id7145703 pdfMachine by Broadgun Software - a great PDF writer! - a great PDF creator! - http://www.pdfmachine.com http://www.broadgun.com BBiiooTTISSNee : 09c7c4 - 7hh435 nnooVlloluoome 1g2g Issuyye 3 An Indian Journal FULL PAPER BTAIJ, 12(3), 2016 [113-120] Evaluation of bioactive compounds of aloe vera extract using sub- critical water method Narsih*, Agato Department of Agriculture Technology, Pontianak State Polytechnic, West Kalimantan, 78124, (INDONESIA) E-mail: [email protected] ABSTRACT KEYWORDS The Aloe vera peel extraction of 10 minutes at 1150C gave the best effect of Aloe vera skin; its chemical composition. Whilst Aloe vera skin using subcritical water Bioactive compounds; method was obtained from extracted with water at 125oC for 60 minutes. It Time and temperature. had following properties: mineral content of 20.654 mg/g; protein content of 120.114 mg/g; vitamin C content of 161.431 mg/g; aloin content of 10.251 mg/g; saponin content of 9.71 mg/g; total phenolic content of 39.851 mg/g; antioxidant activity of 87.651 %. The FTIR analysis showed that CH Stretch in alkana vibration at 2941.24 cm-1 to 2991.39 cm-1 wave was found for the methilene compound as antioxidant agent. 2016 Trade Science Inc. - INDIA INTRODUCTION tection to the bioactive compound on Aloe vera can be conducted by subcritical water method, such as Aloe vera have been attributed to the extraction process using water with temperature be- polysaccarides contained in the gel and skin. Re- tween boiling point and critical point. Subcritical cently, Aloe vera gel have been diversification into water extraction has several advantages of being processing product mostly from gel[1]. Although there readily available, cheap and efficient[5]. The result is a high interest in gel, the interest of skin into a of subcritical water exctraction, extraction water processing is very limited information. However, become short time, cheap and safe for health[6] and the Aloe vera skin has a big potention to developed subcritical water at 100 to 1100C appears to be an into a diversification productAloe vera (L.) skin con- alternative to organic solvent due to efficiency at tained organic and bioactive compound that act as low polarity condition[7]. However, there is limited antioxidant[1-2]. information on the effect of time and temperature of phenolic compound (flavonoid, phenolic acid, extraction on subcritical water to decrease and phenolic deterpene) and antroquinon compound antinutritive compound and bioactive compound of (aloin)[3]. Aloe vera also contained alkoloid and also Aloe vera keep protected. saponin that act as antinutrive[4]. The objectives of current study was to find out The reduction of antinutritive compound and pro- the bioactive compound from the effect of time and 114 Evaluation of bioactive compounds of aloe vera extract using subcritical water method BTAIJ, 12(3) 2016 FULL PAPER temperature of subcritical water which result from acid solution which has been in the mixed indicator. the previous study. Subsequently titrated with a solution of HCl 0.1 N. Blank determination is also performed. MATERIALS AND METHODS Sample (1 g) was dissolved in 1 mL chloroform: µL was drop on TLC methanol 95:5 (v/v) and 10 Aloe vera of 10 months old and weight of 3 kg plat distance of 2 cm. Poured TLC plat to the glass were obtained from Kalimantan Barat, Indonesia. beaker for 40 minutes. Measured the top edge and After harvesting the Aloe vera peel were then washed the plats need to dried for 10 minutes. Plat was heated by aquades, weight, peeled and separated from the at 900C for 10 minutes. To visualize the spots, plat gel. Blended raw materials were extracted with was sprayed with the solution mixed between 25 water at 125oC for 60 minutes. Sample was filtered mL concentrated sulfate acid and 25 mL aquadest using vacuum filtration until produce filtrate and resi- (1:1), then heated at 1400C for 40 minutes. The due. amount of spot were counted and measured by the Evaporator rotary vacuum was used to disap- Rf value. pearance solvent in filtrate at 40oC for 1 h. Filtrate The Aloe vera (L.) skin were analyzed for its was centrifuged with a speed of 5500 rpm for 10 antioxidant activity using the DPPH (2,2-diphenyl- minutes. The supernatant obtained was used to con- 2-picrylhydrazyl) radical scavenging assay. Sample –HCl buffer cerned parameter analysis. (200 g) was dissolved in 100 mM Tris ìl, Total phenolic content was determined using 0.5 (800 pH 7.4) followed by the addition of 1 mL ìM mg/g of gallic acid as a standard. Briefly, 1 mL 500 DPPH. The solution was homogenized us- sample extract in methanol solution was transferred ing a shaker and storage in dark room for 20 min. ’s reagent