{"id":48952,"date":"2023-06-30T11:26:52","date_gmt":"2023-06-30T11:26:52","guid":{"rendered":"https:\/\/biomedpharmajournal.org\/?p=48952"},"modified":"2023-07-11T05:56:14","modified_gmt":"2023-07-11T05:56:14","slug":"antioxidant-activity-phenolic-and-flavonoid-content-of-passion-fruit-seed-oil","status":"publish","type":"post","link":"https:\/\/biomedpharmajournal.org\/staging\/vol16no2\/antioxidant-activity-phenolic-and-flavonoid-content-of-passion-fruit-seed-oil\/","title":{"rendered":"Antioxidant Activity, Phenolic and Flavonoid Content of Passion Fruit Seed Oil"},"content":{"rendered":"\n

Introduction<\/strong><\/p>\n\n\n\n

Passion\nfruit belongs to the family Passifloraceae <\/em>and one\nof the species with high economic value1<\/sup>. Passion fruit is one of the important\ncommodities in South Sulawesi, especially in Makassar, since it is one of the present popular icons of passion fruit syrup. The syrup-making\nprocess produces waste in the form of skins and seeds. Passion fruit contains\nas much as 13.6% seeds; therefore, the more syrup produced, the more seed waste will be produced. The\npassion fruit syrup industry makes about 40% of seed waste, and 100% is unusable.<\/p>\n\n\n\n

Meanwhile,\npassion fruit seeds contain high fiber and nutrients and plant-based vegetables that can\nbe used for cosmetics and food.2,6<\/sup> However, passion fruit waste is only disposed of and unusable; therefore, this is an opportunity to be managed and\ndeveloped into an economically valuable product. Passion seeds oil (PSO) contains flavonoids and piceatannol, which can inhibit the tyrosinase enzyme and\nmelanin biosynthesis.2<\/sup><\/p>\n\n\n\n

The tyrosinase enzyme converts tyrosine into 3,4-dihydroxyphenylalanine (DOPA) and\ndopaquinone which is further synthesized into melanin pigment characterized by\nblack\/brown spots on the skin.13<\/sup> In addition, PSO contains\n87.59% unsaturated fatty acids and other compounds, like tocopherol (499.30 mg\/Kg), phenolic (314.13 mg GAE\/kg), and\nvitamin C (1.90 mg\/L).12<\/sup> The PSO has a high antioxidant activity with a\nradical scavenging mechanism of up to 82.81% and an EC50<\/sub> value of 10.62 g oil\/g DPPH.10,15 & 17<\/sup> Therefore, PSO can\npotentially prevent free radicals that trigger premature skin aging. Environmental\ninfluences such as ultraviolet light, cigarette smoke, pollutants, temperature,\nnutrition, and an unhealthy lifestyle can form free radicals and Reactive Oxygen\nSpecies (ROS). This stimulates skin inflammation, triggering a series of biochemical\nreactions in the skin that damage the collagen network in the epidermis layer, leading to premature aging\n(photoaging\/premature skin aging). It can depigment skin by directly inhibiting\ntyrosinase activity in the process. The binding of flavonoids to copper and\ntheir antioxidant activity have been reported to play an important role in\ninhibiting the action of tyrosinase enzymes.2<\/sup> Tocopherol compounds in passion fruit are known as antioxidant agent. In\nthis section, it is necessary to describe the specific specifications related to the\nscheme. It is strong and able to reduce skin damage due to UV B light. It can also inhibit photocarcinogenesis by\npreventing the formation of cyclopyrimidine dimmers in the epidermal P-53 gene and inhibiting the process of\nmelanogenesis.19<\/sup> Combining tocopherol and phenolic compounds in passion fruit oil will effectively\novercome skin damage leading to premature aging (photo-aging) and hyperpigmentation (melasma). Furthermore,\nHuda et al<\/em> <\/sup><\/em>(2017) reported that passion fruit (Passiflora\nedulis<\/em>) seed extract prevented an increase in the amount of skin melanin\nequivalent to 4% hydroquinone in guinea pigs (Cavia porcellus<\/em>) exposed\nto UV B4 light.<\/p>\n\n\n\n

