Analytical Method Development and Validation Methods for Estimation of Drugs in Pharmaceutical Dosage Forms: A Review

Dr. Shiv Bhadra Singh; Syed Tahir Hussain Rizvi

doi:10.5281/zenodo.20117089

Review Paper | Open Access
Volume 04 | Issue 05 | Article Id IJPS/260404834

Analytical Method Development and Validation Methods for Estimation of Drugs in Pharmaceutical Dosage Forms: A Review
Dr. Shiv Bhadra Singh* Syed Tahir Hussain Rizvi
Department of Pharmaceutical Chemistry, Hygia Institute of Pharmaceutical Education and Research

Abstract

Estimating medicines in pharmaceutical dosage forms has significance for quality control and regulatory compliance. The establishment of reliable, precise and cost-effective analytical procedures is critical to ensuring both the security and efficiency of single and multiple- component compositions of drugs. The review mainly focuses on RP-HPLC techniques emphasizing the method validation as per ICH guidelines parameters and method development parameters. The review presents a relative and comparative evaluation of reported methods in terms of sensitivity, accuracy, precision and cost-effectiveness. Various categories of drugs including cholinergic drugs, anticholinergic drugs, neuromuscular blocking agents, adrenergic agents, antiadrenergic agents, antihistaminic & proton pump inhibitors, migraine drugs, NSAIDs & antipyretics, anti-rheumatoid and antigout are discussed with emphasis on their method optimization parameters such as s wavelength, flow rate, retention duration, mobile phase, and stationary phase. The assessment of reported methods is offered in tabular form to help with the selection of appropriate analytical techniques for standard quality analysis. In order to help researchers and analysts choose and create trustworthy analytical techniques for regular quality control and regulatory compliance, this review has come together to offer a streamlined resource.

Keywords

Method development, RP-HPLC, ICH guidelines, Method validation, Analytical Techniques, Method optimization

Introduction

Chromatography is a collection of methods for separating mixed substances by continuously distributing them between two phases, one of which moves in relation to the other. Chromatography is arguably the determiner analytical method for modern analysts. This technique is able to determine quantitatively numerous distinct elements existing in a blend with just one analytical technique. Differential rates of elution are used in chromatography to resolve solutes as they go through a chromatographic column [1]. Drug discovery, method development, and the manufacturing of different pharmaceutical dosage forms all depend heavily on analytical technique development and validation. These techniques are used to verify the identification, efficacy, transparency, and safety of pharmaceutical product. Separating and calculating the primary active drugs, any response impurities all possible synthesis intermediate and, any degradants are the objective of the HPLC process [2]. One of the most crucial instruments in analytical chemistry today is HPLC, which developed from conventional column chromatography. HPLC is an significant analytical method used in every phase for drug development like research, manufacturing etc. in the contemporary pharmaceutical industry. It is the method of choice for figuring out how pure novel chemical entities can be, monitoring changes in reactions during synthesis or scaling up processes, evaluating innovative formulations, and carrying out quality control and assurance on the final drugs [3]. The primary goals of establishing analytical techniques are the identification, purification, and final quantification of any necessary drugs, etc. The separation and characterisation of impurities and degradation products, analytical investigations, identification studies, and, lastly, parameter optimisation to particular requirements are the primary tasks involved in the analytical development of a technique. Therefore, an analyst can greatly benefit from the fundamental concepts presented in the review article above when estimating pharmaceutical formulations and bulk medications [4].

An important step in analytical chemistry is the development of RP-HPLC methods, which entail optimising many parameters to separate and quantify analytes in complicated mixtures as needed. Polar and non-polar compounds are separated using the well-known chromatographic technique known as RP-HPLC [5]. Reversed phase chromatography can separate molecules with a high degree of recovery and resolution if they have some hydrophobic characteristics. In RP-HPLC, the separation process is driven by the interaction of the stationary phase which is immobilised hydrophobic binders interact hydrophobically to the solute particles in mobile phase. Although there is much disagreement on the precise mechanism, most people believe that a positive entropy effect is responsible for the hydrophobic binding contact. According to reversed phase chromatography, the solute particles and immobilized binder are enclosed in a highly organized water structure due to the mostly aqueous nature of the initial mobile phase binding conditions [6]. The duration of the time taken for the sample to get across the column and get to the detector is also considered as the retention time of the certain material. As part of the HPLC process, a sample consisting of a combination of chemicals is run over a chromatographic column. An adsorbent material covers a packing material, such as silica particles, in this column. Different components of the sample will interact with the coating in different ways, causing the sample to flow along the column at varied speeds (7). RP-HPLC has recently shrunk, new stationary and mobile phases have been developed, and hyphenated techniques like LC-MS, which are shown in Figure No. 1, have been introduced.

Figure No.1. RP-HPLC Advancement

Newly developed stationary phases and mobile phases with improved selectivity and efficiency may enable more accurate separation and analysis of complicated substances. Smaller sample volumes can be analysed by miniaturised RP-HPLC, which shortens run times and uses less solvent. Hybrid approaches offer more precise and sensitive analyte detection by fusing the advances of mass spectrometry and RP-HPLC [7]. The word "method development" is broad. A necessary step in both quantitative and qualitative analysis is to design a novel method for either estimating the amount of material or verifying the presence of the necessary component. Validation is the process of determining the method's parameters. It also aids by highlighting the method's limitations and their scope. For any medicinal component to be beneficially utilised in any pharmaceutical sector, both method development and validation must be developed and established [8].

"Green chemistry" refers to chemistry-related practices and processes that facilitate the removal or bargain of the use and exposure to the substances that pose a risk to the human health or environment. Examples of acute and long-term toxicity hazards from chemicals include health and environmental effects. Green technology ideally refers to avoiding poisonous or hazardous solvents or reagents during analytical techniques, using safe, energy-efficient equipment and devices to complete analyses quickly. Owed to the relatively low cost of the used mobile phase and the relative accessibility of the HPLC-UV equipment in many laboratories, particularly those that specialise in quality control, the HPLC technique stayed evaluated as an inexpensive procedure overall. Additionally, the equipment is reasonably priced when compared to methods like UPLC and LC-MS that help produce sensitive and highly accurate results. It is crucial to emphasise that the method's short chromatographic run time facilitates the routine procedures of analysing several samples, one element at a time, for the ensuing optimisation of the created HPLC approach [9].

Factor influencing the development of RP-HPLC

An RP-HPLC procedure's performance could be impacted by a number of important issues. The following factors can affect the quantification and partitioning evaluations of complicated mixtures.

