GLYCERIN

This is a safety assessment of glycerin as used in cosmetics. Glycerin functions as a denaturant, fragrance ingredient, hair conditioning agent, humectant, oral care agent, oral health-care drug, skin protectant, skin conditioning agent-humectant, and viscosity-decreasing agent. The Cosmetic Ingredient Review Expert Panel (Panel) reviewed relevant animal and human data. The Panel concluded that glycerin is safe as a cosmetic ingredient in the practices of use and concentration described in this safety assessment.

The human stratum corneum (SC) serves as a key barrier against exogenous molecules and is a living, heterogeneous system whose composition and microstructure vary along its depth. Noninvasive in vivo monitoring of its spatiotemporal microstructural changes is thus crucial yet challenging in many areas, such as transdermal drug delivery and cosmetics. Herein, we directly employed confocal Raman microspectroscopy on forearm skin to characterize the SC and its dynamic response to glycerin. To overcome low spectral resolution from signal overlap, two-dimensional correlation spectroscopy (2DCOS), moving window 2DCOS (MW2D), and perturbation-correlated MW2D (PCMW2D) were used to monitor the (1) trends and (2) sequence of Raman signal variations with depth. These served as unique Raman indicators to describe the multiscale structural features of SC, determine its thickness, and clarify glycerin's effects on lipid, keratin, and water molecular structures within SC. Further, a three-layer stratification of SC was proposed, revealing its spatially differential swelling upon adding glycerin. These glycerin-induced changes, from molecular structure to microscale swelling, would enhance the penetration of retinyl palmitate. Finally, the recovery of SC over time was assessed by the depth-dependent variations in the molecular conformations of SC components and the SC thickness, uncovering an unexpected application of glycerin as a temporary and mild skin penetration enhancer (SPE). Consequently, two-dimensional correlation Raman spectroscopy is promising in characterizing the SC and exploring qualified SPEs.

Within the framework of green chemistry and wild fruit valorization, this study optimizes the extraction of bioactive compounds from Prunus spinosa L. fruits using glycerin-based ultrasound-assisted extraction (UAE). Response Surface Methodology (RSM) and Artificial Neural Networks (ANN) were comparatively employed to model the process. Significant improvements in extraction efficiency were achieved, with total phenolic content increasing from 9.28 to 23.22 mg GAE/g DW, total flavonoid content from 6.53 to 21.65 mg CE/g DW, and antioxidant activity (DPPH assay) from 57.04% to 86.34%. While both models performed well, ANN demonstrated slightly higher predictive accuracy, highlighting its potential for capturing complex, non-linear relationships in the extraction process. We identified the optimal extraction conditions as 9 min extraction time, 100% ultrasonic amplitude, and 40% water in glycerin, and these conditions were experimentally validated. UHPLC-DAD-HRMS/MS profiling revealed a rich phytochemical fingerprint dominated by phenolic acids, caffeoylquinic acid derivatives, and flavonol glycosides, and revealed largely overlapping qualitative phytochemical profiles between hydroglyceric and ethanolic extracts. Comparative extraction using 70% ethanol under identical conditions resulted in lower TPC, TFC, and antioxidant activity, indicating the improved efficiency of glycerin under the investigated conditions. Overall, the optimized glycerin-based UAE provides a sustainable, food-safe approach for extracting bioactive compounds from underutilized P. spinosa fruits. These results support its application in functional foods and in nutraceutical and cosmetic formulations.

This study evaluated the clinical bactericidal efficacy and safety of a novel chlorine dioxide teat disinfectant compared to a traditional iodine glycerin disinfectant in dairy cows. The randomized controlled trial included long-term natural exposure (100 cows) and teat surface disinfection (40 cows) experiments. Key metrics assessed were somatic cell count (SCC), teat skin health (dryness, roughness, hyperkeratosis), and bacterial reduction rates against Staphylococcus aureus , Escherichia coli , and Streptococcus spp. Results demonstrated that the chlorine dioxide teat disinfectant achieved comparable to iodine glycerin disinfectant in controlling the rate of SCC exceeding the threhold (3.57% vs. 4.50% at day 10; p > 0.05), teat skin dryness, end roughness, and hyperkeratosis severity showed no significant differences over time or between iodine glycerin (control) and chlorine dioxide teat disinfectant groups (all p > 0.05). Notably, quantitative bacteriological assessment revealed significantly higher log 10 reduction values for chlorine dioxide teat disinfectant (2.14) versus iodine glycerin controls (1.93; p < 0.05). Microbiological evaluation further demonstrated complete pathogen eradication (100.00%) by chlorine dioxide across all isolates ( S. aureus , E. coli , Streptococcus spp.), whereas iodine glycerin achieved 99.84-100.00% bactericidal rates. The findings suggest that chlorine dioxide teat disinfectant is a sustainable and effective alternative to iodine glycerin disinfectant, offering robust antimicrobial activity, improved teat condition, and reduced residue concerns. Further multicenter studies are warranted to validate these outcomes under diverse herd management conditions.

