Phytochemical and Antioxidant Activity of Avocado Leaf Extract
BACKGROUND AND OBJECTIVES: Antioxidants are a necessary nutrient in the body to prevent and deal with oxidative stress. The avocado leaf is claimed to contain natural antioxidants that can meet the needs of still limited antioxidants, especially for people with degenerative diseases. This study aims to identify and analyze phytochemical content and antioxidant strength in avocado leaf extract. Materials and methods: Research type is pure experience. Avocado leaf extract was prepared using 96% ethanol solvent at 1:10 (w / vol) for 3 x 24 hours. The types of phytochemicals analyzed were flavonoids, saponins, polyphenols, tannins, alkaloids and steroids. Measurement of antioxidant activity using the DPPH method (1,1-di-phenyl-2 Becquerel hydrazel) on an avocado leaf extract and vitamin C (control group). Unreadable data analysis was t, unidirectional ANOVA test and linear regression test of significant value P <0.01. Results: The positive avocado plant extract contains flavonoids, saponins, tannins and stimulants. Avocado starch extract for antioxidant activity (DPPH absorption = 0.797) is strong with IC50 from 72.61 mg L-1 and vitamin C the positive control is also strong with IC50 (mg L-1) from 23.03. Abstract: The avocado leaf extract contains phytochemicals that contain powerful antioxidants that can be used to prevent and deal with oxidative stress.
Introduction to Avocado leaves extract research.
Antioxidants are compounds that are able to inhibit the oxidation of other molecules 1. The body does not have an excessive antioxidant defense system, so if there is exposure to excessive free radicals, the body needs external antioxidants. Concerns about the side effects of synthetic antioxidants make natural antioxidants Preferred Alternative . In recent years, there has been growing interest in obtaining natural antioxidants . Studies show that phenolic compounds such as flavonoids have antioxidant activity from free radical maskers present in plants and fruit leaves that are commonly used as a traditional drug
The World Health Organization indicates that traditional medicines coming from plants have been used by the community at about 80% as a personal healthcare effort 8 Use of medicinal plants includes prevention, treatment of disease and health care. Certain plant or plant extracts are believed to contain biologically active compounds and effective in treating diseases 8,9. Most natural antioxidants come from fruits and vegetables, spices, grains and spices such as ginseng, turmeric, ginkgo, rosemary, green tea, grapes, ginger, garlic and avocado (American Americana mill) 10-13.
The avocado is only known for the fruits that people usually consume. Avocado leaf is clearly one of the natural ingredients that can be used as a traditional drug 14. This paper has been used experimentally as a diuretic, analgesic, anti-inflammatory, hypertension, blood sugar, diarrhea, sore throat and curative bleeding 14. Avocado is one Of the groups of fruit plants that are nutritious as preservatives and antioxidants. Avocado meat can be used as an anti-hyperlipidemic, has antioxidant activity and reduces the risk of metabolic syndrome.
One part of the avocado plant that is potentially a natural anti-oxidant is the avocado leaf. Previous research has shown that avocado leaf has the potential as a natural antioxidant and positively contains alkaloids, flavonoids, saponins, imps and stimulants using methanol solution to dissolve and extract avocado leaves. However, the use of methanol in the extract is less safe compared to ethanol 21. This research aims to identify On phytochemical content using 96% ethanol solution and antioxidant strength in avocado leaf extract using the DPPH method (1,1-diphenyl-2 Becquerel hydrazel). Extracting an antioxidant compound into the leaf using ethanol would be safer for future experimental studies on animal or human settings.
The study was conducted at the Analytical Chemistry Laboratory at the Faculty of Mathematics and Science at Tadulako University from June to September 2017. This type of research is experimental with a sample consisting of two groups, the treatment group and the control group. The avocado paper used both in the treatment and control group was the butter type. The treatment group was an avocado leaf extract consisting of 5 series of concentrations which are 10, 30, 50, 70 and 90 mg-1, while the control group was a vitamin C that consists of 5 series of concentrations such as 10, 30, 50, 70 and 90 mg-1 liter.