Table 1. Factors Affecting Method Development of RP-HPLC

Factor	Description	Impact
Stationary Phase	Column Type	Determines Analyte Resolution [10]
Mobile Phase	Organic Solvent (e.g. acetonitrile, menthol) buffer type/pH, gradient/isocratic mode	Influence elution time and peak shape [11]
Column Temperature	25-50°C; impact viscosity and selectivity	Improves Reproducibility [12]
Flow Rate	Classically, 0.5-2 mL/min; affects analysis time and pressure	Balances speed and efficiency [10]
Detection Wavelength	UV-Vis based on analyte absorbance maxima	Enhances sensitivity [13]

Table 2. Factor Affecting Validation [14], [15], [16]

Factor	Description	ICH Parameter
Specificity	Ability to separate analysed from impurities/degradants	Ensures no interferences
Linearity	Correlation between concentration and response over range	Range: 50-150 % of target
Precision	Inter-day accuracy and intra-day repeatability	%RSD <2% typically
Accuracy	Recovery studies at different levels.	98-102% recovery
Robustness	Tolerance to minor changes in limitations (e.g. flow ±0.1 ml/min).	Confirms method reliability
LOD/LOQ	Lowest detectable/quantifiable concentrations.	Signal-to-noise ratios

This review article's main objective is to gather various analytical method development and validation methodologies that have been published for the estimate of a single drug or the estimation of many drugs at once and consolidate them for a thorough overview. The review primarily concentrates on RP-HPLC procedures, with an emphasis on method development parameters and validation in accordance with ICH criteria. The review presents a relative and comparative evaluation of reported methods regarding cost-effectiveness, sensitivity, accuracy, and precision. In order that help researchers and analysts choose and create trustworthy analytical techniques for regular quality control and regulatory compliance, this review has come together to offer a streamlined resource.

Cholinergic Drugs

Cholinergic medicines target acetylcholine, the primary transporter of the PNS. These medications are classified into two types: indirect and direct acting. Direct cholinergic agonists activate muscarinic receptors directly like choline esters (carbachol, bethanechol etc.) and alkaloids (cevimeline, pilocarpine). Indirectly cholinergic medicines upsurge the accessibility of ACH at cholinergic receptors like neostigmine [17]. Figure 2 depicts the structures of pharmaceuticals for which the validation and method development are elaborated in Table 3.

Table 3. Method validation and development of Cholinergic Agents

S. No	Drugs	Phase Stationary	Phase Mobile	Rate of flow (ml/min)	Wavelength (nm)	Ref.
1.	Pilocarpine in Pilocarpus microphyllus (Rutaceae) (1a)	RP-18 column (250 x 4.6 x 5 µm id)	Phosphoric Acid, MeOH and Triethylamine	1.0	215	[18]
2.	Bethanechol (1b)	Phenyl Column	0.05M ethanol-phosphate buffer (pH 6) combination with sodium 1-heptanesulfonate (98:2 v/v)	1.0	190	[19]
3.	Neostigmine (1c) and Methylsulfate	Kromasil C₁₈Column	Phosphate buffer: Acetonitrile (10:90)	1.0	215	[20]
4.	Glycopyrrolate (1d) and Neostigmine	Chromolith High Resolution RP-18e (4.6 × 100mm)	Buffer Solution (pH 3.0) A: Acetonitrile and water (90:10) Mode: Gradient	0.5	220	[21]

Figure No. 2. Structure of Cholinergic Drugs

Anticholinergic Drugs

The neurotransmitter acetylcholine is prevented from being used by the brain by a class of pharmaceuticals called as anticholinergics or anticholinergic drugs. For a variety of conditions, including allergies, Parkinson's disease, depression, heart disease, urine incontinence, and asthma, anticholinergic drugs are often prescribed to the elderly. Acetylcholine is a neurotransmitter that produces cholinergic effects by binding to muscarinic receptors in the CNS and peripheral tissues. The use of anticholinergic drugs has been associated with reduced purposeful and cognitive act, lower worth of lifetime, worsened Activities of Daily Living (ADL), and cognitive weakening in elder adults [22]. Figure 3 depicts the pharmaceutical method validation and method development which are elaborated in Table 4.

Table No.4 Method validation and method development of Anticholinergic Medication

S. No	Drugs	Phase Stationary	Phase Mobile	Rate of Flow (ml/min)	Wavelength (nm)	Ref
1.	Atropine Sulphate in Bulk Drugs (3a)	C₁₈ Column	Potassium dihydrogen phosphate buffer (50:50): Methanol: 5 mmol	1	264	[23]
2.	N-butyl bromide hyoscine and paracetamol (3b)	C₁₈Column (25 × 0.46cm 5µm particle size)	Methanol (50:50, v/v) pH Attuned to 3.9 with CF₃COOH acid	1	210	[24]
3.	Ipratropium Bromide (3c) with Glycopyrrolate	XDB-C₈ Column (150 × 4.6 mm, 3.5 µm)	MeOH: ACN: Trifluoracetic Acid: H₂O as A and 0.3% TFA in water	1.2	220	[25]

Figure No. 3. Structure of Cholinergic Drugs

Neuromuscular Blocking Drugs

The primary effects of neuromuscular blocking drugs (NMBDs) are as antagonists and agonists, although they too act at other locations at the neuromuscular junction. The only depolarising NMBD that is currently available is succinylcholine, which has a number of unfavourable side effects.
Similar to succinylcholine, less powerful non-depolarizing NMBDs function more quickly. Organ function is necessary for the metabolism and excretion of amino-steroid NMBDs.
Organ-independent breakdown occurs in benzylisoquinolinium compounds, although histamine is frequently released. To replace succinylcholine, a short-acting, non-depolarizing NMBD with a quick onset and short-lived DOA is needed [26]. Figure 5 contains the structure of the neuromuscular blocking drugs which are elaborated in the table 5.

Table No.5 Method development and validation of Neuromuscular Blocking Drugs

S. No	Drugs	Phase Stationary	Phase Mobile	Rate of flow	Wavelength	Ref
1.	Suxamethonium (4a)	C18 Column	Water at 100%	0.6 ml/min	218 nm	[27]
2.	Rocuronium Bromide (4b)	Inertsil Silica Ammonium Column	Solution A of Sodium Perchlorate: Solution B of (Ammonium chloride + Ammonium) 75:25	1.0 ml/min	215 nm	[28]
3.	Sugammadex (4c)	C₁₈ Column	Acetonitrile: Double Distilled Water (20:80 v/v%)	1.0 ml/min	210nm	[29]

Figure No. 4. Structure of Neuromuscular Blocking Agents

Adrenergic Drugs

The term "adrenergic medicines" refers to a wide range of drugs that attach to adrenergic receptors all over the body. These receptors include Alpha-1, Alpha-2, Beta-1, Beta-2, and Beta-3. Adrenergic medications have the variety of physiological effects by directly binding to one or more of these receptors. Certain drugs have specific effects by indirectly engaging with specific receptors. Adrenergic medications need to be categorised according to the particular receptors they bind. This article's main focus is on direct-acting medications like bronchodilators and vasopressors. Cocaine and amphetamines are two instances of indirect drugs (31). The structures of the adrenergic medications, which are detailed for pharmaceutical technique development and validation in Table 6, are displayed in Figure 5.