Glycerin is widely used in pharmaceuticals, cosmetics, and foods. In recent years, it has been mainly derived from plant-based materials, particularly palm oil. The most abundant polyphenol in palm oil is 3-O-caffeoylshikimic acid (CSA). Polyphenols reportedly react with oxygen to generate hydrogen peroxide (H 2 O 2 ). In this study, we investigated the generation of H 2 O 2 from CSA and palm-derived glycerin using the bis(2,4,6-trichlorophenyl)oxalate (TCPO) chemiluminescence method and electron spin resonance spectroscopy. H 2 O 2 concentrations in the range of 10 -4 to 10 -6  mol/L were detected in commercial glycerin samples. In addition, the H 2 O 2 concentrations in a commercial glycerin varied widely between batches, even among samples of glycerin produced by the same company. However, the concentrations were approximately 1/1000 of that permitted in disinfectants used in pharmaceuticals, indicating extremely low biological toxicity. Furthermore, the TCPO chemiluminescence method proved to be highly sensitive and precise, even in viscous glycerin samples, showing potential for application in quality control of glycerin formulations and related industrial products.

Glycerol is a trihydroxy alcohol that has been included for many years in topical dermatological preparations. In addition, endogenous glycerol plays a role in skin hydration, cutaneous elasticity and epidermal barrier repair. The aquaporin-3 transport channel and lipid metabolism in the pilosebaceous unit have been evidenced as potential pathways for endogenous delivery of glycerol and for its metabolism in the skin. Multiple effects of glycerol on the skin have been reported. The diverse actions of the polyol glycerol on the epidermis include improvement of stratum corneum hydration, skin barrier function and skin mechanical properties, inhibition of the stratum corneum lipid phase transition, protection against irritating stimuli, enhancement of desmosomal degradation, and acceleration of wound-healing processes. Even an antimicrobial effect has been demonstrated. Topical application of glycerol-containing products improves skin properties in diseases characterized by xerosis and impaired epidermal barrier function, such as atopic dermatitis. The increase of epidermal hydration by glycerol is critical in skin conditions aggravated by dry and cold environmental conditions, e.g. winter xerosis. This paper provides a review on effects of glycerol on the skin, the mechanisms of its action, and the potential applications of glycerol in dermatology.

Glycerol and glycerol-3-phosphate (Gro3P) are key metabolites at the intersection of carbohydrate, lipid, and energy metabolism. Their production and usage are organismal and cell-type specific. Glycerol has unique physicochemical properties enabling it to function as an osmolyte, protein structure stabilizer, and an antimicrobial and antifreeze agent, important to the preservation of many biological functions. Glycerol and Gro3P are implicated in many physiological and disease processes relating to energy metabolism, thermoregulation, hydration, skin health, male fertility, aging, and cancer. Glycerol has countless applications in the food, pharmaceutical, and cosmetics industries. It is used as a sweetener, preservative, thickening agent, humectant, osmolyte, and cryoprotectant. It is widely used in skin and wound care products, laxatives, in cell and tissue preservation, and in medicines for numerous conditions. Here, we review the multiple uses and functions of glycerol and Gro3P and associated transporters, enzymes, and target genes in health, senescence, and disease. We discuss the evidence that glycerol may be present at much higher levels in tissues and cells than in the blood. We bring particular focus to the newly identified glycerol shunt in the direct formation of glycerol independent of lipolysis and as a pathway allowing cells to adapt to various stresses. Relevant to chronic metabolic diseases, cancer and aging, glycerol and Gro3P present important translational implications and thus warrant much more attention.