Materials: about 96% ethanol, DPPH, huts, filter paper, concentrated hydrochloric acid, 5% ferric chloride, Dragendorff reagent, chloroform, acetic hydride and concentrated sulfuric acid.
Tools: Mixer, 60 mesh sieve, Erlenmeyer 1000ml, measuring flask, 500ml measuring cup, analytical balance, glass funnel, chamber, Buchner funnel, vacuum pump, rotary evaporator, UV-VIS spectrometer.
Data were collected through a series of measurements. photo not relevant
Avocado leaf extract preparation: The avocado leaves used were fresh green, separated from the stem. The avocado leaf sample was cleaned, then dried in an oven at 50 ° C for 24 hours. After drying, the sample was mixed to obtain a dry powder. The samples were prepared using 96% ethanol solvent at 1:10 (w / volume) for 3 x 24 hours. The solution of the extract used was 96% ethanol. The ethanol in ethanol is 20% higher, non-toxic, and neutral, and requires less heat for the concentration process, limited soluble binding substances and sub-polar ethanol to attract polar and non-polar compounds. Maceration results are filtered using Buchner vacuum filter paper with the help of a filter. The candidate was absorbed into Erlenmeyer. The extract was vaporized with an evaporator to obtain a concentrated avocado leaf extract. Furthermore, the concentrated extract of avocado leaves has been tested for its phytochemicals (combinations of flavonoids, saponins, alkalis, stimulants, tannins) and antioxidant capacity.
Phytochemical Test 22 – Flavonoid Test: A flavonoid test is that 1.0 ml of sample solution is fed into the test tube and then a little magnesium powder is added and a few drops of concentrated hydrochloric acid (Chenola reagent), so when it reacts positively, it will lead to an orange solution Or pink or red.
Saponin Test: A total of 2.0 ml of sample solution is inserted into the test tube and shaken for a few minutes, when it reacts positively, it will form a stable foam for 15 minutes.
Tannins test: Tannins test is that 1.0 ml of sample solution is fed into the test tube and then a drop of 5% ferric chloride solution is added, when it reacts positively this will lead to brownish precipitation and a dark blue solution.
Alkaloid Test: An alkaloid test is 1.0 mL that is fed from the sample into the test tube and then 2-3 drops of Dragendorff reagent are added, so when it reacts positively, this will lead to an orange precipitation.
Steroid Test: The steroid test is that 1 ml extract is added with 3-5 drops of chloroform, then added again with 3-5 drops of acetic acid hydride and 10 drops of concentrated sulfuric acid. A positive test is characterized by a change in the color of the blue or green solution.
Antioxidant activity test using the DPPH method (1,1-di-phenyl-2 Becquerel hydrazyl) 23: concentrated extract from the sample in which the antioxidant activity was determined, use the spectrophotometric method using the DPPH reagent. The extracted sample was weighed 25 mg and then placed in a 25 ml vial, then adjusted with a solvent of ethanol to obtain a concentration of 1000 ppm solution. Then a series of dilutions were performed to obtain a 10, 30, 50, 70 and 90 mg-1 solution. The solution was prepared, with a diameter of 0.2 ml, and added with a 3.8 ml DPPH solution 50 متر m. The mixture is smoothed and left for 30 minutes in the dark. Next, the absorption is measured with a wavelength of 517 nm. Tests were also performed on the DPPH solution.
The absorbance value obtained is used to determine the inhibition (%) using the following Eq.24:Next, an inhibition curve (%) was prepared and IC50 (inhibition concentration) was determined based on the regression equation obtained. The parameter for interpretation of test results by DPPH method is IC50. The smaller the IC50 value, the higher the antioxidant activity. The value of IC50 is obtained in several stages, before calculating the value of IC50, first calculate the focus record value and the probit value for each percentage of free radical capture activity. Antioxidant power levels based on IC50 values are: Very Strong (<50), Strong (50-100), Medium (100-250), Weak (250-500) and Inactive (> 500) 25.