Table No.6 Method development and validation of Adrenergic Transmission

S. No	Drugs	Stationary Phase	Mobile Phase	Flow Rate	Wavelength	Ref
1.	Epinephrine (5a)	C₁₈ Column Luna Phenomenex	Combination of H₂O: Methanol: Acetic Acid (85:10:5), pH 3.1 with addition of Ammonium Acetate	1 ml/min	280 nm	[30]
2.	Zotepine (5b)	C₁₈G Column	Potassium Dihydrogen Phosphate (pH 3.0) and Ortho Phosphoric Acid: Acetonitrile (45:55)	1 ml/min	264 nm	[31]
3.	L. Broad Beans L-Dopa in Vicia Beans	C₁₈Column	0.2 % and 1% MeOH: 99 % Formic Acid	0.6 ml/min	264 nm	[32]
4.	Isoproterenol HCl (5d)	Phenomenex Luna Column	Methanol: 0.1% Triethylamine (pH 7.0)	1 ml/min	279 nm	[33]

Figure No. 5. Structure of Adrenergic Drug

Antiadrenergic Drugs

A medication that prevents adrenergic receptors from functioning is known as an adrenergic antagonist. Two groups comprise the five adrenergic receptors. The β adrenergic receptors are the first class of receptors. β1, β2, and β3 receptors are present. α-adrenoreceptors are found in the second group. Only α1 and α2 receptors are existing. The heart, kidneys, lungs, and digestive system are all close to adrenergic receptors[34]. Figure 6 depicts the structures of antiadrenergic drugs which are elaborated in table 7.

Table No.7 Method development and validation of Antiadrenergic Drugs

S. No	Drugs	Stationary Phase	Mobile Phase	Flow Rate	Wavelength	Ref.
1.	Methamphetamine (6a) and Propranolol (6b)	RP18	50mM pyrrolidine: Acetonitrile (50:50, v/v) (pH 11.5)	1ml/min	214nm	[35]
2.	Propranolol and Valsartan (6c)	Hypersil C₁₈ Column	Acetonitrile: Methanol: 0.1 M dihydrogen phosphate) (50:35:15), (pH 3.5)	1ml/min	250 nm	[36]
3.	Etizolam (6d) and Propranolol Hydrochloride	Puritus C₁₈Column	Ammonium Phosphate (pH 3): Acetonitrile	1 ml/min	245 nm	[37]
4.	Atenolol (6e)	ODS-3 Column (250mm × 4.6mm, 5µm)	Acetonitrile: Water	1 ml/min	276nm	[38]
5.	Brinzolamide (6f) and Timolol (6g)	(15 cm × 0.46 cm, 5µm) Zorbax Eclipse Plus	Acetonitrile: MeOH: Triethylamine Phosphate Buffer (20:10:70)	1ml/min	246 nm	[39]
6.	Latanoprost (6h), Timolol and Benzalkonium Chloride (6i)	Inertsil C₁₈Column (300 × 3.9 mm, 5µ)	Acetonitrile: Buffer (40:60 v/v)	1ml/min	-	[40]
7.	Brimonidine Tartrate (6j)	Diamonsil C₁₈Column	Triethylamine and MeOH: 10 mM Phosphate Buffer (pH 3.5) (15:85 v/v)	1 ml/min	246 nm	[41]
8.	Cetirizine (6k), Fexofenadine (6l) with Pseudoephedrine (6m)	Zorbax C₈	Acetonitrile: MeOH: 0.5 % (pH 4.5) (30:20:50)	1 ml/min	218 nm and 222 nm	[42]

Figure No. 6. Structure of Anti-adrenergic Drugs

Antihistaminic & PPIs

Antihistamines are a family of medications that lessen pathophysiologic symptoms mediated by histamine. The primary targets of therapeutic targeting are the histamine H1 and H2 receptor subtypes, which mediate several physiological and pathological processes. While H2 receptor antagonists are used to suppress gastric acid secretion in conditions like gastro-oesophageal reflux disease and to further suppress gastric acid control in conjunction with proton pump inhibitors, H1 receptor antagonists are recommended for the treatment of IgE- and non-IgE-mediated allergic disorders, such as allergic rhinitis and chronic urticaria [43]. A class of drugs known as proton-pump inhibitors significantly and permanently reduces the stomach acid production. This occurred due to the permanently blocking of the proton pump of the stomach. The body eventually produces new proton pumps to replace the irreversibly blocked ones as a result of normal cellular turnover, gradually restoring acid production [44]. Figure 7 have all the structures which are mentioned in the table 8 for method validation and method development of anti-histaminic & PPIs.

Table No.8 Method validation and Method development of Anti-histaminic & PPIs

S. No	Drugs	Stationary Phase	Mobile Phase	Flow Rate	Wavelength	Ref.
1.	Cetirizine HCl	Phenomenex Luna 5µ C₁₈	H₂O: ACN (40:60 v/v)	1 ml/min	229 nm	[45]
2.	Fexofenadine and Montelukast Sodium (7a)	Hypersil BDS	Phosphate Buffer: Acetonitrile	0.7 ml/min	233 nm	[46]
3.	Ranitidine (7b)	C₁₈Column (4.6×250mm, 5 µm)	ACN: 0.1 M ammonium dihydrogen phosphate (50:50 v/v) (pH 5.2)	1.0 ml/min	225 nm	[47]
4.	Ranitidine Hydrochloride and Domperidone (7c)	Princeton SPHER C₁₈Column	Phosphate buffer: Acetonitrile: Methanol (40:30:30)	0.6 ml/hr	225 nm	[48]

Figure No. 7. Structure of Antihistaminic & Proton Pump Inhibitors (PPIs)

Anti-Migraine Drugs

The FDA has approved seven triptans that are sold to treat acute migraines. Naratriptan, Almotriptan, Sumatriptan, Zolmitriptan, Eletriptan, Frovatriptan and Rizatriptan are among them. As compared to NSAIDs as a class, triptans are markedly more costly. They are often used as an option when NSAIDs or paracetamol don't work or the headache is really bad. Triptans are serotonin receptor agonists that have a varied empathy for 5-HT1F receptors and a high affinity for 5-HT1D and 5-HT1B receptors. The suggested mode of action entails binding postsynaptic 5-HT1B receptors on blood vascular smooth muscle cells and presynaptic 5-HT1D receptors on dorsal horn neurones and trigeminal nerve terminals [49]. Figure 8 contains the structures of all the anti-migraine drugs which are elaborated in the table 9.

