1. INTRODUCTION
Skin aging is an integral aspect of overall aging, showing distinct patterns across organs, tissues, and cellular structures over time. It results from a synergy of intrinsic factors (genetics, metabolism, hormones) and extrinsic influences such as prolonged ultraviolet (UV) exposure, pollutants, ionizing radiation, and various chemicals and toxins [1,2]. UV is classified into UV-A (315–400 nm), UV-B (280–315 nm), and UV-C [3]. UV-A degrades collagen and elastin, causing wrinkles and premature aging [4]. UV-B stimulates excessive melanin production, contributing to visible skin aging and carcinogenesis [5]. It increases reactive oxygen species (ROS) and induces pro-inflammatory and mitogenic mediators, such as α-Melanocyte-stimulating hormone [6]. Various skin cells, including keratinocytes, melanocytes, fibroblasts, and endothelial cells, interact to drive melanogenesis and age-related pigmentation [7].
Management strategies for skin aging and hyperpigmentation include sunscreens, topical retinoic acid, niacinamide, hydroquinone (HQ), and more invasive interventions like chemical peels, microdermabrasion, botulinum toxin, fillers, and lasers, which carry side effects. HQ 4%–5% remains the gold standard for melasma treatment [8]. However, long-term use is linked to exogenous ochronosis in up to 55.5% of users [9], permanent corneal damage [10], respiratory irritation [11], impaired wound healing, reduced elasticity, neuropathy, and potential carcinogenicity through DNA damage [12]. Therefore, safer alternatives to inhibit melanin synthesis are needed.
Recent studies highlight steady progress in the search for natural agents that can reduce hyperpigmentation by targeting multiple steps in melanogenesis [13]. These include blocking tyrosinase, scavenging ROS, binding metal ions, and attenuating signaling pathways such as MC1R–cAMP–CREB–MITF [14]. Reviews published in the last few years point to a wide range of plant-derived compounds such as phenolics, flavonoids, stilbenes, and coumarins that have shown activity in both cell-based and animal models, along with advances in formulations that improve skin penetration and efficacy [15]. Encouragingly, clinical data on botanicals are also beginning to accumulate. A recent scoping review of 21 clinical trials on herbal remedies for melasma found that several plant products improved pigmentation scores and patient quality of life [16]. More recent trials even suggest that certain botanical or naturally derived regimens can perform on par with standard treatments like HQ. For instance, topical Raphanus sativus seed powder 3% cream has comparable effectiveness for treating melasma with 4% HQ cream [17]. Together, these findings strengthen the case for continuing to explore nature-based options as safe and effective alternatives for managing hyperpigmentation.
The genus Passiflora, comprising ~520 tropical species, includes Passiflora edulis (passion fruit), widely used in traditional medicine, with an annual global production of about 1.5 million tons [18]. Its anti-aging properties stem from potent antioxidant activity due to secondary metabolites such as trans-ferulic acid, chlorogenic acid, p-coumaric acid, quercetin, rutin, naringenin, gallic acid, cryptoxanthin, α- and β-carotene, provitamin A, and kaempferol [19,20]. These compounds neutralize free radicals generated by metabolism and environmental factors, preventing oxidative stress-induced hyperpigmentation [21]. Incorporation into dermatological formulations can help mitigate visible signs of aging [22,23]. Enriched P. edulis fruit extract, containing antioxidants, vitamins, and flavonoids, has been scientifically shown to prevent UV-B-induced skin aging. A study reported that 3% P. edulis seed extract cream improved clinical signs of facial aging in 40 participants with minimal side effects and high satisfaction [24].
Extracts from P. edulis, particularly the seeds, have been studied for their antioxidant, photoprotective, and anti-photoaging properties. However, evidence on the fruit extract is limited, especially regarding its effects on UV-B–induced hyperpigmentation and underlying mechanisms. This study, therefore, offers incremental novelty, not by introducing an entirely new concept, but by extending existing knowledge to P. edulis fruit extract formulated as a cream. Moreover, existing studies often rely on a single experimental approach, limiting mechanistic insights. By integrating in vitro, in vivo (C57BL/6 mice model), and in silico approaches, the present work provides a more comprehensive evaluation of its depigmentation potential compared to HQ, thereby supporting its candidacy as a safer natural alternative for managing hyperpigmentation.
2. METHODS
2.1. Sources of P. edulis fruit extract and cream formulation
The standardized P. edulis fruit extract was purchased from Perseroan Terbatas. Genero Pharmaceuticals in the form of 70% hydrosol. Then, the formulation was suspended in the formulation containing Sepigel 305, 2% lanolin, 2% dimethicone, and 0.5% phenoxyethanol to a final concentration of 5% and 10% of P. edulis fruit extract. The controlled formulations did not contain the P.edulis extract (0%). The final pH of the formulations was adjusted to 5.5–6.0 (skin-compatible range). The formulations were visually inspected to ensure homogeneity (no phase separation, uniform color, and texture) and were stored at 4°C until use. Although formal stability testing was not performed, no changes in color, odor, or texture were observed during the experimental period.
2.2. Mushroom tyrosinase assay
The tyrosinase inhibitory activity was evaluated using a microplate-based spectrophotometric method. Briefly, reaction mixtures were prepared in 96-well microplates in a total volume of 150 µl. For each test concentration, 10–40 µl of the P. edulis fruit extract solution (dissolved in phosphate buffer with up to 0.1% Dimethyl Sulfoxide) was combined with 80–100 µl of sodium phosphate buffer (20–50 mM, pH adjusted between 6.5 and 6.9) and 10–30 µl of mushroom tyrosinase enzyme solution (approximately 2.5–100 units/ml, as appropriate for the desired reaction rate). The mixture was pre-incubated for 5 minutes at 25°C–30°C to equilibrate. Enzymatic oxidation was initiated by adding 80–100 µl of freshly prepared L-DOPA substrate solution (final concentration 5–10 mM). The formation of dopachrome was monitored by measuring absorbance at 475 or 490 nm using a microplate reader (e.g., Thermo Scientific or Bio-Rad), with readings recorded either every 5 minutes for up to 60 minutes to calculate reaction velocity. All reactions were carried out in triplicate, and experiments were independently repeated three times. Kojic acid served as the positive control reference inhibitor in all experiments. The percentage inhibition of the enzyme was calculated as follows:
IA: percentage inhibition of tyrosinase enzyme, C: control without inhibitor, and S: plant extract.
