined Formulation. Archives of Pharmacy Practice. 2022; 12(4): 106-10.

25.   Bhavyasri K, Surekha T, Sumakanth M. A Novel MethodDevelopment and Validation of Dapagliflozin and Metormin Hydrochloride using Simultaneous Equation Method by UV–Visible Spectroscopy in Bulk and Combined PharmaceuticalFormulation including Forced Degradation Studies. International Journal of Pharmaceutical Sciences and Research. Sci Res. 2020; 12(8): 1100-5.

26.   Jani S, Shukla R, Patel P, Mehta B, Detholia K. Quantitative estimation of sitagliptin and dapagliflozin propanediol monohydrate in synthetic mixture using 1st order derivative spectroscopy simultaneous spectrophotometric analysis. International Journal of Pharmaceutical Chemistry and Analysis. 2022; 9(1): 28-34.

27.   Manasa M, Aanandhi VM. Stability indicating simultaneous method development and validation of dapagliflozin and saxagliptin by RP-HPLC. Research Journal of Pharmacy and Technology. 2021; 14(2): 1045-9.

28.   Patel YD, Patel PR, Bhatt J, Mehta B, Detholia K. Quantitative computation and stability evaluation of phase III composition

29.   Patel YD, Patel PR, Bhatt J, Mehta B, Detholia K. Quantitative computation and stability evaluation of phase III composition. By RP-HPLC. Journal of Applied Pharmaceutical Science. 2022; 12(06): 148–155.

30.   Boggula N, Pandiyan PS. Development and validation of RP HPLC method for the simultaneous estimation of dapagliflozin and saxagliptin in bulk and pharmaceutical dosage forms. International Journal of Pharmaceutical Sciences and Research. 2021; 12(1): 314-320.

31.   Kommineni V, Chowdary KP, Prasad SV. Development and validation of a new HPLC method for the simultaneous estimation of saxagliptine and dapagliflozin and its application in pharmacokinetic studies. International Research Journal of Pharmacy and Medical Sciences. 2018; 1(6).

32.   Gundala A, Prasad KV, Koganti B. Application of quality by design approach in RP-HPLC method development for simultaneous estimation of saxagliptin and dapagliflozin in tablet dosage form. Brazilian Journal of Pharmaceutical Sciences. 2019.

33.   Vankalapati KR, Alegete P, and Boodida S. Stability-indicating HPLC method development and validation for simultaneous estimation of metformin, dapagliflozin, and saxagliptin in bulk drug and pharmaceutical dosage form biomedical Chromatography. 2022; 36(7): 5384

34.   Vinu thakom mineni, K.P.R. Chowdary, S.V.U.M. Prasad. Formulation of Dapagliflozin and Saxagliptin Tablets and In vitro Evaluation by RP-HPLC Method. Asian Journal of Pharmaceutical Analysis. 2019; 9(2): 93-98. Doi: 10.5958/22315675.2019.00018.8

35.   Sayali S. More, Sandeep S. Sonawane, Santosh S. Chhajed, Sanjay J. Kshirsagar. Development and Validation of RP-HPLC Method for Simultaneous Estimation of Saxagliptin and Dapagliflozin in Tablets. Asian Journal of Pharmacy and Technology. 2018; 8(3): 145-148. Doi: 10.5958/2231-5713.2018.00023.5

36.   Hiral S. Popaniya, Dinesh K. Dangar Eco-Friendly LC-MS/MS Method for quantification of dapagliflozin and pioglitazonevin combined dosage form: Development, validation and agree assessment. Indian Journal of National Science. 2025.

37.   Pavan K. Regeti, B. Sunitha, C. Parthiban, M. Sudhakar. Method Development and Validation for Simultaneous Estimation of Dapagliflozin and Vildagliptin in Pharmaceutical Dosage Form by RP-HPLC. Asian Journal of Pharmaceutical Analysis. 2024; 14(4): 229-3.

38.   Disha H. Joshi, Jimish R. Patel. Stability Indicating RP-HPLC Method Development and Validation for Simultaneous Estimation of Dapagliflozin Propanediol Monohydrate and Bisoprolol Fumarate in Synthetic Mixture. Asian Journal of Pharmaceutical Research.2025; 15(4): 355-3.

