INTRODUCTION:

Agarwood (Indonesian: Gaharu) is a non-timber forest product that has the potential to be developed in Indonesia. It has a very high economic value. The price of agarwood at the local markets in Samarinda (East Kalimantan), Tarakan, and Nunukan (North Kalimantan) for super quality reach IDR 40-50 million/kg¹. The export value of agarwood increased from US$ 708 293 with a volume of 124 959 kg in August 2019 and reached US$ 993 426 with a volume of 35 817 kg in December 2019². The utilization of agarwood is not only as a traditional medicine but can also be developed as a therapeutic fragrance, for religious purposes, and as an aromatic food ingredient.

This increase the agarwood demand which has an impact on the hunting of agarwood in natural forests which is increasingly uncontrolled so that the existence of agarwood is threatened and rare. Based on CITES 2004, agarwood is included in Appendix II so that the export of agarwood from natural forests is restricted³. Therefore, Indonesia has cultivated agarwood-producing trees.

There are seven species of agarwood producing trees from 26 species that are popularly cultivated in Indonesia, including Aquilaria malaccensis, A. beccariana, A. microcarpa, A. filaria, Gyrinops versteegii, G. rosbergii, and G. moluccana. The number of agarwood-producing trees that already exist in Indonesia is 3 249 599 trees and as many as 67 221 trees of which the G. versteegii species are planted in 29 districts in East Java. However, the cultivation of this plant requires a large investment and takes a long time to harvest agarwood⁴. Therefore, the utilization of other parts of this plant is very necessary to increase the added value of forest products. One part that is prospective to be developed is the leaf of G. versteegii as a source of natural active ingredients in cosmetics. The methanol soluble G. versteegii leaf extract has very high antioxidant, sunscreen, and antidiabetic activities^5,6,7,8. This extract has a very strong sunscreen with a sun protection factor (SPF) value of 16.28 (the ultra protection)^9,10. However, methanol exposure can cause serious disorders and damage to body organs such as the central nervous system, eyes, and even death. Therefore, ethanol is safer as a solvent than methanol to produce commercial extracts that are safe to use¹¹. Reported that the extraction of G. versteegii leaves using 50% ethanol as solvent (E50) resulted in extracts that could be developed as active ingredients for day cream formulas based on yield, sunscreen, antioxidant activities, and extraction cost¹².

The type and composition of extractives in plants are influenced by age, place of growth, genetics, position in the tree, and growth rate. Proves that there are differences in the total flavonoid content of the medicinal plant Sambang Colok at growing sites with different nutrients¹³. The type and composition of the extract can affect the antioxidant and sun protection activities of the extract. Therefore, it is necessary to test the antioxidant and sun protection activities of G. versteegii leaf extracts from different growing places.

Bio ingredients in ethanol extract are generally polar so their bioavailability and absorption into the skin membrane are low. Phytosomes incorporate hydrophilic bioactive phytoconstituents from herbal extracts to phospholipids for exhibiting better pharmacokinetic and pharmacodynamic profiles than non-modified herbal extracts^14,15. The phytosome combines standard water-soluble phytoconstituents into phospholipids to produce complex molecules compatible with lipids. Soy lecithin was chosen to make phytosomes because it is less carcinogenic and easier to flow in the membrane than pure lecithin and egg lecithin. In addition, soy lecithin can also improve taste and odor perception, so it is recommended to apply it to topical dosage formulations^16,17. Therefore, this study aims to evaluate the extract yield of G. versteegii leaf from Central Java (CJ) and West Java (WJ) from the extraction using 50% ethanol (E50) solvents, determine the antioxidant and sunscreen activities of the extracts and its phytosome, and to analyze the phytochemical profile of the extracts.

MATERIALS AND METHODS:

Materials:

The samples used were old leaves of G. versteegii cultivated from West Java (Cilodong) and Central Java (Purworejo), Indonesia. The chemicals used were ethanol (analytical grade), Folin-Ciocalteu reagent, Na₂CO₃, AlCl₃, 2,2-diphenyl-2-picrylhydrazyl (DPPH), CuCl₂·2H₂O, NH4Ac buffer (1 M, 1 mL, pH 7.0), neocuproine (7.5 mM, 1 mL,), K₂S₂O₈, aquadest, soy lecithin, and deionized water.

