{"id":54872,"date":"2023-12-31T11:34:34","date_gmt":"2023-12-31T11:34:34","guid":{"rendered":"https:\/\/biomedpharmajournal.org\/?p=54872"},"modified":"2024-01-05T06:13:34","modified_gmt":"2024-01-05T06:13:34","slug":"a-comparative-study-and-characterization-of-green-solvent-and-hexane-extracted-bioactive-lipids-from-heterospathe-elata-fruit","status":"publish","type":"post","link":"https:\/\/biomedpharmajournal.org\/staging\/vol16no4\/a-comparative-study-and-characterization-of-green-solvent-and-hexane-extracted-bioactive-lipids-from-heterospathe-elata-fruit\/","title":{"rendered":"A Comparative Study and Characterization of Green Solvent and Hexane Extracted Bioactive lipids from Heterospathe elata Fruit"},"content":{"rendered":"\n

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

Hexane has been widely used for oil and fat extraction. From petroleum and other hydrocarbons, hexane is produced. Hexane reacts with pollutants when it is discharged into the atmosphere, creating ozone and photochemicals1<\/sup>. The highly lipophilic nature of hexane makes it soluble in neural lipids. So, it became toxic for neural system when inhaled by humans. According to the REACH regulation (European Directives and Registration, Evaluation, Authorization, and Restriction of Chemicals), hexane is a category 2 compound that is prohibited from being used in cosmetic items and is both aquatic chronic toxic and reprotoxic2<\/sup>. Finding solvents other than those derived from petroleum is now a major concern for chemists because of the advent of green chemistry and the increased focus on environmental and safety concerns. Green solvents are made from renewable materials such as wood, starch, fruit, and oils. Green solvents are biodegradable, non-toxic, and non-flammable, and they have a strong salvaging power2<\/sup>. Consumer interest in foods with health benefits and foods with physiological activity has grown recently. Essential fatty acids are required for good health but cannot be produced by the body; they can only be obtained through dietary sources. Vital nutrients for humans, fats and oils supply both calories and essential fatty acids3<\/sup>. These fatty acids are widely employed in sectors like pharmaceuticals, cosmetics, and medicine. The amount and type of fatty acids in a particular diet have a significant impact on its lipid profile. The food business has been heavily influenced by consumers’ concerns about the ability of the diet to maintain the ratio of saturated to unsaturated fatty acids and identify good fats that are crucial for nutraceutical purposes. It is believed that polyunsaturated fatty acids are crucial adaptive mediators for enhancing and preserving human health. Omega-3 fatty acids are helpful in human pathologies such as diabetes, stroke, atherosclerosis, and cardiovascular disorders4<\/sup>. For the structural elements of cells, tissues, and organs as well as for the creation of several biologically active chemicals, fatty acids are regarded as a vital building block. Fatty acids are viewed as a crucial building block for the structural components of cells, tissues, and organs as well as for the creation of some physiologically important substances5,6<\/sup>.While few studies have also been reported on the characterization of fixed oils (the non-volatile fraction) from these Arecaceae family species like Caryota mitis, Borassus flabellifer, Areca catechu<\/em> nut, and Cocus nucifera7<\/sup><\/em>. To the best of our knowledge, there has been no publication on fixed oils from Heterospathe elata fruit. <\/p>\n\n\n\n

The present study investigated and evaluated GC-FID of fixed oil extracted from Heterospathe elata<\/em> fruit using petroleum\u2013based solvent hexane and green solvent dimethyl carbonate (DMC), physicochemical parameters, and biological activities using antioxidant, anti-inflammatory, and photoprotective activities.<\/p>\n\n\n\n