The AQPs (aquaporins) are a family of homologous water transporting proteins expressed in many mammalian epithelial, endothelial and other cell types. Phenotype analysis of mice lacking individual AQPs has been informative in elucidating their role in mammalian physiology. For example, phenotype analysis has indicated an important role of AQPs in the renal urinary concentrating mechanism (AQP1-AQP4), brain water balance and neural signal transduction (AQP4), exocrine gland secretion (AQP5) and ocular fluid balance (AQP1, AQP5). In skin, the aquaglyceroporin AQP3 is expressed in the basal layer of epidermal keratinocytes. Mice deficient in AQP3 have dry skin with reduced SC (stratum corneum) hydration, decreased elasticity and impaired biosynthesis. Mechanistic analysis of the altered skin phenotype in AQP3 deficiency suggested that the glycerol rather than the water transporting function of AQP3 is important in skin physiology. The glycerol content of SC and epidermis of AQP3 deficient mice is reduced, whereas that of dermis and serum is normal. The dry, relatively inelastic skin in AQP3 null mice is probably related to the humectant properties of glycerol, and the impaired SC repair to impaired epidermal biosynthetic function. The key role of AQP3 in epidermal physiology might be exploited in the development of improved cosmetics and new therapies for skin diseases associated with altered skin water content.

The treatment and management of acute cough due to common cold costs billions of dollars of healthcare expenditure and there is a growing opinion that a simple linctus containing glycerol with flavourings such as honey and lemon is a safe and effective treatment for acute cough in children and adults. Glycerol is a component of most cough syrups, and although it is often thought of only as a solvent or thickening agent in cough syrups, it may be a major component for the efficacy of cough syrups due to its special properties of lubrication, demulcency, sweetness, and acting as a humectant. The major benefit of cough syrups in soothing cough is likely due to the properties of the syrup rather than the active ingredients and this review discusses the special properties of glycerol in relation to the treatment of acute cough.

Two studies were performed to evaluate the influence of glycerol on the recovery of damaged stratum corneum barrier function. Measurements of transepidermal water loss and capacitance were conducted in a 3-day follow-up after tape stripping (study 1) and a 7-day follow-up after a barrier damage due to a repeated washing with sodium lauryl sulphate. In study 1 a faster barrier repair (transepidermal water loss) was monitored in glycerol-treated sites. Significant differences between glycerol open vs. untreated and glycerol occluded vs. untreated were observed at day 3. Stratum corneum hydration showed significantly higher values in the sites treated with glycerol+occlusion, compared with all other sites. In study 2 a faster barrier repair was seen in glycerol-treated sites, with significant differences against untreated and base-treated sites 7 days after the end of the treatment. Stratum corneum hydration showed highest values in the glycerol treated sites after 3 days of treatment. Glycerol creates a stimulus for barrier repair and improves the stratum corneum hydration; stratum corneum hydration is not strictly related to barrier homeostasis and can be optimized by different mechanisms and pathways. The observed effects were based on the modulation of barrier repair and were not biased by the humectant effect of glycerol. As the glycerol-induced recovery of barrier function and stratum corneum hydration were observed even 7 days after the end of treatment, glycerol can be regarded as a barrier stabilizing and moisturizing compound.

