INTRODUCTION:

Ultra-high performance liquid chromatography (UHPLC) uses columns with particles smaller than 2.5-5μm, unlike traditional HPLC methods. Using columns with smaller particles (sub-2μm) leads to higher efficiency per unit of time. Although UHPLC was initially shown with custom equipment, the market introduction of the first UHPLC system in 2004 enabled widespread adoption of this technology. Maximizing the benefits of sub-2-μm particle-containing columns requires optimizing both the columns and their equipment. The system required minimum extra-column dispersion, a column compartment with minimal radial temperature gradients, and the ability to function reliably at pressures up to 1000 bar^1-5.

UPLC outperforms HPLC in terms of speed, sensitivity, and resolution. The stationary phase uses sub-2μm particle technology to reduce solvent use and run time. The technique used in Ultra Performance Liquid Chromatography offers peak capacity and speed during analysis be enhanced and elevated to new heights using microscopic particles, according to the Van Demeter equation H = A + B/μ + Cμ^6,7. UPLC is less efficient than gas chromatography and capillary electrophoresis due to the sluggish diffusion of analytes into the stationary phase. UPLC can be used to transfer and optimize a standard test, which reduces analysis time and improves analysis sensitivity^8,9. Six Based on the developments in particle size and column packing material mentioned above, the water firm was awarded UHPLC, formerly known as UPLC, as its trade name.

Principle:

The van Deemter connection serves as the separation principle for HPLC and UPLC. This explains the relationship between flow rate and plate height^10.11.

The equation is:

H = A + B/μ + Cμ.

Where:

H= Plate height

A= Eddy diffusion

B= Longitudinal diffusion

C= Equilibrium mass transfer

μ= Flow rate

A smaller plate height value equates to greater peak efficiency, because more plates can occur. Over a constant length of column.

Instrument of UPLC:

The apparatus used in ultra-performance liquid chromatography is virtually identical to that used in HPLC. The system is designed to withstand high strain with minimal disruption and regular maintenance. New hardware and firmware are used to support a tunable UV/Visible detector for UPLC detection. When the data rate is high. Volume of the tunable the UV/Vis detector, with a 10mm flow cell path length, holds only 0.5 liter¹². The instrumentation for UPLC includes:

Sample Injection:

The injector is used to add a little amount of solution containing the precisely measured sample in the mobile phase. A consistent and precise injection technique is necessary. To avoid significant pressure instabilities in the column, the injection method must be somewhat pulse-free. Conventional injection valves can be either manually or automatically controlled. To decrease the risk of band spreading, keep the device's swept volume to a minimum. To effectively benefit from UPLC's speed, a short injection cycle time is required. For maximum sensitivity, smaller volume injections with minimal carryover are necessary. UPLC sample volumes typically range from 2 to 5 L. Direct injection techniques are becoming more common for biological materials¹³. Figure 1 shows a UPLC flowchart.

Figure 1. UHPLC flow chart⁴

Pumping Systems:

Small particle, high peak capacity separations cannot be accomplished with the pressure range that current HPLC technology can reach. The anticipated pressure decreases at the flow rate that maximizes efficiency over a 15 cm long column loaded with 1.7µm particles are 15,000 psi. A pump that can smoothly and reliably supply solvent at these pressures, adapt to solvent compressibility, and operate in both gradient and isocratic separation modes is also required.

Quality of mobile phase and buffers:

UHPLC columns have much smaller frits and particles than traditional HPLC columns—for example, UHPLC frit pore sizes can be as small as 0.2µm compared to 2 µm in HPLC. This means tiny particles in the mobile phase that don’t affect HPLC can cause issues in UHPLC. Therefore, it’s crucial to ensure solvents are free of insoluble particles by following strict preparation guidelines for the mobile phase in UHPLC.

UHPLC Columns:

A bonded phase that provides selectivity and retention is required in order to separate the components of the sample. Because larger diameter columns need higher flow rates—which in turn require larger amounts of mobile phase—to achieve the required linear velocity, UHPLC columns have a lower diameter. Today's most sophisticated UHPLC column technology is packed with sub-2micron particle technology and comes in 2.1mm column variants¹⁴. Although 4.6mm columns were thought to be the industry norm for many years, 2.1mm column shapes had to be used in order to lessen the impacts of viscous heating during ultra-high-pressure operations. Heat is distributed through the column wall, creating a 25A radial temperature gradient. The resulting parabolic flow profile will have a detrimental effect on chromatographic efficiency.