Material and Methods<\/strong><\/p>\n\n\n\n

Sample Collection and Authentication<\/strong><\/p>\n\n\n\n

Samples\nwere collected in July 2021 from the northern Indian state of Uttar Pradesh.\nThe sample authentication was carried out, and the identification of the plant\nmaterial based on the morphological criteria was confirmed at the Department of\nBotany, Banaras Hindu University (voucher specimen no. 1\/7A, 22\/07\/21). The\nfruit of Heterospathe elata was washed under distilled water to remove dust.\nThe fruit samples were dried in the air for 20 days. Then, the fruit was ground\nusing electrical grinders (Pulverizer, HR 1500) and then sieved (0.25 mm) to\nobtain a uniform particle size. The sample was stored in a refrigerator for\nfurther examination.<\/strong><\/p>\n\n\n\n

Chemicals and Reagents<\/strong><\/em><\/p>\n\n\n\n

2,2-Diphenyl-1-picrylhydrazyl\n(DPPH), 2,2\u2032-azino-bis(3-ethylbenzothiazoline-6-sulpfonic acid (ABTS),\n2,4,6-tri(2-pyridyl)-s- triazine (TPTZ), ascorbic acid, phenolic standards\nnamely gallic acid, quercetin,  were procured\nfrom Sigma-Aldrich. Folin-ciocalteu reagent, ammonium acetate, sodium\ncarbonate, potassium persulphate, sodium acetate trihydrate, manganese chloride\n(II) and iron (III) chloride hexahydrate, and Benzophenone were purchased from\nHimedia, India. Bovine albumin serum (CDH), Hexane and Dimethyl carbonate from\nSRL Pvt. Ltd. All other chemicals used were analytical grades.<\/p>\n\n\n\n

Extraction of Fixed Oil <\/strong><\/p>\n\n\n\n

The Soxhlet apparatus was used to extract the oil according to standard method4<\/sup>. To prevent solvent loss by evaporation, extraction was done close to the dimethyl carbonate (DMC) solvent boiling point. When the first drop of the extraction solvent was recycled back into the thimble, the extraction period began. The solvent was collected and reused in the subsequent extraction batch in a rotary evaporator operating under a vacuum. After cooling, the oil was weighed. Oil was extracted into amber-colored vials, sealed with Teflon tops, and kept in the refrigerator at 4\u00b0C for later analysis. Similarly, oil was extracted from the same fruit sample using n-hexane The following expression was used to compute the oil yield: <\/p>\n\n\n\n

\"\"<\/figure>\n\n\n\n

Determination of Physical Properties<\/strong><\/p>\n\n\n\n

Physical properties color, odour, peroxide value, acid value, refractive\nindex, Iodine value, ester value, saponification value were estimated using\nstandard reported methods8<\/sup>.<\/p>\n\n\n\n

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

Using\nthe Folin-Ciocalteu reagent, the total phenolic contents of the extracted oil\nwere calculated with slight modification9<\/sup>. Briefly, 250\u03bcl of freshly\nmade Folin-Ciocalteu reagent, 0.75 ml of 20% sodium carbonate, and 3 ml of pure\nwater made up the reaction mixture, along with 50\u03bcl of each plant extract. The\nabsorbance at 765 nm was measured after 2 hours of reaction at room\ntemperature, and the phenolic content was calculated using gallic acid as a\nreference.<\/strong><\/p>\n\n\n\n

Chemical Composition of Fixed Oil by GC-FID<\/strong><\/p>\n\n\n\n

Fatty acids composition investigation was\ncarried out by means of GC-FID (gas chromatography-flame ionization detection\nafter derivatization of fatty acid methyl esters (FAMEs). TQ8040 Shimadzu GC, HS20\nheadspace sampler, mobile phase He at 1 ml\/min, SH-Rxi-5Sil MS capillary column\n(stationary phase: 5% diphenyl-95% dimethyl polysiloxane, length: 30 m, inner\ndiameter ID: 0.25 mm, film thickness df: 0.25 mm), split ratio 1:10, injection\ntemperature 280, column oven temperature increased from 80 \n \n to 280  \n \n at a rate of 5 \/min. The\nbioactive compounds were identified based on retention time with those of\ncertified FAME mix. The results were expressed as the percentage of each fatty\naci in the total10<\/sup>.  <\/p>\n\n\n\n

