INTRODUCTION:

Desidustat is a hypoxia-inducible factor prolyl hydroxylase (HIF-PH) inhibitor that works by blocking prolyl hydroxylase domain enzymes. This action stabilizes hypoxia-inducible factors (HIF), particularly HIF-1α, which increases the expression of genes involved in red blood cell (RBC) production, iron transport, and erythropoietin (EPO) synthesis. Marketed as Oxemia™, it is an orally active drug developed by Zydus Cadila and is used to treat anemia associated with chronic kidney disease (CKD). Desidustat helps improve hemoglobin levels in affected patients.¹ Anemia in CKD is mainly caused by reduced EPO production and impaired iron metabolism, leading to poor quality of life and increased health risks. Conventional treatments include iron supplements and erythropoiesis-stimulating agents. Recently, HIF-PH inhibitors have emerged as a newer class of drugs that improve natural EPO production and regulate iron metabolism.^2,3

Drug Profile of Desidustat:

Drug Name: Desidustat

Chemical formula:

Category: Hypoxia-Inducible Factor Prolyl Hydroxylase Inhibitor (HIF-PHI)

Therapeutic Use: Treatment of anemia associated with chronic kidney disease (CKD)

Molecular Formula: C16H16N2O6

Molecular Weight: 332.31g/ml

Solubility: It is soluble in organic solvents such as ethanol, DMSO, and Dimethyl Formamide (DMF), and is sparingly soluble in aqueous buffer.

Melting Point: 211–214°C.

pKa Value: 3.17.

Pharmacological Class:

Desidustat belongs to a new class of drugs known as hypoxia-inducible factor prolyl hydroxylase inhibitors (HIF-PHIs). These agents stimulate the body’s natural response to low oxygen levels (hypoxia).⁶

Pharmacodynamics:

Desidustat decreases hepcidin levels and increases erythropoietin (EPO), serum iron, hematocrit, and hemoglobin levels. It also increases reticulocyte and red blood cell (RBC) counts in normal and nephrectomies rats and in rodent models of chemotherapy- or inflammation-induced anemia. In healthy volunteers, Desidustat increases hemoglobin and EPO levels. In patients with non-dialysis-dependent chronic kidney disease (CKD), it decreases hepcidin levels to a greater extent than darbepoetin alfa. In patients with dialysis-dependent CKD, it reduces hepcidin levels to a similar extent as epoetin alfa.

Pharmacokinetics:

The time to reach peak plasma concentration (Tmax) is approximately 2.5hours after a single dose of 50–150mg in dialysis-dependent patients. No drug accumulation has been observed after multiple dose administrations. The mean elimination half-life is approximately 6–15 hours after a single 50–150mg dose in dialysis-dependent CKD patients and 6–14hours after multiple doses of 100–200 mg administered on alternate days for six weeks in pre-dialysis CKD patients.

Mechanism of Action:

Desidustat works by inhibiting the enzyme prolyl hydroxylase, which leads to stabilization of hypoxia-inducible factor (HIF). Stabilized HIF increases endogenous erythropoietin production, enhances iron absorption, and improves iron utilization, resulting in increased red blood cell formation and correction of anemia.^4,5

Different chromatography methods for drug analysis:

1) High-Performance Thin-Layer Chromatography (HPTLC):

HPTLC is extensively used for the identification, estimation, and profiling of pharmaceutical compounds. This advanced technique offers rapid separation and flexibility, making it suitable for analyzing multiple drug components. It provides advantages such as quick analysis, simple handling, minimal sample preparation, and the ability to evaluate chromatograms for multiple parameters.^6,7

2) High-Performance Liquid Chromatography (HPLC): -High-Performance Liquid Chromatography is a fundamental technique for separating complex mixtures of chemical and biological substances. Introduced widely in the 1980s, HPLC became a standard method for bulk drug analysis according to USP guidelines. It ensures high accuracy and precision across various sample types. Typically, a UV detector is employed for sample estimation after appropriate wavelength scanning.^8,9

3) Thin-Layer Chromatography (TLC): -Thin-layer chromatography is a conventional technique used in pharmaceutical analysis. It operates using a stationary phase (usually silica gel coated on glass or aluminum plates) and a mobile phase. TLC is applied for analyzing both organic and inorganic substances. It is preferred due to its simplicity, low cost, flexible mobile phase selection, minimal maintenance, and suitability for bulk drug analysis.^10,11

4) Gas Chromatography (GC): -Gas chromatography is mainly used for separating volatile and organic compounds. It enables quantitative determination of trace-level components, including impurities present in parts per trillion. GC plays a crucial role in pharmaceutical drug analysis and contamination detection.^12,13

Method validation:

The process of analytical method validation is carried out to confirm that the developed chromatography method is suitable for the accurate and precise estimation of a drug in bulk and pharmaceutical dosage forms. Validation is performed according to ICH guidelines and involves the following steps:¹⁴

Validation Parameters:

1) Specificity: The ability of the RP-HPLC method to accurately measure Desidustat in the presence of excipients, impurities, or degradation products without any interference.

