Degradation Products Research Articles

Astral-based DIA proteomics explored the flavor enhancement mechanism of Chinese traditional smoked bacon by staphylococcal co-fermentation

The proteolysis pattern during mixed fermentation of Staphylococcus cohnii WX-M8 and S. saprophyticus MY-A10 on Chinese bacon was still unknown. In this study, the changing laws of protein degradation products during staphylococcal mixed fermentation were analyzed, followed by an investigation of endogenous enzymes and cellular components, and finally an examination of flavor profiles. Results indicated that mixed fermentation improved protein degradation and promoted the production of peptides and free amino acids (FAAs). Proteolysis of S. saprophyticus MY-A10 was non-specific, and it promoted protein degradation by cooperating with cathepsin L1. S. cohnii WX-M8 was specific and acted mainly with calpain-3 in the thin filament. The fulfillment of S. cohnii WX-M8 function was enhanced in the presence of S. saprophyticus MY-A10. Mixed fermentation showed synergism with endogenous peptidases in degrading peptides to small-molecule peptides or FAAs and complementarity with endogenous dehydrogenases in converting FAAs to volatile organic compounds (VOCs).

Degradation kinetics, influencing factors, and mechanisms of polycyclic aromatic hydrocarbons in a Fe3O4-nZVI Fenton-like system

To improve the catalytic performance of nano zero-valent iron (nZVI) in a Fenton-like system, Fe3O4 supported nZVI (Fe3O4-nZVI) was produced by the liquid-phase reduction method and co-precipitation method, then characterized by SEM, EDS, XRD, XPS, and VSM. Fe3O4-nZVI was used as the catalyst in a Fenton-like system to degrade polycyclic aromatic hydrocarbons (PAHs) such as naphthalene (Nap), phenanthrene (Phe), and pyrene (Pyr) in simulated wastewater. The results showed that under the optimization conditions, which were 25℃, 0.6g/L of Fe3O4-nZVI dosage, 3 of the pH in the reaction system, and 10-15mmol/L of the concentration of H2O2, the sequence of degradation efficiency in this system was Phe (99.76%)>Pyr (99.71%)>Nap (98.01%). The reused experiments on Fe3O4-nZVI showed that the degradation efficiency for PAHs remained above 87% after 4 reuses of Fe3O4-nZVI. These results indicated that nanoscale magnetic composites of Fe3O4-nZVI had excellent reactivity, stability, and reuse performance. The fitting parameters of kinetics showed that the degradation of PAHs in the Fenton-like system with the Fe3O4-nZVI catalyst fitted well with the Behnajady-Modirshahla-Ghanbery kinetic model. The radial quenching experiment proved that ·OH was the dominant free radical for the degradation of PAHs in the Fe3O4-nZVI Fenton-like system. Based on the degradation products of PAHs and the related literature, the possible degradation paths of the three PAHs in the Fe3O4-nZVI Fenton-like system were inferred. The above results could provide a reference and basis for the treatment of wastewater containing PAHs by Fe3O4-nZVI Fenton-like technology.

Polycystic Ovary Syndrome Accompanied by Hyperandrogenemia or Metabolic Syndrome Triggers Glomerular Podocyte Injury

Objective: To determine whether the urinary excretion of podocyte degradation products varies according to PCOS phenotype and metabolic syndrome (MetS). Methods: The concentrations of podocalyxin (PDX) and nephrin, chronic markers of podocyte damage, and neutrophil gelatinase-associated lipocalin (NGAL), a marker of acute glomerular damage, were analyzed in the morning urine samples of 50 PCOS patients and 50 healthy controls matched by age and BMI. Albuminuria was assessed by calculating the urine albumin–creatinine ratio (uACR). Results: The PDX, nephrin and NGAL concentrations of PCOS participants were significantly higher than those of the control group. While PDX, nephrin and NGAL levels of classic phenotypes were similar, they were higher than ovulatory and non-hyperandrogenic phenotypes. Significant increases in urinary levels of each podocyte protein were detected in PCOS patients with MetS compared to patients without MetS. A positive significant correlation between podocyte proteins and BMI, systolic blood pressure, testosterone, glucose, HOMA-IR and uACR. After adjusting for age and BMI, podocyte proteins were an independent risk factor for microalbuminuria. The incidence of microalbuminuria in PCOS increased 6-fold compared to controls. The frequency of microalbuminuria was higher in classical phenotypes than in ovulatory phenotype. The frequency of microalbuminuria in PCOS patients with MetS was 6.5 times higher than in PCOS patients without MetS. Conclusions: In PCOS accompanied by hyperandrogenemia or metabolic syndrome, leakage of acute and chronic podocyte breakdown products into the urine becomes more pronounced.