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

1-Diphenyl-2-picryl-hydrazyl (DPPH) assay<\/strong><\/p>\n\n\n\n

The radial scavenging activity of the Heterospathe elata<\/em> oil was assessed using DPPH with some modifications11<\/sup>. Different test tubes were filled with ascorbic acid extracts at various concentrations. Petroleum ether was added to bring the volume to 100 mL. These tubes were filled with five mL of a 0.1 mM petroleum ether solution of DPPH and vigorously shaken. For 30 minutes, the tubes were left to stand at 27 0<\/sup>C. Without any extract, the control was made in the same manner. At 512 nm, the produced samples variations in absorbance were measured. The inhibition percentage was used to estimate the amount of radical scavenging activity, and the following formula was used to do so:<\/p>\n\n\n\n

\"\"<\/figure>\n\n\n\n

2\u2032-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid (ABTS) assay<\/strong><\/p>\n\n\n\n

The\nABTS radical cation scavenging assay was used to calculate the total\nantioxidant activity with slight modification12<\/sup>.\nIn an ABTS (stable radical) solution with 2.4 mM potassium persulfate, the ABTS\nradical cation was produced in the absence of light for 12 to 16 hours. The ABTS\nsolution was diluted in petroleum ether (1:89 v\/v) prior to the experiment to\nproduce an absorbance of 0.70 \u00b10.02 at 734 nm. Ascorbic acid was added to 1 mL\nof diluted ABTS solution along with triplicates of the 10 \u00b5L samples. After 30\nminutes of incubation at 30 0<\/sup>C, the reaction mixtures absorbance was\nmeasured at 734 nm in comparison to petroleum ether (blank).<\/strong><\/p>\n\n\n\n

Anti-inflammatory assay by\nbovine albumin serum denaturation<\/strong><\/p>\n\n\n\n

The ability of extracted oil to induce protein denaturation was determined according to the method described with some modification13<\/sup>. The reaction mixture consists of 2.8 mL of phosphate buffered saline (pH 6.4), 0.2 mL of fresh bovine albumin serum, and 2 mL of various extracts at variable concentrations. The reaction mixtures were then incubated in the BOD incubator for 15 minutes at ambient temperature before being heated for 5 minutes at 72 0<\/sup>C. Their absorbance at 660 nm (Systronic 118, UV-VIS) was measured using a blank after cooling. In order to determine absorbance, sodium diclofenac was used as a reference medication and handled similarly to plant extracts. The following equation was used to determine the percent inhibition of protein denaturation:<\/p>\n\n\n\n

\"\"<\/figure>\n\n\n\n

Where, Vs = absorbance of test sample, Vc = absorbance of control.<\/p>\n\n\n\n

Photoprotective Activity<\/strong><\/p>\n\n\n\n

Determination of the sun protection factor (SPF)<\/strong><\/p>\n\n\n\n

Using a UV-VIS spectrophotometer, the absorbance values of each extract dilution were calculated at 5-nm intervals between 290 and 320 nm, using MeOH\/H2O (80:20, v\/v) as a blank. SPF was calculated using an equation with slight modification14<\/sup>. <\/p>\n\n\n\n

 SPF \nspectrophotometric  = C F \u00d7 \u03a3 EE(\u03bb) \u00d7 I(\u03bb) \u00d7 Abs(\u03bb)<\/strong><\/p>\n\n\n\n

Where,\n EE (\u03bb)  indicates  erythemal\neffect spectrum;  <\/p>\n\n\n\n

I\n(\u03bb) indicates the solar intensity spectrum; <\/p>\n\n\n\n

Abs\n(\u03bb) indicates absorbance of the sunscreen  product <\/p>\n\n\n\n

CF\nstands for the correction factor (=10). <\/p>\n\n\n\n

The values\nof EE \u00d7 I are constant.<\/p>\n\n\n\n

Statistical Analysis<\/strong><\/p>\n\n\n\n

All  the \nexperiments  were  done \nin  triplicates  & \nthe  results   were \nexpressed  as  Mean\u00b1 SD. \nThe  data  were \nstatistically  analyzed  using \none  way  ANOVA \nfollowed  by  Duncan\u2019s test. Mean values were considered\nstatistically significant when p>0.05.<\/p>\n\n\n\n