2) Linearity: The ability of the method to produce results that are directly proportional to the concentration of Desidustat over a specified range.

3) Accuracy: The closeness of the test results to the true value, usually determined by recovery studies at different concentration levels.

4) Precision: The degree of agreement among individual test results when the method is applied repeatedly, expressed as intra-day and inter-day precision.

5) Range: The interval between the upper and lower concentration levels of Desidustat for which the method has been demonstrated to be accurate, precise, and linear.

6) Limit of Detection (LOD): The lowest amount of Desidustat that can be detected but not necessarily quantified under the stated experimental conditions.

7) Limit of Quantification (LOQ): The lowest amount of Desidustat that can be quantitatively determined with acceptable accuracy and precision.

8) Robustness: The ability of the method to remain unaffected by small deliberate changes in analytical conditions such as flow rate, mobile phase composition, or detection wavelength.

⁹⁾System Suitability: Tests performed to ensure that the chromatographic system is functioning properly before analysis, including parameters such as retention time, theoretical plates, and tailing factor.¹⁵

Reported Analytical Methods on Desidustat:

1) Dr. S. Naazneen et al. (2023): A validated RP-HPLC method was developed for the determination of Desidustat in pharmaceutical dosage forms. The best separation was achieved using an Inertsil® Octadecylsilyl-3V reverse-phase C18 column (250mm × 4.6mm i.d., 5µm particle size) with acetonitrile as the organic solvent. The mobile phase consisted of 0.03M potassium dihydrogen orthophosphate in water (pH 3.2, adjusted with orthophosphoric acid) and acetonitrile in the ratio of 55:45. The chromatographic separation was carried out in isocratic mode at a flow rate of 0.8 mL/min. UV detection was performed at 246nm. The retention time of Desidustat was found to be 4.67 minutes. The method showed good linearity with a correlation coefficient of approximately 0.999 over the concentration range of 40–120µg/mL. The percentage assay of Desidustat was found to be 98.6%. The limit of detection (LOD) and limit of quantification (LOQ) for Desidustat were 0.05µg/mL and 0.15µg/mL, respectively.¹⁶

Table No.1: RP-HPLC condition of Desidustat

Parameters	Description
Column name	Inertsil ODS C-18-3V (250 x 4.6mm, 5µm particle size)
Mobile phase	Orthophosphoric acid: Acetonitrile (55:45 v/v)
Flow rate	0.8ml/ min
Detection wavelength	Photo diode array (PDA) Wavelength (λmax): 246 nm
Retention time	4.7 mins
Limit-of-Detection (LOD)	0.05µg/mL
Limit-of-Quantification (LOQ)	0.15µg/mL
Assay-in-Percentage (%)	98.6%
Percentage of peak areas	99.74
Resolution	3.89

2) Ameya Lanjewar et al. (2024):

An RP-HPLC method was developed and validated for the assay of Desidustat in pharmaceutical formulations. Desidustat is an orally bioavailable hypoxia-inducible factor prolyl hydroxylase inhibitor used for the treatment of anemia associated with chronic kidney disease. An accurate, precise, and robust reverse-phase HPLC method was developed and validated according to ICH guidelines. Chromatographic separation was carried out using an Eclipse XDB C18 analytical column (150 × 4.6 mm, 5µm particle size). The mobile phase consisted of 0.1% orthophosphoric acid and methanol in the ratio of 38:72, with a flow rate of 1.0mL/min. Detection was performed at a wavelength of 235nm. The developed method was found to be suitable for the development, validation, and assay of Desidustat in pharmaceutical formulations.¹⁷

Table No.2: RP-HPLC condition of Desidustat

Parameters	Description
Column	Eclipse XDB C18 column [(150x4.6) mm i.d, 5µm]
Mobile phase	Methanol :0.1% ortho phosphoric acid (72:38)
Flow rate	1 ml/min
Detection wavelength	235 nm
Retention time	4.75min
Sample size	10 µl

3) Rutvik Pandya et al. (2023):

A reversed-phase HPLC-PDA method was developed for the quantification of Desidustat. The method was optimized using a Hypersil C18 column (250 × 4.6mm, 5µm) as the stationary phase. The mobile phase consisted of methanol and acetonitrile in the ratio of 80:20 (v/v), which was delivered at a flow rate of 1 mL/min. Detection of Desidustat was carried out at an analytical wavelength of 230nm using a photodiode array (PDA) detector. The developed method showed good linearity over the concentration range of 1–6 µg/mL, with a correlation coefficient of 0.9989.¹⁸

Table No.3: HPLC-PDA condition of Desidustat

Parameters	Description
Column	Hypersil C18 (250 × 4.6mm, 5µm)
Mobile phase	methanol: acetonitrile (80:20 v/v)
Detection wavelength	230 nm
Retention Time	3.02 min
Detection Limit (µg/mL)	0.04
Quantitation Limit (µg/mL)	0.12
Assay (%)	99.04
Flow rate	1 mL/min