39.   Hiral S. Popaniya, Dinesh K. Danger, stimulation quatiification of Dapagliflozin and Linagliptin in pharmaceutical formulation: A stability-indicating RP-HPLC approach, International Journal of Bilogy, Pharmacy and Allied Sciences (IJHBPAS), January, 2026; 15(1): 416-428.

40.   Priyabarbude, mukundtawar, Prashant Burange. Method Development using a UV Visible Spectrophotometer for the Simultaneous Estimation of Metformin (MET), Saxagliptin (SXG), and Dapagliflozin (DGF) in Marketed Formulation. Asian Journal of Pharmaceutical Analysis. 2022; 12(4): 243-7. Doi: 10.52711/2231-5675.2022.00039

41.   Amanlou M, Zarei-Ghobadi M, Rofouei MK, Saremi S, Kebriaeezadeh A. Extractive Spectrophotometric Method for Determination of Pioglitazone Hydrochloride in Raw Material and Tablets Using Ion‐Pair Formation. Journal of Chemistry. 2010; 7(3): 915-21.

42.   Mahadik SP, Senthilkumar GP. Method development & validation of pioglitazone in bulk and pharmaceutical dosage forms by using spectrophotometric method. Asian J. Biochem. Pharm. Res. 2012; 2: 159-65.

43.   Ali MY, Swamy PV, Borgaonkar PR, Raju SA. UV-spectrophotometric determination of pioglitazone in pharmaceutical dosage forms. Int. J. Chem. Sci. 2008; 6(4): 2062-5.

44.   Shakya P, Singh K. Determination of pioglitazone hydrochloride in bulk and pharmaceutical formulations by UV spectrophotometric method. International Journal of Pharmaceutical Sciences and Research. 2010; 1(11): 153.

45.   Shakya P, Singh K. Determination of pioglitazone hydrochloride in bulk and pharmaceutical formulations by UV spectrophotometric method. International Journal of Pharmaceutical Sciences and Research. 2010; 1(11): 153.

46.   Singh SC, Kushnoor A. Development and validation of a HPTLC method for estimation of pioglitazone in bulk and tablet dosage form. J Pharm Res. 2011; 4(11): 3919-21.

47.   Ramulu K, Kumar TT, Krishna SR, Vasudev R, Kaviraj M, Rao BM, Rao NS. Identification, isolation and characterization of potential degradation products in pioglitazone hydrochloride drug substance. Die Pharmazie-An International Journal of Pharmaceutical Sciences. 2010; 65(3): 162-8.

48.   Ramulu K, Kumar TT, Krishna SR, Vasudev R, Kaviraj M, Rao BM, Rao NS. Identification, isolation and characterization of potential degradation products in pioglitazone hydrochloride drug substance. Die Pharmazie-An International Journal of Pharmaceutical Sciences. 2010; 65(3): 162-8.

49.   Sharma S, Sharma MC, Chaturvedi SC. Study of stressed degradation behavior of pioglitazone hydrochloride in bulk and pharmaceutical formulation by HPLC assay method. Journal of Optoelectronics and Biomedical Materials. 2010; 1(1): 17-24.

50.   Varma. D. S., Dighe P. R.. Analytical Methods of Antidiabetic Drugs – Sitagliptin, Saxagliptin, Linagliptin, Alogliptin, Gemifibrozil, Troglitazone, Pioglitazone and Rosiglitazone: A Review. Research Journal of Pharmaceutical Dosage Forms and Technology. 2022; 14(4): 324-0.

51.   M Yashpal Naidu, K P Channa Basavaraj, T Tamizh Mani, K Roopa. Validated RP-HPLC Method for the Quantitation of Pioglitazone an Anti - Diabetic Drug in Bulk and Pharmaceutical Dosage Forms. Research J. Pharm. and Tech. 2010; 3(3): 885-887.