Hydrodistillation of aromatic plants for essential oil production generates substantial amounts of solid and liquid residues that are commonly discarded despite their potential value as sources of bioactive compounds. In this study, the essential oil and post-distillation residues of Rhododendron tomentosum Harmaja were evaluated within a waste-to-value framework to recover phenolic compounds with antioxidant and antimicrobial properties. Dry extracts obtained from liquid (DEA) and solid (DEE) residues were characterized in terms of total phenolic and flavonoid contents, antioxidant capacity (DPPH assays), and antimicrobial activity against selected microorganisms. Quantitative HPLC-PDA analysis revealed multiple phenolic compounds. Extracts derived from solid residues exhibited significantly higher phenolic and flavonoid contents and stronger antioxidant activity than those obtained from liquid residues, indicating that solid by-products constitute a richer phenolic matrix. Antimicrobial assays revealed pronounced activity for extracts prepared from plant material harvested in October, particularly those based on propylene glycol and glycerin, which were effective against both Gram-positive bacteria and selected Gram-negative clinical isolates. The essential oil showed broad-spectrum antimicrobial activity, including inhibition of Aspergillus niger . Stability studies demonstrated that the phenolic composition and bioactivity of the dry extracts were largely preserved after one year of storage. These findings demonstrate that R. tomentosum hydrodistillation residues represent a promising source of natural antioxidants and antimicrobial agents, supporting their potential utilization as value-added ingredients in food and cosmetic applications and contributing to circular economy strategies. When used epidurally or intravenously during labor or for a short time immediately postpartum, amounts of fentanyl ingested by the neonate are usually small and are not expected to cause any adverse effects in breastfed infants. Preterm infants are the most susceptible to fentanyl’s effects because their clearance is substantially reduced.[1] The results of studies on the effect of epidural fentanyl on breastfeeding initiation and duration are mixed, because of the many different combinations of drugs, dosages and patient populations studied as well as the variety of techniques used and deficient designs of many of the studies. It has been suggested that a cumulative dose of 80 to 150 mcg of fentanyl during labor and delivery reduces breastfeeding success,[2-4] but another study found no marked decrease in breastfeeding success with doses above 150 mcg in motivated women with previous breastfeeding success.[5] In infants placed skin-to-skin after a normal vaginal delivery, epidural fentanyl given during labor may delay the infant's first suckling in a dose-dependent manner,[6] perhaps because it can persist in the infant's serum for over 24 hours after discontinuation.[7] However, it appears that with good breastfeeding support, epidural fentanyl plus bupivacaine has little overall effect on breastfeeding success.[8-12] No waiting period or discarding of milk is required before resuming breastfeeding after fentanyl is used for short procedures (e.g., for endoscopy).[13,14] After general anesthesia, breastfeeding can be resumed as soon as the mother has recovered sufficiently from anesthesia to nurse. When a combination of anesthetic agents is used for a procedure, follow the recommendations for the most problematic medication used during the procedure. Limited information indicates that transdermal fentanyl in a dosage of 100 mcg/hour results in undetectable fentanyl concentrations in breastmilk. Mothers with opioid use disorder who are using fentanyl have lower breastfeeding initiation rates than other mothers with opioid use disorder.[15] Breastfeeding 72 hours after the birthing parent’s last use of nonprescribed fentanyl h

Gemmotherapy is a branch of phytotherapy based on the use of extracts derived from plant meristematic tissues, including buds, young shoots, and sprouts. Due to their embryonic nature and high metabolic activity, these tissues constitute a concentrated source of bioactive compounds such as polyphenols, phytohormones, amino acids, vitamins, and enzymes. The unique phytochemical profile of bud extracts, together with synergistic interactions within the phytocomplex, contributes to their enhanced biological activity compared to mature plant materials. This review provides a comprehensive overview of the current state of knowledge on the chemical composition, extraction technologies, and biological properties of gemmotherapeutic preparations. Particular attention is given to both traditional and modern extraction methods, including glycerin maceration and pulsed ultrasound-assisted extraction, as well as factors affecting the quality and variability of the obtained extracts. Available evidence indicates that gemmotherapeutic preparations exhibit a broad spectrum of biological activities, including antioxidant, anti-inflammatory, immunomodulatory, antimicrobial, and anticancer effects. These properties suggest their potential application as supportive agents in the management of chronic inflammatory diseases, metabolic disorders, and infections, as well as in functional foods and natural cosmetics. However, the lack of standardized production protocols, variability of raw materials, and limited clinical evidence remain significant challenges. Further research focusing on advanced analytical techniques, metabolomic profiling, and clinical validation is essential for the integration of gemmotherapy into evidence-based medicine.

Due to biodegradability, functionalization, and sustained release, polymer-based films are widely used in different industries. This study explores a bioactive emulsion-based film obtained using high-methoxy pectin (HMP), Origanum onites L. essential oil, and a hydroalcoholic extract of Thymus vulgaris L., prepared using various emulsion recipes. The emulsions obtained were applied to cellulose supports intended for topical applications. Bioactive textiles were analyzed using SEM-EDS elemental mapping, ATR FT-IR spectroscopy, biocompatibility assessment, antimicrobial activity assays, and analysis of comfort indices. SEM images of textile supports treated with bioactive emulsions confirmed the creation of a film surface and that the homogeneity of the film increases with increasing amount of glycerin, which acts as a plasticizer. Infrared spectra combined with their second derivatives and PCA indicate the presence of oregano essential oil, thyme extract, and pectin on the surface of the cotton. The biocompatibility evaluation of functionalized cotton supports revealed minimal cytotoxic effects on HaCaT human keratinocytes after 24 h of exposure. The results of the analyses showed that bioactive textile supports also exhibit antimicrobial activity. Therefore, the active emulsions with pectin, oregano essential oil, and hydroalcoholic extract of thyme provide biocompatible and antimicrobial active films by applying on cellulosic supports.