Chemical Analysis of Soil and Climate:

The analysis of the conditions where the trees grew as samples were carried out by analyzing climate (rainfall) and soil. Rainfall data was taken from data from the Meteorology, Climatology and Geophysics Agency (MCGA) of Semarang for places to grow in Central Java, and MCGA of Bogor for places to grow in West Java. Analysis of soil samples from locations where plants grow includes soil physical properties, namely soil texture and soil chemical properties including pH, C-organic, N-total, P-bray, K, Ca, Mg, Na, and soil texture.

Extraction:

The simplicias of G verstigii leaf from the different locations were made into a powder measuring 40-60 mesh. 200 g of this powder with known moisture content was macerated with 2000 mL of E50 solvent for 24 hours. The extraction was repeated 3 times. The extracted filtrate was concentrated with a vacuum rotary evaporator and dried in an oven at 40 ℃.

Antioxidant Activity and Sunscreen Assay:

The antioxidant activity was carried out in vitro using the DPPH and cupric ion reducing antioxidant capacity (CUPRAC) assays. The DPPH method refers to¹⁸ and CUPRAC refers to¹⁹. The antioxidant activity was expressed in moles of trolox/g extract. The higher the trolox/g mole value indicates the higher the antioxidant activity. The assay of a sunscreen refers to²⁰. The higher the SPF value indicates the stronger the sunscreen activity.

Production of Phytosome:

The making of phytosomes was carried out by²¹ research. The extract used for phytosome production was the extract with the best yield, antioxidant activity, and sun protection activity. Phytosome formula is made with different ratios of extracts and soy lecithin at the level of F1 (1: 2), F2 (1: 1), and F3 (2: 1). The phytosome formulas had tested the antioxidant activity and SPF value.

Analysis of Phytochemical:

The phytochemical analysis was carried out through the total phenolic content (TPC) of the extract. Determination of TPC refers to²².

Data Analysis:

Data analysis used in this study was a randomized group design with the grouped by geographic location of plant for leaves collection. Data analysis using analysis of variance (ANOVA)

RESULTS:

Soil Chemistry and Climate Where it Grows G. versteegii:

The chemical analysis showed that the soil chemistry of the two locations where G. versteegii grew was different. Soil from West Java contains C-organic, Phosphate (P), and potassium (K) which was higher than the soil in Central Java (Table 1).

Table 1. Chemical properties of the soil in the geographic location of the plant

The geographic location of the plant	C-Org (%)	N-Tot (%)	P (ppm)	Ca	Mg	K	NA
The geographic location of the plant	C-Org (%)	N-Tot (%)	P (ppm)	Ca	Cmol/kg
CJ	2.97	0.29	33.90	15.95	2.86	1.24	0.20
WJ	3.19	0.24	82.31	14.51	1.68	2.02	0.13

Extract yield:

The variance analysis showed that the geographic location of the plant group influenced the extract yield of G versteegii leaves. The yield of extract leaves from WJ was significantly higher (α = 0.05) than CJ (Table 1).

Table 2 The yields of E50 extract of G. versteegii leaf (% w/w)

The geographic location of the plant	Extract Yield^1,2(%)
Central Java (CJ)	18.59±0.35a
West Java (WJ)	25.43±0.06b

Note: 1: the mean of triplicate. 2: Different letters in the column of yield indicate significantly different yields at α = 0.05.

Antioxidant Activity of Extract:

The value of the antioxidant activity of the G. versteegii leaf extracts from the DPPH method was lower than the CUPRAC method. The variance analysis showed that the geographical location of the plant had a significant effect on the antioxidant activity of the two test methods. The antioxidant activity of the extract from WJ was higher than the extract from CJ (Table 3).

