Antifungal drugs play a vital role in combating life-threatening fungal infections, yet their therapeutic effectiveness and safety critically depend on purity, stability, and reliable analytical assessment. This review comprehensively discusses impurity profiling, forced degradation studies, and bioanalytical method development of key antifungal agents within a single integrated framework - an approach not previously consolidated in the literature. The review covers major antifungal classes including azoles, polyenes, allylamines, and pyrimidine analogues. It summarizes impurity types - organic, inorganic, and residual solvents - as classified by ICH guidelines, along with their origins, formation mechanisms, and control strategies. Advanced chromatographic and spectroscopic techniques such as HPLC, LC-MS/MS, and GC-MS are highlighted for impurity detection. Forced degradation studies under ICH-recommended stress conditions are evaluated for drugs including flucytosine, terbinafine, tolnaftate, benzoic acid, and azoles. Recent advances in bioanalytical method validation using LC-MS/MS and UPLC-MS/MS for pharmacokinetic and bioequivalence studies are also outlined. The review underscores the importance of these analytical strategies in ensuring safety, efficacy, and quality control of antifungal formulations - from raw material characterization to finished product evaluation - in alignment with international pharmacopeial and ICH standards.

This study explores the use of AI-assisted data handlingin spectrophotometric method development, providing a flexible and globally accessible alternative to traditional manual software algorithms.Quadriderm cream combines four active ingredients: Clioquinol (CLIO), Betamethasone (BETA), Tolnaftate (TOL), and Gentamicin (GEN) with the preservative Chlorocresol (CC). Building on our previous research on complex pharmaceutical mixtures with challenging ratios, this study applied established protocols for CLIO and GEN while focusing on the more analytically demanding ternary subsystem (TOL, BETA, and CC).The integration of AI-enhanced spectral handling and interpretation reduces operator-dependent variability and streamlines the analytical workflow. This includes generating calibration graphs and regression equations, as well as effectively handling scanned spectral data via consecutive prompts. Validation data such as accuracy and precision are assessed to ensure reliability. Furthermore, the system enables intelligent, simultaneous analysis of laboratory mixtures and pharmaceutical formulations, enhancing both efficiency and accuracy. The AI strategy, trained on spectral data supplied and monitored by the expertiseanalyst, can automatically predict optimal wavelengths with minimal interference, while manual handling strategy rely on analyst-driven selection. Two novel approaches were developed: the factorized derivative ratio extraction using double divisor (MAN-[DD- DDE])via Spectra Manager® software and the automated double divisor derivative ratio (AUTO-[DD-DD]) via AI tools and for resolving ternary mixtures with severely overlapping UV spectra and comparing the results with those of(MAN-[DD- DD])at coincidence points. Linear working ranges were 0.5–5.0 µg/mL (TOL), 3.0–30.0 µg/mL (BETA), and 2.0–20.0 µg/mL (CC); LODs were 0.09, 0.09, and 0.26 µg/mL, respectively. AI-driven data processing strategy matched the accuracy and reproducibility of traditional strategy manipulation while reducing subjective steps and effort. Finally, the UV-spectrophotometric method for pharmaceutical cream analysis was evaluated using the MA Tool (2025) to assess sustainability across green, white, and AI-driven criteria. AI-assisted scoring via Microsoft Copilot enabled rapid, reproducible assessment, yielding a Whiteness Score of 60.9% and providing actionable recommendations for greener and more efficient workflows.

A series of novel phenylpiperazine-based carbodithioate derivatives (3a-j) were designed, synthesized, formulated as PEG-Niosomes and evaluated for their potential hydrogen sulphide (H2S) donating properties and their antifungal activities. Structural modifications were inspired by the antifungal drug tolnaftate, incorporating a dithiocarbamate moiety to enhance biological activity and target engagement. H2S-releasing capacity was quantified via the methylene blue assay. Compound 3j (S-phenacyl p-chloro derivative) exhibiting the highest release (89.71%), followed by compounds 3a and 3i. Antifungal efficacy against Candida albicans was determined using the microbroth dilution method, where compounds 3a, 3b, 3f, 3i and 3j demonstrated potent inhibitory activity (MIC = 4.68 μg/mL), exceeding that of fluconazole. Further investigations revealed that these compounds also effectively suppressed hyphal formation and biofilm development, two major virulence factors of C. albicans. Notably, compound 3j exhibited superior performance across all assays. In a preliminary safety evaluation, compound 3j displayed low cytotoxicity toward mammalian fibroblasts (WS-1) (IC50 = 65.73 ± 4.20 μg/mL) compared to doxorubicin (IC50 = 26.72 ± 2.20 μg/mL), providing an 14-fold selectivity index relative to its antifungal MIC (4.68 μg/mL). Molecular docking studies corroborated these findings by showing favorable binding of compound 3j to the lanosterol 14α-demethylase enzyme (CYP51), highlighting π-sulfur and hydrophobic interactions critical for binding affinity. Additionally, compound 3j directly inhibited CYP51 in vitro (IC50 = 0.331 ± 0.025 μM), showing stronger inhibition than the reference azole fluconazole (IC50 = 1.388 ± 0.099 μM. H2S release was also verified by scavenger reversal. In the presence of MgO (1.5 mg/mL), the antifungal activity of compound 3j decreased, with the MIC rising to 32 μg/mL. These results underscore the potential of phenylpiperazine dithioates, particularly compound 3j, as promising antifungal agent with multi-target activity. Additionally, the PEG-Niosomes of compound 3j represent a promising delivery system for enhancing the dissolution and potential bioavailability of compound 3j.