Liquid chromatography-tandem mass spectrometry analysis of nitrogen mustard degradation products after derivatization with benzoylimidazole

Nitrogen mustards are highly toxic compounds and strictly controlled by the Chemical Weapons Convention. When those agents are used, exposure must be confirmed by analyzing biological samples from victims to allow prompt medication and reduction of secondary damage. In this study, a derivatization-liquid chromatography (LC)-tandem mass spectrometry (MS/MS) method to analyze nitrogen mustard degradation products (ethanolamines; EAs) in human urine was developed. Benzoylimidazole (BzI) was used as a derivatization reagent, and two highly hydrophilic EAs (methyldiethanolamine and ethyldiethanolamine) were analyzed by using a reversed-phase column with sufficient retention. For urine samples, after solid-phase extraction with cation exchange cartridge and derivatization, EAs were analyzed at low limits of detection (5 ng/mL). The calibration curves had a wide linear range (50–1000 ng/mL), and the quantitation results were sufficient for screening. Compared with our previous derivatization-LC-MS/MS method using pentafluorobenzoyl chloride as a derivatization reagent and hydrophilic interaction liquid chromatography (HILIC)-MS/MS method, not only the limits of detection but also the operability and throughput were improved. The performance of the newly developed derivatization reagent was also evaluated using density functional theory (DFT) calculations.

Green stability-indicating RP-HPTLC technique for determining croconazole hydrochloride

Abstract The objective of the proposed investigation is the development and validation of a green stability-indicating reverse-phase high-performance thin-layer chromatographic method to determine croconazole hydrochloride (CCZ). The developing system used was an 80:20 v/v mixture of acetone and water. The measurement of CCZ was done at 198 nm. With the use of the Analytical Eco-Scale (AES), ChlorTox, and Analytical GREEnness (AGREE) tools, greenness was evaluated. The linearity was demonstrated by the present method in the 25–1,200 ng/band range. The present approach was additionally reliable, accurate, sensitive, precise, and green. An exceptional greenness profile was demonstrated by the AES, total ChlorTox, and AGREE scales, which were determined to be 89, 1.08 g, and 0.82, respectively. The greenness metrics of the present method were much better than the reported high-performance liquid chromatography approach. Under acid and oxidative degradation circumstances, CCZ was shown to be unstable, while under alkaline and thermal-stress settings, it was sufficiently stable. Furthermore, the stability-indicating component determined by analytical method identified CCZ in the presence of its degradation products. Commercial CCZ cream was found to contain 0.98% w/w of CCZ. The investigation’s results suggested that CCZ in commercially available creams might be regularly examined with the help of the recommended green technology.

Accurate Visualization of C4d Complement Fragment in Immunohistochemistry by C-Terminal Linear Neoepitope-Specific Antibodies

C4d is the end degradation product of activated complement component C4b that appears during the early steps of the classical and lectin complement pathways. Within the primary sequence of C4d, there is a reactive thioester group that binds covalently to nearby surfaces, thus labeling the locations of complement activation. This feature makes C4d a target for immunohistochemical staining aimed to aid the diagnosis of, among others, the antibody-mediated rejection of transplanted organs, membranous glomerulonephritis, bullous pemphigoid, or inflammatory myopathies. However, the credibility of C4d immunostaining is debatable, as a high background in surrounding tissues and body fluids and diffused patterns of deposits in target structures are experienced with some of the available anti-C4d antibodies. Herein, we present an improved version of a rabbit anti-C4d antibody, originally raised against the C-terminal linear neoepitope of this complement fragment. Minor cross-reactivity with C4b and native C4 proteins, measured by ELISAs, as well as relatively low concentrations necessary for obtaining a specific signal in immunohistochemical analyses of formalin-fixed paraffin-embedded material, makes the improved antibody superior to commercially available rabbit monoclonal anti-C4d antibody SP91 dedicated to ex vivo diagnostics, as demonstrated by the staining of a panel of kidney transplant biopsies.

Evolution of the biochemistry underpinning purine alkaloid metabolism in plants.