Table No.9 Method validation and development of Migraine Drugs

S. No	Drug	Phase Stationary	Phase Mobile	Rate of flow	Wavelength	Ref.
1.	Sumatriptan (8a)	C₁₈ODS Column (250×4.6mm, 5µm)	ACN: Buffer: MeOH (10:80:10 v/v/v) with ortho phosphoric acid (pH 4.5)	1.0 3.ml/min	221 nm	[50]
2.	Rizatriptan (8b), Sumatriptan and Zolmitriptan (8c)	Stainless Steel Column (4.6×250mm), C₁₈Silica	Acetonitrile: Sodium Phosphate Buffer	1.0 ml/min	280 nm	[51]
3.	Paracetamol Metoclopramide Hydrochloride (8d) and Sumatriptan Succinate	C₁₈Column	(60:40 v/v) KH₂PO₄Buffer: Methanol	1.0 ml/min	-	[52]

Figure No. 8. Structure of Anti-migraine drugs

NSAIDs & Antipyretics

Non-steroidal anti-inflammatory medicines (NSAIDs) belong to a class of therapeutic drugs that lower fever, reduce inflammation, lessen pain, and prevent blood clots. Heart attacks, kidney problems, and an increased risk of gastrointestinal bleeding and ulcers are among the most common side effects, which vary depending on the particular medication, dosage, and length of usage. Aspirin, ibuprofen, diclofenac, and naproxen are the most common NSAIDs and may be purchased over-the-counter (OTC) in the majority of nations [53]. Due to its weak anti-inflammatory properties, paracetamol (acetaminophen) is typically not regarded as an NSAID [54]. Figure 9 depicts the structures of pharmaceuticals for which the method development and validation are elaborated in Table 11.

Table No.11 Method development and validation of NSAIDs & Antipyretics

S. No	Drugs	Stationary Phase	Mobile Phase	Wavelength	Flow Rate	Ref.
1.	Aspirin (38) and Omeprazole (39)	C₁₈ Column (4.6 x 150 mm)	Methanol and 0.5% OPA (pH = 3.5)	231 nm	0.7 ml/min	[55]
2.	Aspirin and Esomeprazole Magnesium (40)	ODS-BP C₁₈ Column	Methanol: 0.05M phosphate buffer (orthophosphoric acid is used to modify pH 3	230 nm	1 ml/min	[56]
3.	Ibuprofen (41) and Famotidine (42)	C₁₈Column	Buffer: Acetonitrile	213 nm	1.5 ml/min	[57]
4.	Diclofenac (43) and Tolperisone (44)	(4.6 × 150 mm, 5µ) XDB C₁₈Column	Phosphate buffer: ACN (70:30) (pH 3.4)	260 nm	1.0 ml/min	[58]
5.	Montelukast	C₁₈Column	0.1 M Potassium dihydrogen phosphate: ACN (30:70) (pH 4.0)	355 nm	1.0 ml/min	[59]

Figure No. 9. Structure of NSAIDs & Anti-pyretic

Anti-rheumatoid & Antigout

The term "disease-modifying antirheumatic drugs" (DMARDs) refers to a class of seemingly unrelated medications that reduce the course of rheumatoid arthritis. The phrase is frequently used in opposition to steroids, which inhibit the immune response but are insufficient to prevent the disease's progression, and nonsteroidal anti-inflammatory medications, which are substances that treat inflammation but not its underlying cause. In such situations, the phrase "antirheumatic" can be used without claiming to have an impact on the progression of the illness. The same class of medications has also traditionally been referred to as "slow-acting antirheumatic drugs" (SAARDs) and "remission-inducing drugs" (RIDs) [60]. Figure 10 shows all the structure of anti-rheumatoid & anti-gout which are concise in table 12.

Table No.12 Method Development and Validation Anti-rheumatoid & Antigout

S. No	Drugs	Stationary Phase	Mobile Phase	Flow Rate	Wavelength	Ref.
1.	Methotrexate (45) and Naringenin (46)	(15 cm × 4.6 mm, 5µ) ODS C18 Column	(pH 6.0) Phosphate Buffer: ACN	1.0 ml/min	302 nm	[61]
2.	Methotrexate and Curcumin (47)	ODS C₁₈Column (15cm × 4.6 mm, 5µ)	Acetonitrile: 2% Acetic Acid	1.0 ml/min	360 nm	[62]
3.	Allopurinol (48)	(150 mm x 4.6 mm, 3 μm) YMC C₁₈ Column	0.1 M ammonium acetate	1.0 ml/min	255 nm	[63]
4.	Alpha Lipoic Acid (49) and Allopurinol in Tablets	C₁₈G Column (250×4.6mm, 5µm)	ACN: (pH 4.6) 0.2 M Ammonium Acetate	0.8 ml/min	210 nm	[64]
5.	Febuxostat (50)	Column C₁₈	Acetonitrile: Sodium Acetate Buffer (pH 4.0) (60:40 v/v)	1.2 ml/min	254 nm	[65]

Figure No. 10. Structure of Anti-rheumatoid & Antigout

CONCLUSION

For the simultaneous estimation of pharmaceutical bulk medicines, RP-HPLC methods provide reliable, verified solutions that provide impurity profiling, peak purity evaluation, and quantification in various dose forms. These methods, which have been refined in accordance with ICH criteria, exhibit great specificity, linearity, accuracy, and robustness, making regular analysis in the pharmaceutical industry easier. Future developments offer improved efficiency and border application in drug research, such as hyphenated methods like LC-MS and greener solvents.

FUTURE PROSPECT

By advancing this technique for a variety of processes, including QbD-driven, green chemistry, application in FDCs, stability indicating assays, and complex formulations like nano and lipid-based dosage forms, there are opportunities to develop and validate analytical methods for both single and simultaneous drug quantification in the future. Process Analytical Technology (PAT) and real-time release testing are used to optimise columns and integrate spectroscopic or at-line/online HPLC with chemometrics for continuous batch production. More reliable method development and validation by RP-HPLC are needed due to the growing number of diseases and the demanding nature of complicated formulation.

CONFLICT OF INTERSECT

The author declares no conflict of interest. No financial support or relationship with entities that could influence the work were received.

ACKNOWLEDGEMENTS

The Hygia Institute of Pharmaceutical Education and Research's Department of Pharmaceutical Chemistry is acknowledged by the author for supplying the materials and literature database that were necessary for this review.