2.3. Antioxidant measurement
Antioxidant activity was evaluated using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging method. A 0.1 mM DPPH stock solution was prepared by dissolving DPPH in ethanol and incubating it in the dark for at least 30 minutes before use. The test sample, a 70% (v/v) hydrosol extract of P. edulis, was first converted to ppm units (700 mg/ml = 700,000 ppm). To prepare the highest working concentration (200 ppm), 1.43 µl of the extract was diluted in 5 ml of distilled water. Lower concentrations (100, 75, 50, and 25 ppm) were prepared by serial dilution. Each sample solution (1 ml) was mixed with 1 ml of the DPPH solution in a test tube, followed by incubation in the dark at 37°C for 30 minutes. After incubation, the absorbance was measured at 517 nm using ethanol as the blank. Each concentration was measured in technical duplicate within each experiment, and the DPPH assay was independently repeated two times and averaged. The percentage of DPPH radical scavenging activity was calculated using the following formula:
where FRSA% is free radical scavenging activity, Abs sample is the absorbance of the sample solution, and Abs control is the absorbance of the control solution.
2.4. Determination of sun protection factor (SPF)
The SPF value of the P. edulis fruit extract (not the cream formulation) was determined using UV spectrophotometry. Briefly, 1 g of the test cream formulation was dissolved in 96% ethanol and diluted to a final volume of 100 ml to prepare a 1% w/v stock solution. This stock solution was further diluted to a final concentration of 0.02% w/v to ensure that absorbance readings remained within the linear range of the spectrophotometer. The absorbance of the diluted sample was measured using a UV-Vis spectrophotometer across the wavelength range of 290–320 nm, with readings taken every 5 nm (i.e., at 290, 295, 300, 305, 310, 315, and 320 nm). The 96% ethanol was used as the blank. Each measurement was performed in triplicate to obtain an accurate mean value. SPF was calculated using the following equation:
where CF is the correction factor [10], EE(λ) is the erythemal effect spectrum, I(λ) is the solar intensity spectrum at each wavelength, and Abs(λ) is the sample absorbance at each wavelength. The products for each wavelength were summed and multiplied by the correction factor to obtain the final SPF value.
2.5. Animals and skin pigmentation modelling
Healthy adult C57BL/6 mice (male, 12 weeks old, weighing 25–30 g) were obtained from iRatCo, Bogor, Jawa Barat. The animals were maintained under controlled conditions at 22°C ± 1°C with a 12-hour light/dark cycle. Following a 1-week acclimatization period, mice were randomly assigned to five groups (n = 6 per group): a control group (no Ultraviolet B (UVB) exposure and no cream treatment), a model group (UVB irradiation + placebo cream 0.4 mg/day), a HQ group (UVB irradiation with topical HQ cream 4% at 0.4 mg/day), a low dose treatment group (UVB irradiation with P. edulis cream 5% at 0.4 mg/day), and a high dose treatment group (UVB irradiation with P. edulis cream 10% at 0.4 mg/day). The topical dose of 0.4 mg/day was selected based on prior murine studies of HQ and plant extract creams, where doses up to 0.5 mg/day were effective in modulating skin pigmentation without causing local irritation or systemic toxicity [25]. The choice of 5% and 10% concentrations was based on our unpublished preliminary study on melanin levels in UVB-induced C57BL/6 mice, which showed that concentrations below 5% had minimal effect, whereas concentrations above 10% showed no further improvement in efficacy.
Hair was mechanically removed from the dorsal region, spanning from the neck to the rump, to ensure uniform UVB exposure. The backs of the mice were irradiated with UVB at a dose of 180?mJ/cm². Vehicle, HQ, or P. edulis fruit extract cream was applied daily, with treatment administered 20 minutes prior to and 4 hours after irradiation. Cream was applied both before and after UVB exposure to evaluate its dual role in prevention and post-exposure recovery. While this approach may yield stronger protective effects than single-application protocols, it was chosen to explore the broader therapeutic potential of the extract [26,27]. UVB exposure lasted 30 second every other day and continued for two weeks. Following the experimental period, mice were euthanized by cervical dislocation method. The tissue samples of dorsal skin were excised from the irradiated area.
2.6. Biochemical analysis
Protein was extracted from 100 mg of frozen skin tissue. In brief, the tissue was homogenized and the lysates were centrifuged at 14,000×g for 15 minutes at 4°C, and the resulting supernatant was collected into fresh tubes [28]. Protein concentrations were determined using a bicinchoninic acid assay (Thermo Fisher Scientific, Waltham, MA). The concentrations of glutathione (GSH) and tyrosinase in skin tissue samples were measured using commercially available ELISA kits: Rat GSH ELISA Kit (RE10155, Reed Biotech Ltd, Wuhan, China) and Rat TYR ELISA Kit (RE1972, Reed Biotech Ltd, Wuhan, China) following manufacturer’s instruction.
2.7. Histological analysis
Skin samples were fixed in cold 4% paraformaldehyde (Sigma-Aldrich, MO, USA). Following fixation, tissues were rinsed for 30 minutes before embedding, and a tissue processor (Thermo Fisher Scientific, MA, USA) was used to prepare paraffin blocks. Sections were sliced at a thickness of 7?μm using a microtome (Leica, Wetzlar, Germany) and dried at 60°C for 24 hours to mount onto coated slides. Fontana–Masson staining was conducted with the Fontana–Masson Stain Kit (ScyTek, Logan, UT) according to the manufacturer’s instructions. In brief, slides were deparaffinized, and sections were incubated at 60°C for 30 minutes in Fontana ammonia silver solution. After three rinses with distilled water, non-melanin staining was removed using 0.2% gold chloride solution and 5% sodium thiosulfate. Nuclei were counterstained with Nuclear Fast Red Solution. Finally, sections were dehydrated, cover-slipped, and examined microscopically. To calculate the percentage of melanin in the skin, melanin pixel area and skin pixel area were compared using ImageJ.
2.8. Statistical analysis
Comparisons among groups were performed using one-way Analysis of Variance, followed by Tukey’s Honestly Significant Difference for post hoc analysis. Prior to ANOVA, data were tested for normality using the Shapiro–Wilk test and for homogeneity of variances using Levene’s test. Data are expressed as mean ± SD. All statistical analyses were carried out with SPSS version 24 (IBM Corporation, Armonk, NY). Only two-tailed p-values less than 0.05 were considered statistically significant.
2.9. In silico studies
To evaluate the cosmeceutical potential of the primary constituents found in P. edulis fruit, physicochemical properties were assessed using freely available cheminformatics tools, including pKCSM (https://biosig.lab.uq.edu.au/pkcsm/prediction) and SwissADME (https://www.expasy.org/resources/swissadme). Descriptors such as molecular weight (MW), the number of hydrogen bond donors (n-OHNH), and hydrogen bond acceptors (n-ON), octanol–water partition coefficient (LogPo/w), and aqueous solubility (LogSw) are the component of transdermal “rule of five.” According to this rule, optimal skin penetrants typically display MW ≤ 300, n-OHNH ≤ 5, n-ON < 5, LogPo/w < 2.6, and LogSw > –2.3. The possible bioavailability problems if three or more properties were violated [29]. Moreover, to gather information on the adsorption and distribution kinetics relevant to skin permeability, four key Dermatopharmacokinetic parameters were calculated. The skin permeation coefficient in the stratum corneum (LogKp) was estimated using equation: [LogPab = 2,74 + 0,71.Log Po/w – 0,0061.MW], The effective diffusion coefficient within human dermis (Dde) was determined model using equation: [Dde = (7.1 × 10−6/MW 0.5)], to predict the maximum dermal flux (Jmax), which is a critical parameter for assessing the potential systemic or toxic effects, the following equation was applied: [Log Jmax = 3.90–0.0190 MW], and the partition coefficient between stratum corneum lipids and water (LogKsc/w) was estimated using: [LogKsc/w = 0.078.(LogP)2 + 0.868.LogMW 2.04]. These critical parameters define how compounds penetrate, absorb, diffuse, and partition within the skin [30].