Purine alkaloids are naturally occurring nitrogenous methylated derivatives of purine nucleotide degradation products, having essential roles in medicine, food and various other aspects of our daily lives. They are generated through convergent evolution in different plant species. The pivotal reaction steps within the purine alkaloid metabolic pathways have been largely elucidated, and the convergent evolution of purine alkaloids has been substantiated through bioinformatic, biochemical and other research perspectives within S-adenosyl-ʟ-methionine-dependent N-methyltransferases. Currently, the biological and ecological roles of purine alkaloids, further refinement of the purine alkaloid metabolic pathways and the investigation of purine alkaloid adaptive evolutionary mechanisms continue to attract widespread research interest. The exploration of the purine alkaloid metabolic pathways also enhances our comprehension of the biochemical mechanism, providing insights into inter-species interactions and adaptive evolution and offering potential value in drug development and agricultural applications. Here, we review the progress of research in the distribution, metabolic pathway elucidation and regulation, evolutionary mechanism and ecological roles of purine alkaloids in plants. The opportunities and challenges involved in elucidating the biochemical basis and evolutionary mechanisms of the purine alkaloid metabolic pathways, as well as other research aspects, are also discussed. This article is part of the theme issue 'The evolution of plant meta-bolism'.

Ethoxy Acetalated Dextran-Based Biomaterials for Therapeutic Applications

A novel class of pH-responsive polymers, acetalated dextran, has emerged in the field of biomaterials. These versatile materials are derived from dextran through a simple acetalation reaction, allowing for the creation of polymers with a tunable release profile which allows the controlled release of encapsulated therapeutics in response to acidic environments. Despite their recent introduction, acetalated dextran has rapidly garnered significant interest due to its potential for various therapeutic applications. This review delves specifically into the recent advancements of ethoxy acetalated dextran or Ace-DEX, a particular acetalated dextran with a distinct advantage: its degradation products (acetone and ethanol) are less toxic compared to other variants that produce methanol. The focus of this review is the diverse range of biomedical applications currently being explored for Ace-DEX-based scaffolds. Finally, this review concludes by addressing the existing challenges associated with Ace-DEX and outlining potential future research directions within this promising field.

Insight into HTPB pyrolysis mechanism under high-temperature: A reactive molecular dynamics study

Hydroxyl-terminated polybutadiene (HTPB) is widely utilized in solid and hybrid rocket propellants due to its mechanical properties and combustion performance. Insight into its pyrolysis process is key to enhancing combustion efficiency in rocket engines. This study employs ReaxFF molecular dynamics (MD) simulations to explore the pyrolysis mechanism of HTPB under extreme conditions, with temperatures ranging from 1000 K to 2000 K. The results identify the primary degradation products and elucidate their formation mechanisms. The simulation reveals that C-C bond cleavage at polymerization sites is the initial step, followed by the formation of linear oligomers and butadiene. Subsequent reactions, including hydrogenation and dehydrogenation, lead to the generation of smaller molecular species. The kinetic analysis confirms that HTPB pyrolysis follows first-order reaction kinetics, with an activation energy of 8.12 kcal/mol. The findings are compared with existing experimental data, highlighting the influence of thermal environments on pyrolysis mechanisms and product distributions.

Elucidation of a specific alginate lyase Aly7Rm: The products demonstrated the strict recognition of G residue at subsites ±2

An alginate lyase Aly7Rm with novel subsite specificity was cloned, expressed and characterized in this article. Aly7Rm was a G-specific alginate lyase, exhibiting optimum reaction conditions at 30 °C and pH 5.0. UPSEC-VWD-MS analysis revealed that Aly7Rm degraded polysaccharides in a random endo-acting manner. The main degradation products of alginate were trisaccharide to octasaccharide and those of PG were disaccharide to hexasaccharide. The product distribution was related to the specificity of enzyme and the structure of substrates. By utilizing the HPAEC-PAD/MS method, the structure of products was identified and the specificities of subsite were defined. Aly7Rm accommodated both G and M at subsites −1 and strictly recognized G at subsite −2 and +2. This is the first report on the alginate lyase exhibiting G specificity at subsites ±2, which shed light on the diversity in subsite specificity of alginate lyases. The Aly7Rm with clear action pattern could serve as an efficient tool in the specific degradation of alginate and targeted preparation of oligosaccharides.