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

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

Hexane\nand the environmentally friendly solvent dimethyl carbonate (DMC) were used to\nproduce Heterospathe elata<\/em> fruit\noil. 18% oil yield was obtained with hexane extraction and 24% with DMC\nextraction (Fig. 1). In the extraction of DMC oil, the yield is greater. The\nmost popular method for extracting lipids from petroleum using petroleum\nsolvents like hexane is known as solvent extraction. Due to their extreme\nvolatility, petroleum solvents are a major threat to both human health and the\nenvironment. Humans that breathe in hexane suffer neurological harm. A less\ntoxic and environmentally friendly alternative, known as a “green\nsolvent,” has begun to be sought due to environmental, health, and safety\nconcerns. According to recent laboratory testing, many green solvents have the\npotential to replace petroleum solvent in the extraction process12,13<\/sup>.\nDimethyl carbonate (DMC) is one of the environmentally friendly solvents that\ncould eventually take the place of hexane. Green solvents disposal would be\nless expensive than petro-based solvent disposal. We can say that DMC is a good\nalternative solvent for the extraction of lipids because our findings showed\nthat it has a higher ability to extract oil than hexane. When Farid Chemat et\nal. evaluated various green solvents for oil extraction, they discovered that\nthey were more effective than hexane12,14<\/sup>.<\/p>\n\n\n\n\n\n
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Figure 1:<\/strong> Oil yield of Heterospathe elata<\/em> fruit obtained with hexane and DMC extractions<\/strong><\/p>\n

<\/p>\n

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

Physicochemical Properties <\/strong><\/p>\n\n\n\n

The refractive index is the degree of deflection of\na beam of light that occurs when it passes from one transparent medium to\nanother. The refractive indices of DMC and hexane-extracted oil were 0.98 and\n1.53, respectively. The extracted oil was also subjected to evaluation of its\nacid value. The acid values for DMC and hexane-extracted oil were found to be\n0.72 mg KOH\/g and 0.93 mg KOH\/g, respectively. Acidity is used as an indicator\nof the edibility of oil. It was found that the measured iodine value for\nDMC-extracted oil was 72.59 g\/100 g and 78.12 g\/100 g for hexane-extracted oil.\nThe low iodine values indicated that this oil sample has greater oxidative\nstorage stability. The oil sample was also subjected to evaluation of its\nsaponification value. Saponification values of DMC-extracted oil were found at\n345.12 mg KOH\/g and 223.48 for hexane-extracted oil. The saponification value\nindicates the average molecular weight of a fat or oil. An ester value of DMC\nand hexane-extracted oil was found to be 344.4 and 222.5, respectively.\nPeroxide value is a very important characteristic of lipid quantity. The\nassessment of hydroperoxide provides an estimate of the overall oxidation\nstatus of lipids and lipid-containing foods, especially in the primary phase of\noxidation, generally known as the induction period. Fruit DMC-extracted oil\nindicated a low peroxide value. 0.84 g\/kg and a lower number of peroxide values\nindicate a good quality of oil and a good preservation status15,16<\/sup>.<\/p>\n\n\n\n

Table 1:  Physicochemical Properties of Fixed oil from Heterospathe elata<\/em> fruit<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n
\n

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

\n

Oil ( extracted with DMC)<\/strong><\/p>\n<\/td>\n

\n

Oil (extracted with Hexane )<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