The three-dimensional structure of the tyrosinase protein (PDB ID: 5M8M, resolution 2.65 Å) was retrieved from the Protein Data Bank in .pdb format [19]. For docking preparation, any pre-existing ligands were removed, water molecules were deleted, hydrogen atoms were added, and the proteins were subjected to 3D protonation followed by energy minimization to optimize their structure using PyMol. The receptor binding pockets were identified using the Molecular Operating Environment Software. The docking grid was centered at X = –32.2950, Y = –4.8549, and Z = –26.5719 with a box size of 14 × 14 × 14 Å. Binding affinities and interaction profiles were subsequently evaluated, and visual analyses of docking poses were carried out with PyRx software and Discovery Studio Visualizer, respectively.
3. RESULTS
3.1. In vitro anti-tyrosinase, antioxidant, and SPF activities of P. edulis fruit extract
In vitro assays demonstrated that P. edulis fruit extract exhibited very weak tyrosinase inhibitory activity compared to kojic acid. The Inhibitory Concentration (IC50 ) value for kojic acid was 45.96 ppm, while the passion fruit extract did not achieve 50% inhibition at the tested concentrations, making it impossible to determine an IC50 value. The maximum inhibition observed with passion fruit extract was 27.88% at 50% concentration, whereas kojic acid reached an IC50 of 45.96 ppm. The evaluation of the antioxidant activity using the DPPH radical scavenging assay showed that P. edulis extract exhibited moderate antioxidant activity, with an IC50 of 123.7 ppm. Vitamin C was used as the positive control in the DPPH assay and demonstrated very strong antioxidant activity, with an IC50 of 15.9 ppm. The SPF evaluation conducted in this study classified passion fruit extract in the “low protection” category (Table 1).
Table 1. In vitro anti-tyrosinase, antioxidant, and SPF activities of P. edulis fruit extract.
| Parameter | Methods | Compounds | Conc. | Inhibition (%) | IC50 |
|---|---|---|---|---|---|
| Anti-tyrosinase activity | Mushroom tyrosinase inhibitory test | Kojic acid | 2.34 ppm | 8.36 ± 0.66 | 45.96 ppm |
| 4.69 ppm | 13.12 ± 0.23 | ||||
| 9.38 ppm | 14.73 ± 1.72 | ||||
| 18.75 ppm | 26.11 ± 0.47 | ||||
| 37.50 ppm | 43.98 ± 1.05 | ||||
| 75.00 ppm | 63.47 ± 1.58 | ||||
| 150.00 ppm | 76.95 ± 0.32 | ||||
| P. edulis fruit extract | 3.13% | 6.61 ± 1.03 | - | ||
| 6.25% | 10.37 ± 0.81 | ||||
| 12.50% | 13.86 ± 0.31 | ||||
| 25.00% | 18.89 ± 0.31 | ||||
| 50.00% | 27.88 ± 0.68 | ||||
| Antioxidant activities | DPPH test | Vitamin C | 1 ppm | 5.92% | 15.9 ppm |
| 5 ppm | 23.91% | ||||
| 10 ppm | 38.61% | ||||
| 20 ppm | 55.71% | ||||
| 50 ppm | 75.90% | ||||
| P. edulis fruit extract | 25 ppm | 18.9% | 123.7 ppm | ||
| 50 ppm | 25.5% | ||||
| 75 ppm | 39.2% | ||||
| 100 ppm | 52.7% | ||||
| 200 ppm | 75.4% | ||||
| SPF activities | Mansur method | P. edulis fruit extract | 5% | 2.11 | - |
3.2. The effect of P. edulis fruit extract on GSH in UVB-induced C57BL/6 mice
Experiments in UV-B–exposed C57BL/6 mice showed that 10% P. edulis fruit extract cream significantly prevented the decrease in GSH levels compared to the control group (p < 0.05). In contrast, 4% HQ cream did not have a significant effect on GSH levels (p > 0.05). These findings indicate that 10% P. edulis fruit extract cream has sufficient antioxidant activity to protect the skin from oxidative stress, while HQ cream showed no antioxidant effect (Fig. 1).
 | Figure 1. Effects of P.edulis fruit extract cream on the levels of skin tissues GSH in a control group (no UVB exposure and no cream treatment), a model group (UVB irradiation + placebo cream 0.4 mg/day), a HQ group (UVB irradiation with topical hydroquinone cream 4% at 0.4 mg/day), a low dose treatment group (UVB irradiation with P. edulis cream 5% at 0.4 mg/day), and a high dose treatment group (UVB irradiation with P. edulis cream 10% at 0.4 mg/day). p-values were calculated with one-way ANOVA with Tukey’s HSD post-hoc test (n = 6 mice). [Click here to view] |
3.3. The effect of P. edulis fruit extract on tyrosinase in UVB-induced C57BL/6 mice
Measurement of tyrosinase enzyme levels in skin tissue showed that 4% HQ cream significantly reduced tyrosinase levels by 49% in UV-B–exposed mice (p < 0.05). The 5% P. edulis fruit extract cream produced only a minimal, non-significant reduction (p > 0.05). However, the 10% P. edulis fruit extract cream significantly lowered tyrosinase levels by 38% (p < 0.05), and this reduction was not statistically different from that of HQ cream (Fig. 2).
 | Figure 2. Effects of P.edulis fruit extract cream on the levels of skin tissues tyrosinase in a control group (no UVB exposure and no cream treatment), a model group (UVB irradiation + placebo cream 0.4 mg/day), a HQ group (UVB irradiation with topical hydroquinone cream 4% at 0.4 mg/day), a low dose treatment group (UVB irradiation with P. edulis cream 5% at 0.4 mg/day), and a high dose treatment group (UVB irradiation with P. edulis cream 10% at 0.4 mg/day). p-values were calculated with one-way ANOVA with Tukey’s HSD post-hoc test (n = 6 mice). [Click here to view] |
3.4. The effect of P. edulis fruit extract on melanin content in UVB-induced C57BL/6 mice
Measurement of melanin levels in skin tissue showed that 4% HQ cream significantly reduced melanin by 30% in UV-B–exposed mice (p < 0.05). The 5% P. edulis fruit extract cream had only a minimal, non-significant effect (p > 0.05). However, the 10% P. edulis fruit extract cream significantly reduced melanin by 69% (p < 0.05) (Fig. 3).