Based\non the description, passion fruit seeds oil has potential in the pharmaceutical\nfield for developing pharmaceutical preparations, especially herbal cosmetics.\nStill, selecting extraction methods is essential for successfully extracting active compounds. Furthermore, the\ntype and amount of solvent used can also affect the number of active compounds\nthat can be drawn, where compounds with polar properties will dissolve in polar\nsolvents, and non-polar compounds will dissolve in non-polar solvents.<\/p>\n\n\n\n

Materials and methods<\/strong><\/p>\n\n\n\n

Materials<\/strong><\/p>\n\n\n\n

The passion fruit seed samples were obtained from the\npassion fruit syrup industry waste in Makassar. The passion fruit seeds were\ndried and powdered. AlCl3<\/sub>, Acetone, ethyl acetate, methanol,\nn-hexane, chloroform, gallic acid, folin-ciocalteau, potassium acetate, sodium\ncarbonate, quercetin, and rutin were purchased from Merck (Merck KGaA,\nDarmstadt, Germany). <\/p>\n\n\n\n

Extraction <\/strong><\/p>\n\n\n\n

The passion seeds oil was extracted by maceration method\nwith assisted ultrasonication. As much as 500 g of powder passion seeds were added\nwith various solvents, including acetone, ethyl acetate, chloroform, methanol, and n-hexane. The\nmaceration assisted-ultrasonicator method was sonicated for 30\nminutes. <\/p>\n\n\n\n

Antioxidant assay<\/strong><\/p>\n\n\n\n

The antioxidant assay was performed according to the method\nby Laura Gonzalez with minor modifications. Briefly, a fresh solution of DPPH\nwas prepared by dissolving 10.0 mg of DPPH powder in 100 mL of methanol. Different\nconcentrations of extraction solution (2.5 ml) and DPPH solution (2.5 ml) were\nmixed together and incubated at room temperature in the dark for 30 minutes. A\nUV-VIS spectrophotometer was used to detect the absorbance at 517 nm. The\ninhibition rates of free radical scavengers were estimated using the following\nformula:<\/p>\n\n\n\n

Total phenolic content<\/strong><\/p>\n\n\n\n

The total phenolic content\n(TPC) of extracts was determined using the folin-ciocalteu (FC) method, which\nrefers to Nugroho, et al. (2012) with a slight modification. Each extract was\ndissolved in distilled water to a concentration of 50.0 g\/mL. Gallic acid (0-60\ng\/mL) and standard curve was created. Diluted gallic acid (1.6 mL) was mixed\nwell with 0.2 mL of FC reagent (diluted 5-fold with distilled water) for 3\nminutes. The mixture was treated with sodium carbonate (0.2 mL, 10% w\/v) and\nallowed to stand at room temperature for 30 minutes. The absorbance of the\nmixture was measured at 760 nm using a UV-VIS spectrophotometer. TPC was\nexpressed as milligrams of Gallic acid equivalents per gram defatted P.\nedulis<\/em> (mgGAE\/g).<\/p>\n\n\n\n

Total Flavonoid Content<\/strong><\/p>\n\n\n\n

The total flavonoid\ncontent (TFC) of each extract was analyzed by a colorimetric method using\naluminium chloride reagent 14<\/sup>. By diluting rutin with methanol\n(0-100 g\/mL), a standard curve was created. Diluted extracts and rutin\nstandards (2.0 ml) were combined with 0.1 ml of 10% (w\/v) aluminum chloride\nsolution and 0.1 mM potassium acetate solution. For 30 minutes, the mixture was\nleft at room temperature. A UV-VIS spectrophotometer was then used to detect\nthe mixture’s maximum absorbance at 415 nm. TFC was calculated using milligrams\nof rutin equivalents per gram of extract.<\/p>\n\n\n\n