Yield<\/p>\n<\/td>\n

\n

24%<\/p>\n<\/td>\n

\n

18%<\/p>\n<\/td>\n<\/tr>\n

\n

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

\n

Light pink<\/strong><\/p>\n<\/td>\n

\n

Light pink<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

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

\n

Characteristic smell<\/strong><\/p>\n<\/td>\n

\n

Characteristic smell<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

Refractive Index<\/p>\n<\/td>\n

\n

0.98<\/p>\n<\/td>\n

\n

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

\n

Iodine Value<\/p>\n<\/td>\n

\n

72.59<\/p>\n<\/td>\n

\n

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

\n

Acid Value(mg KOH\/g)<\/p>\n<\/td>\n

\n

0.72<\/p>\n<\/td>\n

\n

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

\n

Saponification value (mgKOH\/g)<\/p>\n<\/td>\n

\n

345.12<\/p>\n<\/td>\n

\n

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

\n

Ester Value<\/p>\n<\/td>\n

\n

344.4<\/p>\n<\/td>\n

\n

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

\n

Peroxide Value(g\/kg)<\/p>\n<\/td>\n

\n

0.84<\/p>\n<\/td>\n

\n

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

 <\/p>\n\n\n

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

Phenolic molecules have a significant impact on the sensory and nutritional quality of oil and guard against lipid oxidation by quenching radical reactions. To prevent heart diseases, the potential for phenol-containing oils to promote health exists. The overall amount of polyphenol content is known to be influenced by the polarity of the extraction solvent as well as other elements such as plant cultivar, seed maturity level, environmental changes, and growing location17<\/sup>. Heterospathe elata<\/em> fruit oil extracted using the green solvent DMC showed a higher total phenolic content of 12 mg GAE\/100 g DW. As a result, the DMC suggested in this work enabled the extraction of Heterospathe elata fruit oil with the highest possible polyphenol content. Based on these findings, it appears that greenly extracted oils contain a significant amount of phenolic compounds, which may contribute to the stability of the oil under accelerated oxidative stress9,17<\/sup>.<\/p>\n\n\n\n

Table 2: Total Phenolic Content of Heterospathe elata<\/em> fruit oil<\/strong><\/p>\n\n\n\n\n\n\n\n
\n

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

\n

Total Phenolic Content (mgGAE\/100gm dw)<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

Oil extracted with DMC<\/p>\n<\/td>\n

\n

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

\n

Oil extracted with hexane<\/p>\n<\/td>\n

\n

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

 <\/p>\n\n\n

Fatty Acid Composition<\/strong><\/p>\n\n\n\n

Essential fatty acids are necessary for healthy\nhealth. Since the human body is unable to synthesize these fatty acids, they\nmust be obtained through diet. Table 3 provides a summary of the fatty acid\nprofile (GC-FID) of the oil derived from the Heterospathe elata fruit using\nhexane and the green solvent DMC. Both DMC and hexane-extracted oil contain\nLauric acid, Myristic acid, Palmitic acid, Stearic acid, Oleic acid, and\nLinoleic acid in significant amounts. Butyric acid (3.515%), caproic acid\n(3.389%), and caprylic acid (1.692%) were also identified only from the oil\nextracted from DMC. Butyric acid can reduce arterial blood pressure and be used\nto treat or prevent cancer, irritable bowel syndrome, diverticulosis, and\ndiarrhea18<\/sup>. A medium-chain fatty acid known as caprylic acid is\nthought to have strong antibacterial, antifungal, and anti-inflammatory\nactivities. According to some previous research, it may help treat high\ncholesterol, skin diseases, digestive problems, and yeast infections. The\nlikelihood of antibiotic resistance may be reduced by using it as a\ndisinfectant. Capric acid has direct uses as antimicrobials, plant growth\nstimulants, and feed additives. Additionally, it can be used as a base for\nseveral products, such as lubricants, perfumes, paint additives, and\nmedications19<\/sup>. The lipid membrane of bacteria can be destroyed by\nsaturated fatty acids. Linoleic acid is an Omega-3 fatty acid that has\nanti-inflammatory properties and lowers the risk of chronic disease. Linoleic\nacid (6.176%), which is present in Heterospathe elata oil, confers\nanti-inflammatory characteristics to the substance20,21<\/sup>. Oleic acid\nis a member of the monounsaturated omega-9 fatty acid family. The development\nof deadly brain disorders is slowed by oleic acid. Oleic acid is a helpful\nsource for improving memory. Additionally, oleic acid can lower blood pressure19,21<\/sup>.\nHeterospathe elata fruit oil can be employed as a source of an ingredient in\noleochemicals, soap, cosmetics, and medicine due to the presence of these fatty\nacids. The ester isopropyl myristate of myristic acid is frequently used as a\ncomponent in soaps and shaving creams. The most prevalent saturated fatty acids\nare lauric acid, myristic acid, stearic acid, and palmitic acid21<\/sup>.<\/p>\n\n\n\n