 | Figure 3. Effects of P.edulis fruit extract cream on the amount of dermal melanin content in a control group (no UVB exposure and no cream treatment), a model group (UVB irradiation + placebo cream 0.4 mg/day), a HQ group (UVB irradiation with topical hydroquinone cream 4% at 0.4 mg/day), a low dose treatment group (UVB irradiation with P. edulis cream 5% at 0.4 mg/day), and a high dose treatment group (UVB irradiation with P. edulis cream 10% at 0.4 mg/day). p-values were calculated with one-way ANOVA with Tukey’s HSD post-hoc test (n = 6 mice). [Click here to view] |
3.5. DPK profile and binding affinity of P. edulis fruit extract to tyrosinase
The results showed that vomifoliol, 4-acetyl-3-hydroxy-6-methyl-2H-pyran-2-one, tetrahydropyran, and 3-(methylthio)-1-hexanol did not violate any transdermal “rule of five” parameters. Prunasin, sambunigrin, and 3-acetylmercaptohexyl acetate violated only one parameter each, while (Z)-3,5-hexadienyl butyrate, harmine, and passiflorine were also within the acceptable range (≤2 violations). In contrast, compounds such as beta-carotene, lycopene, and various triterpenoids had more than three violations, indicating they are not suitable for transdermal use without formulation modification or penetration enhancers. Further in silico evaluation of the active compounds in passion fruit extract was conducted by analyzing DPK parameters. The results showed that these compounds have excellent skin penetration profiles, supported by favorable Log Kp, Dde, and Log Ksc/w values. Further prediction on toxicological properties did not show any toxic potential in all three tests: acute dermal toxicity, skin sensitization, and skin irritation or corrosion (Table 2).
Table 2. In silico dermalpharmacokinetic indices and toxicity for the main constituents in P. edulis extract.
| Metabolit | MW (≤ 300) | n-OHNH (≤ 5) | n-ON (< 5) | Log Po/w (< 2.6) | Log Sw (> −2.3) | Transdermal Rule of Five | Predicted dermatopharmacokinetic indices | Toxicity Profile | |||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Log Kp (cm/s) | Dde (cm²/s) | Log Jmax (µg/cm²/h) | Log Ksc/w | Acute Dermal Toxicity | Skin Sensitization | Skin Irritation and Corrosion | |||||||
| beta–Citraurin | 432.648 | 1 | 2 | 7.75 | –5.26 | 3 | 0.12 | 3.41E–07 | –12.12 | 4.93 | No | Yes | Yes |
| 15,15'-cis-Phytoene | 544.952 | 0 | 0 | 15.11 | –6.22 | 3 | 4.66 | 3.04E-07 | –14.25 | 18.14 | No | Yes | Yes |
| Prolycopene | 536.888 | 0 | 0 | 14.29 | –6.99 | 3 | 4.13 | 3.06E-07 | –14.10 | 16.26 | No | Yes | Yes |
| Lycopene | 536.888 | 0 | 0 | 14.25 | –7.06 | 3 | 4.10 | 3.06E-07 | –14.10 | 16.17 | No | Yes | Yes |
| Neurosporene | 538.904 | 0 | 0 | 14.45 | –6.89 | 3 | 4.23 | 3.06E-07 | –14.14 | 16.62 | No | Yes | Yes |
| 15-cis-Phytofluene | 542.936 | 0 | 0 | 14.86 | –6.41 | 3 | 4.50 | 3.05E-07 | –14.22 | 17.56 | No | Yes | Yes |
| beta-Carotene | 536.888 | 0 | 0 | 13.26 | –7.52 | 3 | 3.40 | 3.06E-07 | –14.10 | 14.04 | No | Yes | Yes |
| beta-Cryptoxanthin | 552.887 | 1 | 1 | 11.43 | –7.31 | 3 | 2.00 | 3.02E-07 | –14.40 | 10.53 | No | No | Yes |
| gamma-Carotene | 536.888 | 0 | 0 | 13.72 | –7.32 | 3 | 3.73 | 3.06E-07 | –14.10 | 15.01 | No | Yes | Yes |
| (Z)-3,5-Hexadienyl butyrate | 168.236 | 0 | 2 | 2.94 | –2.53 | 2 | –1.68 | 5.47E-07 | –7.10 | 0.57 | No | Yes | No |
| Amygdalin | 457.432 | 7 | 12 | -2.16 | –1.76 | 3 | –7.06 | 3.32E-07 | –12.59 | 0.63 | No | No | No |
| Prunasin | 295.291 | 4 | 7 | -0.61 | –1.61 | 1 | –4.97 | 4.13E-07 | –8.51 | 0.73 | No | No | No |
| Harmine | 212.252 | 1 | 3 | 3.05 | –4.29 | 2 | –1.87 | 4.87E-07 | –7.93 | 0.71 | No | No | No |
| Antheraxanthin | 584.885 | 2 | 3 | 9.33 | –7.05 | 3 | 0.32 | 2.94E-07 | –15.01 | 7.15 | No | No | Yes |
| Mutatochrome | 552.887 | 0 | 1 | 12.66 | –7.45 | 3 | 2.88 | 3.02E-07 | –14.40 | 12.84 | No | No | No |
| Neoxanthin | 600.884 | 3 | 4 | 7.92 | –6.58 | 3 | –0.78 | 2.90E-07 | –15.32 | 5.26 | No | No | No |
| Violaxanthin | 600.884 | 2 | 4 | 8.42 | –6.77 | 3 | –0.43 | 2.90E-07 | –15.32 | 5.90 | No | No | Yes |
| 6-C-Chinovopyranosylluteolin | 432.381 | 7 | 10 | 1.29 | –4.38 | 4 | –4.46 | 3.41E-07 | –12.12 | 0.38 | Yes | Yes | No |
| 6-C-Fucopyranosylluteolin | 432.381 | 7 | 10 | 1.37 | –4.39 | 4 | –4.40 | 3.41E-07 | –12.12 | 0.39 | Yes | Yes | No |
| Callistephin | 433.389 | 7 | 10 | -1.48 | –2.88 | 4 | –6.43 | 3.41E-07 | –12.13 | 0.42 | Yes | No | No |
| Cyanidin 3-(6''-malonylglucoside) | 535.434 | 8 | 14 | -1.39 | –3.13 | 4 | –6.99 | 3.07E-07 | –14.07 | 0.48 | Yes | No | No |
| Vomifoliol | 224.3 | 2 | 3 | 0.56 | –0.99 | 0 | –3.71 | 4.74E-07 | –8.16 | 0.72 | No | No | No |
| Sambunigrin | 295.291 | 4 | 7 | -0.54 | –1.62 | 1 | –4.92 | 4.13E-07 | –6.51 | 0.63 | No | No | No |
| Cyclotricuspidoside C | 861.032 | 12 | 17 | -0.07 | –2.01 | 3 | –8.04 | 2.42E-07 | –20.26 | 0.51 | No | No | No |
| Cyclopassifloic acid A | 536.75 | 6 | 7 | 3.07 | –3.95 | 5 | –3.83 | 3.06E-07 | –14.10 | 1.06 | No | No | No |