Results and Discussion<\/strong><\/p>\n\n\n\n

Extraction<\/strong><\/p>\n\n\n\n

Passion oil extracts were obtained using solvents such as acetone, ethyl acetate, chloroform, methanol, and n-hexane (Figure 1). The results of extraction with different solvents are presented in the following order: chloroform 18.19%, acetone 14.02%, n-hexane 10.79%, methanol 5.45 %, and ethyl acetate 4.93% (Table 1). <\/p>\n\n\n\n\n\n
\"\"<\/td>\n\n

Figure 1: <\/strong>The extracted amount of PSO in different solvent.<\/strong><\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n

Table 1: Extract the amount of PSO in different solvent<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n
\n

Solvents<\/strong><\/p>\n<\/td>\n

\n

Extract amount (%)<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

Acetone<\/p>\n<\/td>\n

\n

14.02<\/p>\n<\/td>\n<\/tr>\n

\n

Ethyl acetate<\/p>\n<\/td>\n

\n

4.93<\/p>\n<\/td>\n<\/tr>\n

\n

Cholorform<\/p>\n<\/td>\n

\n

18.19<\/p>\n<\/td>\n<\/tr>\n

\n

Methanol<\/p>\n<\/td>\n

\n

5.45<\/p>\n<\/td>\n<\/tr>\n

\n

n-Hexane<\/p>\n<\/td>\n

\n

10.79<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n

Based on the data, the extraction yield of chloroform\n(18.19%) was higher than other solvents. These results indicated that\nincreasing the extraction yield correlates to the chemical compounds of the\npassion fruit seed. The maximum yield was found in compounds with intermediate\npolarity. This could be due to the increased solubility of moderate polarity in\nethyl acetate. Extraction is the primary method of obtaining and separating\nphytochemicals from plant material referred to Laura Gonzalez et al. (2019). The effectivity of\nextraction is affected by the chemical nature of phytochemicals, extraction\nmethod, sample particle size, solvent, and the presence of interfering\nsubstances. The extraction yield depends on the solvents used, pH, temperature,\nand extraction time. The same solvent and composition of the sample are known\nas the most important parameters.<\/p>\n\n\n\n

Antioxidant activity<\/strong><\/p>\n\n\n\n

The antioxidant activity of each extract using the DPPH method is shown in Figure 2. The most potent antioxidant activity was methanolic extract with an IC50<\/sub> value of 71.67 \u00b5g\/mL, followed by acetone at 147.29 \u00b5g\/mL, chloroform at 147.65 \u00b5g\/mL, ethyl acetate with 158.66 \u00b5g\/mL, n-hexane extract with 962.5 \u00b5g\/mL, respectively. <\/p>\n\n\n\n\n\n
\"\"<\/td>\n\n

Figure 2<\/strong>: The IC50<\/sub> value of PSO.<\/strong><\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n

The antioxidant activity of PSO is a potential source of antioxidants from natural oil. However, the activity is lower than quercetin as standard with IC50<\/sub> 3.67 \u00b5g\/mL. In 2011, Ferreira et. al. reported the antioxidant activity of PSO extracted by petroleum ether with EC50<\/sub> > 1000 \u00b5g\/mL. Gonzalez et. al. (2019), reported antioxidants using the DPPH method and found IC50<\/sub> was 82.81 TEAC mmol\/100 g. This finding showed that the higher activity of PSO extracted by methanol has higher antioxidant activity than other solvents. The activity has a direct correlation with phenolic content in methanol. This was different from acetone extract with lower antioxidants compared to methanol extract.<\/p>\n\n\n\n