in tubes containing 5 mL Folin-Ciocalteu Spectrophotometry was used to determine the ab- and also 4 mL Na2CO3 (75% w/v) was transferred sorbance at 517 nm. in tubes. The tubes were then mixed and then al- The Aloe vera (L.) skin was analyzed for its func- lowed to stand at room temperature for 30 min be- tional compounds using FTIR (Fourier Transform fore absorbance at 765 nm was measured. The total Infra Red). The IR spectra were recorded on FTIR- phenolic content was expressed as gallic acid 8400S (Shimadzu Deutchland GmbH) spectropho- equivalents (GAE) in mg/100 mL of sample. tometer in KBr and polyethylene pellets. Samples Extract of Aloe vera skin was weight of 2 g into were weigh-in at 0.01 g and homogenized with 0.01 the vitresile crucible overnight in an electric muffle g KBr anhydrous by mortar agate. The sample and furnace maintaining the temperature between 400- KBr mixture were pressed by vacuum hydrolic 410 0C until obtain the ash. This ashing destroys all (Graseby Specac) at 1.2 psi to obtained transpar- the organic material from the sample. The ash was ency pellet. Scanned sample passed through infra removed from crucible and allowed to dry in red, where its continuing wave by detector connected desicator. The yield of ash was approximately 5 g/ to computer with set values of tested sample spec- 100 g. trum. Samples were usually scanned in the absorp- Sample was weight of 3.5 g and entered into tion area of 500-4000 cm-1. The results of analysis Kjeldahl apparatus. 10 g of Na2SO4 anhydrate and consisted of chemical structure, molecular binding 15-25 mL of H2S04 concentrated were added and form and certain functional group of tested sample then heated on burner flame until obtain clear green- as basic of spectrum type. ish. Cooled, then diluted and put into a 200 mL volu- metric flask, matched to the dash. A total of 5 mL of RESULTS AND DISCUSSION solution pipetted and introduced into the distillation device, add 5 mL 45% NaOH and several drops of Mineral content phenolphthalein indicator. Distilled for about 10 Minerals found in Aleo vera are calcium (118,77 mBiniiuoteTs,e acsh thneo rlleosgeryvoir using 10 mL of 2% boric ppm), zinc (0,45 ppm), chromium (0,28 ppm), po- An Indian Journal BTAIJ, 12(3) 2016 Narsih and Agato 115 FULL PAPER tassium (640,51 ppm), copper (1,28 ppm), manga- in Aloe vera is known as tocopherol[17]. Vitamin C nese (0,36 ppm) and iron (0,23 ppm) (extract by and E are two important nutrients that may reduce Atomic Absorption Spectrophotometer (AAS))[8]. free radicals and a strong line of defense to slow They are essential for the proper functioning of vari- Reactive Oxygen Species (ROS)-induced cellular ous enzyme systems in different metabolic pathways damage. Vitamin C prevents the prooxidant activity [9] and few are antioxidants . of vitamin E by decreasing the activity of Mineral content of Aloe vera skin was 20.654 tocopheroxyl radical to tocopherol, thereby contrib- mg/g. Minerals are not affected significantly by uting to increased total antioxidant status and reduc- chemical and physical treatment during processing ing oxidative stress[18]. as a presence of oxygen. Some minerals lead an oxi- Vitamin C content of Aloe vera skin was 161.431 dized become a higher valent. Although several com- mg/g, which is similar to the one reported by[19], who ponents damaged as a heating process, this method found vitamin C content from Aloe vera extracted at was not affected to the amount of mineral and also 0 the value of its nutrition. Heat treatment had no sig- 80 C for 60 min was 154.64 mg/g. Properties of nificant effect on the ash content[10]. vitamin C is easily changed due to oxidation yet if a stable crystalline (pure)[15]. Degradation of vitamin Protein content C depends upon many factors such as oxygen, heat, Aloe vera gel provides 20 of the 22 necessary light storage temperature and storage time[20]. Anti- amino acids required by the human body, there are 7 oxidant compound such as vitamin C may occur the of the 8 non-essential amino acids and 12 essential degradation at 35 0C. Antioxidant such as vitamin C [8] amino acids . Protein of Aloe vera consist of lectins will lead the degradation about 38% during contact [11] and lectin-like substance . with a high heat[21] and decrease of vitamin C about Protein content of Aloe vera skin was 120.114 40 to 60% may occured due to heating at 82-92 0C[22]. mg/g. The protein content was increase with the in- Vitamin C was decrease about 15% affected both 0 crease in extractio n temperature up to 240 C by increasing temperature and duration of storage[23] and showed no obvious degradation to the extracted and Pi noted that the long duration of extraction may [12] 0 material .