Development and Validation of RP- HPLC Method for Simultaneous Estimation of Telmisartan and Chlorthalidone in Pharmaceutical Dosage Form

 

V. Sowmya*, G. Bhavana, M. Venkataramana, A. Ganesh, Y. Soudarya

Department of Analysis, Surabhi Dayakar Rao College of Pharmacy, Rimmanaguda, Gawel, Telangana, India.

*Corresponding Author E-mail: vengaladasusowmya78@gmail.com

 

ABSTRACT:

Telmisartan and Chlorthalidone in bulk and combined dosage form were analysed and validated using RP-HPLC. A new technique was developed for the simultaneous estimation of this to medications using this method. Telmisartan and chlorthalidone were successfully separated using the following chromatographic conditions. Phenomenex luna c18 4.5mm×250mm 5µm particles size, flow rate 1.0ml/min mobile phase ratio 40:60v/v, acetonitrile TEA buffer pH 4.2, pH was adjusted with orthophosphoric acid detection wavelength 258nm, waters Alliance 2685 separation module, software power 2986 PDA detector. There results showed that the retention times were respectively, 2 and 246 mints. It was discovered that the purity percentage of chlorthalidone and telmisartan were 99.76 and 101.26 percent respectively. The tailing factor and theoretical plates, two system suitability parameters for telmisartan and chlorthalidone were determined to be 5377.

 

KEYWORDS: Telmisartan and Chlorthalidone, Method Development, Validation, Accuracy.

 

 

INTRODUCTION:

To ascertain the identity, strength, quality and purity of the drug substance and drug products pharmaceutical analysis include the necessary procedures. Pharmaceutical analyses are essential to all industry quality control divisions. In analytical chemistry components in a simple matrix are separated, identified and their relative numbers are calculated1-3

 

Drug estimation techniques can be classified as physical, chemical, physicochemical or biological of these, physical and physicochemical techniques are mostly frequently employed studying a substance physical, characteristics is part of physical methods of analysis4-6

 

High surface aera particles are used in adsorption chromatography to absorb the solute molecules. Adsorption chromatography typically uses a non-polar mobile phase like hexane, octane, chloroform along with a polar solid like silica gel, alumina or porous glass beads If vital information is unavailable e.g.: Pka, solubility, independent research ought to be started right away7-8

 

Samples come in various forms. Solutions ready for injection

The solute distribution factor, which represents the various interactions between the solute mobile phase and the solute-stationary phase, controls the solute interaction9-10

 

MATERIALS AND METHODS:

Table 1: Instruments used

S. No.

Instruments and Glass wares

Model

1

HPLC

WATERS Alliance 2695 separation module, Software: Empower 2, 996 PDA detector.

2

pH meter

Lab India

3

Weighing machine

Sartorius

4

Volumetric flasks

Himedia

5

Pipettes and Burettes

LACHOI

6

Beakers

PYREX

7

Digital ultra sonicate

Enertech

 

Chemicals Used:

Table2: Chemicals used

S. No.

Chemical

Brand Names

1

Telmisartan

Sura labs

2

Chlorthalidone

Sura labs

3

Water and Methanol for HPLC

Lichrosolv (MERCK)

4

Acetonitrile for HPLC

Merck

5

Triethylamine

Merck

 

HPLC METHOD DEVELOPMENT:

Preparation of standard solution:

Accurately weigh and transfer 10 mg of Telmisartan and Chlorthalidone working standard into a 10ml of clean dry volumetric flasks add about 7ml of Methanol and sonicate to dissolve and removal of air completely and make volume up to the mark with the same Methanol.

 

Preparation of Buffer and Mobile Phase:

Preparation of Triethylamine (TEA) buffer (pH-4.2):

Dissolve 1.5ml of Triethyl amine in 250ml HPLC water and adjust the pH-4.2. Filter and sonicate the solution by vacuum filtration and ultra sonication.

 

Preparation of mobile phase:

Accurately measured 400ml (40%) of Acetonitrile and 600ml of buffer (60%) a were mixed and degassed in digital ultra sonicate for 15 minutes and then filtered through 0.45µm filter under vacuum filtration.

 

Diluent Preparation:

The Mobile phase was used as the diluent.

 

RESULTS AND DISCUSSION:

Optimized Chromatogram (Sample)

 

Fig:1: Optimized Chromatogram (Sample)

 

Table 3: Optimized Chromatogram (Sample)

S. No.

Peak name

Rt

Area

Height

USP Resolution

USP Tailing

USP plate count

1

Telmisartan

2.256

775674

13124

3.12

0.93

7165.0

2

Chlorthalidone

5.456

2658478

937405

4.06

1.24

7458.0

 

Validation:

Table 4: Results of system suitability for Telmisartan

S. No.

Name

Rt

Area

Height

USP plate count

USP Tailing

1

Telmisartan

2.247

764566

69587

5434

1.4

2

Telmisartan

2.323

764532

69854

5567

1.3

3

Telmisartan

2.321

754677

70211

4657

1.2

4

Telmisartan

2.252

765928

69213

5345

1.4

5

Telmisartan

2.248

765426

69558

5432

1.5

Mean

 

 

765855

 

 

 

Std. Dev

 

 

366.652

 

 

 

% RSD

 

 

1.06093

 

 

 

 

Table 5: - Results of system suitability for Chlorthalidone

S. No.

Name

Rt

Area

Height

USP plate count

USP Tailing

USP Resolution

1

Chlorthalidone

5.345

2545671

190010

5366

1.1

2.04

2

Chlorthalidone

5.432

2343685

190052

5347

1.2

2.05

3

Chlorthalidone

5.345

2525875

190324

5388

1.3

2.01

4

Chlorthalidone

5.482

2531564

190035

5340

1.4

2.02

5

Chlorthalidone

5.495

2533214

190085

5360

1.5

2.03

Mean

 