Types of column¹⁵

a) Charged Surface Hybrid

b) Ethylene-bridged Hybrid

c) High Strength Silica

d) Peptide Separation Technology

Detector:

The UPLC detector features a high sampling rate, narrow peaks, and low peak dispersion to reduce solute loss on the column. It offers 2–3 times greater sensitivity than traditional HPLC due to advanced detector technology. UPLC uses Tunable Vis-UV (TUV) and photodiode array (PDA) detectors with Teflon AF coatings to improve light transmission by reducing internal absorption. It has a 500 nanoliter internal volume, acquisition speeds of 20 Hz (PDA) and 40 Hz (TUV), and 10nm path lengths. UPLC is also combined with mass spectrometry for enhanced detection¹⁶.

Advantages of UHPLC:

· The mobile phase volume consumption is reduced by at least 80% as compared to HPLC.

· Reduced operating expenses and duration¹⁷.

· The injection volume must be reduced¹⁸.

· Reduced band broadening results in a higher signal to noise ratio (S/N), which raises sensitivity²⁰.

· Improved chromatographic peak resolution significantly lessens the issue of ion suppression from co-eluting peaks.

· Quicker power resolution¹⁹.

· Provides analysis in real time while keeping up with manufacturing procedures.

· Ensures the quality of the final work, including testing before release.

Disadvantages of UPLC:

· Higher back pressures shorten the columns' lifespan when compared to traditional HPLC.

· Because they are primarily non-regenerable, particles smaller than 2μm have limited applications²¹^,22

UHPLC Method of Various Drug and Their Research Outcome:

Table 1: UHPLC Method Of Various Drug and Their Research Outcome³⁷

Drug	Mobile phase	Research outcome	References
Teriflunomide (anti inflammatory)	Mobile phase of Acetonitrile: Water in the ratio of 60:40 v/v.	In this work developed a novel and trustworthy technique for quantifying teriflunomide. It has been demonstrated that the established RP-UHPLC method for estimating Teriflunomide API and its commercial preparation is straightforward, cost-effective, precise, linear, sensitive, and accurate.	25
Tedizolid and linezolid (Antibiotics Agent)	(A) 0.1% formic acid in water and (B) Acetonitrile 0–0.3 min, 20–95% (B); 0.3–1.5 min,95–95% (B); 1.5–1.6 min, 95–20% (B);1.6–3.0 min, 20– 20% (B)	In this study, we developed LC-MS/MS to establish a technique for quantifying tedizolid and linezolid in human plasma. A straightforward procedure was used to prepare the sample. In addition to method simplicity, cost concerns are necessary for routine TDM measurement, and the current approach is thought to be beneficial.	26
Cephalomannine (Antibiotics Agent)	0.1% formic acid in water (A) and acetonitrile (B)0–0.6 min, 20–85% (B); 0.6–1.8 min,85% (B); 1.8–2.0 min, 80–30% (B)	To the best of our knowledge, this was first publication that uses the UPLC–MS/MS technology to determine the level of cephalomannine in rat plasma. The technique provided sample preparation with a shortened run time of 2.0 minutes and a straightforward one-step precipitation of plasma protein by perchloric acid–methanol (1:9, v/v).	27
Daclatasvir (Antiviral Agent)	10 mm Ammonium formate, pH 3.5and acetonitrile (50:50% v/v)	It has been described as DAC in pure form and tablets may now be determined utilizing innovative, easy, quick, practical, and affordable spectrophotometric techniques that use cerium (IV) as an oxidizing agent and have been validated in accordance with current ICH criteria.\	28
Oleanolic acid (Antiviral Agent)	10 mm Ammonium Acetate with 0.1% formic acid in water: acetonitrile (10: 90% v/v)	In this study we developed and validated a high-throughput (UPLC-MS/MS) method for determining oleanolic acid (OA) in rat plasma and liver tissue, utilizing glycyrrhetic acid as an internal reference. Solid-phase extraction was used to prepare samples of plasma and liver homogenates.	29
Apixaban (Anticoagulant)	A (MP-A) was prepared with buffer and acetonitrile 90:10 v/v, while (MPB) contained water and acetonitrile 10:90 v/v	In this study, a novel UHPLC method has been developed for apixaban in-process control analysis was established. Innovative chromatographic modeling software was used to facilitate method development. The quality of the separation (resolution map) was investigated in an extended knowledge space by integrating three complementing design spaces.	30
Amphotericin – B (Antifungal Agent)	(A) Methanol: acetonitrile (50: 50% v/v) containing 0.1% formic acid; and (B) 10 mM ammonium formate (pH 3 ± 0.2), containing 0.2% formic acid and 1% acetonitrile 0.0–2.0 min, 35– 90% (A); 2.0–2.7 min, 90–35%	The first-line treatment for potentially fatal invasive fungal infections is amphotericin B (AmB). To determine the levels of amphotericin B in these patients, a sensitive, accurate, and selective UPLC MS/MS method was developed.	31
Ibuprofen (Anti – Inflammatory)	Water: methanol (35:65% v/v) both containing 10 mm ammonium acetate and 0.1% formic acid. 0–12 min, 35% (A) and 65% (B); 12.01–14 min 100% (B); 15.1 min, 35% (A) and 65% (B)	A modified UPLC-MS/MS method has been developed and validated to quantify the (R) and (S) ibuprofen enantiomers in human plasma.	32
Oxcarbazepine (Antiepileptic Agent)	Acetonitrile–10 mm ammonium acetate (85:15% v/v)	A quick and sensitive liquid chromatography tandem mass spectrometry assay was developed to analyze oxcarbazepine and its major metabolite in human plasma. The assay utilizes a straightforward solid-phase extraction method.	33
Letrozole (Anti-cancer Agent)	(A) Acetonitrile and 0.1% formic acid in water (B). 0 min, 20% (A): 80% (B);0.3 min, 20% (A): 80% (B); 2 min, 95% (A): 5% (B); 2.5 min, 95% (A): 5% (B); 2.6 min, 20 % (A): 80% (B)	In this work we has been developed a sensitive and selective UPLC-MS/MS method to determine letrozole in rat plasma. Protein was precipitated using acetonitrile-methanol (9:1, v/v) after adding midazolam as an internal standard (IS).	34
Dendrobine (Anti-cancer Agent)	(A) 0.1% formic acid and (B)acetonitrile. 0–1.0 min, 20–40 (B); 2min, 40–95% (B), 0.5 min, 95% (B); then decreased to 20% (B) within 0.1 min, maintained at 20% (B) for 0.4 min	Dendrobine, a significant alkaloid chemical, it has been utilized in the Chinese Pharmacopoeia to control and differentiate Dendrobium.	35
Solasonine (Anti-cancer Agent)	(A) 0.1% formic acid and (B)acetonitrile. 0–0.5 min, 15% (B); 0.5–3.5 min, 15–75% (B); 3.5–4.0 min, 75–85% (B); 4.0–5.0 min, 85% (B)	In this study developed and validated an ultra-performance liquid chromatography with tandem mass spectrometry (UPLC-MS/MS) method to quantify solasonine in rat plasma. Plasma samples were treated using protein precipitation.	36

Recent Application of UHPLC:

Development and Validation of Methods:

Method development and validation involve optimizing factors like mobile phase, pH, and column chemistry, making it a time-consuming process. However, UHPLC greatly shortens this time, allowing method optimization in just one to two hours and analysis in as little as one minute³⁸.

Production and QA/QC:

Identity, purity, quality, safety, and efficacy are key factors in pharmaceutical production. Both raw materials and final products must meet strict purity standards. Continuous stability monitoring is also vital for quality control. UHPLC plays a crucial role in QA/QC labs, enabling precise and regulated quantitative analysis to ensure reliable, high-quality pharmaceutical products³⁹.

Identifying FDC Products:

A fixed dose combination (FDC) contains two or more drugs in set doses. When drugs have different solubilities, method development is complex, requiring advanced techniques like UHPLC-PDA and UHPLC-Q-TOF/MS. These methods offer fast analysis, enabling many samples to be processed daily⁴⁰.

Degradation Products and Impurity Identification:

Identifying degradation products and impurities is essential but often time-consuming with traditional HPLC-UV or HPLC-MS methods. UHPLC-PDA provides a faster, more effective alternative, using 3D peak profiles to easily detect impurities. Combined with UHPLC/Q-TOF-MS, it allows accurate identification of degradation products, fragmentation patterns, and mechanisms, thanks to adjustable collision energies and precise mass data.^41,42.