Table 3:  Fatty acid composition of fixed oil of Heterospathe elata <\/em>Fruit<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n
\n

Fatty acids <\/strong><\/p>\n<\/td>\n

\n

Oil extracted using    DMC (%)<\/strong><\/p>\n<\/td>\n

\n

Oil extracted using Hexane (%)<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

Butyric acid<\/p>\n<\/td>\n

\n

3.515<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n<\/tr>\n

\n

Caproic acid<\/p>\n<\/td>\n

\n

3.389<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n<\/tr>\n

\n

Caprylic acid<\/p>\n<\/td>\n

\n

1.692<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n<\/tr>\n

\n

Lauric acid<\/p>\n<\/td>\n

\n

15.131<\/p>\n<\/td>\n

\n

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

\n

Myristic acid<\/p>\n<\/td>\n

\n

45.063<\/p>\n<\/td>\n

\n

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

\n

Palmitic acid<\/p>\n<\/td>\n

\n

12.526<\/p>\n<\/td>\n

\n

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

\n

Stearic acid<\/p>\n<\/td>\n

\n

1.046<\/p>\n<\/td>\n

\n

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

\n

Oleic acid<\/p>\n<\/td>\n

\n

11.462<\/p>\n<\/td>\n

\n

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

\n

Linoleic acid<\/p>\n<\/td>\n

\n

6.176<\/p>\n<\/td>\n

\n

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

 <\/p>\n\n\n\n
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Figure 2<\/strong>: GC-FID chromatogram of Hexane extracted fixed oil from Heterospathe elata<\/em> Fruit<\/strong><\/p>\n

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n\n
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Figure 3: <\/strong>GC-FID chromatogram of DMC extracted fixed oil from Heterospathe elata<\/em> Fruit<\/strong><\/p>\n

<\/p>\n

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

Biological Activities<\/strong><\/p>\n\n\n\n

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

The\nantioxidant properties of Heterospathe elata<\/em>\nfruit oil produced using conventional and environmentally friendly solvents\nwere examined in this work. Oil extracted using DMC green solvent had an IC50<\/sub> of 227.14 \u00b10.89 \u00b5g\/mL) when tested using\nthe DPPH assay and (139.25\u00b10.47 \u00b5g\/mL ) when tested using the ABTS assay.\nHexane-extracted oil has an IC50<\/sub>\nof (457.20\u00b11.34\u00b5g\/mL and 263.33\u00b10.25\u00b5g\/mL) when tested with DPPH and ABTS,\nrespectively. A lower IC50<\/sub> value\nand much better antioxidant activity were observed in Heterospathe elata<\/em> oil extracted with DMC\ncompared to oil extracted with hexane, which is an interesting finding.\nLinoleic and oleic acids can be found in Heterospathe\nelata<\/em> fruit oil, according to the GC-FID. The antioxidant action is\ncaused by these fatty acids. Fixed oils, compared to ascorbic acid used as a\nreference, showed significant antioxidant activity. According to our research,\nDMC green extracted oil has a greater overall phenolic content value. Phenolic\ncompounds are powerful antioxidants and efficient intercellular ROS scavengers17<\/sup>. Overall, our findings imply that DMC\nmight be a great hexane replacement for recovering Heterospathe\nelata<\/em> with increased antioxidant activity. Therefore, Heterospathe elata<\/em> improved bioactivity may\nhelp to reduce lipid peroxidation at both low and high temperatures, and it may\nbe employed as a functional oil in dietary supplements22<\/sup>.<\/strong><\/p>\n\n\n\n

