Published on International Journal of Food & Nutrition
Publication Date: June 5, 2019
Ibrahim Saleh, Mia Anggraeni & Tia Agustina
Department of Food Technology, Palapa Institute of Science and Technology, Malang
Department of Food and Science, Widya Karya Catholic University, Malang
Department of Agriculture, Lumajang University, Lumajang
Journal Full Text PDF: Inhibition Xanthin Oxidase Fruit Extract Salak Variety Meats Humpbacked (Salacca edulis Reinw.).
Inhibition of xanthin oxidase of extracts of snake fruit flesh of Hunchback had been tested. Powder of snake fruit of Hunchback was extracted by n-hexane, ethyl acetate, ethanol and water. Extracts were tested by xanthin oxidase enzyme inhibitors in vitro. Value of IC 50 of substant inhibition of xanthin oxidase enzyme was measured using the spectrophotometer at 293 nm. N-hexane, ethyl acetate and ethanol extracts capable to inhibition of xanthin oxidase enzyme, except water extract. Allopurinol is used such as comparison solution. Inhibition concentration (IC50) of ethyl acetate extract was of 24.75 mg / ml and ethanol extract was of 44.95 mg / ml, and allopurinol was of 0.92 ug / ml..
Keywords: Snake fruit of Hunchback, extracts, xanthin oxidase, IC50.
Salak village Hunchback Hunchback derived from Sumedang. These fruits are not so appreciated by the public because it is not so sweet, sour, tart and slightly bitter. Prices of fruits Hunchback only Rp 500 per kilogram, because it does not provide economic value and are not used as food. Based on field surveys haisl, farmers in the village began to cut down the tree bark Hunchback Hunchback. In 2001 production was 63 256 quintals Hunchback bark and in 2002 only 47 469 quintals, the Ministry of Agriculture . This can lead to the extinction of fruits Hunchback. It is necessary for efforts to increase the economic value of fruits Hunchback by taking the active compound into a product that can be healthful.
Study of the active compounds in the fruits are generally not well known. Research conducted on several varieties of fruits showed ascorbic acid, malic acid, adipic acid, citric acid, carotene, and lycopene, Muchtadi D., , Setiawan et al.,  Suter, IK .
Uric acid is the end product of purine metabolism of food or from the breakdown of purine nucleic acids the body. Serum urate in the form of sodium urate, whereas in the urinary tract, veins in the form of uric acid. Gout is a metabolic disorder in its development bermanisfestasi towards increased concentration of serum uric acid. Hyperuricemia occurs when sodium levels of serum uric serum exceed the solubility will be saturated and can stimulate the formation of sodium urate crystals to precipitate. The solubility of sodium urate in serum at 37 ° C is 7 mg / dl, Kong et al., .
Enzymes involved in the synthesis of uric acid is xanthin oxidase that is actively working on the liver, intestines and kidneys. Without the help of these enzymes, uric acid can not be formed. To inhibit the formation of uric sam used allopurinol to inhibit xanthin oxidase enzyme that can reduce the production of uric acid, Vogel & Wolfgang .
2. RESEARCH METHODS
Materials used in this study are: Fruit bark Hunchback varieties, distilled water, n-hexane, ethyl acetate, ethanol, water, Xanthine from Sigma (USA), the enzyme Xanthine oxidase from Sigma (USA). And for chemicals and reagents used in the analysis of laboratory SINDO.
While the tools used are: Balance analytical, tunnel drier, spectrophotometry, maceration tube, semi-micro cuvette, micropipette, stop watch, pipettes, evaporator.
Meat dried fruits Hunchback, made of powder simplicia. Simplicia extracted with a solvent n-hexane, ethyl acetate, ethanol and water. Each extract was collected and evaporated to concentrated at 40 ° C.
2.1.2 Test dampen the activity of xanthine oxidase in vitro
The concentration of each extract maisng and allopurinol are 0:01, 0:02 and 0.2 mg / mL. Samples were incubated with 0.1 units of enzyme Xanthine oxidase, 40 mL and 1750 mL EDTA phosphate buffer solution (pH 7.8). Control solution without the sample extract. Then add 100 mL xanthin to obtain the volume of 2 ml. Test extracts in reducing Xanthine oxidase activity performed by spectrophotometry with a wavelength of 293 nm. Percent reduction obtained konsnetrasi calculated regression equation to the test solution, following equation:
Y = ax + b
Where: Y =% reduction,
x = the concentration of the test solution
Of all the experimental results were then determined IC50, namely the concentration inhibition test solution that can reduce 50% Xanthine oxidase enzyme
To dampen the activity of xanthine oxidase by each maisng extracts and allopurinol was shown by the results of the relationship of absorbance against extract concentration. Extracts were able to abate, xanthine oxidase activity will provide the most low absorbance.
Table 1. Relation between Absorbance of the extract concentration (n-hexane, ethyl acetate, ethanol and water) and allopurinol (standard raw materials)
Picture 1. Relation between Absorbance of the extract concentration (n-hexane, ethyl acetate ethanol and water) and Allopurinol (standard raw materials)
Each Hunchback bark extracts that reduce the activity of the enzyme xanthine oxidase can be seen in Figure 2. In Table 2, not all of the extract can reduce the activity of xanthine oxidase, allopurinol as standard raw materials is still better in reducing the activity of xanthine oxidase to 67% compared to extract only able to reduce 50%.
Table 2. Damping percent Xanthine oxidase enzyme activity from Extract (N-hexane, ethyl acetate, ethanol and water) and allopurinol
50% inhibition concentration of the ethyl acetate extract can be seen in Figure 2, the linear regression equation of the ethyl acetate extract had R2 = 0.91 with IC50 was 24.75 ug / ml, with the concentration of ethyl acetate was effective can reduce 50% of enzyme activity xanthin oxidase. Extract the linear regression equation ethanol extracts obtained R2 = 0.82 with IC50 was 44.95 ug / ml. IC50 is a water extract of more than 100 pg / ml, whereas the IC50 of allopurinol was 0.92 ug / ml with R2 = 0.98.
Uric acid is a normal part of the blood and urine. Uric acid is produced from the breakdown and disposal of the remnants of certain foods containing purine nucleotides or derived from purine nucleotide produced olehtubuh. High levels of uric acid in the blood of patients with gout caused many remnants of disposal proceeds purine metabolism while the excretion of uric acid purine xanthine oxidase too sedikit.Enzim important role in the formation of uric acid, allopurinol is a drug commonly used to relieve the pain of gout. In this study, all the extracts can reduce the activity of xanthine oxidase in vitro. Thus Hunchback extracts of fruits can be used as herbal preparations that may inhibit xanthine oxidase.
Ethyl acetate and ethanol extracts of fruits can reduce the Hunchback xanthin oxidase activity except extract n-hexane and water.