Immunomodulatory Effect of Vitamin D3 in Rats with Induced Osteoarthritis

Background: The (OA) caused by imbalance between anabolic and catabolic processes within the cartilage ,one of the effects of increased catabolic changes is presence of products of extracellular matrix degradation in the synovial fluid which lead to the joint microenvironment alteration which attempt. Aim of study: Evaluate the influence of induce osteoarthritis (OA) on knee joint and looking on (OA) effect as local immune response in addition to the possible role of vitamin (D3) as protective and repairing agent against (OA) complications. Methods: Total number of rats male (89) were used and divided into three groups, first group (control group) (19) rats, second group (OA induced) (35) rats injected with papain enzyme at dose of (10 mg), treated group (35) rats which firstly injected with papain enzyme at dose (10 mg) then after (2) weeks of injection, treated the rats with vitamin D. Results and Dicussion: Macroscopic observations revealed to cartilage surface erosion , fibrillation , hypertrophied condyles, degeneration. The rats knee joints that treated (2) weeks with vitamin (D3) post (OA) induced showed smooth, intact cartilage surface. Radiographic images indicated to narrowing joints cavity (space) at knee joint of (OA) rat model, destruction of epiphyseal plate, while the results related to (OA) rats treated with vitamin (D3) showed no osteophyte growth , the synovial cavity nearest to the normal, no defect. Conclusion: Vitamin (D) prevent the stability and life of chondrocytes which lead to cartilage protection and synthesis of (ECM) , and inhibite the inflammation resulted from cartilage damaged.

An S-Scheme CeO2/Bi2O2S heterojunction photoanode for the photoelectrocatalytic degradation of sulfamethoxazole in synthetic and real wastewater

We prepared an S- scheme heterojunction photoanode using cerium oxide (CeO2) and bismuth oxysulfide (Bi2O2S) for the photoelectrocatalytic degradation of sulfamethoxazole. The CeO2 /Bi2O2S photoanode was synthesised via an in-situ hydrothermal method, ensuring strong contact and efficient charge transfer between the CeO2 and Bi2O2S. The materials and photoanode were characterized with XRD, XPS, photoluminescence, and photoelectrochemistry. The S-scheme configuration observed in the formation of heterojunction in CeO2 /Bi2O2S photoanode was responsible for the improved photoelectrocatalytic performance for the visible light-assisted degradation of sulfamethoxazole. Operational parameters such as the effect of pH and current density were examined. The extent of sulfamethoxazole mineralisation was calculated to be 72 % using the total organic carbon (TOC) measurement. The LC-MS analysis was used to predict the degradation pathway and products. Furthermore, the photoelectrocatalytic efficiency of the CeO2/Bi2O2S photoanode was investigated in real wastewater matrices with TOC removal of 54 %. Therefore, the S scheme CeO2/Bi2O2S photoanodes lends itself to photoelectrochemical water treatment applications.

Intradermal and transdermal absorption of beta-naphthylamine and N-Phenyl-beta-naphthylamine in a viable human skin model

Technical products containing N-Phenyl-beta-naphthylamine (PBNA) are contaminated with beta-naphthylamine (BNA), a known carcinogen. Both amines penetrate the skin to different degrees, but little is known about their dermal-depot formation. This study investigated the dermal penetration of PBNA and its degradation product BNA using a viable human-skin model. PBNA (259 μg) or BNA (0.52 μg) in n-hexane and industrial grease were applied to freshly excised human skin (n = 6, 0.64 cm2) for 2-72 h. After temporary/continuous and single/repeated exposure, samples were taken (stratum corneum, epidermis/dermis, receptor fluid) and analyzed for their amine content by GC–MS. Continuous exposure led to a PBNA dermal depot of ∼47 μg/cm2 over 72 h. Temporary applications also resulted in lower but consistent PBNA dermal depots. A single 2-h application resulted in a dermal depot of ∼16 μg/cm2 after 72 h, while this was ∼25 μg/0.64 cm2 with repeated applications. BNA behaved differently; with repeated 2-h applications, intradermally retained BNA initially increased 3–6 fold, then dropped to ∼200–250 ng/cm2. This incomplete decline upon repeated short-term exposure to PBNA suggests that a BNA dermal depot is formed either due to contamination of PBNA with BNA or to enzymatic conversion of PBNA to BNA. Additionally, PBNA dermal depots were saturable under the given conditions. These findings highlight the importance of understanding the dermal-exposure dynamics of potential carcinogenic compounds in industrial settings.