Table 3 Antioxidant activity of E50 extract of G. versteegii leaf *)

Kind of Antioxidant assay	The geographic location of the plant	Antioxidant activity^1,2(µmol trolox/g)
DPPH	CJ	45.36±0.01a
	WJ	38.92±0.01b
CUPRAC	CJ	393.09±0.04x
	WJ	768.93±0.06y

Note: *): Note: 1: the mean of triplicate. 2: Different letters in the column of antioxidant activity indicate significantly different antioxidant activity at α = 0.05.

Sunscreen of Extract:

The variance analysis showed that the location where the G. versteegii grows affects the SPF value (α = 0.05). The sunscreen of extracts from WJ was higher than CJ (Table 4).

Table 4 Sunscreen of E50 extract of G. versteegii leaf

The geographic location of the plant	SPF Value ^1,2
Central Java (CJ)	12.33±0.02a
West Java (WJ)	14.84±0.30b

Note: *): Note: 1: the mean of triplicate. 2: Different letters in the column of SPV value indicate significantly different SPF at α = 0.05.

Yield of Phytosome:

The E50 extract was developed as an active ingredient for day cream antiaging formulas based on the yield, sun protection activity, antioxidant capacity, and extraction costs. For that, the phytosomes were made from the E50 extract from WJ. Table 5 showed that the yield of making phytosome from the G. versteegii leave extract is 80.13-86.45%. The variance analysis results showed that the phytosome formulation affected the yield of phytosome. The yield F1 was highest than other formulas.

Table 5. The yield of making phytosome from the G. versteegii leaf extract

Phytosome Formula	Yield (%)^1,2
F1 (E:L=1:2)	86.45±0.10^a
F2 (E:L=1:1)	85.47±0.08^b
F3 (E:L=2:1)	80.13±0.10^c

Note: 1: the mean of triplicate. 2: Different letters in the column of phytosome yield indicated significantly different yields at α = 0.05.

Antioxidant Activity of Phytosome:

The variance analysis showed that the phytosome formulation affected its antioxidant activity. The antioxidant activity of the F2 formula (ratio of extract: lecithin was 1:1) was highest and followed by F3 (ratio of extract: lecithin was 2:1).

Figure 1. The antioxidant activity of phytosome formulas

The Sunsreen Activity of Phytosome:

The variance analysis showed that the phytosome formulation affected its sun protection activity. Table 6 showed that the phytosomization of the extract decreased the sunscreen activity because the SPF value of the phytosome was lower than the extract. However, the sunscreen of F2 was higher than F1 and F3.

Table 6. The sunscreenof the phytosome

Phytosome Formula	Sun Protection Factor (SPF) value ^1,2
F0 (E50=1)	14.84±0.30^d
F1 (E:L=1:2)	8.39±0.15^a
F2 (E:L=1:1)	13.74±0.00^c
F3 (E:L=2:1)	11.51±0.58^b

Note: 1: the mean of triplicate. 2: Different letters in the column of SPF value indicate significantly different SPF at α = 0.05.

The Total Phenolic Content of Extract:

The analysis of variance results showed that the geographic location of the plant had a significant effect (α = 0.05) on the total phenol content (TPC) of the extract. The TPC of the extract from WJ was higher and different than the extract from CJ (Table 7).

Table 7. Total phenolic content of G. versteegii leaf extract *)

The geographic location of the plant	Average ^1,2E50
CT	56.210±0,006a
WJ	77.012±0.002b

*): Expressed in mg GAE/g sample; 1): average of 3 replications, 2) different letters indicate significantly different TPC values (α = 0.05).

Total Phenol Content of Phytosome:

The total phenolic content of the phytosome (F2 and F3) was higher than the F0 (only E50 extract) (Figure 2).

Figure 2. The phenolic content of phytosome formulas

DISCUSSION:

Soil from West Java contains C-organic, Phosphate (P), and potassium (K) which was higher than the soil in Central Java. Differences in soil chemical content influence secondary metabolites produced by plants. Some of the soil elements that affect secondary metabolites are P and K. P content affects the production of secondary metabolites (anthocyanins, flavonoids, and alkaloids). Differences in phytochemical content can affect the bioactivity of the extract²³.