Testing for Dissolution:

Dissolution testing is essential for ensuring drug quality and consistency, especially in sustained-release formulations. UPLC streamlines this process with accurate, automated sample collection, handling everything from tablet drop to data analysis and result reporting⁴³.

Sustained Release Tablet Analysis:

UPLC-PDA techniques are used for in vitro dissolution studies of sustained-release tablets, offering precise detection of drug and excipient peaks. These studies often use a mobile phase of acetonitrile and 2mM ammonium acetate (40:60, v/v) for effective UPLC analysis, ensuring accurate monitoring of drug release profiles^44,45.

Studies on pharmacokinetics and bioequivalence:

Drug quantification in biological samples is vital for pharmacokinetic, toxicological, and bioequivalence studies. UPLC-PDA offers high sensitivity and selectivity at low detection levels, providing accurate and reliable data for tasks like pharmacokinetic analysis⁴^4,45.

Metabolite Identification:

Metabolite identification is essential after drug development to understand therapeutic effects. Using MS/MS with HPLC and UHPLC columns plus TOF-MS, human paracetamol metabolites were identified, with UHPLC showing three times greater sensitivity than HPLC. UHPLC/Q-TOF and MetaboLynx™ software enable detailed metabolite profiling, a critical, regulated step in drug discovery and development⁴⁶.

Studies on Metabonomics:

Metabonomics studies biochemical changes in drugs within the human body, helping identify new metabolites and assess their effects. It aids rapid in-vitro and in-vivo drug testing and has been used to analyze components in ginseng. UPLC provides precise and reliable amino acid analysis for protein characterization, cell culture, and nutrition studies⁴⁷.

Identification of Phytoconstituents:

A variety of analytical procedures are used to isolate and characterize phytoconstituents in order to determine their analysis. UPLC-MS was mostly used in primitive procedures to isolate and identify phytoconstituents; in other words, characterizations and quantifications of phytoconstituents have been documented in Piper betle⁴⁸.

Natural Product and Herbal Medicine Analysis:

High-quality separations and detection capabilities are offered by UPLC to help find active chemicals in extremely complex samples derived from conventional herbal remedies and natural goods. Utilizing UPLC-Q-TOF-MS/MS, the Traditional Chinese Formula fingerprint was created. SiJunZiTang⁴⁹.

Mapping of Peptides:

Peptide mapping is the process of determining the chemical structure of the peptides that are linked within proteins. When used to analyze complicated compounds like proteins, this approach yields reliable results. The use of time-of-flight mass detection can facilitate the structural identification of proteins⁵⁰.

Analysis of Amino Acids:

A tried-and-true method for identifying all 20 common amino acids in proteins or peptides, as well as any changed residues that may result from posttranslational changes, is amino acid analysis. In biotechnology, amino acid analysis is a widely used and recognized analytical technique that has been modified to function inside the UHPLC framework⁵¹.

Glycan analysis or glycoprofiling:

Glycans, or oligosaccharides (sugars), are typically found in glycoproteins; connected monosaccharides are typically absent. Any glycoprotein must be characterized by identifying the sugars present, measuring how they are arranged as glycans, identifying the protein's site or sites of attachment, and then analyzing the distribution of glycoforms. The most common method for measuring the percentage of glycoforms is intact-protein UPLC-MS⁵².

Analysis of vitamins:

For the examination of the vitamin B complex (thiamin, riboflavin, biotin, nicotinic acid, pyridoxine, pyridoxamine, pyridoxal, pantothenic acid, FAD, and nicotinamide) in human milk, a number of UPLC-MS techniques have been documented. The analysis of vitamin B from milk samples is done using UPLC-MS in conjunction with ESI methods⁵³.

Determination of alkaloids:

Using UPLC-MS/MS, Ortega et al. reported the identification and measurement of caffeine, theobromine, and alkaloids in a cocoa sample. Alkaloid profiling of therapeutic herbs with cytotoxic qualities has also been documented. It is employed in the examination of several alkaloids, including chelidonine, protopine, berberine, and sanguinarine⁵⁴.

Antibiotic Screening in Surface and Wastewater:

Antibiotics like ofloxacin and ciprofloxacin have been detected in surface water and wastewater from pharmaceutical production. These drugs often enter wastewater through equipment washing, and their residues have been confirmed in industry effluents⁵⁵.

