The materials used in this study consisted of ramania leaves (Bouea macrophylla Griffith), filter paper, 96% p.a. ethanol, 96% technical ethanol, methanol p.a, technical methanol, quercetin (positive control), aquadest, Mg powder, concentrated HCl, HCl 2N, gelatin 1%, FeCl3 1%, Mayer, Dragendorff, Wagner, and Lieberman Burchard reagents.

The tools used are glass tools, blenders, maceration vessels, evaporative cups, horn spoons, drip pipettes, rotary evaporators, cuvettes, analytical balances, waterbaths, micropipettes, mesh 40 sieves, and UV-Vis spectrophotometers.

Ramania leaves (Bouea macrophylla Griffith) were collected from Guntung Manggis Subdistrict, Landasan Ulin District, Banjarbaru City, South Kalimantan. The plant determination was carried out at the Basic Laboratory of the FMIPA Lambung Mangkurat University, Banjarbaru, in November 2024. A total of 6 kg of young ramania leaves underwent wet sorting, washing, slicing, and drying using an oven at 40°C for approximately 24 hours, followed by dry sorting, grinding, and sieving with a 40-mesh sieve (Handoyo & Pranoto, 2020).

A total of 250 grams of ramania leaves simplicia powder was extracted by maceration using 2,500 mL of 96% ethanol as the solvent, with a simplicia-to-solvent ratio of 1:10. The extraction process was carried out 1x24 hours with three remacerations. Every 24 hours, the filtrate is filtered. Subsequently, the filtrate was replaced with an equivalent volume of the same solvent. The obtained filtrate was subjected to evaporation using a rotary evaporator at 50 °C, then further concentrated in a water bath until a constant weight was reached (Nguyen et al., 2022)(Wahyuni et al., 2024)(Wijianto et al., 2024).

A total of 250 grams of ramania leaves simplicia powder was extracted by maceration using 1,250 mL of methanol as the solvent, with a simplicia-to-solvent ratio of 1:5. The extraction process was carried out for 1 × 24 hours with two remacerations. Every 24 hours, the filtrate is filtered. Subsequently, the filtrate was replaced with an equivalent volume of the same solvent. The resulting filtrate was evaporated using a rotary evaporator at 50 °C, followed by concentration in a water bath until a constant weight was achieved (Conitaty et al., 2022).

A quercetin stock solution with a concentration of 1000 ppm was prepared by dissolving 25 mg of quercetin in 25 mL of analytical-grade methanol. Serial dilutions were subsequently carried out to obtain working solutions with concentrations of 10, 20, 30, 40, and 50 ppm in 10 mL of the same solvent (Pramesti, 2023). Test solutions were prepared at a concentration of 1000 ppm (50 mg in 50 mL). Serial dilutions were performed to obtain solutions containing 50, 100, 150, 200, and 250 ppm in 10 mL of each solvent. These procedures were conducted to measure absorbance in the wavelength range of 290–320 nm for determining the SPF. Meanwhile, the wavelength range of 292.5–317.5 nm was used to calculate the %Te, and the range of 322.5–372.5 nm was used to calculate %Tp. Measurements were carried out in triplicate, using each corresponding solvent as a blank at every 5 nm interval. The absorbance values obtained were recorded to calculate SPF, while the transmittance values were used to calculate %Te and %Tp (Pramesti, 2023). The SPF value was determined from the sample’s absorbance data using the Mansur equation as follows:

SPF=CF x ∑320 290 EE x I x (Abs)

Description:

CF = Correction Factor (10)

EE = Erythemal Effect Spectrum

I = Solar Intensity Spectrum

Abs = Absorbance of the sample

(Juliadi et al., 2023)

The %Te and %Tp were calculated using the following equations:

Te = T x Fe

Tp = T x Fp

Description:

T = Transmittance

Fe = Erythema flux

Fp = Pigmentationflux (Hanif et al., 2025)

Plant identification was conducted at the Basic Laboratory of FMIPA, Lambung Mangkurat University. The results confirmed that the specimen was ramania, belonging to the family Anacardiaceae and identified as the species Bouea macrophylla, with certification number 237a/LB. LABDASAR/X/2024.