The percentage of yield in this research was in line with research conducted by²⁴ which reported that there is a difference in the yield of 70% ethanol extract of garlic from WJ (21.86%) and samples from CJ (19.74%). This was consistent with the statement of¹³ that the location of plant growth, i.e. soil and climate conditions affects the kind and composition of chemical compounds produced by secondary metabolites contained in plants. Table 2 showed that the chemical properties of the soil in the plant which the leaves collection was different. The soil in the growing site in WJ contained higher Potassium (K) and phosphor (P) than CJ (Table 2). The K and P content in the soil is an activator that can support the formation of secondary metabolites²⁵.

The value of antioxidant activity of all extracts from the DPPH assay were lower than the CUPRAC assay. This is becaused the DPPH method only detects antioxidant capacity in polar or anti-radical compounds dissolved in organic solvents, especially alcohol (hydrophilic compounds). But, the CUPRAC assay can measure the antioxidant capacity of hydrophilic and lipophilic compounds, more stable reagent than chromogenic reagent, and more selective in measuring antioxidant activity. In addition, the CUPRAC assay can work at physiological pH, is stable, is selective (has a low reduction potential value of 0.17 V), and is easy to detect thiol-type antioxidants²⁶.

The variance analysis showed that the antioxidant activity of the extract was affected by the plant's geographic location based on the antioxidant activity assay using the CUPRAC and DPPH assays. The antioxidant activity of the extract from WJ was higher than the extract from CJ. Mentha piperita leaves (one of the herbs known as a source of antioxidants). State that based on tests with CUPRAC, the antioxidant activity of M. piperita leaves amounted to 386 μmol trolox/g sample and the value was greater than the value obtained from this study (Table 3)²⁷. Using the same antioxidant measurement method, the antioxidant activity of G. versteegii leaf extracts produced from this study was higher than the ethanol extract of the Guazuma ulmifolia leaves reported by²⁸. The results of Meitary's research²⁸ show that the antioxidant activity value of ethanol extract of G. ulmifolia leaves is only 21.36 µmol trolox/g powder based on the DPPH test method. This value was lower than the value obtained from this study (Table 3). This shows that the E50 extract of G. versteegii leaf has a high potential as a source of natural antioxidants.

The sunscreen activity of extracts from WJ was higher than CJ. The total phenolic content of the extract from WJ was different and was higher than the extract from CJ. The phenolic compounds such as tannins, flavonoids, and isoflavonoids affect the SPF value. According to²⁹, based on qualitative observations, flavonoids in leaves and flowers have a pharmacological activity that can absorb light and help protect photosensitive substances. This group of compounds can absorb high UV light because it has a chromophore group (conjugated double bond) that can absorb UV light. According to³⁰, all extracts were classified as having maximum sunscreen activity. Using the same SPF value method, reported the result of the research on the SPF value of Hibiscus rosa-sinensis L. extract and Dragon’s blood resin which are widely used as raw material for making sunscreen activity. The result showed that H. rosa-sinensis extract and the E50 extract of dragon’s blood resin from Daemonorops acehensis had a lower SPF value (SPF: 12.54³¹ and 7.54³²) than the G. versteegii leaf extract used in this study. This showed that agarwood leaf extract has the potential to be developed as a sunscreen.

The variance analysis showed that the phytosome formulation affected the yield of phytosome. The yield F1 was highest than other formulas. These results indicate that the ratio of G. versteegii leaf extract with lecithin of 1: 2 could produce more phytosomes than other formulas. The difference in yield of making phytosomes of G. versteegii leaf extract was caused by the differences in the content of lecithin soya. According to²¹, the higher the lecithin caused the higher the yield. This was caused by residual lecithin contained in the phytosome.