Total Phenolic and Flavonoid Content<\/strong><\/p>\n\n\n\n

Table 1. shows the TPC of the extracts determined by the FC technique. TPC values were calculated using the calibration curve y = 0.0047x + 0.123 (R2=0.9894), where x represents absorbance and concentration of a gallic acid solution (mg\/mL) reported as mg GAE\/g. The TFC of the extracts is shown in Table 2.  All extracts were measured by aluminum chloride method and rutin as a standard. <\/p>\n\n\n\n\n\n
\"\"<\/td>\n\n

Figure 3<\/strong>: Phenolic and Flavonoid contents of PSO<\/strong><\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n

Table 2: Phenolic and Flavonoid contents of PSO<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n
\n

Extract<\/p>\n<\/td>\n

\n

Phenolic content (mgGAE\/g extract)<\/p>\n<\/td>\n

\n

Flavonoid content (mgRE\/g extract)<\/p>\n<\/td>\n<\/tr>\n

\n

Acetone<\/p>\n<\/td>\n

\n

193.60<\/p>\n<\/td>\n

\n

23.08<\/p>\n<\/td>\n<\/tr>\n

\n

Ethyl acetate<\/p>\n<\/td>\n

\n

79.00<\/p>\n<\/td>\n

\n

35.40<\/p>\n<\/td>\n<\/tr>\n

\n

Chloroform<\/p>\n<\/td>\n

\n

109.80<\/p>\n<\/td>\n

\n

5.48<\/p>\n<\/td>\n<\/tr>\n

\n

Methanol<\/p>\n<\/td>\n

\n

158.90<\/p>\n<\/td>\n

\n

15.89<\/p>\n<\/td>\n<\/tr>\n

\n

N-hexane<\/p>\n<\/td>\n

\n

26.10<\/p>\n<\/td>\n

\n

21.00<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Note:<\/p>\n

RE  = Rutin Equivalent<\/p>\n

GAE = Gallic Acid Equivalent<\/p>\n\n\n

The results showed that the phenolic compounds were\nmainly dissolved in acetone and methanol, and the flavonoids were dissolved in\nethyl acetate. This could be because the water extract contains more\nnon-phenolic components like carbohydrates and terpenes than other extracts. It\ncan also be caused by the complicated synthesis of some phenolic compounds,\nwhich can have more phenolic groups or a larger molecular weight than phenol in\nwater extract. Previous reports indicated that the main compounds were fatty\nacids, phenols, and carotenoids.1,4,7,8,18<\/sup><\/p>\n\n\n\n

The\nresult showed that ethyl acetate with 35.40 mgRE\/g extract had the highest flavonoid\ncontent. It was observed that the medium solvent polarity was more\neffective in TFC extraction. The effectivity of the ethyl acetate solvent was followed by the acetone extract, n-hexane,\nmethanol, and chloroform, respectively. Gonzalez, et., al. (2019), reported that PSO contains 5.32 gRE\/100 g samples.<\/strong><\/p>\n\n\n\n

Conclusion<\/strong><\/p>\n\n\n\n

We conclude that PSO has the potential as an antioxidant\nand source of phenolic and flavonoid constituents. Methanolic extract of\npassion seed has the most potent antioxidant activity with an IC50 <\/sub>value\nof 71.67 \u00b5g\/mL.<\/p>\n\n\n\n

Conflict of Interest<\/strong><\/p>\n\n\n\n

There is no conflict of interest<\/p>\n\n\n\n

Acknowledgement<\/strong><\/p>\n\n\n\n

The authors thank Universitas Muslim Indonesia for\ntheir financial support.<\/p>\n\n\n\n