Monitoring of Therapeutic Drugs:

The tracking of β-lactam antibiotic levels in patients' plasma with varying pharmacokinetics. Seven β-lactam antibiotics and two β-lactamase inhibitors were simultaneously measured in human plasma using the UPLC-MS/MS technique⁵⁶.

Synthetic Compound Screening:

This method is essential for high-throughput quality control of synthetic pharmaceuticals. Unlike traditional techniques like NMR, it offers greater sensitivity, requires less pure samples, and reduces operator skill and solvent costs. Automation and precise mass measurement software further streamline the analysis⁵⁷.

Analysis of Phenolic Compounds and Antioxidants:

Free radicals contribute to many diseases, and antioxidants help inhibit their effects. Due to their antibacterial properties, antioxidants are increasingly used in the food industry. UPLC-ESI-MS/MS has been used to analyze the phenolic profiles and antioxidant activities of Salvia species from South West Anatolia, Turkey, including S. potentillifolia, S. albimaculata, and S. nydeggerit⁵⁸.

Identification of unknown pesticides:

The technique is applied when identifying pesticides in fruits and vegetables. This sensitive technology can be used to identify these hazardous compounds from vegetables, which is crucial for public health. Pesticide residue found in extremely low concentrations that was extracted from fruit packaging materials was examined using a method⁵⁹.

Doping Agent Analysis:

The method is used to detect and quantify doping drugs like opioids, stimulants, diuretics, and β-blockers. It was first applied for screening and later for precise measurement. The World Anti-Doping Agency (WADA) confirmed its reliability, and the technique meets the standards of the World Anti-Doping Code⁶⁰.

Water Organic Pollutant Screening:

Identifies organic pollutants in waste and natural water using solid-phase extraction for sample preparation. It applies the conventional addition technique to analyze water samples containing various contaminants. The approach detects multiple organic pollutants, including antibiotics, anti-inflammatory, and analgesic drugs⁶¹.

The UHPLC Applications with their examples of drugs analyzed are presented in Table 2

Table-2: UHPLC Applications with Details of Column, Mobile Phase Composition and Detection Technique Used⁶²