Background and objective Multiple-head skin prick test (SPT) devices are essential tools for percutaneous allergy testing; however, significant interdevice variability exists that may affect diagnostic accuracy, clinical performance, and interpretation. Intradevice variability, characterized by inconsistent responses across individual test heads, may lead to false-positive or false-negative results, potentially compromising allergen identification and treatment planning. This prospective study compared four FDA-cleared multiple-head SPT devices with respect to pain perception, histamine sensitivity, glycerin specificity, and intradevice variability to identify clinically meaningful performance differences. Methods Thirty healthy adults participated in this institutional review board-approved study (approval 33-098). Participants were tested with two 10-head devices (AllerTest-10 (MedScience Research Group, Inc., West Palm Beach, FL, USA; distributed by ALK-Abelló, Inc., Port Washington, NY, USA) and Skintestor OMNI (Greer Laboratories, Inc., Lenoir, NC, USA)) and two eight-head devices (AllerTest-8 (MedScience Research Group, Inc.; distributed by ALK-Abelló, Inc.) and Multi-Test II (Lincoln Diagnostics, Inc., Decatur, IL, USA)). Pain was assessed using a Visual Analog Scale (VAS; 0-10). Histamine (1 mg/mL) and glycerin controls were applied to volar forearm surfaces. Sensitivity was defined as a histamine wheal ≥3 mm, and specificity as a glycerin wheal <3 mm. Intradevice variability was expressed as the coefficient of variation (CV) across test heads. Results Pain scores were low across all devices (mean VAS, 0.92-1.77), with no statistically significant differences. Among the 10-head devices, AllerTest-10 demonstrated superior sensitivity (88% vs. 68%; P = 0.002) and significantly lower intradevice variability (median CV, 0.18 vs. 0.72; P = 0.00024) compared with Skintestor OMNI. Among the eight-head devices, AllerTest-8 showed higher sensitivity (96% vs. 88%; P = 0.026) than Multi-Test II. Both AllerTest devices achieved 100% specificity, whereas comparators achieved 97%-98%. AllerTest devices demonstrated consistently low CV values (0.18-0.19), indicating superior test-head uniformity. Conclusions All evaluated devices demonstrated acceptable patient comfort and clinical performance. However, AllerTest devices exhibited superior clinical performance, characterized by higher histamine sensitivity, perfect glycerin specificity, and markedly lower intradevice variability. These findings suggest that device selection may meaningfully affect diagnostic accuracy, with lower intradevice variability reducing the likelihood of physician misinterpretation and improving the reliability of allergen identification.

Despite the extensive use of synthetic and semi-synthetic lipids in nanostructured lipid carrier (NLC) systems, the utilization of Shorea spp. (Illipe butter or tengkawang fat), an indigenous natural solid lipid from Indonesia, remains unexplored. This study aimed to develop and optimize a Nanostructured Lipid Carrier (NLC) formulation using Illipe butter (Shorea spp.) as a natural solid lipid source. The NLC system was designed through a Box-Behnken design to evaluate the influence of solid lipid (Illipe butter and glyceryl monostearate), liquid lipid (oleic acid), and surfactant mixture (Tween 80 and glycerin) on physicochemical characteristics. The optimized formulation Nanostructured Lipid Carrier-Illipe Butter (NLC-IB) was incorporated into a gel base to produce an NLC gel formulation suitable for topical anti-inflammatory application. Characterization of the optimized NLC revealed a particle size of 276.9 nm, PDI of 0.393, and zeta potential of - 53.5 mV, indicating good stability. Transmission electron microscopy showed spherical particles with an imperfect crystal (Type I) matrix. GC-MS analysis confirmed the retention of major fatty acids, particularly stearic acid (31.9%) and palmitic acid (18.2%), in the NLC system. In vitro anti-inflammatory evaluation using the BSA denaturation assay demonstrated moderate activity, with IC₅₀ values of 197.23 ppm (Illipe butter), 146.46 ppm (NLC-IB), and 238.49 ppm (E-NLC-IB). To our knowledge, this study represents one of the first systematic investigations of Illipe butter (Shorea spp.) as a solid lipid matrix in nanostructured lipid carrier systems.