52.   Ramulu K, Kumar TT, Krishna SR, Vasudev R, Kaviraj M, Rao BM, Rao NS. Identification, isolation and characterization of potential degradation products in pioglitazone hydrochloride drug substance. Die Pharmazie-An International Journal of Pharmaceutical Sciences. 2010; 65(3): 162-8.

53.   Deepa P, Laxmanbhai P, Madhabhai P, Advaita PB. Simultaneous estimation of glimepiride, pioglitazone HCl and metformin HCl by derivative spectrophotometry method. Int. Res. J. Pharm. 2011; 2: 111-4.

54.   Game MD. First order derivative spectrophotometric method for simultaneous estimation of glimepiride and pioglitazone HCl in combined dosage form. J. Pharm. Res. 2011; 4(11).

55.   Sonali D. Rathod, P.M. Patil, S. B. Jadhav, P.D. Chaudhari. UV Spectrophotometric Simultaneous Determination of Metformin Hydrochloride and Pioglitazone Hydrochloride in Combined Dosage Form. Asian J. Pharm. Ana. 2012; 2(1): 05-09.

56.   Ajow Swapna*, Chandaka Madhu, Mallepelli Srivani, M. Sumalatha, Y. Nehalatha, Y. Anusha analytical Method Development and Method Validation for the Simultaneous Estimation of Metformin hydrochloride and Pioglitazone hydrochloride in Tablet Dosage Form by RP-HPLC. Asian J. Pharm. Ana. 2012; 2(3): 85-89.

57.   B. Venkateswara Rao, P. Vijetha, S. Vidyadhara, K. Kavitha. A Novel RP-HPLC Method Development and Validation for the Determination of Pioglitazone and Glimepiride in Bulk and Pharmaceutical Formulations. Asian J. Pharm. Ana. 2017; 7(3): 145-150.

58.   Kapil Rana, Pushpendra Sharma. Development and Validation of a HPLC method for the Determination of Metformin hydrochloride, Nateglinide and Pioglitazone hydrochloride in Multicomponent Formulation. Asian J. Research Chem. 2021; 14(1): 7-12.

59.   Ismail, R Rajavel, M Ganesh, M Jagadeeswaran, K Srinivasan, J Valarmathi, T Sivakumar. RP-HPLC Method for the Simultaneous Determination of Aspirin, Atorvastatin and Pioglitazone in Capsule Dosage Form. Asian J. Research Chem. 2008; 1(1): 40-42.

60.   Varma. D. S., Dighe P. R.. Analytical Methods of Antidiabetic Drugs – Sitagliptin, Saxagliptin, Linagliptin, Alogliptin, Gemifibrozil, Troglitazone, Pioglitazone and Rosiglitazone: A Review. Research Journal of Pharmaceutical Dosage Forms and Technology. 2022; 14(4): 324-0.

61.   Karthik A, Subramanian G, Rao CM, Bhat K, Ranjithkumar A, Musmade P, Surulivelrajan M, Karthikeyan K, Udupa N. Simultaneous determination of pioglitazone and glimepiride in bulk drug and pharmaceutical dosage form by RP-HPLC method. Pakistan journal of pharmaceutical sciences. 2008 Oct 1; 21(4).

62.   Elzayat EM, Sherif AY, Attwa MW, Altamimi MA. A Green Approach: Optimization of the UPLC Method Using DoE Software for Concurrent Quantification of Pioglitazone and Dapagliflozin in a SNEDDS Formulation for the Treatment of Diabetes. ACS omega. 2024; 9(45): 45011.

63.   Mr Tarang P, Ronak P. Method development, validation and forced degradation studies of Dapagliflozin and pioglitazone hydrochlorides in synthetic mixtures by RP-HPLC. International Journal of Trend in Scientific Research and Development. 2022; 6(6): 1858-69

 

 

Received on 19.12.2025      Revised on 11.02.2026 Accepted on 20.03.2026      Published on 16.04.2026 Available online from April 18, 2026 Asian Journal of Pharmaceutical Analysis. 2026; 16(2):137-144. DOI: 10.52711/2231-5675.2026.00021 ©Asian Pharma Press All Right Reserved
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