Table 4: Antioxidant activity of Heterospathe elata<\/em> Fruit Fixed Oil<\/strong><\/p>\n\n\n\n\n\n\n\n
\n

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

\n

           DPPH IC50 <\/sub>\u00b5g\/mL<\/strong><\/p>\n<\/td>\n

\n

        ABTS IC50 <\/sub>\u00b5g\/mL<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

Fixed oil ( extracted with hexane)<\/p>\n<\/td>\n

\n

          475.34<\/p>\n<\/td>\n

\n

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

\n

Fixed oil (extracted with DMC)<\/p>\n<\/td>\n

\n

           275.30<\/p>\n<\/td>\n

\n

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

 <\/p>\n\n\n\n
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Figure 4: <\/strong>DPPH free radical scavenging activity of Heterospathe elata<\/em> fruit fixed oil.<\/strong><\/p>\n

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n\n
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Figure 5: <\/strong>ABTS radical scavenging activity of activity of Heterospathe elata<\/em> fruit fixed oil<\/strong><\/p>\n

<\/p>\n

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

Anti-inflammatory activity<\/strong><\/p>\n\n\n\n

Fixed\noils of Heterospathe elata <\/em>fruit\nwere evaluated against denaturation of bovine albumin. This study represents a\nconcentration-dependent inhibition of protein (albumin) denaturation by fixed\noil throughout the concentration range of 100 to 1000\u00b5g\/mL. Sodium diclofenac,\nwhich is used as a standard drug, also exhibited concentration-dependent\ninhibition of protein denaturation; however, fixed oil of Heterospathe elata <\/em>showed significant inhibition\n(Fig. 6). Denaturation of tissue proteins is one of the authenticated reason of\ndiseases like inflammatory and arthritic23<\/sup>.\nThe formation of antibodies in some arthritic diseases may be due to\ndenaturation of proteins. The increments in absorbances of test samples with\nrespect to control indicated stabilisation of protein, i.e., inhibition of\nheat-induced protein (albumin) denaturation by fixed oil and the reference drug\nsodium diclofenac. From the IC50<\/sub>\nvalues, it becomes evident that fixed oil was amply active after sodium\ndiclofenac, being effective at lower concentrations. It is reported that many\nnon-steroidal anti-inflammatory drugs have the ability to stabilise (forbid\ndenaturation) heat-treated egg albumin at pH 6.2\u20136.5. Hence, from this experiment,\nit can be concluded that the fixed oil of Heterospathe\nelata <\/em>fruit showed a significant in vitro anti-inflammatory effect\nagainst the denaturation of protein. The IC50<\/sub>\nof oil extracted with DMC green solvent was 336.84 \u00b11.09) and hexane-extracted\noil has an IC50<\/sub> value of\n496.80\u00b10.37. The anti-inflammatory efficacy of Heterospathe elata<\/em> oil\nextracted with DMC was remarkably higher than that of oil extracted with\nhexane, which is interesting. Linoleic acid (6.176%) and other polyunsaturated\nfatty acids are present, according to fatty acid composition (GC-FID). The\nanti-inflammatory effects are caused by polyunsaturated fatty acids.<\/p>\n\n\n\n\n\n
\"\"<\/td>\n\n

Figure 6: Anti-inflammatory activity of Heterospathe elata<\/em> fruit fixed oil<\/strong><\/p>\n