Yield calculation was conducted as an initial step to identify the effectiveness of solvents in extracting active compounds from the plant material. The extract yield data are presented in Table 1.

A phytochemical analysis was conducted to determine the types of secondary metabolites present in the extracts. The outcomes of this analysis are presented in Table 2.

The phytochemical screening results of both extracts, using 96% ethanol and methanol as solvents, showed similarities in the presence of flavonoids, phenols, saponins, tannins, and steroids and triterpenoids. However, a significant difference was observed in the alkaloid test results. The methanol extract showed positive results with all three alkaloid reagents Mayer, Dragendorff, and Wagner. In contrast, the 96% ethanol extract only yielded a positive result with Dragendorff reagent, while Mayer and Wagner reagents showed negative results.

SPF activity determination was conducted to evaluate the ability of the extracts to absorb ultraviolet (UV) radiation. Quercetin was used as a comparator in this test due to its well-known sunscreen activity. The results are presented in Table 3. The results of the sunscreen activity test demonstrated that both quercetin and ramania leaves extracts exhibited increasing UV protection in line with higher concentrations. Quercetin, used as a comparator, showed the highest SPF value of 15.97 at a concentration of 50 ppm, which falls under the ultra protection category. The 96% ethanol extract of ramania leaves reached a maximum SPF value of 17.62 at 250 ppm, while the methanol extract yielded an even higher SPF value of 19.80 at the same concentration, also categorized as ultra protection. Secondary metabolites, including flavonoids, phenolic compounds, saponins, steroids, and tannins, exhibit photoprotective potential owing to their capacity to absorb, reflect, and block ultraviolet (UV) radiation on the skin's surface. This activity is associated with their chemical structures, which generally contain aromatic rings or conjugated double bonds, enabling effective absorption of UVA and UVB radiation(Tesalonika et al., 2024).

The %Te values were measured at wavelengths ranging from 292.5 to 317.5 nm, while %Tp values were measured at 322.5 to 372.5 nm, each at 5 nm intervals. The %Te and %Tp values for quercetin and Bouea macrophylla Griffith leaves extracts are presented in Table 4. The percentages of %Te and %Tp represent the amount of ultraviolet (UV) radiation transmitted through the sunscreen that may contribute to erythema and skin pigmentation. Lower %Te and %Tp values indicate a higher efficacy of the sunscreen in providing skin protection(Widhihastuti et al., 2024). The results showed that quercetin, used as a positive control at a concentration of 50 ppm, exhibited a % Te of 0.85% and a %Tp of 0.34%. These values fall within the Sunblock category for UVB and UVA rays, indicating that quercetin possesses very high photoprotective activity. In the 96% ethanol extract of ramania leaves, the %Te value decreased with increasing concentration from 26.64% at 50 ppm to 1.59% at 250 ppm indicating enhanced protection, shifting from the tanning category to extra protection. The %Tp value also decreased from 49.90% to 20.74%, suggesting improved protection against UVA radiation, moving from the tanning to the sunblock category.

Similarly, the methanol extract showed a decreasing trend in %Te from 30.27% at 50 ppm to 1.24% at 250 ppm, and %Tp from 83.05% to 43.46%, indicating an increase in protection level from tanning to extra protection. Based on these findings, both 96% ethanol and methanol extracts of ramania leaves demonstrate potential as natural sunscreen agents, particularly at concentrations of 200 and 250 ppm.

The results showed that the methanol extract had a lower erythema percentage (%TE) than the 96% ethanol extract, suggesting greater effectiveness in reducing UV-induced redness. This may be due to methanol’s higher polarity, which enhances the extraction of phenolic and flavonoid compounds responsible for antioxidant and anti-inflammatory activity.

In contrast, the 96% ethanol extract produced a lower pigmentation percentage (%TP), indicating better protection against melanin formation. Ethanol’s moderate polarity allows it to extract both polar and semi-polar compounds, yielding a balanced composition that supports photoprotection and skin tone stability.

These findings highlight that solvent polarity influences the types of bioactive compounds extracted, resulting in different photoprotective effects.