4) Jaivik Prajapati Mansukhbhai et al. (2023):

A stability-indicating RP-HPLC method was developed and validated for the analysis of Desidustat in tablet dosage form. The method was validated according to ICH guidelines over a concentration range of 3–7ppm. Chromatographic separation was carried out using a Hypersil BDS C18 analytical column (25cm × 0.46cm). The mobile phase consisted of methanol and acetonitrile in the ratio of 70:30 (v/v), delivered at a flow rate of 1.0 mL/min. Detection was performed using an SPD-20A detector at a wavelength of 315nm. The retention time of Desidustat was found to be 9.520 minutes. The method demonstrated acceptable accuracy, precision, specificity, sensitivity, and selectivity in accordance with ICH guideline Q1A (R2).¹⁹

Table No.4: Stability-indicating condition of Desidustat

Parameters	Description
Column	C18 (25cm × 0.46 cm) Hypersil BDS
Mobile phase	Methanol: Acetonitrile (70:30 % v/v)
Detection Wavelength	315 nm
Flow rate	1.0 ml/min
Retention time	9.520 min.
Melting point	211-213°C
Limit of detection (LOD)	0.165 ppm
Limit of quantification	0.539 ppm

5) Ameya Lanjewar et al. (2024):

An RP-HPLC analytical method was developed for the forced degradation study of Desidustat. Chromatographic separation was carried out using an Eclipse XDB C18 analytical column (150 × 4.6mm, 5 µm particle size). The mobile phase consisted of methanol and 0.1% orthophosphoric acid in the ratio of 72:38(v/v), delivered at a flow rate of 1mL/min. Detection was performed at a wavelength of 235nm. The retention time of Desidustat was found to be approximately 5 minutes.²⁰

Table No.5: RP-HPLC condition for forced Degradation of Desidustat

Parameters	Description
Column	C18 (150X4.6mm, 5µ) eclipse SDB
Mobile phase	methanol0.1% orthophosphoric acid (72% :38% v/v)
Flow rate	1ml/min.
Detection wavelength	235nm
Retention time	5min

6) Riya Patel et al. (2023):

A stability-indicating RP-HPLC method was developed and validated for the estimation of Desidustat in tablet dosage form. Chromatographic separation was performed using a Thermo Fisher C18 column (150mm × 4.6mm, 5µm particle size) with a flow rate of 1.0 mL/min. The mobile phase consisted of acetonitrile (ACN) and 0.1% formic acid in the ratio of 80:20 (v/v). The injection volume was 20µL, and the total run time was 15 minutes. Detection was carried out at a wavelength of 235nm. The stability-indicating capability of the method was established through forced degradation studies of both bulk drug and pharmaceutical dosage forms. The stability study indicated that major degradation of the bulk drug was observed under thermal conditions, while degradation in the formulation was observed under oxidative conditions. The correlation coefficient for the calibration curve of Desidustat was found to be 0.999. The %RSD for precision and robustness was found to be less than 2%, indicating good reliability of the method.¹

Table No.6: Stability-indicating RP-HPLC of Desidustat

Parameters	Description
Column	C18 (150mm*4.6mm)
Mobile phase	ACN + 0.1% Formic Acid (80:20%v/v)
Flow rate	1.0ml/min
Detection wavelength	235nm
Retention time	5.8 min
Limit of detection (LOD)	1.607
Limit of quantification (LOQ)	4.872
Melting point	213-216°C.
Assay (%)	99.859

CONCLUSION:

This systematic review provides a comprehensive overview of the analytical methods developed and validated for the estimation of Desidustat in bulk drug and pharmaceutical dosage forms. The findings reveal that Reverse Phase High Performance Liquid Chromatography (RP-HPLC) is the most widely employed and reliable technique due to its high sensitivity, specificity, precision, and reproducibility. UV–Visible spectrophotometric methods have also been reported as simple and cost-effective alternatives for routine analysis. Most of the developed methods comply with ICH validation guidelines, demonstrating acceptable linearity, accuracy, precision, robustness, limit of detection (LOD), and limit of quantification (LOQ). Stability-indicating methods incorporating forced degradation studies under various stress conditions confirm the ability to effectively separate Desidustat from its degradation products. Overall, the reviewed literature highlights significant progress in establishing validated and regulatory-compliant analytical procedures. However, further research is encouraged to develop greener, more economical, and advanced analytical techniques to enhance sustainability and efficiency in the pharmaceutical analysis of Desidustat.

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Received on 04.04.2026 Revised on 06.05.2026

Accepted on 03.06.2026 Published on 10.07.2026

Available online from July 25, 2026

Asian Journal of Pharmaceutical Analysis. 2026; 16(3):239-243.

DOI: 10.52711/2231-5675.2026.00037

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License. Creative Commons License.