The antioxidant activity of phytosome formulas was higher than the extract. The increase in antioxidant activity was caused by the increase in phenol groups formed in the phytosomes. According to the research of ³³, the polar extract and soy lecithin form a phytosome complex through hydrogen bond formation between OH groups of phenolic groups from polar extracts with P=O groups from soy lecithin. The antioxidant activity of the F2 phytosome was lower than extract E50. According to³⁴, the addition of lecithin which exceeds the doubing of the extract weight can reduce its bioavailability.

Phytosomization of the extract decreased the sunscreen because the SPF value of the phytosome was lower than the extract. However, the sunscreen of F2 was higher than F1 and F3. F1 had the lowest SPF value. This could be due to the F1 formulation containing high lecithin. The high lecithin content caused a decreased bioavailability of the compound in reducing UV.

The total phenolic contents of the phytosome formulas (F2 and F3) were higher than the F0 (only E50 extract). It was caused by the increase in phenol groups formed in phytosomes. According to the research of³³, polar extracts and soya lecithin form phytosomal complexes through hydrogen bond formation between OH groups and phenolic groups from polar extracts and P = O groups from soya lecithin. According to³⁵ total phenolic content is highly dependent on its chemical structure. Phenolic compounds with many hydroxyl functional groups or free conditions (aglycones) will produce high total phenolic content. The measurements results of phenolic content and antioxidant activity showed that the presence of phenols was positively correlated to the antioxidant activity value. This was supported by the results of the correlation test between the two seen in Figure 3. This correlation was stronger than the correlation between antioxidant activity with total phenolic of Portulaca oleracea extract (DPPH, R2 = 0.75; H₂O₂, R2 = 0.71). The presence of phenolic compounds can act as catchers of hydroxyl free radicals (*OH) to not oxidize fats, proteins, and DNA. The ability of polyphenols to capture free radicals is 100 times more effective than vitamin C and 25 times more effective than vitamin E³⁶. Phenolics are known to correlate with antioxidants^37,38.

CONCLUSIONS:

The yield, total phenol content, antioxidant activity, and sunscreen activity of E50 extract of G. versteegii leaf from WJ was higher than from CJ. The yield of the phytosome formulas of E50 extract from WJ was 80.13-86.45%. The yield of F1 was highest and followed by F2, and F3. Phytosimization of the extract increased the antioxidant activity (F2 and F3) but decreased the sunscreen activity. The antioxidant and sunscreen activities of F2 (ratio of extract: lecithin was 1:1) was higher than other formulas. The total phenolic contents of the phytosome formulas (F2 and F3) were higher than the F0 (only E50 extract). The presence of phenols was positively correlated to the antioxidant activity of the extract and phytosome formulas.

CONFLICTS OF INTEREST:

The authors declare no conflict of interest

ACKNOWLEDGMENTS:

The Directorate of Higher Education of the Ministry of Education, Culture, Research, and Tercnology of the Republic of Indonesia with The IPB Higher Education Research Grant scheme 2021 (contract No. 1/EI/KP.PT.PTNBH/2021) for funding support for some parts of this research.

REFERENCES:

1. Susmianto A, Turjaman M, Setio P. Track Record of Inoculation Agarwood. FORDA Press, Bogor. 2014.

2. Central Bureau of Statistics. Bulletin of foreign trade export statistics based on a harmonized system of December 2019. PT Citra Mawana Patamoro Jakarta, Jakarta. 2019.

3. Barden A, Anak NA, Mulliken T, Song M. Heart of the matter: agarwood use and trade and cites implementation for Aquilaria malaccensis. Traffic Network Report: Cambridge. 2000.

4. Santoso E, Turjaman M, Irianto RSB, Agustini L, Efiyanti L, Faulina SA. Forestry science and technology book series V. Puslitbang Bogor: Bogor. 2014.

5. Wahyuningrum M, Sari RK, Rafi M. Antioxidant activity and sunscreen of Gyrinops versteegii leaf extract. Jurnal Ilmu dan Teknologi Kayu Tropis. 2018 July 2;16(2):141-149.