A TNF-α-induced inflammatory model using the human keratinocyte HaCaT cell line was developed and validated as an alternative in vitro method for evaluating the anti-inflammatory potential of cosmetic ingredients. In comparison with the conventional lipopolysaccharide (LPS)-stimulated RAW264.7 murine macrophage model, the HaCaT-based system demonstrated enhanced sensitivity, detecting significant cytokine inhibition at concentrations up to 1000-fold lower for certain actives. The model reliably distinguished established anti-inflammatory agents (ectoine, troxerutin, and dipotassium glycyrrhizinate) from non-active controls (glycerin, butanediol, and propylene glycol) through the suppression of interleukin-6 (IL-6). Notably, the HaCaT model also identified a potential pro-inflammatory shift at high concentrations of some active ingredients-an effect not observed in the RAW264.7 system. Multi-laboratory verification using the novel cosmetic ingredient β-nicotinamide mononucleotide (NMN) confirmed both the anti-inflammatory activity of NMN and the high reproducibility of the assay. These results support the TNF-α-HaCaT model as a sensitive, human-relevant, and reproducible alternative for screening cosmetic ingredients, contributing to the growing toolbox of nonanimal methods for safety and efficacy assessment.

This study investigated the development of poly(vinyl alcohol) (PVA)/sericin hydrogels incorporating humectant-plasticizers glycerin, propylene glycol, and mannitol for wound dressing applications. Although PVA/sericin hydrogels exhibit excellent cytocompatibility and promote fibroblast proliferation and collagen production, making them promising candidates for wound care, they suffer from rapid moisture loss and dimensional instability. To address these limitations, glycerin, propylene glycol, and/or mannitol were incorporated to enhance the properties of the hydrogels. The physicochemical and mechanical properties, together with the in vitro cytocompatibility of L929 fibroblasts, were evaluated. The results demonstrated that incorporating propylene glycol and/or mannitol improved dimensional stability, skin adhesion, moisture retention, exudate absorption, flexibility, and softness. Among all the formulations, the hydrogel containing 1 % v/v propylene glycol and 1 % w/v mannitol exhibited the most balanced performance, combining superior physicochemical and mechanical properties (time to initial upward bending: 2.83 h; time for complete upward bending of all four edges: 10.50 h; gel fraction: 60 %, highest absorption at 3 h: 34.41 %; tensile strength: 46.93 kPa, elastic modulus: 33.53 kPa, compressive strength: 19.94 kPa; compressive modulus: 15.58 kPa) with enhanced fibroblast viability (102 % cell viability) and sustained sericin release (∼90 % cumulative release at 168 h). In vitro assays confirmed the cytocompatibility of the optimized hydrogel. Taken together, these findings indicate its potential for effective management of moderately exudating wounds. This study introduces a simple and practical plasticization approach for PVA/sericin hydrogels, offering a promising candidate for wound dressing applications. Further clinical studies are warranted to evaluate its therapeutic efficacy and safety.

Piper nigrum L. (PNL) is a rich source of piperine, a bioactive alkaloid with pharmaceutical, cosmetic, nutritional supplement, and agricultural applications, yet efficient and sustainable extraction methods remain underexplored. This study compared ultrasonic bath extraction (UBE) and ultrasonic probe extraction (UPE) using natural deep eutectic solvents (NADES) for isolating piperine from PNL fruits. Six NADES formulations were screened, with NADES-5 (choline chloride:glycerin:urea, 1:1:1) showing superior performance. Response surface methodology with a Box-Behnken design optimized extraction parameters, including liquid-to-solid ratio, extraction time, temperature, and water content, for both UBE and UPE. Optimized UPE consistently outperformed UBE, yielding 49.97 mg/g of piperine versus 25.67 mg/g under identical NADES conditions. Comprehensive characterization using TLC, HPTLC, UV, FTIR, Raman, HPLC, NMR, XRD, SEM, and EDX confirmed the successful isolation and structural integrity of piperine, with samples obtained via UPE exhibiting higher purity (98.7% vs. 95.2%) and enhanced crystallinity. In vitro cytotoxicity assays demonstrated that piperine extracted by UPE showed stronger activity against C2C12 myoblasts (IC 50 : 24.3 μg/mL vs. 40.6 μg/mL) and greater anticancer effects in MCF-7 and HT-29 cells compared to piperine extracted by UBE. Antioxidant evaluation via DPPH, ABTS, FRAP, and TAC assays, along with intracellular reactive oxygen and nitrogen species suppression in THP-1 and RAW 264.7 macrophages, further confirmed the superior biological potential of the UPE-derived piperine sample. These findings indicate that UPE using NADES is a sustainable approach for high-yield piperine extraction with enhanced purity and bioactivity, supporting its potential for pharmaceutical applications.