| Cyclopassifloic acid B | 520.751 | 5 | 6 | 4.19 | –4.76 | 4 | –2.94 | 3.11E-07 | –13.79 | 1.69 | No | No | No |
| Cyclopassifloic acid C | 536.75 | 6 | 7 | 3.12 | –3.93 | 5 | –3.80 | 3.06E-07 | –14.10 | 1.09 | No | No | No |
| Cyclopassifloic acid D | 504.708 | 4 | 6 | 3.5 | –4.67 | 4 | –3.33 | 3.16E-07 | –13.49 | 1.26 | No | No | No |
| Cyclopassifloic acid E | 552.749 | 7 | 8 | 2.3 | –3.15 | 4 | –4.48 | 3.02E-07 | –14.40 | 0.75 | No | No | No |
| Cyclopassifloic acid G | 536.75 | 6 | 7 | 3.3 | –3.78 | 5 | –3.67 | 3.06E-07 | –14.10 | 1.18 | No | No | No |
| Cyclopassifloside I | 698.891 | 9 | 12 | 1.22 | –2.78 | 4 | –6.14 | 2.69E-07 | –17.18 | 0.55 | No | No | No |
| Cyclopassifloside II | 682.892 | 8 | 11 | 2.14 | –3.06 | 4 | –5.39 | 2.72E-07 | –16.87 | 0.78 | No | No | No |
| Cyclopassifloside III | 845.033 | 11 | 16 | 0.58 | –2.52 | 4 | –7.48 | 2.44E-07 | –19.96 | 0.53 | No | No | No |
| Cyclopassifloside IV | 698.891 | 9 | 12 | 1.27 | –2.68 | 4 | –6.10 | 2.69E-07 | –17.18 | 0.55 | No | No | No |
| Cyclopassifloside V | 861.032 | 12 | 17 | -0.24 | –2.08 | 3 | –8.16 | 2.42E-07 | –20.26 | 0.51 | No | No | No |
| Cyclopassifloside VI | 666.849 | 7 | 11 | 1.49 | –3.01 | 4 | –5.75 | 2.75E-07 | –16.57 | 0.58 | No | No | No |
| Cyclopassifloside VII | 714.89 | 10 | 13 | 0.6 | –2.21 | 3 | –6.67 | 2.66E-07 | –17.48 | 0.47 | No | No | No |
| Cyclopassifloside X | 698.891 | 9 | 12 | 1.52 | –2.59 | 4 | –5.92 | 2.69E-07 | –17.18 | 0.61 | No | No | No |
| Cyclopassifloside XI | 861.032 | 12 | 17 | -0.07 | –2.01 | 3 | –8.04 | 2.42E-07 | –20.26 | 0.51 | No | No | No |
| 3-Acetylmercaptohexyl acetate | 218.318 | 0 | 3 | 2.29 | –2.43 | 1 | –2.45 | 4.81E-07 | –8.05 | 0.60 | No | No | Yes |
| Cyclotricuspidogenin C | 536.75 | 6 | 7 | 3.3 | –3.78 | 5 | –3.67 | 3.06E-07 | –14.10 | 1.18 | No | No | No |
| 4-Acetyl-3-hydroxy-6-methyl-2H-pyran-2-one | 168.148 | 1 | 4 | 0.24 | –1.09 | 0 | –3.60 | 5.48E-07 | –7.09 | -0.10 | Yes | No | No |
| Passiflorine | 182.226 | 1 | 2 | 3.04 | –3.99 | 2 | –1.69 | 5.26E-07 | –7.36 | 0.64 | No | No | No |
| Tetrahydropyran | 86.134 | 0 | 1 | 1.2 | 0.22 | 0 | –2.41 | 7.65E-07 | –5.54 | 0.75 | No | No | Yes |
| 3-(Methylthio)-1-hexanol | 148.271 | 1 | 1 | 2.09 | –1.76 | 0 | –2.16 | 5.83E-07 | –6.72 | 0.69 | No | No | No |
Based on the DPK and toxicity predictions, nine secondary metabolite compounds from passion fruit extract were selected for further molecular docking studies against tyrosinase (Table 3, Fig. 4). Almost all tested ligands showed lower ΔG values compared to HQ (−6.29 kcal/mol), except for 3-(methylthio)-1-hexanol, which had a ΔG of −6.21 kcal/mol. Notably, (Z)-3,5-hexadienyl butyrate, prunasin, harmine, sambunigrin, and passiflorine displayed even lower ΔG values than the native tyrosinase ligand (kojic acid), indicating stronger binding affinity to the enzyme.
 | Figure 4. 2D representation of molecular docking poses of selected compounds from the fruit extract of P. edulis ((Z)-3,5-Hexadienyl butyrate [A], Prunasin [B], Harmine [C], Vomifoliol [D], Sambunigrin [E], 3-Acetylmercaptohexyl acetate [F], Passiflorine [G], Tetrahydropyran [H], 3-(Methylthio)-1-hexanol [I], Hydroquinone [J], Kojic Acid [K]) with tyrosinase (5M8M) as a receptor. [Click here to view] |
Table 3. Best binding energy (kcal/mol) based on AutoDock scoring of the P. edulis fruit constituents into the active sites of tyrosinase enzymes (5M8M).
| CAS ID | Metabolite | Binding energy (kcal/mol) | Interacting amino acids |
|---|---|---|---|
| 69925-34-4 | (Z)-3,5-Hexadienyl butyrate | -7.21 | TyrA:362, AsnA:378, and LeuA:382 |
| 99-18-3 | Prunasin | -7.49 | AspA:212, GluA:216, TyrA:362, ArgA:374, HisA:381, and ThrA:391 |
| 442-51-3 | Harmine | -8.33 | ArgA:374, HisA:381, LeuA:382, and GlyA:389 |
| 23526-45-6 | Vomifoliol | -6.45 | SerA:394 |
| 99-19-4 | Sambunigrin | -7.81 | ArgA:321, ArgA:374, HisA:381, GlyA:389, GlnA:390, and ThrA:391 |
| 136954-25-1 | 3-Acetylmercaptohexyl acetate | -6.48 | HisA:215, HisA:377, and HisA:381 |
| 1392-82-1 | Passiflorine | -8.25 | TyrA:361, LeuA:381, and ThrA:391 |
| 67715-80-4 | Tetrahydropyran | -6.54 | HisA:381 |
| 90180-89-5 | 3-(Methylthio)-1-hexanol | -6.21 | HisA:192, HisA:215, GluA:360, HisA:377, HisA:381, and LeuA:381 |
| 123-31-9 | Hydroquinone | -6.29 | LeuA:382 and GlyA:389 |
| 501-30-4 | Kojic Acid | -7.12 | AsnA:378, HisA:381, GlyA:389, GlnA:390, ThrA:391, and SerA394 |
4. DISCUSSION
In vitro assays demonstrated that P. edulis fruit extract exhibited very weak tyrosinase inhibitory activity compared to kojic acid. Prior studies have shown that piceatannol, a stilbene derivative, has potent tyrosinase inhibitory activity (IC50 = 149.73 µg/ml) but is present at only 0.02% in crude extracts [31]. Other reports similarly observed high IC50 values (195 µg/ml) for passion fruit extracts [32]. Optimizing chromatographic fractionation is, therefore, necessary to improve purity and enhance bioactivity.