References<\/strong><\/p>\n\n\n\n

  1. Ch\u00f3ez-Guaranda, I., Ortega, A., Miranda, M., & Manzano, P. Chemical composition of essential oils of Passiflora edulis<\/em> f. flavicarpa agroindustrial waste. Emirates Journal of Food and Agriculture., 2017; 29 (6): 458\u2013462. https:\/\/doi.org\/10.9755\/ejfa.2016-10-1542
    CrossRef<\/a><\/li>
  2. Charissa, M., Djajadisastra, J. and Elya, B. Uji aktivitas antioksidan dan penghambatan tirosinase serta uji manfaat gel Extract kulit batang taya (Nauclea subdita<\/em>) terhadap kulit. Jurnal Kefarmasian Indonesia., 2016; 6 (2): 98-107.
     CrossRef<\/a><\/li>
  3. De Paula, R. C. M., Soares, A. G., & Freitas, S. P. Volatile compounds in passion fruit seed oil (Passiflora setacea BRS P\u00e9rola do Cerrado and Passiflora alata BRS Doce Mel). Chemical Engineering Transactions, 2015; 44: 103\u2013108. https:\/\/doi.org\/10.3303\/CET1544018<\/li>
  4. Ferreira, B. S., De Almeida, C. G., Faza, L. P., De Almeida, A., Diniz, C. G., Da Silva, V. L., Grazul, R. M., & Le Hyaric, M. Comparative properties of amazonian oils obtained by different extraction methods. Molecules, 2011; 16 (7): 5874\u20135885. https:\/\/doi.org\/10.3390\/molecules16075875
    CrossRef <\/a><\/li>
  5. Huda, S. M. N., Wiraguna, AAGP., dan Pangkahila, W.. Krim Extract Biji Markisa (Passiflora edulis<\/em>) Sama Efektifnya dengan Krim Hidrokuinon 4% dalam Menghambat Peningkatan Jumlah Melanin pada Kulit Marmut 88 Jantan (Cavia porcelus<\/em>) yang Dipapar Sinar UV-B. Jurnal Biomedik (JBM), 2017; 9 (1): 1-6.
    CrossRef <\/a><\/li>
  6. Karsinah, Hutabarat R.C dan Manshur A. Buah Eksotik Kaya Manfaat. Balai Litbang Pertanian, Kementrian Pertanian, Iptek hortikultura. Aripan Solok: Balitbu, 2010.<\/li>
  7. Laura Gonzalez, Andree \u00c1l Varez, Elizabeth Murillo, Carlos Guerra, & Jonh Mendez. Potential Uses of the Peel and Seed of Passiflora edulis<\/em> Sims F. Edulis (Gulupa) From Its Chemical Characterization, Antioxidant, and Antihypertensive Functionalities. Asian Journal of Pharmaceutical and Clinical Research, 2019; 12 (10): 104\u2013112. https:\/\/doi.org\/10.22159\/ajpcr.2019.v12i10.33828
     CrossRef <\/a><\/li>
  8. Lima-Neto, A. B. M., Marques, M. M. M., Mendes, F. N. P., Vieira, \u00cd. G. P., Diniz, D. B., & Guedes, M. I. F. Antioxidant activity and physicochemical analysis of passion fruit (Passiflora glandulosa<\/em> Cav.) pulp native to Cariri region. Acta Scientiarum – Biological Sciences, 2017; 39 (4): 417\u2013422. https:\/\/doi.org\/10.4025\/actascibiolsci.v39i4.34045.
    CrossRef <\/a><\/li>
  9. Lucarini, M., Durazzo, A., Raffo, A., Giovannini, A., & Kiefer, J. Passion Fruit (Passiflora<\/em> sp.) Seed Oil. Fruit Oils: Chemistry and Functionality, 2019; 577\u2013603. https:\/\/doi.org\/10.1007\/978-3-030-12473-1_29
     CrossRef <\/a><\/li>
  10. Malacrida, C.R. and Jorge, N. Yellow passion fruit seed oil (Passiflora edulis<\/em> f. flavicarpa): physical and chemical characteristics. Brazilian Archives of Biology and Technology, 2012; 55 (1): 127-134.
     CrossRef <\/a><\/li>
  11. Mamede, A. M. G. N., Soares, A. G., Oliveira, E. J., & Farah, A. Volatile Composition of Sweet Passion Fruit (Passiflora alata<\/em> Curtis). Journal of Chemistry, 2017. https:\/\/doi.org\/10.1155\/2017\/3497216