Data analysis: Data analysis t was non-test, unidirectional ANOVA test and linear regression test of significant value was p <0.01.
image not relevant
Qualitative analysis of phytochemical compounds on avocado leaf extract contains flavonoids, saponins, tannins, triterpenoids, steroids and does not contain alkaloids (Table 1). Table 2 shows that the absorption of DPPH by the presence of avocado leaf extract and vitamin C decreased with increased avocado leaf extracts and vitamin C concentrations and all concentrations were significantly different with P <0.01. The value of DPPH absorption by avocado leaf extract was significantly different with DPPH absorption by vitamin C with p <0.01, where the value of DPPH absorption by vitamin C was on average smaller which was 0.288 compared to DPPH absorption by avocado leaf extract which was 0.452. DPPH inhibition increased the presence of avocado leaf extract and vitamin C with increased avocado leaf extracts and vitamin C concentrations and all concentrations were significantly different with a value of p <0.01. The percentage of DPPH inhibition with avocado leaf extract was significantly different with DPPH absorption by vitamin C at a value of p <0.01, where DPPH inhibition with vitamin C was on average larger which was 63.83% compared to the percentage of DPPH inhibition with avocado leaf extract which was 43.26 %
Table 3 shows that the absorption of DPPH with 70 mg L-1 of avocado leaf extract concentration was not significantly different from 90 mg L-1.
This means that the optimal concentration of avocado leaf extract to inhibit DPPH activity is 70 mg L -1. IC50 as an indicator in the evaluation of the antioxidants in avocado leaf extract as a powerful antioxidant power with an IC50 value of 72.61 mg-1, while vitamin C contains a very strong antioxidant strength with an IC50 value of 23.03 mg -1. Antioxidant capacity levels based on IC50 values are: Very Strong (<50), Strong (50-100), Medium (100-250), Weak (250-500) and Inactive (> 500) .
The results showed that the positive avocado leaf extract contains phytochemicals such as flavonoids, saponins, tannins and stimulants. Avocado leaf extract that uses 95% ethanol solution is a global solvent and is able to dissolve polar compounds 26, so that many polar and non-polar compounds such as alkaloids, flavonoids, saponins, tannins and steroids present in avocado leaves can be attracted to solvents, although there is research showing Ethanol is less effective in examining antioxidants 27. Methanol is the most effective solvent for extracting antioxidants, followed by water, ethanol and acetone 26. However, ethanol use is safer compared to methanol because methanol is toxic .
Qualitatively, the results of certain phytochemicals in the avocado leaf extract demonstrate that the avocado leaf can be used as a food source that contains antioxidants. Numerous studies have shown that phytochemicals of phytosterol, saponin, polyphenol, flavonoids and ascorbic acid have the ability to regulate cholesterol metabolism and improve antioxidant status in highly cholesterol mice Flavonoids contained in avocado leaf extracts can be used in the treatment of oxidative stress because flavonoids can freeze free radicals by donating hydrogen atoms or by transferring a single electron
The requirements for flavonoids in daily consumption are still higher than vitamin C (70 mg / day), vitamin E (7-10 mg / day) and carotenoids (autotene, 2-3 mg / day) 33. Flavonoids may range between 50 and 800 mg / Day, depending on the consumption of vegetables, fruits and some drinks, such as unfiltered red wine, tea and beer. In particular, red wine and tea contain high levels (about 200 mg per cup of red wine or a cup of tea) of total phenol. But the Indonesian people, red wine consumption is still limited, so this avocado leaf extract can be a solution to meet the needs of flavonoids. In addition, the use of avocado leaves as vegetables is still very limited compared to other nutrients such as tree nuts and palm fruits that contain important nutrients and phytochemicals, including carotenoids, polyphenols and tocopherols that have antioxidant and other functions of vital activity 34.