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Shirsat VA, Parab M, Kodgule YM, Kodgule M. A RP-HPLC Method for the Analysis of Neostigmine Methylsulfate and Process-Related Impurities, Forced Degradation Studies, in the Injection Formulation. Int J Anal Chem. 2021 May 12;2021:1–14. doi:10.1155/2021/5570173
Agarwal B, Jagdale S, Kadam P, Sakpal P, Nalawade S, Maske S, et al. Development and Validation of Stability Indicating RP-HPLC Method for the Estimation of Glycopyrrolate and Neostigmine in Bulk and Injection. J Chromatogr Sci. 2024 Mar 1;62(3):213–21. doi:10.1093/chromsci/bmad035 PubMed PMID: 37207317.
Nishtala PS, Salahudeen MS, Hilmer SN. Anticholinergics: Theoretical and Clinical Overview. Expert Opin Drug Saf. 2016 Jun 2;15(6):753–68. doi:10.1517/14740338.2016.1165664
Raghuwanshi AS, Jain UK. RP-HPLC Method Development for Estimation of Atropine Sulphate in Bulk Drug. Oriental Journal of Chemistry. 2009 Jun 4;25(3):621–4.
Ali NW, Gamal M, Abdelkawy M. Simultaneous Determination of Hyoscine N-butyl bromide and Paracetamol in their binary mixture by RP-HPLC method. Arabian Journal of Chemistry. 2017 May 1;10:S1868–74. doi:10.1016/j.arabjc.2013.07.015
Li J, Li G, Zhao L, Wang H, Sun C. Simultaneous Analysis of Ipratropium Bromide and its Related Substances Using HPLC. Curr Pharm Anal. 2021 Jan 25;17(2):293–300. doi:10.2174/1573412917666201001115322
Appiah-Ankam J, Hunter JM. Pharmacology of neuromuscular blocking drugs. Continuing Education in Anaesthesia Critical Care & Pain. 2004 Feb;4(1):2–7. doi:10.1093/bjaceaccp/mkh002
Beck W, Kabiche S, Balde IB, Carret S, Fontan JE, Cisternino S, et al. Stability of Suxamethonium in Pharmaceutical Solution for Injection by Validated Stability-Indicating Chromatographic Method. J Clin Anesth. 2016 Dec 1;35:551–9. doi:10.1016/j.jclinane.2016.09.025 PubMed PMID: 27871592.
Sri Nataraj K, Rao AS, Venkata S, Kumar S, Satya P, Ganesh S. Method Development and Validation for the Determination of Stero Isomeric Purity of Rocuronium Bromide Bulk Drug by HPLC. WAFFEN-UND KOSTUMKUNDE JOURNAL. 2020 May;11(5):7–18.
Mashru R, Parekh H, Chokshi P. Analytical Method Development and Validation for the Estimation of Sugammadex. Journal of Drug Delivery and Therapeutics. 2020 Jan 15;10(1):52–9. doi:10.22270/jddt.v10i1.3842
Ciccarone D. Stimulant Abuse: Pharmacology, Cocaine, Methamphetamine, Treatment, Attempts at Pharmacotherapy. Primary Care: Clinics in Office Practice. 2011 Mar;38(1):41–58. doi:10.1016/j.pop.2010.11.004
Mishra AK, Mishra A, Chattopadhyay P. A Reversed-Phase High Performance Liquid Chromatographic Method for Determination of Epinephrine in Pharmaceutical Formulation. Vol. 2. 2010;2(2):251–6.
Thota S, Kolagani S, Venisetty RajK, Venumadhav N. Development and Validation of Stability Indicating RP-HPLC Method for the Estimation of Neuroleptic Drug Zotepine in Bulk and Tablet Dosage Form. Int J Pharm Pharm Sci. 2014 Feb 4;6(2):456–9.
Tesoro C, Ciriello R, Lelario F, Di Capua A, Pascale R, Bianco G, et al. Development and Validation of a Reversed-Phase HPLC Method with UV Detection for the Determination of L-Dopa in Vicia faba L. Broad Beans. Molecules. 2022 Nov 1;27(21). doi:10.3390/molecules27217468 PubMed PMID: 36364292.
Muneer S, Ahad HA, Kothapalli Bonnoth C. Stability Indicating Method Development and Validation for the Quantification of Isoproterenol HCL in Bulk and Its Formulation by RP-HPLC Using PDA Detection. Journal of Pharmaceutical and Scientific Innovation. 2018 Oct 30;7(5):183–7. doi:10.7897/2277-4572.075105
Wiysonge CS, Volmink J, Opie LH. Beta-blockers and the treatment of hypertension: it is time to move on. Cardiovasc J Afr. 2007;18(6):351–2. PubMed PMID: 18092107.
Shabir GA. Development and Validation of RP-HPLC Method for the Determination of Methamphetamine and Propranolol in Tablet Dosage Form. Indian J Pharm Sci. 2011 Jul;(4):430–5. doi:https://doi.org/10.4103/0250-474X.95632 PubMed PMID: 22707828.
Imam SS, Ahad A, Aqil M, Sultana Y, Ali A. A Validated RP-HPLC Method for Simultaneous Determination of Propranolol and Valsartan in Bulk Drug and Gel Formulation. J Pharm Bioallied Sci. 2012 May 21;5(1):61–5. doi:10.4103/0975-7406.106573
Kate P, Patel P, Patel N, Kulkarni G, Kumar Patel B. Stability Indicating RP-HPLC Method Development and Validation of Etizolam and Propranolol Hydrochloride in Pharmaceutical Dosage Form. World Journal of Pharmaceutical Sciences. 2015 May 15;3(6):1113–24.
Mohammed NS. Method Development and Validation of Atenolol using two HPLC Systems. Int J Pharm Sci Res. 2017 Jun 1;8(6):1000–6. doi:10.13040/IJPSR.0975-8232.8(6).1000-10
Khatun R, Ashraful Islam S. Development and Validation of Analytical Method for Simultaneous Estimation of Brinzolamide and Timolol by HPLC from Ophthalmic Preparation. Int J Pharm Sci Res. 2014 Mar 1;5(3). doi:10.13040/IJPSR.0975-8232.5(3).1001-07
Agarwal A, Tiwari S, Nagariya K. Method Development and its Validation for Quantitative Simultaneous Determination of Latanoprost, Timolol and Benzalkonium Chloride in Ophthalmic Solution by RP-HPLC. Journal of Drug Delivery & Therapeutics. 2011;2013(3):26–30.
Sun J, Zhang X, Huang T. A validated stability-indicating HPLC method for determination of brimonidine tartrate in BRI/PHEMA drug delivery systems. Chem Cent J. 2017 Jul 11;11(1). doi:10.1186/s13065-017-0292-2