Although the P. edulis fruit extract cream showed only low SPF protection by standard classifications, this level of photoprotection remains important in context. While insufficient as a standalone sunscreen, even modest SPF values can contribute to reducing oxidative stress when used alongside broader protective regimens, mainly due to their antioxidant effects. Studies in sunscreen formulations reveal that adding antioxidants significantly decreases ROS formation by approximately 1.7-fold in SPF 4 products and up to 2.4-fold in SPF 15–50 formulations, demonstrating how adjunctive UV absorption enhances cellular protection [33]. Furthermore, botanical extracts with combined functions such as mild UV protection plus antioxidant and melanogenesis-modulating activities may provide clear advantages over traditional UV filters alone [34]. Rather than replacing conventional sunscreens, the P. edulis cream is best suited as a complementary depigmenting. This framing defines its clinical role as a multifunctional adjunct in photoprotection and skin depigmentation strategies.
Topical application of the extract cream effectively prevented the decline of GSH in UV-B-exposed C57BL/6 mouse skin, whereas HQ cream did not affect GSH levels. This protective effect aligns with evidence that piceatannol can increase GSH concentrations in UV-B-irradiated keratinocytes [35]. Flavonoids such as isoorientin may also upregulate antioxidant defense through the Nrf2 pathway, inducing expression of GSH peroxidase and reductase [36]. In contrast, the pro-oxidant properties of HQ may exacerbate ROS production [37]. These findings suggest that the antioxidant activity of P. edulis extract can mitigate oxidative damage, which contributes to melanogenesis and skin aging, though formulation optimization is necessary to ensure sufficient skin penetration and stability of active compounds.
In vivo measurements demonstrated that both 4% HQ cream and 10% P. edulis extract cream significantly reduced tyrosinase activity in UV-B-exposed skin, with comparable efficacy. It should be noted that P. edulis fruit extract did not achieve 50% tyrosinase inhibition under the in vitro conditions tested, which indicates weak direct enzyme inhibition. Therefore, its depigmenting activity cannot be attributed to a strong tyrosinase-blocking effect. Instead, the significant in vivo reductions in melanin and tyrosinase activity are more likely explained by indirect mechanisms. These may include enhancement of antioxidant defenses, preservation of GSH, and downregulation of melanogenesis-related signaling pathways such as Microphthalmia-Associated Transcription Factor, which collectively reduce tyrosinase expression and melanin synthesis in vivo [38–40]. This distinction underlines the importance of evaluating both in vitro and in vivo outcomes when assessing natural depigmenting agents.
Analysis of melanin content revealed that both treatments suppressed UV-B-induced melanin accumulation, with the highest reduction observed in the group treated with 10% extract cream. This effect, although statistically comparable to HQ, may be attributed to antioxidant mechanisms and modulation of melanogenesis-related gene expression rather than direct enzyme inhibition. Previous studies have also reported that passion fruit extract can inhibit melanogenesis and support collagen synthesis [31], while clinical trials have confirmed its depigmenting effects in humans [24]. The favorable safety profile of P. edulis fruit extract positions it as a promising alternative or adjunct to HQ, which is associated with risks such as ochronosis and irritation [9].
In silico analyses indicated that several constituents of P. edulis extract—such as harmine, sambunigrin, prunasin, and passiflorine—have favorable dermatopharmacokinetic properties and strong predicted binding affinities to tyrosinase, surpassing those of HQ and kojic acid. Harmine, for example, has demonstrated melanin-suppressing effects via DYRK1A/NFATC3 pathway regulation [41], and prunasin exhibits antioxidant and anti-inflammatory properties [42,43]. Passiflorine may also contribute to melanogenesis inhibition [44]. These molecular docking results provide a mechanistic rationale for the extract’s observed in vivo activity, although further in vitro validation is essential.
Given the relatively modest direct inhibitory effect of P. edulis fruit extract on tyrosinase activity observed in vitro, optimizing the formulation to enhance dermal delivery represents a critical avenue for improving its depigmenting efficacy. The intrinsic limitation in penetration of bioactive compounds through the stratum corneum often restricts the clinical effectiveness of botanical agents [45]. Advanced formulation strategies such as nanoencapsulation, liposomal carriers, and the inclusion of safe chemical penetration enhancers have been demonstrated to significantly improve the transdermal absorption and bioavailability of phytochemicals [46]. These delivery systems not only facilitate enhanced skin permeation but also protect labile compounds from enzymatic and oxidative degradation, allowing for sustained release and prolonged pharmacological effects on melanogenic pathways [47]. In light of these considerations, the incorporation of such cutting-edge delivery technologies could potentiate the clinical performance of P. edulis extract cream by ensuring adequate topical bioavailability.
While promising, this study has several limitations. First, the sample size was limited to six mice per group, which, although adequate for detecting significant differences in biochemical outcomes, may not capture the full biological variability. Second, the findings are based on an animal model with thinner skin than humans, which limits direct extrapolation to clinical practice. Third, no human trials were performed, and further studies are needed to confirm efficacy and safety in patients with hyperpigmentation. Finally, mechanistic molecular assays, such as the analysis of melanogenesis-related gene expression (e.g., MITF and Tyrosinase) or Nrf2 antioxidant signaling, were not conducted. Such assays would provide deeper insights into the molecular pathways underlying the observed effects. Future work should therefore include larger sample sizes, clinical validation in human subjects, and incorporation of molecular assays to elucidate precise mechanisms of action.
5. CONCLUSION
In summary, this study indicates that a 10% P. edulis fruit extract cream could serve as an effective alternative to HQ for managing UV-B–induced hyperpigmentation and oxidative stress in skin, with potentially fewer side effects. However, these findings should be interpreted as an experimental proof-of-concept. Further preclinical optimization and clinical validation are necessary before it can be considered a clinical candidate; at present, it should be viewed as a promising research lead in the development of nature-based depigmenting therapies. Future work should focus on optimizing formulation strategies (e.g., nanoencapsulation for enhanced skin penetration), conducting mechanistic molecular assays, and performing well-designed clinical trials to establish efficacy, safety, and tolerability in humans.