 

2531034

 

 

 

 

S.D

 

 

1183.301

 

 

 

 

% RSD

 

 

0.04664

 

 

 

 

 

Limit of Detection:

Result:

Telmisartan: 0.56µg/ml

Chlorthalidone: 1.7µg/ml

Limit of Quantitation:

Result: Telmisartan: 1.2µg/ml

Chlorthalidone: 3.6µg/ml

 

Specificity:

Table 6: Peak results for Assay sample

S. No.

Name

Rt

Area

Height

USP Resolution

USP Tailing

USP plate count

Injection

1

Telmisartan

2.247

756985

689342

 

0.98

7276

1

2

Chlorthalidone

5.432

2569856

196543

2.04

1.23

8863

2

3

Telmisartan

2.212

758745

698367

 

1.05

6503

3

4

Chlorthalidone

5.234

2598654

195678

2.03

0.99

7260

2

5

Telmisartan

2.543

7563451

69324

 

1.7

7589

2

6

Chlorthalidone

5.234

256783

193425

2.02

1.6

8323

3

 

Linearity:

Telmisartan:

Table 7: Results for linearity

Concentration mg/ml

Average Peak Area

30

54676

40

67698

50

84431

60

10102

70

11213

 

Table 8: - Results for linearity

Concentration (mg/ml)

Average Peak Area

60

2245671

80

3045658

100

3861257

120

4723517

140

560458

 

Fig 2: Calibration graph for Telmisartan                                                  Fig 3: Calibration graph for Chlorthalidone

 

Repeatability:

Table 9: Results of Repeatability for Telmisartan:

S. No.

Name

Rt

area

height

USP count

USP Tailing

1

Telmisartan

2.269

766854

702543

5645

1.5

2

Telmisartan

2.234

765884

698567

5565

1.3

3

Telmisartan

2.213

765842

701231

5512

1.5

4

Telmisartan

2.265

768982

700123

5521

1.8

5

Telmisartan

2.261

765811

695671

5572

1.6

mean

 

 

766622

 

 

 

S. D

 

 

1357.912

 

 

 

RSD

 

 

0.177124

 

 

 

 

Table 10-: Results of method precision for Chlorthalidone:

S. No.

Name

Rt

area

Height

USP count

USP Tailing

1

Chlorthalidone

5.265

2565677

2232131

5365

1.5

2

Chlorthalidone

5.245

2578453

2645690

5425

1.6

3

Chlorthalidone

5.232

2556754

2223145

5368

1.5

4

Chlorthalidone

5.212

2584567

2421230

5359

1.5

5

Chlorthalidone

5.231

2545786

2323214

5498

1.6

mean

 

 

2571230

 

 

 

S. D

 

 

15309.45

 

 

 

  RSD

 

 

1.2595695

 

 

 

Accuracy:

Table 11: The accuracy results for Telmisartan

Concentration at specification Level

area

Amount Added ppm

Amount Found ppm

percentage Recovery

Mean Recovery

50%

42594.67

24

25.070

100.270%

100.04%

100%

84867

51

49.965

99.940%

150%

127654

65

75.164

100.28%

 

Table 12: - The accuracy results for Chlorthalidone

Concentration at specification Level

area

Amount Added ppm

Amount Found ppm

percentage Recovery

Percentage mean Recovery

50%

2079453

50

50.434

100.80%

100.46%

100%

4082234

102

100.456

100.71%

150%

607345

140

150.312

100.26%

 

Robustness

Table 13: - Telmisartan Results for Robustness

Parameter used for sample analysis

Peak Area

Retention Time

Theoretical plates

Tailing factor

Actual Flow rate of 1.0 ml/min

765456

2.231

5376

0.93

Less Flow rate of 0.9 ml/min

758346

2.541

5443

0.92

More Flow rate of 1.1 ml/min

7689545

2.032

5667

0.84

Less organic phase

7584132

2.512

5585

0.92

More organic phase

769762

2.031

5356

0.93

 

Table 14: - Chlorthalidone Results for Robustness

Parameter used for sample analysis

peak Area

Retention time

Theoretical plate

Tailing factor

Actual Flow rate of 1.0 ml/min

2532154

5.451

5378

1.24

less flow rate of 0.9 ml/min

2458623

5.589

5328

1.24

more Flow rate of 1.0 ml/min

2653561

4.564

5252

1.20

less organic phase

2452132

5.589

5213

1.21

more organic phase

2653874

4.584

5521

1.20

 

CONCLUSION:

A straightforward, sensitive, accurate and precise RP-HPLC method created for the current study in order to quantify the levels of chlorthalidone and telmisartan in pharmaceutical dosage forms and bulk drugs.in contrast to being nearly insoluble in water, telmisartan was found to be slightly soluble in ethanol, dichloromethane, toluene, benzene ethyl acetate methanol and acetone and dimethyl formamide. Almost insoluble in water and soluble in alcohol. TEA buffer ph-4.2(40:60 v/v) acetonitrile was selected as mobile phase. This method made use of an economical solvent system.

 

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Received on 09.03.2026      Revised on 14.04.2026

Accepted on 11.05.2026      Published on 10.07.2026

Available online from July 25, 2026

Asian Journal of Pharmaceutical Analysis. 2026; 16(3):167-170.

DOI: 10.52711/2231-5675.2026.00025

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