6. Fayyadh AA, Ibrahim H, Zain HHM, Al-Qubaisi. The Effect of Agarwood Leaf Extracts on Blood Glucose Level of Type II Diabetes Mellitus in ICR Male Mice. Research Journal Pharmacy and Technology. 2020 Jan;13(1):237-242.

7. Jangde R, Daharwal SJ. Herbal sunscreen: an overview. Research Journal Pharmacy and Technology. 2011 Jul;2(2):35-39.

8. Kale SS, Rajmane AH, Urunkar VC, Gaikwad MK, Bhandare SB. Formulation and In-Vitro Evaluation of Sun Protection Factor of Methanolic Extract of Zanthoxylum rhetsa DC. Sunscreen lotion. Research Journal Pharmacy and Technology. 2011 Sep;3(5): 206-210.

9. Khan MA and Engla G. Comparative studies on sun protection factor of some sunscreen formulations used in cosmetics. Research Journal Pharmacy and Technology. 2012 Jul;3(2): 34-36.

10. Wilkinson JB and Moore RJ. Harry’s Cosmeticology (7’th edition). Chemical Publishing Company, New York. 1982.

11. Romadanu, Rachmawati SH, Lestari SD. Testing the antioxidant activity of lotus flower extract. Fishtech. 2014;3(1): 1-7.

12. Sari RK, Wahyuningru M, Rafi M, Wientarsih I. Effect of ethanol polarity on extraction yield, antioxidant, and sunscreen activities of phytochemicals from Gyrinops versteegii leaves. 2020; IOP Conf Series: Materials Science and Engineering. 2020. doi:10.1088/1757-899X/935/1/012038

13. Ministry of Helath. Acuan Sediaan Herbal. Ministry of Health: Jakarta. 2000.

14. Rayate Y, Shewale S, Patil A, Nitalikar M, Mohite S. Phytosomes–A Novel Approach in Herbal Drug Delivery System. Asian Journal of Research in Pharmaceutical Sciences. 2018 Sept;8(3): 151-154.

15. Selvaraju K, Arulkumaran KSG, Karthick K, Prabhu KV, Preetha JP, Arrivulkkarasu R. Asian Journal of Research in Pharmaceutical Sciences. 2011 Sept;3(5): 191-194.

16. Gnananath K, Nataraj KS, Rao BG. Phospholipid complex technique for superior bioavailability of phytoconstituents. Adv Pharm Bull. 2017 Apr 7;7(1):35-42. doi: 10.15171/apb.2017.005.

17. Mund AK, Guru AC, Singh RP, Pradhani KK, Pangi B, Pasayat MK. Preparation and characterization of phytosome of norbixin. World Journal of Pharmacy Pharmaceutical Sciences. 2018 ;8(1): 1045-1057.

18. Salazar AR, Perez LLA, Loppez AJ, Alanis GBA, Torres NW. Antimicrobial and antioxidant activities of plants from Northeast of Mexico. Evid Complement and Alterntat Medic. 2009 Mar 9; 1-6. doi: 10.1093/ecam/nep127

19. Ozturk M, Ermin MD, Kivrak S, Mercan N, Turkoglu A, Ozler MA. In vitro antioxidant, anticholinesterase and antimicrobial activity studies on three Agaricus species with fatty acid compositions and iron contents: a comparative study on three most edible mushrooms. Food Chem Toxicol, 2011 Mar;49: 1353-1360. doi:101016/j.fct.2011.03.019.

20. Kawira JA. Laboratory procedures for determining sun protection factors. University of Indonesia: Depok. 2005.

21. Singh RP, Narke R. Preparation and evaluation of phytosome of lawsone. Internat Journal Pharmaceut Sciences Research. 2015 Dec 1;6(12): 5217-5226.

22. Irawati I. Comparison of methods for determining the antioxidant activity of ginger rhizome. Indonesia: Bogor Agricultural University, Bogor. 2008.

23. Plaxton CW, Carswell MC. Metabolic Aspect of the Phosphate Starvation Response in Plants. Marcel Dekker, New York. 1999.