6. ACKNOWLEDGMENTS
The authors gratefully acknowledge iRatCo, Bogor, Jawa Barat, for their support in conducting the in vivo experiments. The in-silico assays and DPPH antioxidant testing were performed at the Biochemistry Laboratory, School of Medicine and Health Sciences, Atma Jaya Catholic University of Indonesia. The in vitro anti-tyrosinase activity assays and Sun Protection Factor (SPF) testing were conducted at PT. Biotek Rekayasa Indonesia, Bogor.
7. AUTHORS’ CONTRIBUTIONS
All authors made substantial contributions to conception and design, acquisition of data, or analysis and interpretation of data; took part in drafting the article or revising it critically for important intellectual content; agreed to submit to the current journal; gave final approval of the version to be published; and agree to be accountable for all aspects of the work. All the authors are eligible to be an author as per the International Committee of Medical Journal Editors (ICMJE) requirements/guidelines.
8. FINANCIAL SUPPORT
This study was partly supported by a Grant-in-Aid from Atma Jaya Catholic University of Indonesia (Grant ID. 0106.17/III/LPPM-PM.10.01/03/2025).
9. CONFLICTS OF INTEREST
The authors report no financial or any other conflicts of interest in this work.
10. ETHICAL APPROVAL
All procedures were reviewed and approved by the Ethical Board of School of Medicine and Health Sciences, Atma Jaya Catholic University of Indonesia (Approval Number 10/02/KEP-FKIKUAJ/2025) and were performed in compliance with the Institutional Animal Care and Use Committee guidelines. This study was conducted in accordance with the ARRIVE guidelines for reporting animal research.
11. DATA AVAILABILITY
All data generated in this study are included in this manuscript.
12. PUBLISHER’S NOTE
All claims expressed in this article are solely those of the authors and do not necessarily represent those of the publisher, the editors and the reviewers. This journal remains neutral with regard to jurisdictional claims in published institutional affiliation.
13. USE OF ARTIFICIAL INTELLIGENCE (AI)-ASSISTED TECHNOLOGY
The authors declare that they have not used artificial intelligence (AI)-tools for writing and editing of the manuscript, and no images were manipulated using AI.
REFERENCES
1. Siswanto FM, Andarisuta PS. Effect of Angelica keiskei leaves extract against overtraining-related oxidative stress and hormonal changes in male mice. J Herbmed Pharmacol. 2025;14:332–8. CrossRef
2. Siswanto FM. Angelica keiskei leaves extract attenuates psychosocial stress in overcrowding-subjected rats. J Appl Pharm Sci. 2024;14:156–64. CrossRef
3. Siswanto FM. Resveratrol protects Caenorhabditis elegans from ultraviolet B-induced photoaging via skn-1. Mol Cell Biomed Sci. 2025;9(1):48–57. CrossRef
4. Gromkowska-K?pka KJ, Pu?cion-Jakubik A, Markiewicz-?ukowska R, Socha K. The impact of ultraviolet radiation on skin photoaging - review of in vitro studies. J Cosmet Dermatol. 2021;20:3427–1. CrossRef
5. Alcantara GP, Esposito ACC, Olivatti TOF, Yoshida MM, Miot HA. Evaluation of ex vivo melanogenic response to UVB, UVA, and visible light in facial melasma and unaffected adjacent skin. An Bras Dermatol 2020;95:684–90. CrossRef
6. Pillaiyar T, Manickam M, Jung SH. Recent development of signaling pathways inhibitors of melanogenesis. Cell Signal. 2017;40:99–115. CrossRef
7. Kim JC, Park TJ, Kang HY. Skin-aging pigmentation: who is the real enemy?. Cells. 2022;11:2541. CrossRef
8. Gan C, Rodrigues M. An update on new and existing treatments for the management of melasma. Am J Clin Dermatol. 2024;25:717–33. CrossRef
9. Ishack S, Lipner SR. Exogenous ochronosis associated with hydroquinone: a systematic review. Int J Dermatol. 2022;61:675–84. CrossRef
10. Hollick EJ, Igwe C, Papamichael E, Gore DM, Angunawela RI, Philippidou M, et al. Corneal and scleral problems caused by skin-lightening creams. Cornea. 2019;38:1332–5. CrossRef
11. Juliano CCA. Spreading of Dangerous Skin-Lightening Products as a Result of Colourism: a Review. Appl Sci. 2022;12:3177. CrossRef
12. Fabian IM, Sinnathamby ES, Flanagan CJ, Lindberg A, Tynes B, Kelkar RA, et al. Topical hydroquinone for hyperpigmentation: a narrative review. Cureus. 2023; 15(11):e48840. CrossRef
13. Do?an A, Akocak S. Natural products as tyrosinase inhibitors. Enzymes. 2024:56:85–109. CrossRef
14. Pang M, Xu R, Xi R, Yao H, Bao K, Peng R, et al. Molecular understanding of the therapeutic potential of melanin inhibiting natural products. RSC MedChem. 2024;15:2226–353. CrossRef
15. Tung XY, Yip JQ, Gew LT. Searching for natural plants with antimelanogenesis and antityrosinase properties for cosmeceutical or nutricosmetics applications: a systematic review. ACS Omega. 2023;8:33115–201. CrossRef
16. Parvizi MM, Hekmat M, Yousefi N, Javaheri R, Mehrzadeh A, Saki N. Clinical trials conducted on herbal remedies for the treatment of melasma: a scoping review. J Cosmet Dermatol. 2024;24:e16741. CrossRef
17. Mendoza CG, Singzon IA, Handog EB. A randomized, double-blind, placebo-controlled clinical trial on the efficacy and safety of 3% Rumex occidentalis cream versus 4% hydroquinone cream in the treatment of melasma among Filipinos. Int J Dermatol. 2014;53:1412–6. CrossRef
18. Kawakami S, Morinaga M, Tsukamoto-Sen S, Mori S, Matsui Y, Kawama T. Constituent characteristics and functional properties of passion fruit seed extract. Life. 2021;12:38. CrossRef
19. Dos Reis LCR, Facco EMP, Salvador M, Flôres SH, de Oliveira Rios A. Antioxidant potential and physicochemical characterization of yellow, purple and orange passion fruit. J Food Sci Technol. 2018;55:2679–91. CrossRef
20. Ribeiro DN, Alves FMS, dos Santos Ramos VH, Alves P, Narain N, Vedoy DRL, et al. Extraction of passion fruit (Passiflora cincinnata Mast.) pulp oil using pressurized ethanol and ultrasound: antioxidant activity and kinetics. J Supercrit Fluids. 2020;165:104944. CrossRef