Karaku? S, Küçükgüzel ?, Küçükgüzel ?G. Development and Validation of a rapid RP-HPLC Method for the Determination of Cetirizine or Fexofenadine with Pseudoephedrine in binary Pharmaceutical Dosage Forms. J Pharm Biomed Anal. 2008 Jan;46(2):295–302. doi:10.1016/j.jpba.2007.10.018
Mainie I, Tutuian R, Castell DO. Addition of a H2 Receptor Antagonist to PPI Improves Acid Control and Decreases Nocturnal Acid Breakthrough. J Clin Gastroenterol. 2008 Jul;42(6):676–9. doi:10.1097/MCG.0b013e31814a4e5c
Fossmark R, Martinsen TC, Waldum HL. Adverse Effects of Proton Pump Inhibitors—Evidence and Plausibility. Int J Mol Sci. 2019 Oct 21;20(20):5203. doi:10.3390/ijms20205203
Barabde G, Kokate A, Ambadekar S. Development and Validation of RP-HPLC Method for Quantitative Estimation of Cetirizine HCL in Pharmaceutical Formulation as Per ICH Guideline. IOSR J Pharm. 2024 Oct;14(10):57–65.
Priyadarshini GI, Reddy S, Pappula N, Mounika V. Analytical Method Development and Validation for the Estimation of Fexofenadine HCL and Acebrophylline and Montelukast Sodium in Bulk and Pharmaceutical Dosage Form by RP-HPLC. J P Sci. 2025 Jun 5;12(06):31–6.
Badgujar VL, Ansari YA, Shaikh TJ, Deore H, Chauhan N, Gomase P, et al. Stability Indicating RP-HPLC Method Development and Validation for Determination of Ranitidine in Bulk and Pharmaceutical Dosage Form. Biochem Cell Arch. 2024 Jan 20;24(1):353–60. doi:10.51470/bca.2024.24.1.353
Singh RK, Ramakrishna S, Gupta P. RP-HPLC Method Development and Validation for Simultaneous Estimation of Ranitidine Hydrochloride and Domperidone in Combined Tablet Dosage Form. IJPSR. 2010 Jul 3;1(8):77–87.
Taylor FR, Kaniecki RG. Symptomatic Treatment of Migraine: When to Use NSAIDs, Triptans, or Opiates. Curr Treat Options Neurol. 2011 Feb 2;13(1):15–27. doi:10.1007/s11940-010-0107-4
Rajendra Kumar J, Srinidhi M, Basha MM, Raj Kumar V. Stability Indicating RP-HPLC Method Development and Validation for the Estimation of Sumatriptan in Bulk and Pharmaceutical Dosage Form. J Appl Pharm Sci. 2016 Jun 28;6(6):20–5. doi:10.7324/JAPS.2016.60604
P. Sagar V, Kumar D, Dey S, Samal HB. Simultaneous Estimation of Rizatriptan, Sumatriptan and Zolmitriptan by RP-HPLC Method in Bulk. J Pharm Res. 2010 Nov 13;3(12):2930–3.
Singh A, Singh B. Simultaneous Estimation of Sumatriptan Succinate, Metoclopramide Hydrochloride and Paracetamol by RP-HPLC Method. Journal of Pharmaceutical Sciences and Research. 2012;4(6):1848–51.
Mallinson TE. A review of ketorolac as a prehospital analgesic. Journal of Paramedic Practice. 2017 Dec 2;9(12):522–6. doi:10.12968/jpar.2017.9.12.522
Hinz B, Cheremina O, Brune K. Acetaminophen (paracetamol) is a selective cyclooxygenase?2 inhibitor in man. The FASEB Journal. 2008 Feb 20;22(2):383–90. doi:10.1096/fj.07-8506com
Sarode TK, Jadhav PB. RP-HPLC Method Development and Validation for Simultaneous Estimation of Aspirin and Omeprazole in Bulk and Dosage Form. Journal of Drug Delivery & Therapeutics. 2018 Sep 15;8(5):322–8. doi:https://doi.org/10.22270/jddt.v8i5.1825
Patel D, Patel N, Vaishy R, Patel V, Solanki C, Patel M. Development and Validation of RP-HPLC Method for Simultaneous Estimation of Aspirin and Esomeprazole Magnesium in Tablet Dosage Form. J Chem. 2012 Nov 12;2013(1):1–5. doi:10.1155/2013/751940
Sandhyarani G, Sarangapani M. Method Development and Validation for the Simultaneous Estimation of Ibuprofen and Famotidine. Original Research Article-Pharmaceutical Sciences| Open Access| UGC Approved | MCI Approved Journal. 2017 Jun;7(2):56–71.
Vemula VRB, Sharma PK. RP-HPLC Method Development and Validation for Simultaneous Estimation of Diclofenac and Tolperisone in Tablet Dosage Form. Asian Journal of Pharmaceutical Research and Clinical Research. 2013 Jul 7;6(3):186–9.
Eldin AB, Shalaby AA, El-Tohamy M. Development and Validation of a HPLC Method for the Determination of Montelukast and its Degradation Products in Pharmaceutical Formulation using an Experimental Design. Acta Pharmaceutica Sciencia. 2010 Apr;53:45–56.
Buer JK. A history of the term “DMARD.” Inflammopharmacology. 2015 Aug 23;23(4):163–71. doi:10.1007/s10787-015-0232-5
Kadiyan R, Gautam S, Kumar B, Kumar Sahoo P. RP-HPLC Method Development and Validation for Simultaneous Estimation of Methotrexate and Naringenin in Pharmaceutical Dosage Form. Res J Pharm Technol. 2024 Jul 24;3050–4. doi:10.52711/0974-360X.2024.00477
Ankalgi AD, Chaudhary NK, Kaushal P, Ashawat MS. Method Development and Validation for Simultaneous Estimation of Methotrexate and Curcumin in Bulk Drug by using RP-HPLC. Journal of Drug Delivery and Therapeutics. 2019 Dec 15;9(6s):127–35. doi:http://dx.doi.org/10.22270/jddt.v9i6-s.3775
Kumar ChJ, Pinnamaneni P, Morla SP, Kanth KNR, Nadendla RR. Application of RP-HPLC for the Estimation of Allopurinol and Its Related Substances in Bulk and Tablet Dosage Form. J Pharm Res Int. 2021 Apr 5;30(20B):11–20. doi:10.9734/jpri/2021/v33i20b31354
Rajkumar B, Bhavya T, Kumar AA. Reverse Phase HPLC Method Development and Validation for the Simultaneous Quantitative Estimation of Alpha Lipoic Acid and Allopurinol in Tablets. Int J Pharm Pharm Sci. 2013 Oct 25;6(1):307–12.
Mukthinuthalapati MA, Bandaru SPK, Bukkapatnam V, Mohapatro C. Development and Validation of a Stability-Indicating RP-HPLC Method for the Determination of Febuxostat (a Xanthine Oxidase Inhibitor). J Chromatogr Sci. 2013 Nov 1;51(10):931–8. doi:10.1093/chromsci/bms192

