A Review on Analytical Method Development and Validation for Simultaneous Estimation of Perindopril and Indapamide in Bulk and Pharmaceutical Dosage Form by RP-HPLC
- Banothu Bhadru
- Dasam Bhagya Lakshmi
- Tadikonda Rama Rao
- 122-126
- Jun 28, 2025
- Health
A Review on Analytical Method Development and Validation for Simultaneous Estimation of Perindopril and Indapamide in Bulk and Pharmaceutical Dosage Form by RP-HPLC
1Banothu Bhadru*, 2Dasam Bhagya Lakshmi, 2Tadikonda Rama Rao
1Associate Professor, Department of Pharmaceutical Analysis CMR College of Pharmacy, Kandlakoya Medchal, Hyderabad, Telangana, India-501401
2CMR College of Pharmacy, Kandlakoya, Medchal, Hyderabad, Telangana, India-501401
*Corresponding Authors
DOI: https://doi.org/10.51584/IJRIAS.2025.10060010
Received: 09 June 2025; Accepted: 13 June 2025; Published: 28 June 2025
ABSTRACT
Perindopril and Indapamide, a fixed-dose combination medication, are utilized for the management of hypertension by integrating the properties of an angiotensin-converting enzyme (ACE) inhibitor with a thiazide-like diuretic. The detection of Indapamide, Perindopril, and its active metabolite perindoprilat in human plasma or whole blood was achieved through hyphenated ultra-performance liquid chromatography-mass spectrometry (UPLC-MS/MS). Internal standards included indapamide-d3, perindopril-d4, and perindoprilat-d4. The chromatography was conducted at a column temperature of 50°C with a flow rate of 0.6 mL/min using isocratic elution, and UV detection was performed at 215 nm. The method demonstrated acceptable linearity, accuracy, and precision across concentration ranges of 1-5μg/mL for Perindopril and 10-15μg/mL for Indapamide. A statistical comparison of the proposed chromatographic method against reference methods was conducted using one-way analysis of variance, with Beer-Lambert’s range established at 0-60 μg/mL.
The most commonly used mobile phases are methanol and acetonitrile, whereas the stationary phase used is a Phenomenex Luna C18 column (250 mm × 4.6 mm, 10 µm). Various parameters are collected, including LOD, LOQ, correlation coefficient and linearity, absorbance maxima, and retention time, which were within the limits specified by the ICH guidelines.
Keywords: RP–HPLC, Perindopril, Indapamide.
INTRODUCTION
Perindopril, also known chemically as (2S,3aS,7aS)-1-[(2S)-2-[[(2S)-1-ethoxy-1-oxopentan-2-yl]amino]propanoyl]-2,3,3a,4,5,6,7,7a-octahydroindole-2-carboxylic acid (fig. no. 1), is the active ingredient of an angiotensin-converting enzyme inhibitor (ACE-I). In addition, heart attacks, strokes, and kidney problems are among the various illnesses that can be treated with this1. It is poorly soluble in methylene chloride but easily soluble in water and 96% ethanol.
Fig No. 1: Structure of Perindopril
Indapamide, also known chemically as 4-chloro-N-(2-methyl-2, 3-dihydroindol-1-yl)-3-sulfamoylbenzamide (fig. no. 2), is a thiazide-like diuretic that has become a standard treatment for edema and hypertension associated with various cardiovascular diseases2. It reduces blood pressure and increases urine output by blocking the reabsorption of sodium by the kidneys distal convoluted tubules3. Water-insoluble, it dissolves in 96% ethanol, methanol, acetic acid, and ethyl acetate, and it dissolves very slightly in ether, chloroform, and benzene4.