     CrossRef <\/a><\/li>
  12. Ngginak, J., Rupidara, A. and Daud, Y. Analisis Kandungan Vitamin C dari Extract Buah Ara (Ficus carica<\/em> L) dan Markisa Hutan (Passiflora foetida<\/em> L). Jurnal Sains dan Edukasi Sains, 2019; 2(2): 54-59.
    CrossRef <\/a><\/li>
  13. Nguyen. Tyrosinase inhibitory activity of flavonoids from Artocarpus heterophyllous. Chemistry Central Journal, 2016; 4\u20139
     CrossRef <\/a><\/li>
  14. Nugroho, A.E, Malik, A, Pramono, S. Total Phenolic and Flavonoid Contents, and in vitro <\/em> Antihypertension Activity of Purified Extract of Indonesian Cashew Leaves (Anacardium occidentale <\/em>L.). IFRJ; 2012; 20(1): 299-305.<\/li>
  15. Pereira, M.G., Maciel, G.M., Haminiuk, C.W.I., Bach, F., Hamerski, F., de Paula Scheer, A. and Corazza, M.L. Effect of extraction process on composition, antioxidant and antibacterial activity of oil from yellow passion fruit (Passiflora edulis<\/em> Var. Flavicarpa) seeds. Waste and Biomass Valorization, 2019; 10 (9): 2611-2625.
     CrossRef <\/a><\/li>
  16. Promono, A. A. dan Rustam, E. Perubahan Kondisi Fisik, Fisiologis, dan Biokimia Benih Michelia champaca pada Berbagai Tingkat Kemasakan. Pros. Sem. Nas. Masy. Biodiv. Indon. 2017; 3 (3): 368-375.
    CrossRef <\/li>
  17. Silva, R.M., Pl\u00e1cido, G.R., Silva, M.A.P.D., Castro, C.F.D.S., Lima, M.S. and Caliari, M. Chemical characterization of passion fruit (Passiflora edulis<\/em> f. flavicarpa) seeds. African journal of biotechnology, 2015; 14 (14): 1230-1233.
     CrossRef <\/a><\/li>
  18. Surlehan, H. F., Noor Azman, N. A., Zakaria, R., & Mohd Amin, N. A. Extraction of oil from passion fruit seeds using surfactant-assisted aqueous extraction. Food Research, 2019; 3<\/em>(4): 348\u2013356. https:\/\/doi.org\/10.26656\/fr.2017.3(4).146
     CrossRef <\/a><\/li>
  19. Woolery-Lloyd H, Kammer JN. Treatment of Hyperpigmentation. Semin Cutan Med Surg. NewYork. 2011; 30 (3): 171-175.
     CrossRef <\/a><\/li>
  20. Zeraik, M. L., Pereira, C. A., Zuin, V. G., & Yariwake, J. H. Maracuj\u00e1: um alimento funcional. Revista Brasileira de farmacognosia, 2010; 20 (3): 459-471.
     CrossRef <\/a><\/li><\/ol>\n\n\n\n

    <\/p>\n","protected":false},"excerpt":{"rendered":"

    Introduction Passion fruit belongs to the family Passifloraceae and one […]<\/p>\n","protected":false},"author":15,"featured_media":0,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[107],"tags":[],"class_list":["post-48952","post","type-post","status-publish","format-standard","hentry","category-vol16no2"],"_links":{"self":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts\/48952","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/users\/15"}],"replies":[{"embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/comments?post=48952"}],"version-history":[{"count":5,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts\/48952\/revisions"}],"predecessor-version":[{"id":50170,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts\/48952\/revisions\/50170"}],"wp:attachment":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/media?parent=48952"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/categories?post=48952"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/tags?post=48952"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}