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Azar ZJ, Afshar M. Development and Validation of a Stability-Indicating Ion-Pair RP-HPLC Method for Determination of Bethanechol with UV Detection: Application to Pharmaceutical Analysis. Pharm Chem J. 2019 Jul 15;53(4):376–82. doi:10.1007/s11094-019-02008-w
Shirsat VA, Parab M, Kodgule YM, Kodgule M. A RP-HPLC Method for the Analysis of Neostigmine Methylsulfate and Process-Related Impurities, Forced Degradation Studies, in the Injection Formulation. Int J Anal Chem. 2021 May 12;2021:1–14. doi:10.1155/2021/5570173
Agarwal B, Jagdale S, Kadam P, Sakpal P, Nalawade S, Maske S, et al. Development and Validation of Stability Indicating RP-HPLC Method for the Estimation of Glycopyrrolate and Neostigmine in Bulk and Injection. J Chromatogr Sci. 2024 Mar 1;62(3):213–21. doi:10.1093/chromsci/bmad035 PubMed PMID: 37207317.
Nishtala PS, Salahudeen MS, Hilmer SN. Anticholinergics: Theoretical and Clinical Overview. Expert Opin Drug Saf. 2016 Jun 2;15(6):753–68. doi:10.1517/14740338.2016.1165664
Raghuwanshi AS, Jain UK. RP-HPLC Method Development for Estimation of Atropine Sulphate in Bulk Drug. Oriental Journal of Chemistry. 2009 Jun 4;25(3):621–4.
Ali NW, Gamal M, Abdelkawy M. Simultaneous Determination of Hyoscine N-butyl bromide and Paracetamol in their binary mixture by RP-HPLC method. Arabian Journal of Chemistry. 2017 May 1;10:S1868–74. doi:10.1016/j.arabjc.2013.07.015
Li J, Li G, Zhao L, Wang H, Sun C. Simultaneous Analysis of Ipratropium Bromide and its Related Substances Using HPLC. Curr Pharm Anal. 2021 Jan 25;17(2):293–300. doi:10.2174/1573412917666201001115322
Appiah-Ankam J, Hunter JM. Pharmacology of neuromuscular blocking drugs. Continuing Education in Anaesthesia Critical Care & Pain. 2004 Feb;4(1):2–7. doi:10.1093/bjaceaccp/mkh002
Beck W, Kabiche S, Balde IB, Carret S, Fontan JE, Cisternino S, et al. Stability of Suxamethonium in Pharmaceutical Solution for Injection by Validated Stability-Indicating Chromatographic Method. J Clin Anesth. 2016 Dec 1;35:551–9. doi:10.1016/j.jclinane.2016.09.025 PubMed PMID: 27871592.
Sri Nataraj K, Rao AS, Venkata S, Kumar S, Satya P, Ganesh S. Method Development and Validation for the Determination of Stero Isomeric Purity of Rocuronium Bromide Bulk Drug by HPLC. WAFFEN-UND KOSTUMKUNDE JOURNAL. 2020 May;11(5):7–18.
Mashru R, Parekh H, Chokshi P. Analytical Method Development and Validation for the Estimation of Sugammadex. Journal of Drug Delivery and Therapeutics. 2020 Jan 15;10(1):52–9. doi:10.22270/jddt.v10i1.3842
Ciccarone D. Stimulant Abuse: Pharmacology, Cocaine, Methamphetamine, Treatment, Attempts at Pharmacotherapy. Primary Care: Clinics in Office Practice. 2011 Mar;38(1):41–58. doi:10.1016/j.pop.2010.11.004
Mishra AK, Mishra A, Chattopadhyay P. A Reversed-Phase High Performance Liquid Chromatographic Method for Determination of Epinephrine in Pharmaceutical Formulation. Vol. 2. 2010;2(2):251–6.
Thota S, Kolagani S, Venisetty RajK, Venumadhav N. Development and Validation of Stability Indicating RP-HPLC Method for the Estimation of Neuroleptic Drug Zotepine in Bulk and Tablet Dosage Form. Int J Pharm Pharm Sci. 2014 Feb 4;6(2):456–9.
Tesoro C, Ciriello R, Lelario F, Di Capua A, Pascale R, Bianco G, et al. Development and Validation of a Reversed-Phase HPLC Method with UV Detection for the Determination of L-Dopa in Vicia faba L. Broad Beans. Molecules. 2022 Nov 1;27(21). doi:10.3390/molecules27217468 PubMed PMID: 36364292.
Muneer S, Ahad HA, Kothapalli Bonnoth C. Stability Indicating Method Development and Validation for the Quantification of Isoproterenol HCL in Bulk and Its Formulation by RP-HPLC Using PDA Detection. Journal of Pharmaceutical and Scientific Innovation. 2018 Oct 30;7(5):183–7. doi:10.7897/2277-4572.075105
Wiysonge CS, Volmink J, Opie LH. Beta-blockers and the treatment of hypertension: it is time to move on. Cardiovasc J Afr. 2007;18(6):351–2. PubMed PMID: 18092107.
Shabir GA. Development and Validation of RP-HPLC Method for the Determination of Methamphetamine and Propranolol in Tablet Dosage Form. Indian J Pharm Sci. 2011 Jul;(4):430–5. doi:https://doi.org/10.4103/0250-474X.95632 PubMed PMID: 22707828.
Imam SS, Ahad A, Aqil M, Sultana Y, Ali A. A Validated RP-HPLC Method for Simultaneous Determination of Propranolol and Valsartan in Bulk Drug and Gel Formulation. J Pharm Bioallied Sci. 2012 May 21;5(1):61–5. doi:10.4103/0975-7406.106573
Kate P, Patel P, Patel N, Kulkarni G, Kumar Patel B. Stability Indicating RP-HPLC Method Development and Validation of Etizolam and Propranolol Hydrochloride in Pharmaceutical Dosage Form. World Journal of Pharmaceutical Sciences. 2015 May 15;3(6):1113–24.
Mohammed NS. Method Development and Validation of Atenolol using two HPLC Systems. Int J Pharm Sci Res. 2017 Jun 1;8(6):1000–6. doi:10.13040/IJPSR.0975-8232.8(6).1000-10
Khatun R, Ashraful Islam S. Development and Validation of Analytical Method for Simultaneous Estimation of Brinzolamide and Timolol by HPLC from Ophthalmic Preparation. Int J Pharm Sci Res. 2014 Mar 1;5(3). doi:10.13040/IJPSR.0975-8232.5(3).1001-07