Fig. No. 2: Structure of Indapamide
Table.No.1: Analytical methods for Perindopril and Indapamide
| S.No. | Author Name | Title | Name of the Journal | Chromatographic Conditions | Results |
| 1 | Jain P s5
|
RP-HPLC Assay Method for Simultaneous Determination of Perindopril Erbumine and Indapamide Combination in Bulk and Tablet Dosage Form | Journal of Pharmaceutical Research,2012
|
Mobile Phase:Acetonitrile: Buffer pH 2.8 (40: 60 % v/v) Stationary phase: Phenomenex Luna C18 column (250mm × 4.6mm, 10µm)λmax- 225nm Flow rate-1 .2ml/min | Perindopril
Rt – 3.150min LOD- 0.120 µg/ml LOQ-0.365 µg/ml Indapamide Rt – 10 min LOD- 0.606µg/ml LOQ-1.8µg/ml R.S.D= <2.0% |
| 2 | P.S.R.CH.N.P. Varma D6
|
Validated Stability Indicating Reverse Phase HPLC Method for The Simultaneous Estimation of Perindopril and Indapamide in Pharmaceutical Dosage Forms | International Journal of Pharmacy,2013
|
Mobile phase:
methanol = 65:35 v/v pH 3.5±0.05 Stationary phase:Hypersil BDS C18 column λmax- 215nm Flow rate-1 ml/min |
Perindopril
Rt – 3.53min LOD- 0.120 µg/ml LOQ-0.365 µg/ml Indapamide Rt – 4.09 min LOD- 0.606µg/ml LOQ-1.839 µg/ml Linearity range 160 to 480 µg/mL |
| 3 | B. Siva Sai Kiran7
|
Stability Indicating isocratic RP-HPLC Method Development and Validation for Indapamide and Perindopril Erbumine in pure and its combined tablet dosage Form | International Journal of Pharmaceutical Sciences and Research,2014
|
Mobile phase: pH 2.5 acetonitrile :60:40 v/v
Stationary phase: YMC Column (150 x 4.6mm, 3µ particle size) λmax- 230nm Flow rate-0.8ml/min
|
Perindopril
Rt -1.5min LOD- 0.07 µg/ml LOQ-0.23 µg/ml Indapamide Rt – 0.5 min LOD-2.8µg/ml LOQ-8.48 µg/ml |
| 4 | Kirtan P. Patel8 | A new RP–HPLC method for simultaneous quantification of perindopril erbumine, indapamide, and amlodipine besylate in bulk and pharmaceutical dosage form | Future Journal of Pharmaceutical Sciences,2020
|
Mobile phase: acetonitrile: methanol: water (30:20:50, v/v/v)
Stationary phase:Phenomenex C-18 column (250 mm × 4.6 mm, 5 μm) pH 3.0 λmax- 215nm Flow rate-1 ml/min |
Perindopril
linearity range 0.400–7.600 μg/ml LOD- 0.065 µg/ml LOQ-0.198 µg/ml Indapamide LOD- 0.019µg/ml LOQ-0.056µg/ml |
| 5 | Naser F AlTannak9 | UHPLC-UV Method for Simultaneous Determination of Perindopril Arginine and Indapamide Hemihydrates in Combined Dosage Form: A Stability-Indicating Assay Method | Scientia Pharmaceutica, 2018 | Mobile phase:0.01% v/v formic acid in water
Stationary phase: BEH C18 (1.7 μm, 2.1 × 50 mm) λmax– 227nm Flow rate -0.3 ml/min. |
Perindopril& Indapamide
linearity range (20–450 g/mL) LOD- 3:1 LOQ-10:1 Relative Standard Deviation (%RSD) -0.63% |
| 6 | Yi Tao10 | Simultaneous determination of indapamide, perindopril and perindoprilat in human plasma or whole blood by UPLC-MS/MS and its pharmacokinetic application | Journal of Pharmaceutical Analysis,2018 | Mobile phase: Thermo BDS Hypersil
Stationary phase: C18 column (4.6mm 100mm, 2.4mm) |
Perindopril
Linearity range 0.2–20ng/ml Indapamide Linearity range 1–250ng/ml |
| 7 | Pawar Seemarani11 | Development and Validation of UV Spectrophotometric Estimation of Perindopril Erbumine and Indapamide in Bulk and Tablet Dosage by using Area Under Curve Method | London Journal of Medical and HealthResearch,2023 | Perindopril
λmax-208-214 nm Indapamide λmax- 239-244 nm |
Perindopril
Concentration range 1-5 µg/ml Indapamide Concentration range 10-50 µg/ml Recovery study 99-101% |
| 8 | Erica Alves12 | Development and Validation of a Novel UV Spectrophotometric Method for Simultaneous Analysis of Amlodipine, Indapamide and Perindopril | Indian journal of pharmaceutical analysis,2020 | Perindopril
λmax=204nm Indapamide λmax=245 nm Beer Lambert’s range0-60 μg/ml |
Perindopril
LOD- 1.860µg/ml LOQ-5.60µg/ml Indapamide LOD-0.630µg/ml LOQ-1.920 µg/ml Correlation coefficient (r2) of ≥0.999. |
| 9 | John Chalmers13 | Effects of Combination of Perindopril, Indapamide, and Calcium Channel Blockers in Patients with Type 2 Diabetes Mellitus | Ahajournals, 2025 | perindopril–indapamide (4/1.25 mg)
CCB at baseline compared with 5% (−12% to 20%) |
CCB at baseline compared with 5% (−12% to 20%)among those without CCB (P homogeneity 0.02) |
| 10 | Xin Zheng14 | Simultaneous determination of indapamide, perindopril and its active metabolite perindoprilat in human plasma using UPLC-MS/MS method | Journal of Chromatography B, 2021 | Stationary phase:
X-terra MS C18 (2.1 mm × 150 mm, 3.5 μm) with isocratic elution Linearity range: 0.250-50.0 ng/mL |
Perindopril
ion transitions at m/z 369.165 → 172.0 Indapamide 366.010 → 132.100 |
| 11 | Nevin Erk15 | Comparison of Spectrophotometric and an LC method for the determination perindopril and indapamide in pharmaceutical formulations | Journal of Pharmaceutical and Biomedical Analysis, 2001 | Mobile Phase:
phosphate buffer pH 2.4 and acetonitrile (7:3 v/v) Linearity range for Perindopril and Indapamide was 5.0–70.0 and 8.0–35.0 μg ml−1 |
Linear calibration graphs of first derivative values at 225.7 and 255.4 nm for Perindopril and Indapamide |
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