24. Prastiwi R, Siska, Marlita N. Physicochemical parameters and analysis of Allyl disulfide levels in ethanol extract of 70% garlic with a comparison of the area where the parameters grow. Pharm Sci Res. 2017; 4(1): 32-47.

25. Nell M, Vötsch M, Vierheilig H. Effect of phosphorus uptake on growth and secondary metabolites of garden sage (Salvia officinalis L.). Journal Science Food and Agricult. 2009 Mar 12;89(6):1090 –1096.

26. Apak R, Guclu K, Demirata B, Ozyurek M, Celik SE, Bektasoglu B, Berker KI, Ozyurt D. Comparative evaluation of various total antioxidant capacity assays applied to phenolic compounds with the CUPRAC assay. Molecules. 2007; 12: 1496-1547.

27. Luminita P. Comparative evaluation of the antioxidant capacity of herbal plants by different methods. J of Horticul. Forest and Biotech. 2015; 19(4): 9-12.

28. Rafi M, Meitary N, Septaningsih DA, Bintang M. Phytochemical profile and antioxidant activity of Guazuma ulmifolia leaves extract using different solvent extraction, Indonesia. Indonesian Journal Pharmacy. 2020 ;31(3), 171–180.

29. Jawarkar SV, Wadkar KA, Kore P, Lokapure SG, Mohite SK, Magdum CS. Study of UV protective activity of ethanolic extract of fruit of Carica papaya L. Research J Pharm and Tech. 2014 7(11): 1262-1263.

30. Wolf R, Wolf D, Morganti P, Ruocco V. Sunscreen. Clin Derm. 2001;19: 252-459.

31. Lokapure SG, Patil SS, Phutane KR, Mohite SK, Magdum CS. In vitro evaluation of sun protection factor and diffusion study of Hibiscus rosa sinensis L. flower extract gel. Research J. Pharm and Tech. 2014; 7(6) 643-647.

32. Sari RK. Prayogo YH, Rozan SA. Rafi M. Wientarsih I. Antioxidant activity, sun protection activity, and phytochemical profile of ethanolic extracts of Daemonorops acehensis resin and its phytosomes. Sci. Pharm. 2022 90 (10):1-11. https:// doi.org/10.3390/scipharm90010010.

33. El-Menshawe SF, Adel AA, Rabeh MA, Khalil NM. Nanosized soy phytosome-based thermogel as topical anti-obesity formulation: an approach for an acceptable level of evidence of an effective novel herbal weight loss product. Int Journal Nanomedicine. 2018 Jan 9;13(1): 307-318.

34. Haruni PS, Priani SE, Aryani R. Preparasi fitosom ekstrak etanol biji kopi robusta(Coffea canephora pierre a. Froehner) menggunakan variasi konsentrasi fosfatidilkolin. Prosiding Farmasi. 2019 ;5(1), 25-27.

35. Deore S, Khadabadi SS, Baviskar BA, Khangenbam RA, Koli US, Daga NP, Gadbail PA, Jain PA. In vitro Antioxidant activity and Phenolic Content of Croton caudatum. International Journal of ChemTech Research. 2009 ;1(2), 174-176.

36. Sibue. Pengolahan Teh. Gambung: Bandung. 2003.

37. Gini EJ, Sivakkumar T, Kuppuswami S. etermination of Antioxidant Activity of various extracts of Pajanelia longifolia (Willd.) K. Schum, isolation and characterization of Flavonoid from Ethanol extract by Column Chromatography. Research Journal of pharmacy and Technology. 2017 Oct;10(10): 3391-33-97.

38. Jain B, Ahirwar D, Jain VK, Ahirwar B. etermination of Antioxidant Activity of various extracts of Pajanelia longifolia (Willd.) K. Schum, isolation and characterization of Flavonoid from Ethanol extract by Column Chromatography. Research Journal of pharmacy and Technology. 2018 Feb;11(2): 717-722.

Received on 09.11.2021 Modified on 06.02.2022

Research J. Pharm. and Tech 2022; 15(11):5106-5111.

DOI: 10.52711/0974-360X.2022.00858