Impact of high-pressure columbite phase of titanium dioxide (TiO2) on catalytic photoconversion of plastic waste and simultaneous hydrogen (H2) production

Photoreforming is a sustainable photocatalytic process that degrades plastic waste while simultaneously producing hydrogen (H2) from water. However, this process has received limited attention due to the scarcity of effective catalysts capable of both plastic degradation and H2 production, such as titanium dioxide (TiO2). In this study, an active catalyst is developed by stabilizing the high-pressure orthorhombic phase of TiO2, known as columbite, using a high-pressure torsion (HPT) method. This material effectively degrades polyethylene terephthalate (PET) plastic under light, converting it into valuable organic compounds such as formic acid, terephthalate, glycolic acid, and acetic acid. Additionally, it produces a significant amount of H2. The findings show that the high-pressure orthorhombic phase, especially in the presence of oxygen vacancies, enhances catalytic H2 production and microplastic degradation by increasing light absorption, reducing electron-hole recombination, and generating hydroxyl radicals. These results highlight the substantial potential of modified high-pressure TiO2 photocatalysts in simultaneously addressing the plastic waste crisis and the demand for H2 fuel.

Ratiometric fluorescent composites united metal-organic frameworks sensing system for detection of methyl-paraoxon

Organophosphorus pesticide residues pose significant risks to human health, but conventional analytical methods are costly and complex. To address this, we developed a new environmentally friendly technique using a simple hydrothermal method to create a ratiometric fluorescent composite (RFC) with two distinct signals: blue and orange. This RFC allowed for rapid detection and quantitative analysis of organophosphorus pesticides. The composite incorporated a cerium-based metal–organic framework (Ce-UiO-66), which could degrade methyl paraoxon (MP) due to the two oxidation states of cerium. The resulting degradation product, p-nitrophenol (p-NP), specifically quenched the blue fluorescence of the RFC via the inner filter effect (IFE), while the orange fluorescence remained unaffected, ensuring consistency under microenvironmental and spectrometer conditions. This innovative approach was sensitive, with a detection limit of 0.55 μM, selective, and repeatable, offering a new strategy for pesticide analysis and safety evaluation of edible and medicinal plant and water samples.

From aromatic rings to aliphatic compounds - degradation of duloxetine with the use of visible light driven Z-scheme photocatalyst

The growing consumption of antidepressants, associated with their high chemical stability and low biodegradability, become a serious problem for aquatic environment. Many of their components are resistant to conventional water treatment technologies. The photocatalytic processes with the use of semiconductors becoming attractive methods of drugs degradation because the highly reactive species: electrons, holes, hydroxyl radicals (⦁OH) and superoxide anion radicals (O2⦁−) generated under illumination may be involved directly or indirectly in decomposition of complicated organic molecules.In this work we have shown for the first time that Z-scheme photocatalyst, consisted of bismuth vanadate (BiVO4), graphitic-like carbon nitride (g-C3N4) and Au nanoparticles (Au NPs) as a charge transfer mediator, is very efficient in degradation of duloxetine (DLX). We performed the comprehensive evaluation focused not only on degradation rate, mechanism of photocatalytic process, but also identification of decomposition products and the pathways of DLX degradation. Due to application of the elaborated Z-scheme photocatalyst, the degradation rate constant was more than 2-3 times higher than that obtained for photolysis, but more important, the final compounds were mainly aliphatic products, formed by opening of aromatic rings. The chemical structure of the degradation products were determined with the use of HPLC and LC-MS. The toxicity of photoproducts with respect to water organisms was tested by means of Spirotox and Microtox assays. The Microtox test indicated that toxicity of the products after 6h of the solution irradiation in the presence of Z-scheme photocatalyst is about 4 times lower than that after photolysis.

How Does Triclocarban Affect Sulfur Transformation in Anaerobic Systems?

Triclocarban (TCC), as a typical antimicrobial agent, accumulates at substantial levels in natural environments and engineered systems. This work investigated the impact of TCC on anaerobic sulfur transformation, especially toxic H2S production. Experimental findings revealed that TCC facilitated sulfur flow from the sludge solid phase to liquid phase, promoted sulfate reduction and sulfur-containing amino acid degradation, and largely improved anaerobic H2S production, i.e., 50-600 mg/kg total suspended solids (TSS) TCC increased the cumulative H2S yields by 24.76-478.12%. Although TCC can be partially biodegraded in anaerobic systems, the increase in H2S production can be mainly attributed to the effect of TCC rather than its degradation products. TCC was spontaneously adsorbed by protein-like substances contained in microbe extracellular polymers (EPSs), and the adsorbed TCC increased the direct electron transfer ability of EPSs, possibly due to the increase in the content of electroactive polymer protein in EPSs, the polarization of the amide group C═O bond, and the increase of the α-helical peptide dipole moment, which might be one important reason for promoting sulfur bioconversion processes. Microbial analysis showed that the presence of TCC enriched the organic substrate-degrading bacteria and sulfate-reducing bacteria and increased the abundances of functional genes encoding sulfate transport and dissimilatory sulfate reduction.