The results show that the higher the concentration of vitamin C and avocado leaf extract, the more antioxidant activity that the lower absorbance value (p = 0.000) increases. This is because the higher the concentration, the less absorption of the antioxidant capacity to reduce DPPH free radicals. Additionally, the value of inhibition (%) also experienced a significant increase (p = 0.000) along with increased vitamin C concentrations and avocado leaf extract. The absorption and inhibition value (%) was found between extracts of vitamin C and avocado leaves significantly different (p = 0.002), as vitamin C remains as an antioxidant higher in the ability to moisturize DPPH free radicals compared to avocado leaf extract. Vitamin C has an average inhibition value (%) of 63.83% while avocado plant extract is 43.26%. The factors that influence the stability of antioxidant activity are pH, temperature, rays, and oxygen, in addition to other factors such as metal ions. The temperature factor affects the level of antioxidant activity in the avocado leaf. Anggorowati et al.42 research has shown that the temperature of making avocado leaf tea at 60, 70 and 80 ° C than the IC50 is higher because of the number of antioxidants that begin to degrade at this temperature, because the more antioxidants decompose, the higher the value of IC50, the value of IC50 rises, the active ingredient in avocado leaf decreased.
Next, a comparison of the IC50 leaf extract of avocado and vitamin C as an indicator of the strength of antioxidants showed that vitamin C had a smaller IC50 value which was 23.03 mg -1 compared to the avocado leaf extract which was 72.61 mg L-1. This means that vitamin C has the power of antioxidants with a very strong class, while avocado leaf extract is a strong class. The small strength of avocado leaf antioxidant activity compared to vitamin C is due to the fact that avocado leaf extract is still a mixture of several compounds such as flavonoids, saponins, tannins and stimulants, while vitamin C is a pure synthetic compound as an antioxidant 13 also contains more Hydroxyl groups, so vitamin C can donate more hydrogen atoms to interact with DPPH’s free radicals.
This research is in line with Owolabi et al. 43, which has suggested that avocado leaves have a strong antioxidant activity, which may help prevent or slow the progression of various diseases associated with oxidative stress. Avocado leaf extract is a natural antioxidant and anti-inflammatory compound that prevents the formation of calcium oxalate crystals by interfering with the process of epithelial cell damage 44. The total phenol content and antioxidant potential of avocado phenols are influenced by the extracted solvents and various avocados 45. In addition to the avocado leaf, the grape cream contains Avocado is also skinned on bioactive compounds and high antioxidants against DPPH (43 and 35%, respectively) of avocado fruit (only 23%) 19.
This study indicates that the avocado leaf is most likely used to avoid certain diseases associated with oxidative stress. Therefore, avocado leaves known as non-economic values can contribute to being a natural drug with the potential to become a source of income for farmers and industries. However, the avocado leaves in this study were limited to one difference. Examination of other varieties would bring a clear insight into the differences of biologically active compounds between avocado leaf shapes. Moreover, the contents of phytochemicals in this study were only examined qualitatively, without providing information on the amount of the compounds. This study also did not separately measure the antioxidant activity of all phytochemical contents while each type of phytochemical could have a different level of antioxidant activity. Further studies should be conducted on the evaluation of the compound of phytochemicals, the extraction of the contents of phytochemicals separately to demonstrate its antioxidant activity, as well as the study in animals and human settings to provide a more detailed understanding of the natural antioxidant chemical process of avocado leaves.
Phytochemical examination of avocado leaves extracted with 90% positive ethanol solution containing flavonoids, saponins, tannins and stimulants. The percent absorption and DPPH inhibition value of avocado leaf extract and vitamin C showed a significant difference. The 70% concentration is the avocado leaf extract that gives optimal absorption and inhibition value percent. The activity of avocado leaf extract with antioxidants (absorption DPPH = 0.797) is strong with IC50 is 72.61 mg L-1 and vitamin C is very strong with IC50 which is 23.03 mg -1. Avocado leaf extract can be used in ccombination with a flavonoids compound and a strong antioxidant activity in the intervention of the disease associated with free radicals.
The study discovered that the effects of phytochemical combinations (flavonoids, saponins, tannins and stimulants) avocado Leaf.
Learn more from a study at
Nour Al-Din and Necmah Otami Dewi and BohariSummary