21. Xing X, Dan Y, Xu Z, Xiang L. Implications of oxidative stress in the pathogenesis and treatment of hyperpigmentation disorders. Oxid Med Cell Longev. 2022;2022:1–2. CrossRef
22. Maruki-Uchida H, Kurita I, Sugiyama K, Sai M, Maeda K, Ito T. The protective effects of piceatannol from passion fruit (Passiflora edulis) seeds in UVB-irradiated keratinocytes. Biol Pharm Bull. 2013;36:845–99. CrossRef
23. He X, Luan F, Yang Y, Wang Z, Zhao Z, Fang J, et al. Passiflora edulis: an Insight Into Current Researches on Phytochemistry and Pharmacology. Front Pharmacol. 2020;11:617. CrossRef
24. Muslim M, Jusuf N, Putra I. The effect of 3% passion fruit purple variant (Passiflora edulis Sims var. Edulis) seed extract cream on facial skin aging. J Pak Assoc Dermatol. 2023;33:566–73. CrossRef
25. Moreno E, Calvo A, Schwartz J, Navarro-Blasco I, González-Peñas E, Sanmartín C, et al. Evaluation of skin permeation and retention of topical dapsone in murine cutaneous leishmaniasis lesions. Pharmaceutics. 2019;11:607. CrossRef
26. Riyanto A, Nasihun T, Sumarawati T. The difference between the effect of green tea cream and tocopherol on decreasing level of tyrosinase enzyme and amount of melanin in Rattus norvegicus exposed to UVB Rays. Sains Med. 2020;11(1):20. CrossRef
27. Harlisa P, Puspitasari IW, Yuliyanti S. Protective effect of corncob extract cream on guinea pig (Cavia porcellus sp) skin pigmentation exposed to ultraviolet B (UVB) rays. J Med Sci (Berkala Ilmu Kedokteran). 2023;55: 205–11. CrossRef
28. Siswanto FM, Wijaya IMT, Handayani MDN, Dewi R, Ekowati AL, Manalu JL, et al. Extract of Angelica keiskei leaves attenuates spatial memory impairment on the D-galactose model of brain aging in mice. Biomed Pharmacol J. 2024;17:1563–73. CrossRef
29. Benet LZ, Hosey CM, Ursu O, Oprea TI. BDDCS, the rule of 5 and drugability. Adv Drug Deliv Rev. 2016;101:89–98. CrossRef
30. Yepes A, Ochoa-Bautista D, Murillo-Arango W, Quintero-Saumeth J, Bravo K, Osorio E. Purple passion fruit seeds (Passiflora edulis f. edulis Sims) as a promising source of skin anti-aging agents: enzymatic, antioxidant and multi-level computational studies. Arab J Chem. 2021;14:102905. CrossRef
31. Matsui Y, Sugiyama K, Kamei M, Takahashi T, Suzuki T, Katagata Y, et al. Extract of passion fruit (Passiflora edulis) seed containing high amounts of piceatannol inhibits melanogenesis and promotes collagen synthesis. J Agric Food Chem. 2010;58:11112–8. CrossRef
32. Damanik DC, Katrin K, Sauriasari R, Elya B, Saputri F. Inhibition of tyrosinase activity test from yellow passion fruit seed oil Passiflora edulis f flavicarpa and oil parameter determination. Jawa Barat: Universitas Indonesia; 2014.
33. Geisler AN, Austin E, Nguyen J, Hamzavi I, Jagdeo J, Lim HW. Visible light. Part II: photoprotection against visible and ultraviolet light. J Am Acad Dermatol. 2021;84:1233–44. CrossRef
34. Li L, Chong L, Huang T, Ma Y, Li Y, Ding H. Natural products and extracts from plants as natural UV filters for sunscreens: a review. Anim Model Exp Med. 2023;6:183–95. CrossRef
35. Shelan Nagapan T, Rohi Ghazali A, Fredalina Basri D, Nallance Lim W. Photoprotective effect of stilbenes and its derivatives against ultraviolet radiation-induced skin disorders. Biomed Pharmacol J. 2018;11:1199–208. CrossRef
36. Urrego N, Sepúlveda P, Aragón M, Ramos FA, Costa GM, Ospina LF, et al. Flavonoids and saponins from Passiflora edulis f. edulis leaves (purple passion fruit) and its potential anti-inflammatory activity. J Pharm Pharmacol. 2021;73:1530–8. CrossRef
37. Mizuno M, Fukuhara K. Antioxidant and prooxidant effects of thymoquinone and its hydroquinone metabolite. Biol Pharm Bull. 2022;45:22. CrossRef
38. Kim SH, Lee J, Jung J, Kim GH, Yun CY, Jung SH, et al. Interruption of p38 MAPK -MSK1-CREB-MITF-M pathway to prevent hyperpigmentation in the skin. Int J Biol Sci. 2024;20:1688–704. CrossRef
39. Sun L, Guo Y, Zhang Y, Zhuang Y. Antioxidant and anti-tyrosinase activities of phenolic extracts from rape bee pollen and inhibitory melanogenesis by cAMP/MITF/TYR pathway in B16 mouse melanoma cells. Front Pharmacol. 2017;8:104. CrossRef
40. Kami?ski K, Kazimierczak U, Kolenda T. Oxidative stress in melanogenesis and melanoma development. Wspó?czesna Onkol. 2022;26:1–7. CrossRef
41. Park CH, Kim G, Lee Y, Kim H, Song MJ, Lee DH, et al. A natural compound harmine decreases melanin synthesis through regulation of the DYRK1A/NFATC3 pathway. J Dermatol Sci. 2021;103:16–24. CrossRef
42. Tahir F, Ali E, Hassan SA, Bhat ZF, Walayat N, Nawaz A, et al. Cyanogenic glucosides in plant-based foods: occurrence, detection methods, and detoxification strategies – a comprehensive review. Microchem J. 2024;199:110065. CrossRef
43. Kim GJ, Choi HG, Kim JH, Kim SH, Kim JA, Lee SH. Anti-allergic inflammatory effects of cyanogenic and phenolic glycosides from the seed of Prunus persica. Nat Prod Commun. 2013;8:1739–40.
44. Zhang J, Koike R, Yamamoto A, Ukiya M, Fukatsu M, Banno N, et al. Glycosidic inhibitors of melanogenesis from leaves of Passiflora edulis. Chem Biodivers. 2013;10:1851–65. CrossRef
45. Hmingthansanga V, Singh N, Banerjee S, Manickam S, Velayutham R, Natesan S. Improved topical drug delivery: role of permeation enhancers and advanced approaches. Pharmaceutics. 2022;14:2818. CrossRef
46. Kang Y, Zhang S, Wang G, Yan Z, Wu G, Tang L, et al. Nanocarrier-based transdermal drug delivery systems for dermatological therapy. Pharmaceutics. 2024;16:1384. CrossRef
47. Albakr L, Du H, Zhang X, Kathuria H, Fahmi Anwar-fadzil A, Wheate NJ, et al. Progress in lipid and inorganic nanocarriers for enhanced skin drug delivery. Adv NanoBiomed Res. 2024;4: 2400003. CrossRef