Comprehensive Analysis of Phycoerythrin 545 Stability and the Apoptotic Impact of Its Degradation Products on HT29 Cells.

This study investigates the properties and potential applications of phycoerythrin 545, a naturally occurring light-harvesting pigment protein from Rhodomonas salina. Phycoerythrin 545, characterized by its bright red color and maximum absorption wavelength at 545 nm, was extracted using freeze-thawing methods, further purified, and analyzed using chromatographic, spectroscopic techniques, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Phycoerythrin 545 consists of two subunits, primarily α and β, but lacks the γ subunit, and is stable at 4 °C within a pH range of 3-10. To further characterize it, its susceptibility to degradation by trypsin was assessed. The biological activity of phycoerythrin 545 and its degradation products were investigated in HT29 human colon cancer cells. The results showed that the degradation products, particularly those within 3-10 kDa, significantly decreased the viability of HT29 cells by inducing apoptosis. Mechanistic studies indicated these effects were mediated through the activation of mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinases and MAPK/c-Jun N-terminal Kinase signaling pathways and involved the regulation of key apoptotic proteins such as p53, Bim, Bad, Bak, and Bax, leading to the activation of the Caspase-3 apoptotic pathway. These findings contribute to understanding the structural and functional properties of phycoerythrin 545, laying a foundation for its exploration in food industry applications and cancer therapy supplementation.

Atmospheric Degradation Mechanism of Isoamyl Acetate Initiated by OH Radicals and Cl Atoms Revealed by Quantum Chemical Calculations and Kinetic Modeling.

Isoamyl acetate is one of the volatile organic compound class molecules relevant to agricultural and industrial applications. With the growing interest in isoamyl acetate applications in industry, the atmospheric fate of isoamyl acetate must be considered. Reaction mechanisms, potential energy profiles, and rate constants of isoamyl acetate reaction with atmospheric relevant oxidant OH radicals and Cl atoms have been obtained from the quantum chemical calculations and kinetic modeling. The geometry optimizations were conducted using M06-2X/6-311++G(3df,3pd) followed by single point-energy calculations at the DLPNO-CCSD(T) method with an extrapolated complete basis set. The rate constants were calculated by solving the master equation. A hydrogen-abstraction reaction dominates the first step of isoamyl acetate degradation, while the addition-substitution reaction plays a small role in the degradation products. The kinetic study was conducted to evaluate the rate constants within a temperature range of 200-400 K. The total rate constants for the isoamyl acetate degradation reactions initiated by the OH radical and Cl atom were determined to be 6.96 × 10-12 and 1.27 × 10-10 cm3 molecule-1 s-1, respectively, under standard temperature and pressure conditions. The product degradation mechanism, ozone formation potential, and atmospheric impacts were discussed.

A Review on Analytical Methods of Anti-fungal Agents used for the Treatment of Onychomycosis

Background: Analytical methods have an essential function in the healthcare sector, particularly when it emanates from the creation and approval of medications. These methods are essential for identifying the active ingredients, ensuring the purity of compounds, and quantifying the existence of any impurities or degradation products. Objective: The objective of the research article is to discuss an analytical method available for the drug Onychomycosis, which is indeed crucial in treating nail infection, to confirm the efficacy and quality of pharmaceutical products. Among various analytical techniques available, UV Spectroscopy, RP-HPLC (high performance-liquid chromatography) and tandem mass spectrometry are normally employed due to their speed and accuracy. Method: Literature surveys typically reveal the predominance of UV, RP-HPLC and LC-MS/MS methods for estimating azole derivatives due to their established reliability, sensitivity, and applicability to an extensive range of compounds. Researchers often select the most suitable method based on factors such as sample complexity, sensitivity requirements, and available instrumentation. Results: This present art states most of the validation parameters like LOQ, Linearity range, Column and mobile phase used in different dosage forms as well as versatile techniques. Conclusion: This review article provides a comprehensive evaluation of various analytical methods employed in the estimation of antifungal agents, particularly those used to treat Onychomycosis. Onychomycosis, a prevalent infection on nails caused by fungi, demands precise and accurate analytical techniques to ensure the safety and effectiveness of antifungal therapies. In this review, we discussed and compared a range of analytical technologies, highlighting their applications and sensitivity in the context of Onychomycosis drug analysis. This critical assessment aims to guide researchers and practitioners in selecting the most suitable methods for their specific analytical needs.