<\/p>\n

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

Photoprotective Activity<\/strong><\/p>\n\n\n\n

The fixed oil of Heterospathe elata had the\nphotoprotective activity evaluated with the method developed by Mansur 198613<\/sup>.\nThe results of the determinations of in vitro SPF values are shown in Table 5.\nAccording to Brazilian law, only SPF greater than or equal to 6 is suitable for\nuse in cosmetic products with photoprotective activity13<\/sup>. This\nactivity can be attributed to the flavonoids and phenolic compounds found in\nthe oil of Heterospathe elata fruit which is efficient in absorbing ultraviolet\nlight, and usually show two maximum peaks of ultraviolet absorption, one\nbetween 240-280 nm and another 300-550 nm Bobin 1995. Several classes of\nnatural compounds were examined for their antioxidant and photoprotective\nactivities. Previous studies reviewed the importance of natural sunscreens in\nthe compositions of commercial sunscreens and therefore analyzed their role in\nthe prevention of skin cancer. Photoprotective properties have been evaluated\nin many varieties of plants; oil proved to be good at reducing agent-induced\nerythema as compared to benzophenone. The calculated values for a sun\nprotection factor of hexane and DMC-extracted oil were 20.38 \u00b1 0.11 and\n44.14\u00b10.19, respectively. DMC-extracted oil from Heterospathe elata<\/em>\nfruit oil has a greater SPF rating than hexane-extracted oil. Palmitic acid\noffers strong skin protection, whereas linoleic acid works well as an\nemollient, so Heterospathe elata<\/em> fruit oil can be used in the cosmetic\nindustry.<\/p>\n\n\n\n

Table 5 Photoprotective activity of Heterospathe elata<\/em> fruit fixed oil<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n
\n

\u039b (nm)<\/strong><\/p>\n<\/td>\n

\n

EE \u00d7 I<\/strong><\/p>\n<\/td>\n

\n

Absorbance of hexane extracted oil<\/strong><\/p>\n<\/td>\n

\n

Absorbance of  DMC extracted oil <\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

290<\/p>\n<\/td>\n

\n

0.0150<\/p>\n<\/td>\n

\n

0.601<\/p>\n<\/td>\n

\n

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

\n

295<\/p>\n<\/td>\n

\n

0.0817<\/p>\n<\/td>\n

\n

0.498<\/p>\n<\/td>\n

\n

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

\n

300<\/p>\n<\/td>\n

\n

0.2874<\/p>\n<\/td>\n

\n

0.377<\/p>\n<\/td>\n

\n

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

\n

305<\/p>\n<\/td>\n

\n

0.2780<\/p>\n<\/td>\n

\n

0.269<\/p>\n<\/td>\n

\n

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

\n

310<\/p>\n<\/td>\n

\n

0.1864<\/p>\n<\/td>\n

\n

0.127<\/p>\n<\/td>\n

\n

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

\n

315<\/p>\n<\/td>\n

\n

0.0837<\/p>\n<\/td>\n

\n

0.099<\/p>\n<\/td>\n

\n

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

\n

320<\/p>\n<\/td>\n

\n

0.0180<\/p>\n<\/td>\n

\n

0.063<\/p>\n<\/td>\n

\n

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

\n

Total<\/p>\n<\/td>\n

\n

1<\/p>\n<\/td>\n

\n

2.034<\/p>\n<\/td>\n

\n

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

\n

SPF<\/p>\n<\/td>\n

\n

 <\/p>\n<\/td>\n

\n

20.38\u00b10.11<\/p>\n<\/td>\n

\n

44.14\u00b10.19<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n\n\n

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

The findings indicate that fixed oil extracted from Heterospathe elata fruit had good qualities for industrial use based on physicochemical parameters. Green solvent DMC is a promising alternative to hexane for recovering enhanced functional oils. The Heterospathe elata oil yield extracted using green solvent was increased compared to conventional solvent hexane. The oil extracted with DMC had the highest total phenolic content, antioxidant, and anti-inflammatory activity. DMC-extracted oils possess potent photoprotective activity also due to the presence of active constituents in the composition of fixed oil. These results showed that using a substitute solvent to extract oil from Heterospathe elata would be helpful to obtain oils with enhanced bioactive components. The fixed oil of Heterospathe elata fruit could be a potential natural source and could have greater importance as a therapeutic agent in preventing or slowing oxidative stress and inflammation-related disorders. <\/strong><\/p>\n\n\n\n

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

None<\/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

Funding Sources<\/strong><\/p>\n\n\n\n

There are no funding source.<\/p>\n\n\n\n

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

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