Onychomycosis is a common fungal nail infection, often presenting with nail discoloration, thickening, and brittleness. Despite the availability of oral and topical antifungal agents, recurrence rates remain high, and systemic therapies can pose risks such as hepatotoxicity and drug interactions. This case report describes an innovative treatment approach using a compounded topical preparation made by combining a crushed 250 mg oral terbinafine tablet with an over-the-counter 1% tolnaftate nail lacquer. A healthy 28-year-old female presented with diffuse toenail onychomycosis affecting all digits. After declining oral therapy, she initiated daily application of the compounded lacquer for 10 months. Weekly complete removal and reapplication were performed, and the vial was shaken before each use to maintain a homogenous mixture. The patient documented progress photographically and reported high satisfaction with a significant improvement in nail appearance. No adverse effects were noted. This topical use of pulverized terbinafine takes advantage of the drug's favorable molecular profile for nail penetration, offering targeted treatment with minimal systemic absorption. The estimated daily delivery of terbinafine per nail was approximately 0.4 mg, a potentially therapeutic dose when applied consistently over time. This case supports the potential of creative compounding in expanding safe and patient-friendly treatment options for onychomycosis, especially for individuals who are unable or unwilling to take oral antifungals. Further studies are needed to evaluate the efficacy, pharmacokinetics, and broader applicability of this approach.

Topical tolnaftate has not been studied during breastfeeding and no data are available on the extent of its absorption after topical application. Because it is probably poorly absorbed after topical application, it is considered a low risk to the nursing infant. Avoid application to the nipple area and ensure that the infant's skin does not come into direct contact with the areas of skin that have been treated. Only water-miscible cream, gel or liquid products should be applied to the breast because ointments may expose the infant to high levels of mineral paraffins via licking.[1]

Psoriasis is a chronic inflammatory disease characterized by clearly marginated silvery plaques that affect men and women equally. Symptoms can vary among individuals; typically, it presents on the scalp, elbows, and knees. We present two cases of patients initially diagnosed with tinea pedis who showed no improvement with medical treatment. The first patient is an African American male in his 50s who arrived at a free clinic for unhoused persons with lesions to both feet initially diagnosed as tinea pedis. Although the patient was compliant with applying topical formulations of tolnaftate and clotrimazole, there was no discernible improvement in his symptoms and the skin lesions. After a thorough examination of the skin throughout the entire body, the diagnosis of psoriasis was considered. The patient started treatment with steroidal cream with improvement of the symptoms and lesions. The second patient is a Caucasian male in his 20s who also presented initially with complaints of a dry, intensely pruritic, and scaly rash on the dorsum of both his feet, as well as in between the digits of his feet for which an initial diagnosis of tinea pedis was also made. The patient remained non-compliant with treatment and, after reevaluation of his lesions along with an extensive survey of his body, was deemed to have psoriasis and prescribed topical hydrocortisone. The patient continued to remain non-compliant with his therapeutic regimen and subsequently developed cellulitis which is yet to resolve with treatment.