S.No.	Applications	Drugs	Column	Mobile Phase Composition	Detection	Ref.
1	Method Development and Validation	Metformin, Glimepiride and Pioglitazone Tablets	BEH C-18	Acetonitrile: 2Mm Ammonium Acetate (50:50, % V/V)	PDA Detector	38
2	Manufacturing and QA/QC	Pharmaceutical Production (GMP)	Monolithic and 3micron particle	Acetonitrile: 2Mm Ammonium Acetate (50:50, % V/V)	UV Detector	39
3	Determination of FDC Products	Aceclofenac and Paracetamol Tablets	BEH C-18	Acetonitrile: 2Mm Ammonium Acetate (40:60, % V/V)	Q-TOF-MS	40
4	Identification of Impurities and Degradation Products	Telmisartan and Hydrochlorothiazide	BEH C-18	Acetonitrile: 2Mm Ammonium Acetate (50:50, % V/V)	Q-TOF-MS	41
4	Identification of Impurities and Degradation Products	Valsartan	BEH C-18	Acetonitrile: 2Mm Ammonium Acetate (50:50, % V/V)	Q-TOF-MS	42
5	Dissolution Studies	Rabeprazole	BEH C-18	Acetonitrile: Phosphate Buffer (35:65, % V/V)	UV Detector	43
6	Analysis of Sustained Release Tablet	Aceclofenac and Paracetamol Tablets	BEH C-18	Acetonitrile:2Mm Ammonium Acetate (40:60, % V/V)	PDA Detector	44
6	Analysis of Sustained Release Tablet	Telmisartan and Hydrochlorothiazide ablets	BEH C-18	Acetonitrile:2Mm Ammonium Acetate (40:60, % V/V)	PDA Detector	45
7	Pharmacokinetic and Bioequivalence Studies	Aceclofenac and Paracetamol Tablets	BEH C-18	Acetonitrile: 2Mm Ammonium Acetate (40:60, % V/V)	Q-TOF-MS	44
7	Pharmacokinetic and Bioequivalence Studies	Telmisartan and Hydrochlorothiazide	BEH C-18	Acetonitrile: 2Mm Ammonium Acetate (40:60, % V/V)	Q-TOF-MS	45
8	Identification of Metabolitees	Acetaminophen	Monolithic C-18	Water: Acetonitrile, with 0.1% formic acid	Q-TOF- MS/MS	46
9	Metabonomic studies	Ginseng	BEH C-18	Water: Acetonitrile, with 0.1% formic acid	Q-TOF- MS/MS	47
10	Determination of Phytoconstituents	Eugenyl acetate & Eugenol in Piper betle leaf	BEH C-18	Water: Acetonitrile, with 0.1% formic acid	ESI-MS/MS	48
11	Analysis of Herbal Medicine	Chinese Formula- SiJunZiTang	BEH C-18	Water: Acetonitrile, with 0.1% formic acid	Q-TOF- MS/MS	49
12	Peptide Mapping	Peptides	BEH C-18	Water: Acetonitrile, with 0.1% formic acid	ESI-TOF- MS/MS	50
13	Amino Acid Analysis	Amino Acids	BEH C-18	25% to 60%Acetonitrile: Water	ESI-TOF- MS/MS	51
14	Glycoprofiling (Glycan Analysis)	Glycoproteins	BEH C-18	Water: Acetonitrile, with 0.1% formic acid	UPLC-MS/MS	52
15	Vitamin analysis	Thiamin, Riboflavin, Biotin, Nicotinic acid and Pyridoxine,	UPLC HSS T3	2.5 mmol/L Ammonium Formate in Water & Acetonitrile	ESI-MS/MS	53
16	Determination of alkaloids	Procyanidins, Theobromine and caffeine	Acquity HSS T3	Water/Acetic acid (99.8/0.2, v/v) and Acetonitrile	Triple- q-ESI- MS/MS	54
17	Screening of Antibiotics in Surface and Wastewater	Ofloxacin, Ciprofloxacin, Erythromycin	BEH C-18	Acetonitrile: Ammonium Acetate (50:50% V/V)	Hybrid QTOF- MS	55
18	Therapeutic Drug Monitoring	Progestin	BEH C-18	Water: Acetonitrile, with 0.1% formic acid	Tandem- MS/MS	56
19	Screening of Synthetic Compounds	Anti-hypertensives Anti-diabetics	BEH C-18	Acetonitrile: 2mM Ammonium Acetate (50:50%V/V)	Q-TOF- MS/MS	57
20	Analysis of Antioxidants and Phenolic Compounds	Phenolic compounds	BEH C-18	0.5% Acetic Acid in water and 0.5% Acetic Acid in methanol	ESI-MS/MS	58
21	Identification of Unknown Pesticides	Carbendazim, Imazalil, and Ethoxyquin	BEH C-18	Acetonitrile: Ammonium Acetate (50:50% V/V)	Q-TOF- MS/MS	59
22	Analysis of Doping Agents	Beta-blockers, Stimulants and Narcotics	BEH C-18	Acetonitrile: Ammonium Acetate (50:50% V/V)	Q-TOF- MS/MS	60
23	Screening of Organic Pollutants in Water	Organic Pollutants	BEH C-18	Acetonitrile: Ammonium Acetate (50:50% V/V)	Q-TOF- MS/MS	61

CONCLUSION:

In the presented in the review study demonstrates that Ultra-Performance Liquid Chromatography outperforms traditional HPLC. In fact, it has become the industry standard for HPLC. The key advantage is a reduction in analysis time and solvent use. This is accomplished by the employment of small particle sizes and a short column. The only disadvantage of UPLC could be high back pressure, which can be reduced by increasing column temperature. Applications, including instances of some of the most advanced work in pharmaceutical analysis, have been documented. Qualitative and quantitative analysis are vital for drug quality since they determine the efficacy of any drug or dosage form. UHPLC has been effectively used to identify and quantify substances in practically every aspect of chromatographic and pharmaceutical analysis. The UPLC technique is generally accepted and offers considerable improvements in speed, sensitivity, and resolution compared to conventional high performance liquid chromatography.

CONFLICT OF INTEREST:

The authors have no conflicts of interest regarding this investigation.

ACKNOWLEDGEMENT:

The authors are grateful to the Principal and Management for providing the necessary facilities for completing this work successfully

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Received on 28.05.2025 Revised on 16.07.2025

Accepted on 22.08.2025 Published on 08.10.2025

Available online from October 15, 2025

Asian Journal of Pharmaceutical Analysis. 2025; 15(4):319-327.

DOI: 10.52711/2231-5675.2025.00050

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