Agarwal A, Tiwari S, Nagariya K. Method Development and its Validation for Quantitative Simultaneous Determination of Latanoprost, Timolol and Benzalkonium Chloride in Ophthalmic Solution by RP-HPLC. Journal of Drug Delivery & Therapeutics. 2011;2013(3):26–30.
Sun J, Zhang X, Huang T. A validated stability-indicating HPLC method for determination of brimonidine tartrate in BRI/PHEMA drug delivery systems. Chem Cent J. 2017 Jul 11;11(1). doi:10.1186/s13065-017-0292-2
Karaku? S, Küçükgüzel ?, Küçükgüzel ?G. Development and Validation of a rapid RP-HPLC Method for the Determination of Cetirizine or Fexofenadine with Pseudoephedrine in binary Pharmaceutical Dosage Forms. J Pharm Biomed Anal. 2008 Jan;46(2):295–302. doi:10.1016/j.jpba.2007.10.018
Mainie I, Tutuian R, Castell DO. Addition of a H2 Receptor Antagonist to PPI Improves Acid Control and Decreases Nocturnal Acid Breakthrough. J Clin Gastroenterol. 2008 Jul;42(6):676–9. doi:10.1097/MCG.0b013e31814a4e5c
Fossmark R, Martinsen TC, Waldum HL. Adverse Effects of Proton Pump Inhibitors—Evidence and Plausibility. Int J Mol Sci. 2019 Oct 21;20(20):5203. doi:10.3390/ijms20205203
Barabde G, Kokate A, Ambadekar S. Development and Validation of RP-HPLC Method for Quantitative Estimation of Cetirizine HCL in Pharmaceutical Formulation as Per ICH Guideline. IOSR J Pharm. 2024 Oct;14(10):57–65.
Priyadarshini GI, Reddy S, Pappula N, Mounika V. Analytical Method Development and Validation for the Estimation of Fexofenadine HCL and Acebrophylline and Montelukast Sodium in Bulk and Pharmaceutical Dosage Form by RP-HPLC. J P Sci. 2025 Jun 5;12(06):31–6.
Badgujar VL, Ansari YA, Shaikh TJ, Deore H, Chauhan N, Gomase P, et al. Stability Indicating RP-HPLC Method Development and Validation for Determination of Ranitidine in Bulk and Pharmaceutical Dosage Form. Biochem Cell Arch. 2024 Jan 20;24(1):353–60. doi:10.51470/bca.2024.24.1.353
Singh RK, Ramakrishna S, Gupta P. RP-HPLC Method Development and Validation for Simultaneous Estimation of Ranitidine Hydrochloride and Domperidone in Combined Tablet Dosage Form. IJPSR. 2010 Jul 3;1(8):77–87.
Taylor FR, Kaniecki RG. Symptomatic Treatment of Migraine: When to Use NSAIDs, Triptans, or Opiates. Curr Treat Options Neurol. 2011 Feb 2;13(1):15–27. doi:10.1007/s11940-010-0107-4
Rajendra Kumar J, Srinidhi M, Basha MM, Raj Kumar V. Stability Indicating RP-HPLC Method Development and Validation for the Estimation of Sumatriptan in Bulk and Pharmaceutical Dosage Form. J Appl Pharm Sci. 2016 Jun 28;6(6):20–5. doi:10.7324/JAPS.2016.60604
P. Sagar V, Kumar D, Dey S, Samal HB. Simultaneous Estimation of Rizatriptan, Sumatriptan and Zolmitriptan by RP-HPLC Method in Bulk. J Pharm Res. 2010 Nov 13;3(12):2930–3.
Singh A, Singh B. Simultaneous Estimation of Sumatriptan Succinate, Metoclopramide Hydrochloride and Paracetamol by RP-HPLC Method. Journal of Pharmaceutical Sciences and Research. 2012;4(6):1848–51.
Mallinson TE. A review of ketorolac as a prehospital analgesic. Journal of Paramedic Practice. 2017 Dec 2;9(12):522–6. doi:10.12968/jpar.2017.9.12.522
Hinz B, Cheremina O, Brune K. Acetaminophen (paracetamol) is a selective cyclooxygenase?2 inhibitor in man. The FASEB Journal. 2008 Feb 20;22(2):383–90. doi:10.1096/fj.07-8506com
Sarode TK, Jadhav PB. RP-HPLC Method Development and Validation for Simultaneous Estimation of Aspirin and Omeprazole in Bulk and Dosage Form. Journal of Drug Delivery & Therapeutics. 2018 Sep 15;8(5):322–8. doi:https://doi.org/10.22270/jddt.v8i5.1825
Patel D, Patel N, Vaishy R, Patel V, Solanki C, Patel M. Development and Validation of RP-HPLC Method for Simultaneous Estimation of Aspirin and Esomeprazole Magnesium in Tablet Dosage Form. J Chem. 2012 Nov 12;2013(1):1–5. doi:10.1155/2013/751940
Sandhyarani G, Sarangapani M. Method Development and Validation for the Simultaneous Estimation of Ibuprofen and Famotidine. Original Research Article-Pharmaceutical Sciences| Open Access| UGC Approved | MCI Approved Journal. 2017 Jun;7(2):56–71.
Vemula VRB, Sharma PK. RP-HPLC Method Development and Validation for Simultaneous Estimation of Diclofenac and Tolperisone in Tablet Dosage Form. Asian Journal of Pharmaceutical Research and Clinical Research. 2013 Jul 7;6(3):186–9.
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Dr. Shiv Bhadra Singh

Corresponding author

Hygia Institute of Pharmaceutical Education and Research

Syed Tahir Hussain Rizvi

Co-author

Hygia Institute of Pharmaceutical Education and Research

Syed Tahir Hussain Rizvi, Dr. Shiv Bhadra Singh, Analytical Method Development and Validation Methods for Estimation of Drugs in Pharmaceutical Dosage Forms: A Review, Int. J. of Pharm. Sci., 2026, Vol 4, Issue 5, 2265-2283, https://doi.org/10.5281/zenodo.20117089

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Analytical Method Development and Validation Methods for Estimation of Drugs in Pharmaceutical Dosage Forms: A Review

Abstract

Keywords

Introduction

Reference

Dr. Shiv Bhadra Singh

Syed Tahir Hussain Rizvi

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