Topical antifungals with activity against dermatophytes include amorolfine, allylamines, azoles, ciclopiroxolamine, and tolnaftate. Polyene antimycotics, such as amphotericin B and nystatin, alternatively, miconazole are suitable for yeast infections of the skin and mucous membranes. For severe yeast infections of the skin and mucous membranes, oral triazole antimycotics, such as fluconazole and itraconazole, are used. Pityriasis versicolor is treated topically with antimycotics, and in severe forms also orally with itraconazole, alternatively fluconazole. Terbinafine, itraconazole and fluconazole are currently available for the systemic treatment of severe dermatophytoses, tinea capitis and onychomycosis. In addition to proven therapeutic regimens, unapproved (off-label use) intermittent low-dose therapies are increasingly being used, particularly in onychomycosis. Oral antimycotics for the treatment of tinea capitis and onychomycosis in children and adolescents can only be used off-label in Germany. In general, any oral antifungal treatment should always be combined with topical antifungal therapy. In tinea corporis and tinea cruris caused by Trichophyton (T.) mentagrophytes ITS (internal transcribed spacer) genotype VIII (T. indotineae), there is usually terbinafine resistance. Identification of the species and genotype of the dermatophyte and resistance testing are required. The drug of choice for T. mentagrophytes ITS genotype VIII dermatophytoses is itraconazole. In individual cases, treatment-refractory onychomycosis may be due to terbinafine resistance of T. rubrum. Here too, resistance testing and alternative treatment with itraconazole should be considered. Therapy monitoring should be carried out culturally and, if possible, using molecular methods (polymerase chain reaction). Alternative treatment options include laser application, and photodynamic therapy (PDT).

Botrytis cinerea is a high-risk pathogen for fungicide resistance development. Within the fungal populations, strains have developed multiple mutations in different target genes leading to multiple resistance (MLR) or mutations associated with overexpression of efflux transporters leading to multidrug resistance (MDR). These types of resistance are a major threat, and their successful management is a major challenge. The current study was initiated to a) determine frequencies of MLR/MDR strains in populations originating from several crops, b) identify the types of MDR that occur in Greece, and c) determine interactions between MLR and MDR at the level of sensitivity to botryticides. The frequencies of MLR/MDR phenotypes were determined in 515 isolates subjected to bioassays using discriminatory concentrations of thiophanate-methyl, iprodione, cyprodinil, fenhexamid, boscalid, fluopyram, fludioxonil, pyraclostrobin, and tolnaftate. Interestingly, 7.8% and 31.3% of isolates from strawberry and rootstock seedlings were resistant to every single fungicide class, while MDR phenotypes from strawberries, rootstocks, and tomatoes accounted for 26%, 87%, and 13.4%, respectively. The MLR and MDR isolates were further molecularly analyzed regarding genes erg27, sdhB, Bcpos5, and Mrr1, responsible for resistance to fenhexamid, boscalid and fluopyram, cyprodinil, and MDR, respectively. The different mutations' presence was determined along with a new mutation in Mrr1 leading to MDR. MDR isolates were characterized as MDR1 or MDR1h based on the presence of a 3-bp deletion in Mrr1. MDR1h was predominant in isolates from rootstocks and MDR1 from tomatoes and strawberries, whereas the most frequent target-site mutations were F412S (erg27), H272R (sdhB), and L412F (Bcpos5). To determine whether the accumulation of target-site mutations along with MDR mutations exhibits an additive effect concerning fungicide resistance, the sensitivity of isolates possessing the predominant target-site mutations was calculated in both the presence and the absence of MDR-associated mutations. EC50 in cyprodinil and boscalid increased to about twofold in the presence of MDR mutations, while there was no difference for fenhexamid. In conclusion, MLR/MDR frequencies are notably high in heavily treated crops in Greece, and the combination of MLR and MDR mutations leads to even higher fungicide resistance levels, highlighting the importance of resistance management.

The eye is a complex organ with a unique physiology and anatomy. Using novel nanosystems is expected to enhance ocular drug permeation and retention. Hence, this work aimed to study the potential of flexosomes as an ocular delivery system to enhance the corneal permeation and antifungal activity of Tolnaftate (TOL). Different flexosomes formulae were formulated using ethanol injection method, employing a 3[1].2[2] full factorial design. The studied formulation variables were: X1: amount of stearyl amine, X2: hydration volume and X3: type of edge activator. Encapsulation efficiency, particle size and zeta potential were selected as dependent variables. FX5 was selected as the optimal TOL flexosomes and showed encapsulation efficiency of 66.08 ± 11.38%, particle size of 154.99 ± 29.11 nm and zeta potential of 42.95 ± 0.64 mV. FX5 was subjected to further ex vivo and in vivo studies which showed that TOL flux was significantly increased through FX5 compared to TOL suspension. Draize test and histopatholoigal tests assured that FX5 is safe to be used for eye.. The in vivo fungal susceptibility testing using Aspergillus niger demonstrated the superior and more durable antifungal activity of FX5 than TOL suspension. Hence, FX5 can be considered as promising nanocarrier for safe and efficient ocular TOL delivery.